2005-2012 Global Science Books

MANUSCRIPTS ACCEPTED OR IN PRESS, INCLUDING ABSTRACTS (2012)

For Current journal content see individual journal page at http://www.globalsciencebooks.info/Journals/GSBJournals.html

th Updated 26 April, 2012 (ordered alphabetically, unless in press)

Bioremediation, Biodiversity and Bioavailability

SPECIAL ISSUE: Induced mutagenesis in plants. ~ 2012 Paolo Ranalli (Italy) The Role of Induced Plant Mutations in the Present Era ABSTRACT Invited Mini-Review: With the imminent threats posed by global climate change to crop production and the ever increasing and more sophisticated demands of agricultural products, crop improvement efforts have to be more powerful and precise in developing new crop varieties. Breeders therefore require tools that permit achieving subtle changes to the genetic make-up of otherwise superior crop varieties e.g. high yielding but lacking in specific quality traits and yet leaving the genome largely intact in order not to disturb already stacked alleles of genes. The availability of genomics information in the public domain coupled with recent advances in molecular and cellular biology techniques have paved the way for transforming old mutation techniques into state of the art technology for both crop improvement and basic genomics research. Cellular biology techniques will address the bottlenecks imposed by the need to rapidly generate large mutant populations of suitable genetic backgrounds (homozygous for the mutation events, and devoid of chimeras). New, space-age technologies are being developed for mutation induction. Thus, mutation assisted plant breeding will play a crucial role in the generation of designer crop varieties to address the uncertainties of global climate variability and change, and the challenges of global food insecurity. Suprasanna Penna, Sonawane Balasaheb Vitthal, Patade Vikas Yadav (India) In Vitro Mutagenesis and Selection in Plant Tissue Cultures and their Prospects for Crop Improvement ABSTRACT Invited Review: Mutation induction has become a powerful tool for developing new and novel plant germplasm. Methods such as gamma ray irradiation, ion beams and chemical mutagens have been applied to induce mutations. Since availability of a large number of mutagenized populations for screening and methods of selection are still a hindrance with conventional mutagenesis, in vitro mutagenesis of cultured explants, cells and tissue cultures represent a feasible method for induction of genetic variability. Selection at the cellular level has been practised for desirable traits and success has been achieved in several crop plants. This article outlines the different aspects of in vitro mutagenesis and selection for varied applications in crop improvement. Subodh K. Datta (India) Success Story of Induced Mutagenesis for Development of New Ornamental Varieties ABSTRACT Invited Review: Induced mutagenesis is well recognized as one of the most important technology for the development of new varieties through genetic manipulations. Mutation techniques, using physical and chemical mutagens, have successfully produced quite a large number of new promising varieties in different ornamental plants. This technique has been most successful in ornamental plants due to some additional advantages. Changes in any phenotypic characteristics like colour, shape or size of flower and chlorophyll variegation in leaves can be easily detected. Heterozygous nature of many of the cultivars offers high mutation frequency. The main advantage of mutation induction in vegetative propagated crops is the ability to change one or a few characters of an otherwise outstanding cultivar without altering the remaining and often unique part of the genotype. Voluminous literature for successful application of classical induced mutagenesis have been generated on radio-sensitivity, selection of materials, methods of exposure to gamma rays, suitable dose, detection of mutations, mutation frequency and spectrum, isolation of mutants and commercial exploitation of mutants. Different treatment methodology like recurrent irradiation, combined treatment, split dose, colchicine treatment, ion beam technology, space breeding, TILLING,

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EMAIL, etc., have been precisely determined for successful development of new varieties. The main bottlenecks in mutation breeding of vegetatively propagated plant are formation of chimeras. Therefore, attempts were made to find out the ways to overcome this situation. Management of chimera and in vitro technique have opened a new way for isolating new flower colour/shape ornamental cultivars through retrieval of mutated cells. Step wise advancement/refinement of practical approaches for application of classical induced mutagenesis and recent techniques for improvement of ornamental crops have been highlighted. Sanjay G. Auti (India) Induced Morphological and Quantitative Mutations in Mungbean ABSTRACT Invited Review: Pulses occupy an important position in world agriculture because of their high protein content, several essential amino acids and their capacity for fixing atmospheric nitrogen. Mungbean (Vigna radiata (L.) Wilczek) is one of the most important pulse crops due to its nutritive value and property of maintaining and restoring soil fertility through biological nitrogen fixation. Genetic variability is one of the prerequisites for crop improvement. Lack of required amount of variability, limits the scope for the selection of better genotypes in mungbean. Artificially induced mutations are the best way to enlarge genetic variability considerably within a short time. Frequency of the natural mutation is very low and hence artificial mutations are induced and genetic variability is best enhanced with the application of mutagens. Based on above fact, the development of high yielding varieties in mungbean is possible by the exploitation of larger range of genetic variability. Mungbean have been traditionally grown in marginal land of lower productivity with the application of little inputs. The selection pressure in case of these crops has been concerned more with adaptation to stress conditions than for yield. Therefore the genetic improvement of such crops, for higher yield requires their genetic reconstitution to evolve different plant types. Induced mutations can help to regenerate and restore the variability, which has been lost in the process of adaptation to various stresses or adaptations during the course of evolution. Thus mutation breeding or induced mutation is having great potential for the improvement of traditional agricultural crops like mungbean. Rodrigo R. Latado, Augusto Tulmann Neto, Antonio Figueira (Brazil) In Vivo and in Vitro Mutation Breeding of Citrus ABSTRACT Invited Mini-Review: Spontaneous mutations are important for the development of new citrus varieties, but they appear at very low frequency in nature. On the other hand, induced mutagenesis using radiation or chemical mutagens can increase many folds the rate of mutation resulting in increased genetic variability to be used for selection. But, up to now, few mutants induced were officially released for cultivation, possibly because citrus plants are vegetatively propagated, with a long reproductive cycle and require much space to be cultivated. Physical mutagens (gamma-rays, X-rays or thermal neutrons) were the most commonly used in mutation induction, usually employing buds or seeds as starting materials. Chemical mutagens were less used, possibly due to the difficulties associated with tissue penetrability and that the target propagules are not seeds. Up to now, most projects using induced mutagenesis in citrus aimed at developing new cultivars with emphasis on obtaining seedless or fewer seeds per fruit; resistance to diseases; improved fruit color (skin and pulp); fruits with alteration in harvest time; and plants with height reduction. Induced mutants of citrus have been described in several countries. The development of new areas of biology (e.g functional genomics and bioinformatics) and new high throughput related technologies, such as high performance sequencing, TILLING (targeting induced local lesions in genomes), microarray, real-time PCR and others, associated with induced mutagenesis, can help in gene discovery programs to elucidate function and mechanisms of gene action. Digambar Dadaji Ahire (India) Induced Mutations for Elevated Oleic Acid and Reduced Linolenic Acid Content in Soybean Seeds ABSTRACT Invited Mini-Review: The high linolenic acid content (7-9%) is associated with poor flavor of soybean oil. Soybean oil with less than 2% linolenic acid would be desirable for improving the quality. Oil in which the linolenic acid (18:3) content has been reduced from 8 to 1% makes it possible to reduce or eliminate the need for chemical hydrogenation and henceforth can achieve the stability and increased shelf life. Oil with elevated levels of oleic acid is less susceptible to oxidative changes during refining, storage and frying. Oil in which the oleic acid (18:1) has been increased from 25 to 80% also have increased stability and longer shelf life. Natural genetic variability for fatty acid composition in commercial soybean seed oil is limited, therefore, additional variability for increasing economical usage is to be created. In this augmentation, induced mutagenesis has been seen to be an

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excellent tool for creating variability for fatty acid contents in soybean. Induced mutagenesis is regarded as a supplement to the conventional breeding method used to alter the concern genes for reduced linolenic acids and elevated oleic acid content in the cultivars currently grown for commercial production. A significant positive correlation of 100-seed weight with oleic acids and negative correlation with linolenic acids is also useful to select elevated oleic acid and low linolenic acid lines indirectly. The purpose of this paper is to review the methods that have been used to develop the fatty acid modifications, the inheritance of the modifications, the impact of the trait on agronomic and seed characteristics, the methods of phenotypic and genotypic selection, and the commercial status of the modified oils. M. A. K. Azad, M. I. Uddin, M. A. Azam (Bangladesh) Achievements in Rice Research at BINA through Induced Mutation ABSTRACT Invited Mini-Review: The Bangladesh Institute of Nuclear Agriculture (BINA) since its inception in 1961 as a radio tracer laboratory under the then Pakistan Atomic Energy Commission (PAEC) has released so far 7 improved varieties of rice through induced mutation technique. Of the 7 varieties Iratom-24 and Iratom-38 were developed by irradiating the seeds of the rice IR 8 and released by the National Seed Board (NSB) of Bangladesh in 1974 for boro (December to May) and aus (March to June) seasons. The mutated characteristics of these varieties are shorter crop duration, resistance against BLB and medium fine grain size. Binashail was also developed by irradiating the seeds of Naizershail and released in 1987 for aman (July to December) season with mutated characters like long panicles, more number of grains, medium fine grains with higher weight, early maturing with photo-insensitivity and higher grain and straw yields. Binadhan-4, Binadhan-5 and Binadhan-6 were developed by irradiating F2 seeds and released in 1998, Binadhani-4 for aman and Binadhan-5 and Binadhan-6 for boro season. Binadhan-7 was developed from M3 seeds of the cv. Tai Nguen of Vietnam. This variety was released in 2007 for aman season with earliness, long fine grain and high yield. Early maturing character of this variety helps to escape drought and insect attack during flowering and milky stages. Moreover, it helps increasing cropping intensity by facilitating timely cultivation of following winter vegetables. Binadhan-9 was developed by hybridizing between the local cv. Kalozira and an exotic mutant line Y-1281. It was released in 2012 for aman season, also be grown in boro season. Unlike the parent Kalozira, it has short duration, long and slender grains. Using recent ion beam irradiation technique BINA has already developed some mutant lines that can be grown after harvest of mustard or rapeseed during the first or second week of February and can be harvested after 120 days. These mutant lines produce high yield. Nasya Tomlekova, Svetla Yancheva, Bistra Atanasova (Bulgaria) Molecular Identification of Tomato Mutant Lines ABSTRACT Original Research Paper: Molecular characterization of 7 tomato breeding lines (6 mutant and a parent line), maintained in the collection of Maritsa Vegetable Crops Research Institute, Bulgaria, was conducted. Irradiations on an initial genotype of Solanum lycopersicum L. performed by 250 Gy 137Cs generated mutation causing late-flowering. Six tomato mutant lines were advanced to M3 by induced mutation. In this study, both applied, AFLP and ISSR techniques were effective in assessing polymorphic patterns between the studied tomato mutant lines and the corresponding initial ones. The AFLP technique, which has the potential to provide valuable information in a number of areas, revealed discriminating polymorphism among tomato mutant lines and enabled their identification from the initial line with three primer combinations only. AFLP polymorphism was not related to the mutant character. Clear discrimination of the mutant in comparison with the initial tomato lines was revealed by using the ISSR technique. The bulk of the observed morphological evidence, corroborated by the molecular data in this study, indicated DNA variability in the tomato mutant lines. Dafedar H. Tejavathi, Kaivaram Gayathramma, Rajashekharaiah Nijagunaiah (India) Genetic Variability by in Vitro Mutagenesis and Arbuscular Mycorrhizal Fungal Symbiosis among Micropropagated Plants of Agave vera-cruz Miller ABSTRACT Original Research Paper: Agave vera-cruz Mill. is naturally found in semi-arid regions of India and provides a high quality fiber as a good source of income to farmers. Samples for the present study were obtained through direct and indirect organogenesis, arbuscular mycorrhizal fungi (AMF)-treated tissue-cultured plants as well as from tissue-cultured plants derived from exposure to gamma irradiation at 15 Gy for up to 20 min at 2-min intervals (physical mutagen) and ethyl methane sulfonate at 0.1 to 1% for 1 to 5 h (chemical mutagen). These, together with untreated plants were analyzed by amplified fragment length polymorphism (AFLP) markers. A phenotypically distinct variant derived from indirect organogenesis showed more

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polymorphism followed by mutagen-treated samples. Jaccards coefficient of similarity was analyzed by the unweighted paired group method with arithmetic average (UPGMA) method. Plants obtained from mutagen-treated and untreated cultures formed two main clusters. Plants obtained through direct organogenesis and subsequently treated with AMF formed a separate sub-cluster indicating an altered molecular marker pattern. It is feasible to use in vitro culture methods coupled with mutagenesis and AMF symbiosis as a potential tool for increasing genetic variability of vegetatively propagated plants. Sobri Hussein, Anna Pick Kiong Ling, Chee Yang Lau, Siew Ling Ong, Abdul Rahim Harun (Malaysia) Morphological and Biochemical Responses of Eurycoma longifolia Callus to Gamma Irradiation ABSTRACT Original Research Paper: Eurycoma longifolia is commonly used as medicinal plant in South East Asia, particularly for its aphrodisiac property. In this study, the mutation of Eurycoma longifolia callus cultures was induced by gamma irradiation at 20, 40, 60, 80 and 100 Gray (Gy). After three weeks of irradiation, the morphological and biochemical changes of the callus were examined. The radiation sensitivity test determined that 60 Gy was the optimum dose to cause a 50% of decline in the fresh weight (FW) of E. longifolia callus. Even though there was no significant difference on the colour and texture of irradiated and non-irradiated calli, the FW of the calli decreased gradually with the increase of gamma doses. The total soluble protein content recorded 17, 57 and 30% higher than the non-irradiated calli in 20, 40 and 60 Gy, respectively. A further increase of gamma dose to 80 and 100 Gy caused a decrease of 64% in total soluble protein content. Contrarily, gamma irradiation generally brought about a decrease in total phenolic and total flavonoids content. The lowest total phenolic content (0.05 0.01 mg gallic acid equivalents/g FW) was recorded in calli exposed to 40 Gy while the lowest total flavonoid content (0.13 0.01 mg catechin equivalents equivalents/g FW) was obtained in calli irradiated at 60 Gy. Dibyendu Talukdar (India) An Induced Glutathione-deficient Mutant in Grass Pea (Lathyrus sativus L.): Modifications in Plant Morphology, Alterations in Antioxidant Activities and Increased Sensitivity to Cadmium ABSTRACT Original Research Paper: A glutathione-deficient mutant, designated as gshL-1, was isolated in 350 Gy gamma ray treated M2 progeny of grass pea (Lathyrus sativus L.) var. BioL-212. Its fitness was tested in control (un-stressed) and in four different concentrations (5, 10, 25 and 50 M) of CdCl2. The mutant, containing only 15% of total glutathione in leaves and 20% of that in roots of mother control (MC) variety, exhibited slow growth, poor root development and significant reduction in biomass production, in comparison to MC plants. The impact was more severe in Cd-treated plants. The huge deficiency in total and reduced glutathione (GSH) content was mainly due to significantly low level of glutathione reductase activity, triggering a massive disruption in antioxidant defense systems in both leaves and roots of the mutant. Interestingly, glutathione biosynthesis was not affected in the mutant. Within ascorbate-GSH cycle, low GSH availability presumably impeded the normal functioning of dehydroascorbate reductase (DHAR), badly hampering the regeneration of reduced ascorbate in the mutant. The cascading effect of low GSH pool was also felt in scavenging of hydrogen peroxide due to below normal activity of ascorbate peroxidase and glutathione peroxidase, although catalase level was significantly high in both untreated and Cd-treated mutant plants. Furthermore, nearly 7-fold decrease in activity of GSH-S-transferases (GSTs) and accumulation of total phytochelatins only 10% of MC plants might led to increased sensitivity of the mutant plant to the imposed Cd stress. The results indicated potentiality of the induced mutagenesis in creation of novel mutants to explore the regulation of antioxidant redox homeostasis, mechanism of reactive oxygen species scavenging and tolerance to metal toxicity in grass pea. Sanjay G. Auti (India) Lhb Mutant A Novel Mutant of Mungbean (Vigna radiata (L.) Wilczek) Induced by Gamma Radiation ABSTRACT Original Research Paper: In the present investigation an attempt was made to induce mutations in mungbean (Vigna radiata (L.) Wilczek) var. Vaibhav by employing gamma radiation with the objective of obtaining novel and desirable mutants. Mungbean seeds were irradiated with 30, 40 and 50 kR doses of gamma radiation and sown in the experimental fields to raise M1, M2 and M3 populations. Mutations were screened at M1, M2 and M3 generations. All three doses of gamma radiation effectively produced various morphological mutations. However, the 50 kR dose effectively produced a novel mutant that showed multiple morphological mutations such as large flowers with dark-yellow petals, dense and thick hairy pods and black seeds. In this study, SDS-PAGE analysis of seed proteins of large flower, hairy pods and black seeds revealed a difference in the banding pattern between the Lhb mutant, other mutants (tall mutant, dwarf mutant, high-yielding mutant and early maturing

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mutant) and control. Avijeet Chatterjee, Sudhir Shukla, Anu Rastogi, Brij K. Mishra, S. P. Singh (India) Induction of Variability and Selection of Elite Lines for Specific Alkaloids from a Population of Induced Mutagenesis in Opium Poppy (Papaver somniferum L.) ABSTRACT Original Research Paper: Induced mutations often produce abnormalities which cause morphological alterations in external form of plants including colour, shape, size etc. In the present study two white flowered varieties of opium poppy i.e. NBRI-1 and NBRI-5 were subjected to physical (gamma rays), chemical (ethyl methane sulphonate; EMS) and combined (gamma rays + EMS) mutagenesis to determine the effective doses of mutagens to create plants deformities and chlorophyll (chl) mutation. The study also includes selection of elite lines for specific alkaloids especially for thebaine and codeine. The frequency of chl mutations was maximum for the treatment 50 kRad (0.42%) followed by 40 kRad (0.24%) in NBRI-1 while frequency was highest for the treatment 50 kRad + 0.8% EMS (0.86%) followed by 50 kRad + 0.4% EMS (0.60%) and 50 kRad + 0.6% EMS (0.56%) in NBRI-5. Among the chlorophyll mutations, albino was the most frequently screened followed by xantha type at all doses. The variety NBRI-5 was more responsive to the mutagens than NBRI-1 for chl factor. A typical chl variant was observed in M1 generation in the combined dose of 10 kRad + 0.4% EMS in plant no.9 of NBRI-5. The treatments 20 kRad in NBRI-1 and 30 kRad, 40 kRad and 10 kRad + 0.4% EMS in NBRI-5 showed high thebaine in both M2 and M3 generations. Barshile Jagannath, Boddu Apparao (India) Genetic Improvement of Chickpea (Cicer arietinum L.) Cultivar Vijay (Phule G 81-1-1) through Induced Mutagenesis ABSTRACT Original Research Paper: In the present attempt three well known mutagens, sodium azide (SA), ethyl methane sulphonate (EMS) and gamma radiation (GR) were employed to induce genetic variability for improvement of locally popular chickpea (Cicer arietinum L.) cv. Vijay (Phule G 81-1-1). Seeds were treated with three different concentrations/doses of SA (2, 3 and 4 mM), EMS (8, 12 and 16 mM) and gamma radiation (400, 500 and 600 Gy). Different mutants were isolated and characterized. They included leaf, pod, seed, flower colour and morphological mutants. Five true breeding mutant lines were studied for various quantitative and qualitative traits in the M4 generation. Induced mutant lines showed both positive and negative increases in quantitative traits. True breeding mutant lines in the M4 generation differed considerably in their quantitative traits from the control. The early mutant lines matured 10 to 11 days earlier than the parent cultivar. Gigas mutant plants obtained after 400 Gy gamma radiation were tallest (44.2 cm), with a 2-3 fold increase in pod and seed size over the control. The protein content in the M4 generation ranged from 259.11 to 282.18 mg/g vs 255.59 mg/g in the control. Lower TI activity was observed in the compact mutant (2465.4 TIU/min/g) as compared to the control (3091.5 TIU/min/g). Analysis of variance showed both positive and negative significant increase in the quantitative traits among the mutant lines. The higher heritability coupled with high genetic advance was observed for quantitative traits like number of pods/plant and number of seeds/plant for the mutants in the chickpea cv. Vijay. Variation was also observed for number of bands electrophoresed in seed protein of mutants. Digambar Dadaji Ahire (India) Correlation Studies and Coefficient of Variation among Fatty Acids and Oil Quality Parameters in Soybean Mutants ABSTRACT Original Research Paper: The interrelationships between fatty acids and oil quality parameters were determined by correlation studies to estimate the best selection criteria of mutants for improving oil quality. Correlation analysis results showed that a highly significant (P < 0.01) negative correlation of oleic (MUFA) with linoleic (r = -0.847) and linolenic acid (r = -0.692) (PUFAs) contents is important for selection of mutants with high oleic and low linolenic acid content to improve the oxidative stability index (OSI) of soybean oil. A strong significant but negative correlation was found between linolenic acid and the 6/3 ratio (r = -0.764) and a strong significant positive correlation between OSI and oleic acid (r = 0.938) content indicating improved oxidative stability of the oil while retaining nutritional quality. Mutagenic treatments produced significant genetic variation in fatty acid composition and oil quantity without altering the population means. Among all quality parameters, OSI (50.0%) and stearic acid (62.8%) exhibited maximum variation compared to other traits among the fatty acids. Sonu Goyal, Mohammad Imran Kozgar, Samiullah Khan (India) Effectiveness and Efficiency of Gamma Rays, Ethylmethane Sulphonate and their Combinations in Vigna mungo (L.) Hepper

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ABSTRACT Short Communication: A systematic study to determine the effectiveness and efficiency of gamma rays, ethylmethane sulphonate (EMS) and their combinations (gamma rays + EMS) were carried out in two urdbean (Vigna mungo (L.) Hepper) varieties namely, T-9 and Pant U-30. Based on effectiveness, the order of mutagens was EMS > gamma rays + EMS > gamma rays in both the varieties. Three criteria viz., pollen sterility, seedling injury and meiotic abnormalities were taken into consideration to determine the efficiency of the mutagens. The efficiency of the mutagens varies with the variety type and the mutagen doses applied depending upon the criteria chosen. Overall, the combination treatments were more efficient followed by EMS and gamma rays. Assessing mutagenic effectiveness and efficiency are beneficial to screen mutagen doses which are potent to increase genetic diversity among self-pollinated crops like urdbean for successful plant breeding strategies. Asnake Woldemedhin Fikre (Ethiopia) Gamma Irradiation-Derived, Methionine-Enriched Mutant Lines of Lathyrus sativus L. ABSTRACT Short Communication: Methionine (Met), an essential sulphur-containing amino acid, is supposed to protect from lathyrism disease caused by over-consumption of Lathyrus sativus (grass pea). There is a strong and positive association between moisture stress and the biosynthesis of the crippling lathyrism causative molecule, 3-(N-oxalyl)-L-2, 3-diamino propionic acid (-ODAP). Seeds of L. sativus local cv. Debre Zeit were irradiated using 40 kR gamma cell irradiation aimed at genetic alteration in favour of overproduction of Met-based seed proteins. The amino acid determination in seeds were analysed using HPLC with pre-column PITC derivatization. Three morpho-phenotypically distinct mutant lines were identified and selected from irradiated clusters, which displayed glittering deep green leaves, stem and pods, smaller leaf and plant size, and reasonable yield compared to the parental line. Observation of root growth response in ethionine media supported the phenotypic and chromatograph findings. Chemical analysis of seeds revealed that the putative mutants had 63% improved Met over the parent (control). This level shifts the Met supply capacity of grass pea from 25% in the parent line to 50% in the altered putative mutant lines. Pasquale Martiniello (Italy), Jaime A. Teixeira da Silva (Japan) Botanical Survey of Floral Species and Animal Feeding Values in Pasturelands of Environments with a Mediterranean Climate ABSTRACT Original Research Paper: The native flora of Mediterranean environments with meso-Mediterranean zones, instead of being protected by European Union and Italian government law, continue to be threatened as a result of the effect of social and anthropological evolution and misleading utilization of herbs causing genetic erosion of flora species. A floral survey was established over a two-year period in 20 pasturelands. The biodiversity discovered in the environmental pastures amounts to 29 botanical families and 361 floral species. The most represented floral species (expressed as a percentage of the mean value of the floral species of pasturelands) belong to the following botanical families: Graminaceae (26%), Leguminosae (17%), Compositae (19%), Labiatae (5%), Liliaceae (5%), Umbelliferae (3%), Cruciferae (4%), Plantaginaceae and Ranunculaceae (3%), and Caryophyllaceae (2%). The floral species less represented (< 1% of total flora) are included in a miscellaneous group composed of 19 botanical families. The floral biodiversity influences the herbage and milk feeding units of pasturelands. Five species from the Graminaceae (Aegilops geniculata L., Dasypyrum villosum (L.) Borbas, Stipa barbata Desf., Lolium perenne L. and Phalaris minor L.), which face the risk of extinction, were evaluated over a separate (but later) two-year period at Foggia for seed production. Agronomic evaluations of seed yield and its components of the five most popular grass species evidenced different bioagronomic characteristics and the possibility to provide a seed source for reseeding degraded pasturelands as a way to recover the natural equilibrium of native species in Mediterranean environments. Agronomic practices for seed production of floral species under the risk of extinction in favourable environments in order to provide seed stock to reseed represent a scientific tool to reduce the genetic erosion of floral species present in the degraded swards of the Mediterranean pasturelands. Tukaram D. Nikam, Janardhan N. Nehul, Yogesh R. Gahile, Bhausaheb K. Auti, Mahendra L. Ahire, Kirti M. Nitnaware, Bimba N. Joshi, Narendra Jawali (India) Cyanobacterial Diversity in Western Ghats Region of Maharashtra, India ABSTRACT Original Research Paper: Cyanobacterial diversity in Ahmednagar, Pune and Satara district of Maharashtra State, Western

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Ghats one of the biodiversity hotspots of Indiawas assessed. Screening of 627 soil samples from different locations in the above regions revealed that 94 cyanobacterial spp. belonged to 38 genera, 14 families and 5 orders. Diversity analysis revealed that Westiellopsis prolifica Janet. was relatively abundant (47.21%) followed by Nostoc calcicola Brebsson ex Born. et Flah. (44.82%). The frequency distribution of Myxosarcina spectabilis Geitler was less. Nostoc (Nostocaceae), Chroococcus (Chroococcaceae) and Anabaena (Nostocaceae) were the most densely populated genera. The information obtained will be useful in exploitation of cyanobacteria for biotechnological, pharmaceutical and other applications. Oluwatosin Ebenezer Atobatele (Nigeria) Pelagic Phytoplankton Succession Pattern in a Tropical Freshwater Reservoir (Aiba Reservoir, Iwo, Osun, Nigeria) ABSTRACT Original Research Paper: Fortnight phytoplankton sampling of Aiba Reservoir was carried out between March 2004 and February 2005 to determine phytoplankton succession pattern. Five major phytoplankton groups identified show two similar ordered directional repeated sequences of abundance peak patterns during an annual cycle; each began with a peak in Cyanophyceae abundance followed in order by peak abundances of Dinophyceae, Chlorophyceae, Bacillariophyceae and Euglenophyceae. The first succession pattern occurred between late April and mid-September (early rainy season) while the second occurred between early September and early February (late rainy season dry season). A brief dry spell (August break) during the rainy season is a major factor determining this biannual pattern.

Dynamic Biochemistry, Process Biotechnology and Molecular Biology

SPECIAL ISSUE: Microbiology. Guest Editor: Chandicharan Rath (North Orissa University, India) 2012 Shewli Bhattacharya, Jnanendra Rath, Samit Ray (India) Composition, Basic Features and Distribution of`Cyanobacteria in Soil Crusts A Review ABSTRACT Invited Review: Many of the cyanobacterial species are capable of growing on the soil and other terrestrial habitats. Soil particles form an intimate association with cyanobacteria and this results in the formation of cyanobacterial crusts that covers the surface of the soil as a coherent layer. The crusts cover the open spaces between the plantations and the soil surface remain covered by these highly specialized organisms. Crusts are found in an astonishing variety of habitats throughout the world in desert and semi-desert plant communities ranging from shrubs and succulent desert to open woodlands, in steppe formation in both northern and southern hemisphere, in the gaps between evergreen shrubs and in the forests, on open ground or between alpine or tundra vegetation. Cyanobacteria were the major component of the blackish-brown crusts on the upper surface of the soil. Major works on cyanobacterial crusts have been reported from Africa, North and South America, Europe, Mediterranean region and Asia. Reports from India are meager. Cyanobacterial soil crusts are an important source of fixed carbon and nitrogen, which enrich the soil. They act as ecological indicator, polysaccharides produced by them prevent desiccation, presence of cyanobacteria in the soil augments mineral upake by vascular plants and seed germination is enhanced. Considering the significance of cyanobacterial soil crusts in improving the soil condition and fertility attempts are being made to develop soil crusts artificially. In this review, crust formation, types of cyanobacterial crusts, physiological and biochemical characters of crust cyanobacteria, importance of cyanobacterial crusts in ecology and their distribution in various parts of the world have been discussed in detail. Ashok Kumar, Shamsher Singh Kanwar (India) Lipase Production in Solid-State Fermentation (SSF): Recent Developments and Biotechnological Applications ABSTRACT Invited Review: Lipases are the most widely used biocatalysts, because they can catalyze several unnatural and remarkable reactions in non-aqueous media, such as bio-fuel production, production of value-added products such as esters, organic acids, food, beverage, cosmetics and pharmaceutical materials. Solid-state fermentation (SSF) represents an interesting alternative to produce industrial enzymes at lower costs due to the possibility of using inexpensive agro-industrial residues as culture media. This review aims to explore various agriculture by-products like husk, straw, agricultural raw materials, waste of the oil industry, among others that are locally available and are also cost-effective requiring low nutrient supplementation to produce microbial

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lipase(s) in SSF. Enzyme production is associated with the growth of the bacterial culture. The physico-chemical fermentation parameters such as pH of the medium, moisture content, particle-size, nature of particles and microbial inoculum level play crucial role(s) in lipase production. SSF has gained renewed interest and fresh attention of researchers to develop processes to achieve large-scale enzyme production by solid waste treatment and in its application in the industry to synthesize the products of commercial value. Madan Lal Verma (India/Australia), Shamsher Singh Kanwar (India) Harnessing the Potential of Thermophiles: The Variants of Extremophiles ABSTRACT Invited Review: Extremophiles are life-forms that thrive under some of the harshest conditions found on earth. In recent years, extremophiles have been discovered in such inhospitable places as active volcanoes, deep sea vents, and the ultra-saline remnants of extinct inland seas. Extensive studies of extremophiles ecology, physiology, and molecular biology have yielded valuable information about life processes at every level, with a number of important industrial applications. This review articles summarize the most outstanding features of thermophilic microorganisms that can survive under extreme conditions. The latest findings on the thermophiles, the protein structure of these exotic organisms, potential applications of extremophiles in biotechnology industries including the production of enzymes, are discussed in the present research study. Lata Kumari, Shamsher Singh Kanwar (India) Cholesterol Oxidase: Role in Pathogenesis ABSTRACT Invited Review: Cholesterol oxidase (COx) catalyzes the oxidation of cholesterol into 4-cholesten-3-one. COx has various clinical and industrial applications. Cholesterol oxidase a bifunctional FAD-containing microbial enzyme belongs to the family oxidoreductases. In recent time, cholesterol oxidase has received great attention due to its wide use in clinical (determination of serum cholesterol) and laboratory practices. The COx has also been implicated in the manifestation of some of the diseases of bacterial (tuberculosis), viral (HIV) and non-viral prion origin (Alzheimers). This review summarize the important pathogenic features of COx enzyme, its protein structure, pathogenic bacteria requiring COx for their virulence in host, and some of its clinical applications. Sanjay Kumar Ojha, Snehasish Mishra, Sarat Kumar Nayak, Mrutyunjay Suar (India) Molecular Biology and Biochemistry for Enhanced Biomethanation ABSTRACT Invited Review: Methanogenic bacteria are one amongst the three classes of Archaebacteria representing the most primitive dwellers of the Earth, reportedly since some 3.5 billion years ago. While their activity is inhibited by oxygen, these bacteria are robust enough to appear in a wide variety of ecological niches, such as, the intestinal tracts of ruminants, sewage digesters, groundwater and deep soil/water. Biomethanation by these is an interesting biotechnology that converts almost all types of organic polymers including the recalcitrant lignocelluloses, to methane and carbon dioxide. This process can be enhanced by manipulating various physical, chemical and molecular factors, though molecular level manipulation needs deeper understandings. Research in genetics, gene regulation and expression of methanogens is rapidly progressing. Relatively proficient genetic manipulation system, including cloning, expression and identification of new species in the last few years is definitely going to provide direction and leads to future investigations. Methyl CoM reductase (MCR), the enzyme responsible for biomethanation, constitutes approximately 10% of the total protein in methanogenic cultures. The significance and abundance of MCR inevitably focused initial attention on elucidating its structure and the mechanisms directing its synthesis and regulation. MCR-coding genes have been cloned and sequenced from various methanogens, though biomethanation process as a whole needs to be further understood and standardised. A plausible solution to biomethanation enhancement at the molecular level seems to lie in metagenomics. The biochemistry and microbiology of anaerobiosis of organic polymers to methane and the roles of the participating microbes are discussed here, along with their molecular biology, application and suggestions for enhanced biogas production. Smita Lata, Smriti Shrivastava, Pratyoosh Shukla (India) An Insight on Recent Advances on Immobilization Methods for Industrial Enzymes and its Relevance to Xylanases

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ABSTRACT Invited Mini-Review: Immobilization is the key for optimizing the operational performance of an enzyme in industry. Immobilization of an enzyme provides greater resistance to changes in physiological conditions such as pH or temperature and leads to the reuse of catalysts, easier reactor operation and separation of products with a wider choice of bioreactors. By virtue of this it provides a cost-effective industrial process. It allows enzymes to be held in place throughout the reaction, following which they are easily separated from the products and can be used again in a far more efficient process and so can be widely used in industries for enzyme-catalyzed reactions. Glycoside hydrolases are enzymes hydrolyzing glycosidic linkages in complex plant polysaccharides and are thus utilized in a number of industries using plant polysaccharides as the raw material. Xylanases are glycoside hydrolases diversely used in various industries including the paper and pulp industry, brewing, animal feed, starch, textiles, etc. The present review describes the state of the art of basic immobilization strategies of enzyme immobilization. We further focus on immobilization of xylanases for commercial applications in various industries. Sarabjeet Singh Ahluwalia (India) Waste Biomaterials for Removal of Heavy Metals An Overview ABSTRACT Invited Mini-Review: Environment fortification must require the use of natural products instead of chemicals to minimize pollution. The detoxification of metal pollutants from wastewater by the development of biomaterials having higher affinity, capacity and selectivity had been found to be as an alternative over conventional technologies that were often inadequate to reduce concentrations in wastewater to acceptable regulatory standards. Plant-derived biomaterials and biomaterials of agricultural by-products for biosorption of heavy metals are an innovative and alternative low cost technology for treatment and/or reuse of waste products. Biotechnological exploitation of biosorption technology to remove heavy metals depends upon the efficiency of the regeneration of the biosorbent after metal desorption. The purpose of this paper is to approach one of the most relevant parts of biotechnology, namely bioremediation with its particular branch biosorption applied for heavy metal removal. In addition to that available information on the various cheaper biomaterials occurring as natural biosorbent, for clean up operation applications along with metal binding mechanism have been highlighted. Santosh Kumar Sethi, Siba Prasad Adhikary (India) Azotobacter: A Plant Growth-Promoting Rhizobacteria Used as Biofertilizer in Organic Farming ABSTRACT Invited Mini-Review: Nitrogen fixation is mainly responsible for improvement of crop yield. In this regard, diazotrophs like Rhizobium, Azotobacter and Azospirillum are important as they enrich nitrogen nutrition in N-deficient soils. Of these, Azotobacter promotes plant growth as well as nitrogen fixation. Thus technology has been developed for making use of Azotobacter biofertilizer for nitrogen and non-nitrogen fixing plants and popularized by educating about their benefits in agriculture to users for practicing integrated nitrogen management. Elsa Marric, Jyotirmayee Mohanta, Bharati Behera, Abhaya Kumar Dalai (India) Effect of Metal Ions on Biphasic Production of Thermostable Amylase by Bacillus sp. Isolated from a Local Hot Spring from Odisha, India ABSTRACT Original Research Paper: The effect of various metal ions on colony growth of five amylase-positive thermo-tolerant Bacillus strains was studied. The metal ions used could be grouped into two categories. One group comprised metal ions that have less than 30% inhibition (Ca2+ and Mg2+) and the second group consisted of metal ions that impart more than 30% inhibition (Pb2+, Ag2+, Cr2+, Zn2+, Hg2+ and Cu2+). However, Mn2+ had an intermediate effect relative to the different strengths of ions used. All strains showed a biphasic pattern of amylase activity with two peaks at 48 and 96 h of culture. To observe the effect of ions on amylase activity at crucial points in the biphasic amylase production curve, i.e. at 48, 72 and 96 h of incubation, two strains, ARBE LlCrg and ARBE UuSs, were selected on the basis of maximum and minimum amylase activity, respectively. Ca2+ and Cu2+ were selected from the two groups of metal ions with respect to their effect on colony growth. The general biphasic trend was marked with Ca2+ and Cu2+ supplementation for both strains, although amylase activity increased with Ca2+ supplementation and decreased in the presence of Cu2+. When starch was added the biphasic trend was more pronounced. However, Ca2+ supplementation blurred the biphasic trend. Sridevi Jagavati, Vimala Rodhe Adivikatla, Nirupama Paritala, Venkateswar Rao Linga (India) Cellulase Production by

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Co-Culture of Trichoderma sp. and Aspergillus sp. under Submerged Fermentation ABSTRACT Original Research Paper: The demand for cellulases is increasing globally because of its potential in the production of cellulosic bioethanol. The major applications of cellulases are in the textile and detergent industries. Cellulases have most promising application in the bioconversion of renewable lignocellulosic biomass into fermentable sugars which can be fermented to ethanol by yeasts. Species of fungi like Trichoderma and Aspergillus are known to be cellulase producers. Fungi of the genus Trichoderma produce relatively large quantities of endo--glucanase (E.C.3.2.1.4) and exo--glucanase (EC.3.2.1.91), but only low levels of -glucosidase (EC.3.2.1.21), while those of the genus Aspergillus produce relatively large quantities of endo--glucanase and -glucosidase with low levels of exo--glucanase production. Furthermore, the -glucosidases of T. reesei are subject to product (glucose) inhibition, whereas, those of Aspergillus species are more glucose-tolerant. Most often, T. reesei cellulase preparations are supplemented with Aspergillus -glucosidase, for cellulose saccharification on an industrial scale. The present investigation aims to demonstrate cellulase production by co-culture of Trichoderma sp. and Aspergillus sp. isolated from a degrading wood source. The results showed an increased filter paper activity of 0.46 U/ml for co-culture of Trichoderma sp. and Aspergillus sp. in the ratio of 1:1, when compared to individual filter paper activities of 0.24 and 0.20 U/ml for Trichoderma sp. and Aspergillus sp., respectively. There was also an increase in CMCase and -glucosidase activities of co-culture, when compared to their monoculture counterparts. The maximum CMCase activity was 13.46 U/ml for co-culture (1:1), when compared to CMCase activities of monocultures, i.e., 8.01 U/ml for Trichoderma sp. and 6.87 U/ml for Aspergillus sp. Maximum -glucosidase activity of 2.02 U/ml was shown by co-culture (1:1). The monocultures of Trichoderma sp. and Aspergillus sp. showed much lower levels of -glucosidase i.e., 0.43 U/ml and 0.98 U/ml respectively. Vimala Rodhe Adivikatla, Sateesh Lanka, Sridevi Jagavati, Venkateswarlu Bandi, Venkateswar Rao Linga (India) A Co-culture Process with Pichia stipitis NCIM 3498 and Thermotolerant Saccharomyces cerevisiae VS3 for Ethanol Production using Acid Hydrolysate of Delignified Sorghum Straw ABSTRACT Original Research Paper: Rising oil prices has attracted the research on bioconversion of lignocellulosic biomass to ethanol as an alternative fuel. Pretreatment of lignocellulosic biomass causes release of fermentable sugars both hexoses and pentoses. An efficient conversion of these two sugars is a prerequisite for a profitable process of bioethanol production from lignocellulose. Considering the approaches available for this conversion, co-culture is a simple process, employing two different organisms for the fermentation of the two sugars. The NaOH delignified sorghum straw released maximum amount of reducing sugars (30.0 g/l) in biphasic dilute acid hydrolysis. The microbial co-cultures of Pichia stipitis NCIM 3498 and thermotolerant Saccharomyces cerevisiae VS3 were employed for efficient bioconversion of mixed sugars present in the hydrolysate into ethanol. The fermentation of detoxified acid hydrolysate with monocultures of P. stipitis, S. cerevisiae VS3 and co-culture produced 10.25 0, 7.40 0.07, and 12.0 0.55 g/L ethanol, respectively. Sravanthi Koti, Sai Prashanti, Jahnavi Gentela, Srilekha Kothagauni, Sridevi Jagavati, Venkateswar Rao Linga (India) Optimization of Pretreatment of Wheat Straw using Alkali and Biphasic Acid Hydrolysis ABSTRACT Original Research Paper: Pretreatment is an important step in the conversion of biomass to biofuels. Alkali pretreatment increases cellulose digestibility and it is the most effective method for lignin solubilization, exhibiting minor cellulose and hemicellulose solubilization compared to acid or hydrothermal processes. The present study describes the potential of pretreatment of wheat straw to degrade lignin and facilitate conversion of polymers to single monomers using alkali treatment followed by biphasic acid hydrolysis. Two alkalis (NaOH and KOH) at various concentrations (0.25, 0.5, 0.75 and 1 M) and at two incubation periods (6 and 12 h at room temperature) were used to delignify wheat straw. 1 M NaOH incubated for 6 h significantly degraded lignin (77%) with minimum sugar loss (1.6%). Further, delignified wheat straw was subjected to two different biphasic acid hydrolyses; firstly, biphasic acid hydrolysis was performed with 2% sulphuric acid at 121C for 1 h followed by 4% sulphuric acid at 121C for 1 h. Secondly, biphasic acid hydrolysis was performed with 3% sulphuric acid followed by 4% sulphuric acid under the same conditions. The conversion of holocellulose (cellulose + hemicellulose) during the above two different biphasic acid hydrolyses was 52 2 and 41 2%, respectively.

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C. U. Agbo, E. C. Okechukwu (Nigeria), Jaime A. Teixeira da Silva (Japan), E. I. Eze (Nigeria) Gongronema latifolia Clones: Genetic Effects on some Phytochemical Composition and Anti-microbial Activity against Salmonella typhi ABSTRACT Original Research Paper: The levels of some phytochemicals (alkaloids, phenols and tannins) and in vitro anti-microbial activity of different genetic resources and plant parts of Gongronema latifolia on Salmonella typhi were determined. There were significant (P = 0.05) differences in genetic effects and plant parts in phytochemical composition and anti-microbial activity against S. typhi. The leaves showed significantly higher phenolic contents (3.81 mg/100 g) than the stems (0.42 mg/100 g). Conversely, the stems contained higher alkaloid and tannin levels (12.50 and 4.67%, respectively) than the leaves (7.80 and 2.99%, respectively). Clone ABS-42-ABA had the highest phenol (4.02 mg/100 g) and alkaloid (13.75%) levels while clone ANS-38-AWKA had the lowest level of phenol (0.24 mg/100 g). The tannin level was significantly higher in clone IMS-20-NJIABA (6.03%) and lower in clone AKS-33-EKPENE EDIENE (2.11%). The leaves expressed significantly higher anti-microbial activity (2.4 mg/ml) against S. typhi than the other plant parts. The antimicrobial activity of the clones on S. typhi was significantly higher in ABS-42-ABA (2.15 mg/ml), and lowest in AKS-33-EKPENE EDIENE (6.43 mg/ml). The phenol concentration had a significantly higher negative correlation with minimum inhibitory concentration (MIC) than other phytochemicals. Similarly, path coefficient analysis indicated that phenol had a higher negative direct effect on MIC. Antibacterial properties shown by the clones and their parts provide a scientific basis and thus validate the use of the plant in treatment of typhoid fever and other bacterial diseases. Also, the varying efficacy by the different genetic resources and the plant parts with respect to phytochemical contents and anti-microbial activity against S. typhi could be the basis for further selection and improvement of the species for pharmacological purposes against typhoid fever. Pushpita Mishra, Sriprakash Mohanty, Mrinal Samanta, Chandi Charan Rath (India) Reestablishment of

Cellulase-Producing Bacteria in the Intestine of Grass Carp (Ctenopharyngodon idella) ABSTRACT Original Research Paper: Cellulase-producing aeorobic bacterial flora in the intestine of phytophagous Chinese grass carp (Ctenopharyngodon idella) were isolated and characterized using selective carboxymethylcellulose agar (CMC-agar) medium. Bacteria were identified and characterized by conventional, biochemical and API kits. Out of 48 bacteria isolated, 26 were cellulase-positive. -glucosidase activity was measured to discover potential cellulase-producing strains. Bacillus, Erwinia and Actinobacillus species were good -glucosidase producers (0.186, 0.181 and 0.180 U/ml/h, respectively). Antibiotic sensitivity tests were conducted for all the isolates against 22 antibiotics based on their spectrum in order to find the most sensitive antibiotic and hence making the intestine devoid of cellulase-producing bacteria and again establishing the potent cellulolytic bacteria by a dip treatment. Antibiogram results revealed that almost all isolates were resistant to clindamycin, erythromycin and metronidazole but sensitive to ciprofloxacin and trimethoprim. This information might contribute to the utilization of these extracellular enzyme-producing bacteria in commercial aquaculture. Subhadip Mahapatra, Debdulal Banerjee (India) Production and Characterization of Thermal Acid Amylase from Aspergillus aculeatus DBF9 ABSTRACT Original Research Paper: Amylase was produced through submerged fermentation of Aspergillus aculeatus DBF9. Initial media pH of 6.0, incubation time of 48 h and an incubation temperature of 30C were found optimum for amylase production. The enzyme was partially purified through acetone precipitation. Amylase activity was found highest at 50C and at pH 5.5. The enzyme was stable within a temperature range of 30-40C and pH 5.0-7.0. Among different metal ions studied, Cu+2 showed maximum induction (46%) of enzyme activity. Significant inhibitory effect of EDTA (80%) on amylase activity was noticed. From SDS-PAGE and native protein gel electrophoresis only one type of amylase was found from Aspergillus aculeatus DBF9. Sama Jagadish Kumar, Goli Jyosthna Khanna, Koppaka Nithya, Linga Venkateswar Rao (India) Production of Xylitol from Alkali Pre-Treated Corn Cobs Hydrolysate ABSTRACT Original Research Paper: Corn cobs consist of cellulose, hemicellulose, lignin and pectin, with about 80% of fermentable sugars of which approximately 48% of sugars are derived from cellulose and 32% sugars derived from hemi cellulose. Both the

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compounds cellulose and hemicellulose are very important as they have wide range of applications in production of various value added products like ethanol and xylitol. When corn cobs (100 g dry weight) were treated with sodium hydroxide in alkali pretreatment method, 43.83 0.13 g (95%) of hemicellulose (contains 70-90% of xylose) and 0.022 0.011 mg/ml of lignin was obtained. During this process, inhibitor compounds like phenolics and furfurals were found to be 0.27 0.01 mg/ml and 0.0031 0.00040 mg/ml, respectively. Hydrolysate obtained by this alkali pretreatment method gave high yield of hemicellulose and less quantity of inhibitors when compared with normal acid pretreatment method. Hemicellulose pellet obtained by alkali pretreatment method yielded 32.83 0.10 g of xylose (90%), which when further, subjected to fermentation with wild and adapted (20 cycles) Candida tropicalis WP, produced 0.63 and 0.69 g xylitol/g xylose, respectively. Ashima Kapoor, Shraddha Sharma, Shveta Prakash (India) Optimization of Culture Conditions for the Production of Lipase from Gliomastix indicus and its Enzymatic Properties ABSTRACT Research Note: Gliomastix indicus is a novel filamentous fungus which was isolated from a wasteland soil sample collected from Tamil Nadu, India. The strain was tested for the production of extracellular lipase by a submerged fermentation technique and was found to produce 10.6 U/ml lipase activity. The culture conditions were optimized for maximum enzyme production. Parameters such as temperature, incubation period, pH, carbon and nitrogen source were studied. Maximum growth and lipase production were observed after 72 h of incubation at 30C in culture medium with an initial pH of 7.5. Among the number of oils tested as a carbon source as well as inducer for lipase production, linseed oil at 1% (v/v) resulted in a marked increase in lipase production. Malt extract at 1% was the best nitrogen source. The optimization of various growth parameters resulted in a 2.62-fold increase in lipase activity (27.8 U/ml). Some properties of lipase which are desirable for its industrial applications were characterized. Crude lipase had broad substrate specificity, exhibiting maximal activity with linseed oil as substrate. The enzyme had an optimum temperature of 45C, with 80% of the maximum activity retained at 45C. Lipase from G. indicus also exhibited appreciable stability in different organic solvents except n-butanol, suggesting that it has potential to be used in the oil and detergent industries. Among the additives, Ca2+ stimulated lipase activity whereas Mn2+ and Hg2+ had an inhibitory effect. Chiliveri Swarupa Rani, Panda Smita Hasini, Linga Venkateswar Rao (India) Isolation and Screening of Polygalacturonase-Producing Bacillus sp. ABSTRACT Research Note: Polygalacturonases (PGases) are the depolymerizing enzymes which cleave the (1,4) glycosidic bonds between two galacturonic acid residues and are being used for the pretreatment of waste water from vegetable food processing industries containing pectinaceous material, processing and degumming of plant fibers such as ramie, sunn hemp, buel, etc. In the present study, 20 bacterial strains were isolated from different samples: soil, spoiled vegetables and fruits, and Godavari River water. All the strains were screened for their ability to utilize pectin as the sole carbon source. Out of the 20 bacterial strains studied, 12 possessed pectinolytic activity as evident from zones of hydrolysis on pectin plates. Selected strains were used for pectinase production by submerged fermentation using 0.25% citrus pectin and the enzyme filtrates were assayed for PGase activity by the DNS method. The effect of temperature, pH on enzyme activity and different media on enzyme production was studied. Maximum enzyme production was shown by strain 11 with medium 1 and optimum PGase activity was found to be at pH 9 at 60C. Based on cultural and morphological studies the strains were preliminarily identified and assigned to the genus Bacillus. Aparajita Majumder, Supradip Sarkar, Subhash Chandra Kole (India) Characterization of Efficient Arsenic-Removing Bacteria from In-vitro Conditions ABSTRACT Research Note: A total of 20 arsenic-tolerant bacterial strains were isolated from arsenic-contaminated soil of eastern India by using basal salt minimal medium (BSMYI) medium supplemented with 150-500 mg l-1 of arsenate (AsV). Among them, 10 isolates exhibited higher arsenic resistance and could grow in up to 12,000 mg l-1 of arsenate (AsV) and 200 mg l-1 of arsenite (AsIII). Those 10 isolates were incubated for 3 days in BSMYI medium containing 25 mg l-1 of both AsV and AsIII separately to assess their ability to remove arsenic. The isolates could remove 1.0-6.4 mg l-1 of AsV and 2.0-7.6 mg l-1 of AsIII from the arsenic-amended media. The bacterial isolate AGH-21 showed the highest arsenic-removing capacity, both for AsV (25.6%) and AsIII (30.4%). The isolated pure culture showed white irregular undulating colonies on a BSMYI agar slant and was an

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encapsulated endospore-forming Gram-positive bacterium. Molecular characterization of the strain based on sequencing of 16SrDNA and subsequent comparison with existing databases identified the organism as Bacillus flexus. SPECIAL ISSUE: Proceedings of Bio-energy from Wastes: Green Chemistry Interventions, 25-26 November, 2010, National Environmental Engineering Research Institute (NEERI)-CSIR, Dept. of Science & Technology, Govt. of India, Nehru Marg, Nagpur, India. Guest Editors: R. A. Pandey and B. K. Sarangi, Environmental Biotechnology Division, National Environmental Engineering Research Institute (NEERI), Nagpur, India ~ 2012 Venkatesh Balan, Leonardo da Costa Sousa, Shishir P.S. Chundawat, James Humpula, Bruce E. Dale (USA) Overview to ammonia pretreatments for lignocellulosic biorefineries ABSTRACT Invited Review: Development of environmentally sustainable and economically viable technologies for plant cell wall deconstruction to fermentable sugars has been impeded due to native plant cell wall recalcitrance to thermochemical and biological based processing. Lower severity alkaline based pretreatments processes like Ammonia Fiber Expansion (AFEXTM) can overcome several limitations of traditional pretreatment approaches (e.g., acidic pretreatments) to producing cellulosic biofuels and biochemicals. Here, we give an overview of chemical reactions taking place during alkaline pretreatments including reactions between ammonia and polysaccharides/lignin (e.g., ammonolysis, hydrolysis and maillard-type reactions). AFEXTM based pretreatments enhance enzymatic digestibility and fermentability of lignocellulosic biomass through various chemical and ultra-structural modifications within the cell wall. An improved mechanistic understanding of the AFEXTM process has led to the development of novel alkaline pretreatments that are briefly discussed in this review. Antje C. Spiess (Germany) Ionic Liquid-Assisted Enzymatic Depolymerisation of Cellulose from Biomass ABSTRACT Invited Review: The recalcitrance of lignocellulose poses a major challenge for its sustainable utilization as source for chemicals, materials and fuels. The capability of some ionic liquids (IL) to dissolve lignocellulose and gain a precipitated amorphous material is exploited within the Cluster of Excellence Tailor-made fuels from biomass (www.fuelcenter.rwth-aachen.de) as an alternative pre-treatment for further (bio)-chemical conversion to fuel components. Based on scattered light intensity measurements (BioLector, Germany) of cellulose suspended in IL, a number of IL capable of dissolving cellulose could be identified. After precipitation from the IL, enzymatic hydrolysis rates and yields are significantly enhanced (~ 20-fold rate and + 10% yield). These results can be partially transferred to wooden biomass. When retaining e.g. 10% (v/v) ionic liquid content in an aqueous system however, the enzymatic activity of commercial cellulase preparations (Celluclast, Novozyme, Denmark) is significantly reduced to between 20 and 30% of its activity in aqueous solution. Ionic strength and viscosity of the IL have been identified as important contributing factors. Interestingly, the enzyme stability was fully maintained. However, the IL interacts differentially for endo- and exo-acting cellulases. The interpretation of these data is facilitated using mathematical models, e.g. those based on population balances (Predici), that allow incorporating both the polymeric nature of the substrate and in case of precipitated cellulose also the particle characteristics of the substrate, i.e. size, crystallinity, and porosity. As a result of the experimental and theoretical studies, improvements for IL-assisted enzymatic hydrolysis should be directed at 1) increasing enzyme activity in presence of IL and 2) tailoring the composition of the enzyme mixture to the resulting cellulosic material using the mathematical models. Madan Junghare, Sanjukta Subudhi, Priyangshu M. Sharma, Ajoy Kumar Mandal, Banwari Lal (India) Mesophilic Hydrogen Production by Clostridium butyricum strain TM-9A, an Alkaline-Tolerant Dark Fermentative Bacterium ABSTRACT Original Research Paper: The present study reports hydrogen production potential by an alkaline-tolerant bacterium Clostridium butyricum strain TM-9A isolated from an estuarine river sediment sample and identified on the basis of 16S rRNA gene sequencing. Different process parameters such as initial pH, temperature and NaCl concentration affected the hydrogen production potential and growth of TM-9A strain in batch dark fermentation experiments. Glucose (10 g L-1) was used as substrate for an optimization study. TM-9A strain was able to tolerate up to 16 g L-1 of NaCl. Strain TM-9A produced maximum hydrogen, 57.8 mmol L-1, at an initial pH 8 under mesophilic conditions, i.e. in the absence of NaCl. Acetic and butyric acid were the major soluble metabolites detected at 12.32 and 11.61 mmol L-1, respectively. Hydrogen yield was 2.02.1 mol H2/mol

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glucose. Furthermore, the strain was also evaluated for its ability to utilize different carbohydrate-rich substrates like molasses (23.44 mmol L-1), corn syrup (25.74 mmol L-1), starch (43.29 mmol L-1), sucrose (31.45 mmol L-1), and cellulose (4.16 mmol L-1), respectively for hydrogen production. Hoysala N. Chanakya, Sreesha Malayil (India) Techno-economic Potential for Value Added Products from Digestion of Urban Solid Wastes and Rural Residues ABSTRACT Original Research Paper: India is urbanizing rapidly accompanied by a gradually increasing rate of municipal solid waste (MSW) production that ranges from 200-500 g/cap/day. The domestic, restaurant, garden and community eatery wastes constitute nearly 90% of the total MSW produced in a city and comprises of >85% decomposable or fermentable components. Decentralized anaerobic digesters are an attractive option that reduces transportation costs and C-footprint but is impeded by difficult to digest components such as banana leaves, stems etc about 15-20% of the total fermentables of USW. This situation requires simultaneous fermentation of banana leaf/stem along with food wastes. Plug flow approaches allow multi-feed digestion without preprocessing and such an option has been tried out for banana leaf. The feasibility and techno-economic potential of multi-product anaerobic digesters using banana leaf is examined. The operation and function of a 50 kg/d plant (equivalent to a large household plant) is described here. The study quantifies recovery of four value-added products (VAP) sensible to this option and describes the processes required, estimates their value and projects commercial viability to finally make a case for decentralized, commercially run zero waste option for this type of waste predominant in south India. Sridharan Jagadeeswari, Manickam Dakshinamoorthy Balakumaran, Padmanabhan Vidya, Pudupalayam Thangavelu Kalaichelvan, Kanesan Panneer Selvam (India) Utilization of Shrimp and Crab Wastes for the Production of N-Acetylglucosamine by Chitinolytic Soil Streptomyces sp. SJKP9 ABSTRACT Research Note: Shrimp, crab and other crustacean shells are the major wastes of sea food processing industries, possessing a rich amount of protein and oligosaccharides such as chitin and chitosan. In this study, shrimp and crab wastes were utilized to produce the amino sugar compound N-acetylglucosamine using chitinolytic Streptomyces sp. SJKP9 isolated from a sea shore soil sample. The chitin flakes obtained from the sources were digested using crude chitinase enzyme derived from Streptomyces sp. SJKP9 and N-acetylglucosamine was precipitated from a concentrated solution of hydrozylate by adding ethanol. Crab chitin produced more N-acetylglucosamine than shrimp chitin. Mithun Raj, Vinayaka Hegde, Muthulekshmi Lajapathy Jeeva, Archana Prathapachandran Vasanthakumari, Pravi Vidyadharan, Vishnu Sukumari Nath, Muthukrishnan Senthil alias Sankar (India) Rapid and Efficient Method for the Extraction of Genomic DNA from Colletotrichum spp. Suitable for PCR Analysis ABSTRACT Techniques Paper: Contemporary approaches for the extraction of genomic DNA from Colletotrichum spp., the filamentous fungal pathogen genera infecting many important crop plants, are often expensive and yield poor quantity and quality of DNA. The high mucilage and polysaccharide content in this fungus add difficulties in genomic DNA isolation, and further downstream applications. We therefore investigated a new and rapid DNA isolation method, which involves inactivation of contaminant proteins by using guanidine hydrochloride/Proteinase K and precipitation of DNA using ice cold isopropanol. This protocol yielded 0.89 0.10 g DNA mg-1 of mycelium with purity ranges from 1.75-2.05 as confirmed by A260/280 spectrophotometric readings. An advantage of this protocol is its compliance even without a refrigerated centrifuge. The new protocol can be successfully used for both mini and maxi preparation of genomic DNA which meet the quality parameters for further downstream processes like PCR-RAPD, AFLP, SSR and ITS amplification of the rDNA-ITS region.

Dynamic Soil, Dynamic Plant

SPECIAL ISSUE: Vermitechnology III. Guest Editor: Natchimuthu Karmegam (Department of Botany, Government Arts College, Tamil Nadu, India) 2012 Takeshi Hirano, Kazuyoshi Tamae (Japan) The Utility of Earthworms as a Monitoring Organism for Soil Pollution

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ABSTRACT Invited Mini-Review: To assess risks to human health from soil contamination, bio-monitoring systems are required. Earthworms are promising candidates as bio-monitoring organisms for soil contamination. However, there have been few studies concerning the utility of earthworms as bio-monitoring organisms. 8-Oxoguanine (8-oxo-Gua), a relatively abundant form of oxidative DNA damage, plays a critical role in carcinogenesis. In our previous study, we found that the levels of 8-oxo-Gua in DNA were increased in cadmium (Cd)-exposed earthworms, suggesting that the analysis of 8-oxo-Gua generated in the DNA of earthworms may be useful for monitoring metal polluted soil. In this review article, we discuss the utility of earthworms as a bio-monitoring organism for soil pollution, with reference to our recent study. Mohammad Javad Zarea, Nasrin Karimi (Iran) Effect of Herbicides on Earthworms ABSTRACT Invited Review: Nature consistently integrates plants and animals into a diverse landscape as part of a major tenet of sustainable agriculture, which is to create and maintain diversity. Chemicals have become the weed control strategy employed most frequently, despite the fact that many important herbicides create water and environment pollution. Earthworms are an important component of the soil system. Earthworms are being increasingly threatened by the excessive application of herbicides to soils. Herbicides can influence earthworms function, growth, reproduction and health. The mortality of earthworms in soils and excessive use of herbicides is still vague. The mortality of earthworms depends on the kind and concentration of herbicide and the duration to which earthworms are exposed to the herbicide. The adoption of conservation tillage has increased worldwide over the past decades. Weeds may become a problem, both in no-tillage systems and in reduced tillage systems. The use of effective herbicides into no-tillage planting systems may provide a feasible option for enhancing weed control, which can become a toxicological risk for invertebrates such as earthworms. This review treats the role of herbicide on the behavior of earthworms. This review will outline the current state of knowledge about fate of earthworm under conservation tillage. Mira Grdisa (Croatia) Therapeutic Properties of Earthworms ABSTRACT Invited Mini-Review: The medical value of earthworms has been known for centuries. This is evident from the history of the ancient Southeastern Asian medicine (China, Japan, Vietnam). The earthworms are the source of proteins, peptides, enzymes and physiologically active substances. Thus, the extracts prepared from the earthworm tissue have been used for the treatment of numerous diseases. Earthworms, like other complex invertebrates, possess several types of leukocytes which synthesize and secrete a variety of immunoprotective molecules. The immunoprotective system is involved in phagocytosis, encapsulation, agglutination, opsonization, clotting and lysis of foreign components. The lytic reactions against several targets are mediated by two major leukocytes, small and large coelomocytes. In the last 10 years a number of earthworms clot-dissolving, lytic and immune-boosting compounds have been isolated and tested in laboratory and clinical studies. In particular, research has been focused on clot-dissolving molecules. Fibrinolytic enzymes, which are regarded as potent and safe, have been purified and studied from several species of earthworms, including Lumbricus rubellas and Eisenia fetida. Its therapeutic and preventive effects on thrombosis-related disease have been clinically confirmed. However, several studies have shown that earthworm extracts contain different macromolecules, which exhibit a variety of activities, such as antioxidative, antibacterial, antiinflammatory, antitumor, etc. Some of these activities are involved in wound healing using an earthworm preparation. Fernando De Len-Gonzlez, Mariela Fuentes Ponce, Fidel Payn-Zelaya (Mexico) Earthworms and Agricultural Systems Management: Emphasis on the Latin American Region ABSTRACT Invited Review: The study of earthworms in Latin America is crucial for promoting the sustainability of this region. In the past 30 years, the tropical forests of Latin America have been exposed to deforestation rates greater than 100,000 ha yr-1 (in the case of Mexico), which will result in the loss of biodiversity of plant species and the micro- and macrofauna of the soil. By biomass, earthworms are the main macrofauna group in tropical forest soils. Although a significant amount of research has been conducted on the ecology of earthworms in the natural ecosystems of the region, there are still large geographical areas that lack adequate descriptions of the diversity of the endemic earthworms. Indeed, there has been an increase in research interest

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with regard to agricultural systems, specifically in relation to earthworm ecology and its connection with agricultural systems that are designed to reduce erosion, increase the organic matter content in the soil and ensure the reproduction of micro- and macro-fauna. However, in this topic the advances in research on earthworms in Latin America have been sporadic. Therefore, this field of research and agricultural development should be steadily promoted. In this review, we outline the further investigation that is required in Latin America with regard to the following topics: (a) the relationship between tillage and cropping systems and earthworm populations, (b) the cumulate effects of pesticides and heavy metals on the earthworm life cycle and (c) the linkage between the organic substrate quality and earthworm dynamics and processing of nutrients. Leticia G. Rodrguez-Canch, Lina Cardoso-Vigueros (Mexico) Vermitechnology: Sustainable Practices in Mexico ABSTRACT Invited Mini-Review: Earthworms remain valid as an object of study and the information generated is diverse. This review summarizes the overall situation of vermitechnology in Mexico, gathering theories that have been offered and techniques that have been developed for urban waste management, agro-industry, manures, toxicity tests, removal of substances such as peptides and soil remediation, as well as other techniques, that show corrective, preventive or improvement actions with environmental, social and economic involvement, promoting sustainable development. Mark Maboeta, Leon van Rensburg (South Africa) Vermicomposting of Industrial Organic Wastes and its Application in Mine Rehabilitation Strategies An Overview from a South African Perspective ABSTRACT Invited Mini-Review: Large areas of land are needed to dispose of wastes produced by mining activities worldwide which poses a myriad of environmental hazards with regards to soil, water and air contamination. This is a problem, especially for a mining country like South Africa, where more than 1500 mines are registered which affects 0.2 million ha of land. Legislation in this country do, however, stipulate that disturbed land should be rehabilitated/revegetated but this is both difficult and expensive because of the unavailability of potential topsoil as well as deficiencies in organic matter, elemental imbalances, and absence of essential nutrients. The use of waste and residual organic matter in soil amendment strategies has been well documented with vermicomposting as a prime example of this. Waste woodchips produced by the platinum mining industry in South Africa have been identified as an organic ameliorant during the rehabilitation of platinum tailings. Based on this, several mines in South Africa, including diamond, iron and platinum, were identified where remediation had to be done on tailings dams. This paper gives an overview of the vermicomposting study undertaken and how successful the use of this product was in real world conditions on different rehabilitation strategies from a South African perspective. The presented studies aim to ascertain the feasibility of utilising vermicomposting of industrial organic wastes and its application in mine rehabilitation strategies under South African conditions. Specific objectives included the evaluating the efficiency of utilising vermicompost on platinum, diamond and iron ore tailings dams by determining rehabilitation success. The results presented in this review are based on these studies; some of which have been published already. Kavitha Balakrishnan, Joseph D. Bagyaraj, Radha D. Kale (India) Microflora in Earthworm Burrow Walls ABSTRACT Invited Mini-Review: Earthworms as ecosystem engineers play an important role in soil ecosystems. The effects of earthworm activity on soil processes differ between ecological categories and species creating distinctive microhabitats for soil microorganisms and other invertebrates. Due to their relatively large size and characteristic feeding behaviour, certain species have a significant impact on soil structure, soil fertility, plant growth and crop yield. This role is achieved inside or in the vicinity of burrows. Since one half of the nitrogenous waste of an earthworm is excreted through the body surface, it presumably accumulates in the burrow walls and affects the soil microbial community in those areas. Moreover, earthworm burrow walls harbour distinctive communities of soil animals like protozoa, nematodes and microarthropods, which presumably control microbial activity in these microhabitats. It is proposed that the most important effect of earthworms on soils may be the stimulations of microbial activity that occurs in casts. This may be the case also with burrows since not all earthworms cast at the soil surface; most species that deposit casts do so in their own burrows. The microorganisms associated with the burrow walls are species specific, different in composition and function and significantly different from soil only a few millimeters away. The present review emphasizes the interaction of microbes and earthworms and the significance of earthworm burrow wall as a hot spot of microbial activity. It elaborates on the types of burrow walls and the microbes associated with it. Further research

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can shed light on the diversity of microbial flora in the burrow wall and surrounding soil. Wael M. Nada (Germany), Leon van Rensburg, Sarina Claassens (South Africa), Oswald Blumenstein (Germany) Vermicomposting as a Sustainable Procedure for the Reduction of Carbon Dioxide Emissions ABSTRACT Original Research Paper: The burning of wood to dispose thereof generates large amounts of carbon dioxide (CO2), one of the critical greenhouse gases. However, vermicomposting of woodchips is a desirable and safe disposal method that requires no combustion and has the additional benefit of being a potential organic ameliorant. This study evaluated the cumulative amount of CO2 produced during vermicomposting of Quercus rubra (QR) and Pinus sylvestris (PS) compared to the amount of CO2 that evolved from burning the wood. The experiment was conducted over a period of 100 days in plastic pots in a greenhouse. The compost temperature, CO2 produced, degree of biodegradation and ash content were measured during the composting period. Also, the amount of CO2 evolved by combustion of wood was determined. The results show that CO2 produced increased over the decomposition period. Higher cumulative amounts of CO2 were produced from QR compared to PS wood compost. The cumulative amount of CO2 produced for both types of wood over the composting period, was lower than that produced by combustion. Ash content, used as an indicator of compost stability, increased over the incubation period; this trend was more pronounced for the QR than the PS wood compost. Prabhat Pramanik, Pil Joo Kim (Republic of South Korea) Effect of Composting and Vermicomposting on Recycling of Three Aquatic Weeds Treated with Rock Phosphate on P-dynamics, Phosphatase Activity and Biomass P ABSTRACT Original Research Paper: In this experiment, three aquatic weeds viz., Lemna, Vallisneria and water hyacinth were mixed with two doses to rock phosphate (RP) i.e., P0 control and P1 200 mg RP per kilogram of waste and these treatments were used for both compost and vermicompost preparation. Results suggested that vermicomposting was faster technique which produced organic amendment comparatively more nutrient enriched than traditional composts. Vermicomposting not only increased phosphorus content of organic substrates, but also increased phosphatase activities and microbial properties, which are responsible for solubilization of insoluble phosphorus. Addition of RP to aquatic weeds significantly (P < 0.05) increased total phosphorus (TP) content of final product. Both composting and vermicomposting increased microbial biomass P (MBP). In both the cases, addition of RP increased MBP content in final product, these along with enhanced phosphatase activity was possibly responsible for higher TP content in these treatments. Natchimuthu Karmegam, Vijayan Karthikeyan, Dhevadas Ambika (India) Vermicomposting of Sugarcane Trash and Leaf Litter in Combination with Pressmud Using the Earthworm, Perionyx ceylanensis ABSTRACT Original Research Paper: In the present study, initial vermicomposting trials were conducted for 100 days using leaf litters of Polyalthia longifolia (LPL) and sugarcane trash (SCT), each in combination with pressmud (PM) (i.e., 0:100, 25:75, 50:50, 75:25 and 100:0) by inoculating an epigeic earthworm, Perionyx ceylanensis. Based on C/N ratio, worm biomass and numbers recorded in vermicomposting trials, organic substrates + PM in 50:50 (1:1) proportion was selected for 60 days vermiconversion studies. Nutrient and microbiological changes during vermicomposting was studied with a special focus on the enzymes in the gut and vermicasts of P. ceylanensis. The NPK contents in worm worked substrates were higher than worm unworked LPL + PM and SCT + PM substrates. The C/N ratio of vermicompost showed decrease over compost (worm-unworked) and percent decrease observed was significant (P < 0.001). The total microbial population (bacteria, fungi and actinomycetes) present at the end of experiment (60th day) was significantly higher (P < 0.05) than the microbial population present at the start of the experiment in both the substrates. Amylase, cellulase, invertase, phosphatase and protease in fresh vermicasts of P. ceylanensis showed higher activities when reared in LPL + PM substrate. The activities of the enzymes cellulase, invertase and protease in vermicasts of P. ceylanensis were significantly higher in LPL + PM. The activity of these enzymes while ageing showed an increase up to 15-20 days and then declined. In the vermicasts recovered from LPL + PM and SCT + PM (1:1), cellulase showed highest activity at the end of 30 days. The present study reveals that the clitellate P. ceylanensis exhibit higher activity of amylase, cellulase, invertase, phosphatase and protease. Keshav Singh, Rabish Chandra Shukla (India) Potential Utilization of Different Wastes through Vermicomposting in

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Agriculture ABSTRACT Original Research Paper: Vermicomposting is an adequate technology for the biooxidation and stabilization of organic material relying on the joint action of earthworms and microorganisms in which organic wastes are converted into rich plant growth media. The aim of this study was to investigate the preparation and potential utilization of vermicomposts (VCs) of different animal dung, agriculture and kitchen wastes and their effect on the growth, flowering period and productivity of three crops namely rice (Oryza sativa), maize (Zea mays) and pearl millet (Pennisetum typhoides). The final VCs obtained from different combinations of three wastes by the help of earthworm Eisenia foetida showed a significant increase in total nitrogen, potassium, phosphorus and calcium and a significant decreased in total organic carbon, the C:N ratio, pH and electrical conductivity in comparison with the initial feed mixture. Among all VCs tested, maximum growth of rice (101.50 cm) and maize (998.90 cm) crop was observed when cow dung + vegetable waste (1: 1, w/w) was used. However, maximum growth of millet (120.20 cm) was recorded in the VCs prepared from buffalo dung + vegetable waste (1: 1, w/w). Rice flowered early (62.76 days) when buffalo dung + gram bran (1: 1, w/w) VCs or the combination of horse dung + rice bran VCs for both maize (82.14 days) and millet (80.18 days) were used. The productivity of rice (0.880 kg/m 2) and maize (0.896 kg/m2) was highest in the cow dung + gram bran and cow dung + vegetable waste VCs, respectively while the productivity of millet was highest (1.92 kg/m2) in buffalo dung + vegetable waste VCs. This study indicates that VCs based on animal dung, agro and kitchen wastes not only produce a value-added product but also act as a better nutrient source for plants. Ishtiyaq Ahmed Najar, Anisa B. Khan (India) Vermicomposting of Fresh Water Macrophytes by Eisenia fetida (Savigny, 1826), Aporrectodea caliginosa trapezoides (Duges, 1828) and Aporrectodea rosea rosea (Savigny, 1826) ABSTRACT Original Research Paper: A study was conducted to evaluate the efficiency of Eisenia fetida, Aporrectodea caliginosa trapezoides and Aporrectodea rosea rosea in recycling of macrophytes (fresh water weeds). E. fetida indicated 100%, A. c. trapezoides 53.66 0.88% and A. r. rosea 33.66 1% recycling potential at the end of a 60-day study period. A significant increase in number (285 7.63%) and biomass (69.16 2.06%) was exhibited by E. fetida compared to A. c. trapezoides (95 7.63% and 11.95 1.12%) and A. r. rosea (45 3% and 8.82 2.53%). Significant variation in cocoon production (P < 0.05) among these species was also observed. Vermicompost obtained by 60 days indicated an increase in potassium (19 0.6 g/g to 37.33 0.90 g/g), available phosphorous (324 4.93 g/g to 600 7.93 g/g) and organic nitrogen (5.53 0.18 g/kg to 8.06 0.17 g/kg), but a decline in organic carbon (576.66 14.52 g/kg to 156.23 8.51 g/kg) and mineralization (C:N ratio) (104.27 5.58 to 19.38 0.14) in the order: E. fetida > A. c. trapezoides > A. r. rosea. Significant variation (P < 0.05) in pH, organic carbon, organic nitrogen, C:N ratio were observed during the same time period. E. fetida has a relatively higher potential of recycling macrophytes among the three earthworm species. Thyagarajan Lakshmi Priya, Rontala Navakoti Uma (India), Subpiramaniyam Sivakumar (Republic of South Korea), Thirumoorthy Meenambal (India), Hyun-Keun Son (Republic of South Korea) Bioremediation of Pulp and Paper Industry Secondary Sludge Spiked with Cow Dung and Effective Microorganisms Using Epigeic Earthworm Eudrilus eugeniae (Kinberg) ABSTRACT Original Research Paper: The aim of this study was to evaluate the vermicompost formed from three different mixing ratios (T1 (1:1), T2 (2:1), T3 (3:1)) of pulp and paper industry secondary sludge with a fixed quantity of saw dust used as a carbon source and spiked with cow dung and effective microorganisms (Rhodopseudomonas, Lactobacillus and Saccharomyces spp.) by using it as an earthworm feed. The epigeic earthworm Eudrilus eugeniae (Kinberg) was used in this study to stabilise the waste under laboratory conditions. Efficacy of the resulting compost in supporting plant growth was also tested with tomato (Lycopersicum esculentum) seeds. At the end of the vermicomposting process, the vermibeds showed a significant decrease in the total organic carbon (10 to 14%) content and increase in total nitrogen (51-68%), phosphorous (170-206%) and potassium (127-180%). The content of heavy metals (Pb, Cd, Ni and Cu) was lower in the final product than in initial feed mixtures. Phytotoxicity tests like germination index and vigour index showed that the resulting composts were less phytotoxic. Among the three treatments, T2 (pulp and paper sludge mixed with sawdust in a ratio of 2:1 and spiked with cow dung and effective microorganisms) showed significantly higher (P < 0.05) seed vigour and germination index than the other two treatments T1 (1:1 ratio) and T3 (3:1 ratio). The vigour index values were: T1 (1225), T2 (1714) and T3 (1228). The order of germination index among three treatments was T2 (77) > T3 (66.5) > T1 (60). Thus the epigeic earthworm E. eugeniae, along with inocula like cow

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dung and effective microorganisms, can easily degrade pulp and paper industry waste. Muthukaruppan Gobi, Paramasamy Gunasekaran (India) Biomanagement of Sugar Factory Pressmud through Vermitechnology for the Growth and Yield of a Pulse Crop, Black Gram (Vigna mungo) ABSTRACT Original Research Paper: Vermicompost (VC) of sugar factory pressmud (PM) using two different species of earthworms, Eudrilus eugeniae and Lampito mauritii was prepared. Plant nutrient contents of these VC and their effects on black gram (Vigna mungo L) growth in relation to the effects of sole soil were investigated. VC produced by the two species of earthworms differed in their nutrient concentrations. But possessed higher concentration of total N and Ca than that of the control. The results showed that effects of VC are more efficient for the vigorous production of black gram. VC had significant positive effects on flower number, leaf growth, shoot length, root length, number of leaves, leaf area index, wet weights and dry weights compared to control. It is also suggested that PM compost is more favorable for vigorous production of black gram and maintenance of soil environment but VC can be economically and environmentally suitable. Kaviraj Singh, Satyawati Sharma (India), Jaime A. Teixeira da Silva (Japan), Santosh Satya, Kalpana Arora (India) Vermicomposting of Municipal Solid Waste Employing Eisenia fetida together with Penicillium spp. and Azotobacter Bioinoculants ABSTRACT Original Research Paper: In the present study, the use of bioinoculants (Azotobacter chroococcum, Penicillium chrysogenum and P. funiculosum) in vermicomposting using an epigeic earthworm, Eisenia fetida, to transform the organic fraction of municipal solid waste (MSW) into valuable vermicompost, was explored. The organic fraction of MSW, when mixed with leaf litter (4:1), was treated with bioinoculants and processed for use as a vermicompost. Various biochemical parameters such as total organic carbon (TOC), total Kjeldhal nitrogen (TKN), EC, pH, available phosphorus (P), available potassium (K), cellulose, hemicelluloses and lignin were analyzed. The number of earthworms and the percentage nitrogen, P and K in vermicompost increased while pH and TOC declined as a function of the vermicomposting period. A 14-60% increase in TKN was observed in different microbial combinations at the end of the vermicomposting period. Available P increased 1.4- to 9.5-fold in different feed mixtures in comparison to the control. TOC was most reduced in the E. fetida treatment inoculated with a combination of Azotobacter chroococcum and Penicillium chrysogenum (37.2%) followed by A. chroococcum (36.1%) and A. chroococcum + P. funiculosum (33.8%). Analysis of three enzymes (-glucosidase, phosphatase and urease) also showed better results in the E. fetida treatment together with all microbial combination than the control. -glucosidase showed a significant increase (51%) while urease decreased sharply during the process. Vermicomposting using E. fetida + bioinoculants (A. chroococcum + P. chrysogenum) is a suitable technology for the decomposition of different types of organic wastes (domestic and industrial) into value-added material. Gorakh Nath, Keshav Singh (India) Combination of Vermiwash and Biopesticides against Aphid (Lipaphis erysimi) Infestation and their Effect on Growth and Yield of Mustard (Brassica campestris) ABSTRACT Original Research Paper: Vermiwash (VW), singly and in combination with different biopesticides (BPs), was used in an agricultural field to control aphid (Lipaphis erysimi) infestation and to increase the growth and yield of mustard (Brassica campestris). A significant reduction in the aphid population was observed on mustard plants after spraying VW obtained from different combinations of animal dung and agro-wastes with 95% neem (Azadirachta indica) oil and 86% custard apple (Annona squamosa) leaves. The combination of garlic (Allium sativum) extract with different VWs could control 97% of the aphid population. VW obtained from animal dung + gram bran with neem oil was also very effective against aphids. A VW spray with BP increased the productivity of the mustard crop up to 3.5 times more than the control. These results clearly demonstrate that the use of VW with plant products is more beneficial for organic farming. It also compensates for deficiencies in essential nutrients, certain plant growth hormones, enzymes and vitamins. VW, singly and in combination with plant products, provides effective control against aphids, which are injurious to mustard plants. Sohela Akhter, Ranjit Sen, Shahana Akter (Bangladesh), Jaime A. Teixeira da Silva (Japan), Azizul Haque, Shamsun Noor (Bangladesh) Efficacy of Vermicompost to Improve Soil Health, Yield and Nutrient Uptake of Cauliflower in Grey Terrace

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Soil of Bangladesh ABSTRACT Original Research Paper: Vermicompost (VC) is a potential input for sustainable agriculture. This paper reports on the ability of VC to efficiently increase the growth, yield and nutrient uptake of cauliflower (Brassica oleracea var. botrytis) and to improve soil health. Field trials were conducted with cauliflower Snow White during 2007-2008 in Grey Terrace Soil (Inceptisol) of Bangladesh, Agro Ecological Zone (AEZ)-28. There were 12 treatments replicated three times. The sources of VC were cowdung and kitchen wastes (3: 1) processed by epigeic earthworm Eisenia fetida. Leaf number, circumference and curd yield of cauliflower were significantly higher when NPKSZnB fertilizers (100% recommended dose of chemical fertilizer, RDCF) were used together with 1.5 t ha-1 VC but were statistically identical to 100% RDCF + 1.5 t ha-1 aerobic compost (AC), NPKSZnB (80% RDCF) + 3 t ha-1 VC and 80% RDCF + 3 t ha-1 AC. VC performed better than AC alone or in combination with chemical fertilizers. In this case, enhanced cauliflower yield was attributed to the elevated levels of NPKSZnB in VC. There was a considerable increase in nutrient uptake by VC-treated cauliflower. The residual effect of VC showed an increase in available nutrients in post-harvest soil. To improve and maintain soil health and crop production, chemical fertilizers need to be reduced. VC (1.5 t ha-1) + 100% RDCF favours higher curd yield of cauliflower but VC (3 t ha-1) + 80% RDCF may be economically and environmentally suitable since it contains 20% less chemical fertilizer and 1.5 t ha-1 more organic manure. Hence, 3 t ha-1 VC + 80% RDCF is recommended for cauliflower cultivation in Grey Terrace Soils of Bangladesh. M. Vikram Reddy, Swati Pattnaik, Sushree Sangeeta (India) Microarthropod Diversity Associated with Vermicomposting Process and Vermicompost of Urban Waste Processed by Three Earthworm Species ABSTRACT Original Research Paper: The present investigation assessed, for the first time, the species diversity of microarthropods associated with the vermicompost (VC) and compost (C) of the organic fraction of three types of urban wastes municipal solid waste (MSW), vegetable market waste (MW) and flower waste (FW) processed by three earthworm species, Eudrilus eugeniae, Eisenia fetida and Perionyx excavatus during the vermicomposting process. The data showed the diversity and abundance of different taxa of microarthropods Acarina (mites) such as Mesostigmata, Prostigmata and Cryptostigmata, and Collembola (springtails) such as Isotomidae, Entomobryidae and Sminthuridae and other miscellaneous microarthropod taxa. Microarthropods were extracted from the VCs, Cs and the substrate samples, MSW, MW and FW using a Tullgren funnel. Dry funnel extractions were examined microscopically, revealing a total of 16 species and morpho-species of Acarina, 19 species of Collembola and 12 species of miscellaneous microarthropods. Collembola was the dominant group followed by Acarina. Among Collembola, Isotomidae dominated while Mesostigmata was the dominant group of Acarina. Species belonging to Formicidae in Hymenoptera were also found in good numbers. The biodiversity of microarthropods was significantly higher in the VCs of all three substrates produced by E. eugeniae followed by that of E. fetida and P. excavatus and that of Cs (P < 0.05). Positive and significant co-relationships were found between microarthropod densities and nutrients nitrogen, phosphorus, potassium, calcium and magnesium of VCs (P < 0.05). Prostigmata and Mesostigmata densities showed positive and significant co-relationship with each other and with the densities of Collembola Isotomidae, Entomobryidae and Sminthuridae (P < 0.05). Interestingly, there was no relationship with the densities of Cryptostigmata and miscellaneous groups. Subhani Rath, M. Vikram Reddy (India) Disposal of Municipal Solid Waste (MSW) of the Millennium City (Cuttack): Bioconversion to Nutrient Rich Vermicompost ABSTRACT Original Research Paper: Vermicomposting is one of the effective ways of disposal and management of municipal organic solid waste (MOSW), and is a process of composting of organic waste by using either or both exotic and indigenous species of earthworms. These worms devour organic wastes rapidly and produce a good quality end-product called vermicompost that becomes enriched with various plant nutrients (available nitrogen, potassium, phosphorus, calcium and magnesium), has more organic matter, and is more porous than soil. This paper deals with the disposal of MOSW of millennium city, Cuttack through vermicomposting using the exotic species Eisenia fetida. However, MOSW vermicompost possessed reduced concentrations of organic carbon, but a higher concentration of P, and K. Moreover, a comparative analysis was made between the major nutrients of the MOSW vermicompost and agricultural waste (AW); vermicompost from AW is better than vermicompost from MOSW.

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Debajyoti Chakrabarty, Sanjib Kumar Das (India) Dissolution of Phosphorus from Rock Phosphate using Earthworms (Eudrillus euginae) ABSTRACT Short Communication: Earthworms (Eudrillus euginae) were used to test their effects in the dissolution of phosphorus (P) from insoluble, naturally occurring, Purulia rock phosphate (PRP) through vermicomposting of two different substrates for 90 days. The same substrates (partially decomposed water hyacinth and cow dung with or without rock phosphate) were also allowed to decompose without earthworm (controls). The substrates were monitored for concentration of P, nitrogen and potash and changes in pH. The observations showed that the treatment combination with earthworm and rock phosphate (RP) was the best among four treatments employed in terms of P and N release. There were significant differences in concentration of available phosphorus among all the treatments and K concentration was not affected by the treatments. The findings suggest that the earthworms facilitated the dissolution of P from RP which affects mineralization of N in the substrates.

Floriculture and Ornamental Biotechnology

Iftikhar Ahmad (Pakistan/USA), Muhammad Hussnain Shah (Pakistan) Trichoderma and Intercropping Impact Production, Quality and Corm Rot Disease of Gladiolus grandiflorus ABSTRACT Original Research Paper: The effects of Trichoderma harzianum Rifai application and intercropping with Gazania rigens L. (treasure flower) or Calendula officinalis L. (pot marigold), members of the Asteraceae family, were studied on growth, yield, quality and controlling corm rot disease of gladiolus. Two experiments were conducted; first using healthy corms of two gladiolus cultivars, Fado and Cantate and second on both healthy and artificially infected (with Fusarium oxysporum) corms of cultivar, Fado in order to compare the individual treatment effects. The addition of Trichoderma to the medium increased growth, production and quality as well as reduced disease incidence when applied to healthy corms, although intercropping had a similar effect. When individual treatment effects were evaluated, use of both Trichoderma and intercrops, applied to infected corms, significantly improved plant height (65%), foliage growth (122%), reduced crop harvest time (23%), and disease (corm rot) incidence (78%), increased leaf area (70%), total leaf chlorophyll contents (162%), number of florets (71%), stem length (69%), and stem diameter (68%), and cormel diameter (158%) and were similar to those of grown from healthy corms. However, when both Trichoderma and intercropping were applied individually to healthy corms, the results were statistically similar. In both experiments, plants treated with Trichoderma and intercropped had up to 67% less corm rot disease on average than control (plants grown without Trichoderma and intercropping). In summary, Trichoderma application to soil and intercropping with members of the Asteraceae family can be effective for not only improving growth, yield and quality of cut gladiolus, but also controlling corm rot disease of gladiolus. Moreover, intercropped species can also be a source of additional income to the growers. Mohamed Elimem, Brahim Chermiti (Tunisia) Color Preference of Frankliniella occidentalis (Pergande) (Thysanoptera; Thripidae) and Orius sp. (Hemiptera; Anthocorridae) Populations on Two Rose Varieties ABSTRACT Original Research Paper: A study of the preferential choice of Frankliniella occidentalis Pergande revealed the presence of a phototropism of this pest towards the petal color of its host plant. The color of roses may influence infestation rates by Western Flower thrips. The white-cream rose variety Ociane attracted more individuals of F. occidentalis than the red variety First-red in the same greenhouse; 29.54 and 39.85 thrips/flower on Ociane while only 12.40 and 29.59 thrips/flower recorded on First-red on May 31st and June 14th 2007, respectively. In fact, those differences were highly significant almost throughout the entire study period. The predatory bug Orius sp. also showed a similar preference for the Ociane than First-red, and thus followed the distribution of its prey.

Food
Chih-Cheng Lin, Lu-Te Chuang, Yuan-Chen Wang, Yan-Hong Tang, Jia-Shang Liao (Taiwan), Boakye Amoako-Atta (Ghana), Robert H. Glew, Robert S. Glew (USA) Polyphenol and Antioxidant Content of Kigelia africana Leaves from Ghana

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ABSTRACT Original Research Paper: In this study we investigate the polyphenol and antioxidant content of Kigelia africana leaves from Ghana, West Africa. K. africana is a semi-deciduous tree that grows wild and its leaves and fruit are used as food and medicine by local residents. The aims of this study are to compare the content and tentatively identify the major polyphenols in methanolic (aq.) versus aqueous extracts of K. africana leaf and the antioxidant activity of these extracts. The polyphenol content of the methanolic (aq.) (20/80, v/v) extract of Kigelia was 1.3-fold greater than that of the aqueous extract and 2-fold greater than that of the methanolic (aq.) extract (80%) of spinach, Chinese cabbage or lettuce. According to the results of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, the methanolic (aq.) extract of K. africana leaves contained 6-fold more antioxidant activity than similar extracts of spinach, cabbage or lettuce. Of these four green leafy vegetables, the methanolic (aq.) extract of K. africana was the most potent in reducing NO production by LPS-stimulated macrophages (RAW 264.7) in culture. HPLC analysis showed that the dominant phenolic compounds in K. africana leaf were ellagic acid and caffeic acid (491 and 70 mg/100 g dry weight, respectively). In conclusion, this study demonstrates that the leaves of K. africana contain large amounts of polyphenolic compounds and antioxidants. These findings provide a basis for encouraging efforts to conserve this endangered species and promote further studies on its potential nutritional benefits to populations in sub-Saharan Africa. Hayam M. Ibrahim (Egypt) Lipid and Color Stability as Affected by Combination of Sodium Ascorbate and -Tocopherol Acetate in Minced Buffalo Meat During Refrigerated Storage ABSTRACT Original Research Paper: The target of this study was to evaluate the inhibition of oxidative changes of minced buffalo meat during refrigerated storage at 4C for 6 days by adding a combination of vitamin C and E salts at two different blend levels on lipid and color stability as well as some other quality parameters. A significant difference was observed between the percentage antioxidant activity as a result of these suggested additions. The minced buffalo meat sample B blended with 600-mg/l sodium ascorbate + 5 mg/l -tocopherol acetate exhibited higher antioxidant activity (P < 0.05) than sample A blended with 400 mg/l sodium ascorbate + 10 mg/l -tocopherol acetate during the storage period. Both the suggested blend levels, especially in sample B, can help to minimizing TBARS and met-myoglobin accumulation during the storage period. Also, the same sample at the end of the storage period had higher redness (a*) color values, more acceptable visual color, moderately pleasant odor score and lower cooking loss compared to other samples. Thus, a combination of 600 mg/l sodium ascorbate + 5 mg/l -tocopherol acetate could be utilized effectively for enhancing the shelf life of minced buffalo meat. Murlidhar Meghwal, T. K. Goswami (India) Effect of Moisture Content on the Physical and Textural Properties of Fenugreek Seed ABSTRACT Original Research Paper: The fenugreek seed (Trigonella foenum-graecum) exhibits marked unpredictability in major engineering properties (physical properties) owing to its varying moisture content (mc). Hence, fenugreek seeds were evaluated for changes in various physical properties viz. seed geometry (length, width, thickness and equivalent diameter), sphericity, roundness, specific surface area, surface area, seed volume, 1000-seed-mass, densities (true and bulk), porosity, angle of repose, specific gravity, coefficient of static friction on different surfaces, terminal velocity, aspect ratio as a function of mc. Some of the textural properties such as hardness, fracturability, cohesiveness and gumminess were also measured. The mc of the seeds was varied from 7.6-20.0% on dry weight basis (db). The physical dimensions i.e., seed length, width, thickness, equivalent diameter, sphericity, volume, seed roundness, specific gravity, surface area, true density, porosity, 1000 seed mass, angle of repose increased with increase in mc, while reverse was true for specific surface area and aspect ratio. Positive linear relationship was observed between terminal velocity and mc. Textural characteristics viz., seed hardness, fracturability and gumminess showed a declining trend with increasing mc, whereas cohesiveness increased with increase in the mc. Abiodun Aderoju Adeola, Ogugua Charles Aworh (Nigeria) A Comparative Evaluation of the Chemical Properties of Wild Tamarind (Tamarindus indica L.) Fruits in Nigeria ABSTRACT Original Research Paper: A comparative study was carried out to evaluate the chemical properties of mature wild tamarind fruits in Nigeria. Samples of the fruits were collected from 19 major towns in the savannah vegetation of Nigeria. Fruit pulp was

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hand-scraped from the seeds and separated from other non-pulp materials. Proximate composition, physicochemical properties, ascorbic acid and mineral composition, total carotenoids and antinutritional factors of mature tamarind fruit pulp were analysed. Proximate and mineral compositions were expressed as g/100 g fresh weight (FW) of the fruit pulp, the ascorbic acid and total carotenoids as mg/100 g FW and g/100 g FW, respectively; the colour as optical density at a wavelength of 325 nm and total acidity as tartaric acid. Moisture content of the mature fruits ranged between 16.8 and 36.2%. The crude protein, crude fat, ash and total crude carbohydrate of the mature tamarind fruit pulp ranged from 3.5 to 7.4, 3.5 to 7.4, 3.0 to 6.9 and 52.0 to 62.7, respectively. The ascorbic acid, colour and soluble solids varied between 3.7 and 11.3, 0.30 and 1.42, and 5.2 and 6.3 oBrix, respectively. The mature tamarind fruits were high in acid (pH 2.3-3.3) but low in total carotenoids, antinutrients and micronutrients. Calcium and sodium were the most abundant macro- and micro-nutrients. Tamarind fruits are good sources of nutrients and could be effectively employed in combating food insecurity in developing sub-Saharan countries where tamarind fruits grow. Jean-Paul K. M. Bouatenin, Theodore N. Djeni, Solange Aka, Kouakou Brou, K. M. Dje (Cte DIvoire) The Contribution of Microorganisms to the Fermentation of Cassava Dough during Attik Processing in Cte dIvoire ABSTRACT Original Research Paper: Biochemical properties of fermentative microorganisms of attik traditional starters were evaluated in vitro for highlighting the contribution of each of them in cassava dough fermentation and selection of potential starter strains. A total of 345 isolates from three types of inocula (Adjoukrou, Alladjan and Ebri) were screening for their abilities to produce -amylase, -glucosidase, pectinases and their acidification rate. Lactic acid bacteria (LAB) and at a lesser extent Bacillus sp. have been recognized as high acidifying microorganisms. Yeasts isolates were most involved in -amylase (47.56%) production. But, LAB were the most involved in the detoxication of cassava by the high rate of isolates producing -glucosidase (34.66%), while Bacillus sp showed high pectinases producing isolates (43.75%). A proportion of 7 to 10% of moulds contributes to the production of amylase, linamarase and pectinase. A total of 42 strains with one or more desirable biochemical properties were pre-selected for the development as starter cultures for attik production. Marina Christelle Nanouman Assohoun, Thodore NNd Djni, Florent Kouadio NGuessan, Marina Koussemon (Cte DIvoire) Preliminary Study on Antimicrobial Properties of Lactic Acid Bacteria Involved in the Fermentation of Corn Dough during Doklu Processing in Cte DIvoire ABSTRACT Original Research Paper: This study aimed to assess the antimicrobial activities of lactic acid bacteria (LAB) isolated from corn dough for doklu production during spontaneous fermentation. Fermentation was conducted for 10 days by mixing corn dough with water at room temperature and biochemical analysis and LAB isolation was performed every 2 days. LAB isolated were then tested for potential antimicrobial activity. Biochemical analyses revealed the presence of 3 main organic acids in corn dough: tartaric acid, oxalic acid and lactic acid. Moreover, pH decreased while titratable acidity increased. Corn dough contains a diversity of LAB including homo- and heterofermentatives. From the 109 isolates, 83.33% showed the ability to inhibit the growth of a pathogen bacterium, Staphylococcus aureus ATCC 25915 with diameters of inhibition ranging from 10.4 0.55 mm to 16 mm. Twenty six of these positive LAB produced inhibitory substances which could be bacteriocins with a large spectrum of action against several human pathogens.

Fresh Produce Fruit, Vegetable and Cereal Science and Biotechnology

SPECIAL ISSUE: Sweet potato. Guest Editors: M. Nedunchezhiyan, G. Byju (Regional Centre of Central Tuber Crops Research Institute, Orissa, India) ~ 2012 Rajasekhara Rao Korada, Samir Kanti Naskar, Archana Mukherjee, Cheruvandasseri Arumughan Jayaprakas (India) Management of Sweet Potato Weevil, Cylas formicarius: A World Review ABSTRACT Invited Review: Sweet potato is infested by many insect pests. Sweet potato weevil (SPW) Cylas formicarius (Fab.) is the

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important insect pest throughout the world, wherever it is grown. The weevil is managed by a package of practices together called integrated pest management (IPM). In India, a few genotypes of sweet potato have shown durable resistance throughout 2006 to 2011. A new screening method of germplasm, volatile-assisted selection (VAS), was developed to identify resistance/susceptibility in sweet potato genotypes based on the volatile chemicals that are released. Transgenic sweet potato was not successful at the field level. Farmers in Asia practice intercropping of sweet potato with ginger, bhendi, taro and yams to reduce the incidence of pests as well as to conserve soil moisture. Entomopathogenic fungi and nematodes are used successfully to control C. formicarius in the West and Latin America. Female sex pheromone (Z)-3-dodecen-1-ol (E)-2-butenoate has changed the pest dynamics in the field and has become an important tool in C. formicarius IPM. It was used to monitor and trap male weevils, thus reducing the reproductive success of female weevils. A number of botanical pesticides are available and their use is limited in developed countries. A few insecticides that were used to control C. formicarius were banned in recent years in many countries and it is essential to identify new molecules with low or no persistence in tubers and soil with toxic effects on weevils. We reviewed the research done on SPW during the last five decades to assess the management practices of SPW and to identify new strategies required to control the pest effectively and economically.

Functional Plant Science and Biotechnology

SPECIAL ISSUE: Sugarcane pathology. Guest Editors: A. Ramesh Sundar, Rasappa Viswanathan (Sugarcane Breeding Institute, Coimbatore, India) ~ 2012 Asha Dookun-Saumtally, Salem Saumtally, Nawshad Joomun (Mauritius) Biotechnological Applications to Sugarcane Pathogens ABSTRACT Invited Review: Sugarcane (Saccharum spp) is an important cash crop accounting for nearly two thirds of the world sugar production. Much effort has and is still being devoted to breeding resistant varieties, as the crop is vulnerable to a number of diseases that affect production. These constraints have triggered the adoption of biotechnological tools to counteract the impact of diseases on sugarcane. The increasing use of biotechnology is enabling early detection of pathogens of fungal, viral, bacterial and phytoplasma origin, allowing preventive measures to be adopted much earlier, before the appearance of disease symptoms as well as enabling the increased use of clean seed for establishment of nurseries and for the exchange of germplasm. The strategy of developing transgenic sugarcane varieties for resistance to several diseases is now well established and several transgenic lines are being evaluated in the field in a number of countries, although no commercial transgenic cane is grown thus far. Recent advances in sugarcane genomics as well as the genomics of sugarcane pathogens will soon provide information on disease resistance genes enabling new measures to be adopted in the breeding for disease resistance. Sandy Snyman, Tania van Antwerpen, Sumita Ramgareeb, Sharon McFarlane (South Africa) Sugarcane Pathogens: Detection and Elimination through in Vitro Culture ABSTRACT Invited Mini-Review: Commercial sugarcane cultivation, which involves: (a) multiple ratoons in the field for several years, (b) propagation by means of vegetative stem cuttings, (c) large areas under sugarcane monoculture, (d) production in tropical and subtropical climates, can exacerbate pathogen infestation and the spread of diseases. This paper reviews important diseases in sugarcane, methodology for detection of pathogens and highlights several interventions for pathogen elimination such as hot water treatment of stem sections and in vitro culture using meristematic cells as the initiation material. These have ensured a multi-faceted approach for disease management and enabled the production of material without pathogens such as Leifsonia xyli subsp. xyli, Xanthomonas albilineans, sugarcane mosaic virus and sugarcane yellow leaf virus. Additionally, international germplasm exchange may benefit by incorporating aspects of in vitro culture at various stage of the process. Govind P. Rao, Smriti Mall (India), Carmine Marcone (Italy) Recent Biotechnological Approaches in Diagnosis and Management of Sugarcane Phytoplasma Diseases ABSTRACT

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Invited Review: Phytoplasmas have been reported to be associated with two major sugarcane diseases viz., grassy shoot disease (GSD) and white leaf disease (WLD). Sugarcane yellow leaf syndrome is a new syndrome of sugarcane caused by a luteovirus and a phytoplasma. Sugarcane green grassy shoot (SCGGS) and ramu stunt are also associated with sugarcane from Thailand and Papua New Guinea, respectively. GSD and WLD are causing significant economic losses to sugarcane yield and sugar recovery in Asian countries. Both these phytoplasmas have been spreading very rapidly to newer locations with the help of infected seed material and leafhopper vectors. Hence it would be important to diagnose and manage these phytoplasmas at an early stage of sugarcane growth to avoid further spread and significant losses caused by them. Because of unreliable and unspecific symptoms, the identification and characterization of the associated phytoplasma at an early stage of plant growth is problematic and unreliable. The introduction of molecular genetic methods into plant mycoplasmology about 15 years ago greatly improved the diagnosis of phytoplasma infections in plant and insect hosts. PCR offers several advantages over other methods including versatility, relative simplicity, specificity and high sensitivity, which can be increased by a two-step PCR (nested PCR). It has also become possible to differentiate, characterize and classify the phytoplasmas on a phylogenetic basis, using mainly sequence analysis of ribosomal DNA (rDNA). The biotechnological and molecular assays developed in recent years have been utilized at a large scale to characterize these pathogens at an early stage of growth and for screening planting seed materials for possible elimination and management of theses phytoplasma at an early stage of their growth and propagation. We summarize research being conducted on characterization and management of these phytoplasma by novel biotechnological and molecular approaches. Rasappa Viswanathan, Muthusamy Poongothai, Palaniyandi Malathi, Amalraj R. Sundar (India) Sugarcane Wilt: New Insights into the Pathogen Identity, Variability and Pathogenicity ABSTRACT Invited Review: Wilt of sugarcane, a fungal disease is known to cause significant damage to sugarcane production and productivity in India and other countries for the past one century. Although sugarcane wilt is known in India for a long time, research work on this important disease is totally lacking. The causal organism was found to vary with time and investigator and could not reproduce the disease under artificial conditions in the field. We have made detailed disease surveys in 13 major sugarcane growing states in the country and 263 Fusarium isolates were isolated. We have established the variation in Fusarium isolates associated with sugarcane wilt, based on cultural, morphological, pathogenic and molecular characterization of 117 isolates. Critical observations in the conventional techniques combined with molecular biological approaches clearly established that Fusarium sacchari as the causal agent of the disease. Other Fusarium sp isolated from wilt infected sugarcane stalks were found to be either secondary invaders or non-pathogenic in nature. We have developed an artificial simulation technique to induce wilt in sugarcane under field conditions and efficient management strategy through biocontrol was also developed. Rasappa Viswanathan (India) Molecular Basis of Red Rot Resistance in Sugarcane ABSTRACT Invited Review: Red rot of sugarcane caused by Colletotrichum falcatum Went is one of the devastating diseases of sugarcane causing significant loss to sugarcane production in India and other Asian countries. Complex polyploidy and lack of information on inheritance to red rot in sugarcane make breeding for red rot resistance more difficult. Hence researchers have studied the mechanism of red rot resistance in sugarcane in detail. Initial studies based on biochemical tools identified oxidative enzymes and red rot pigments in disease resistance. Further studies revealed the role of pathogenesis-related (PR) proteins and 3-deoxyanthocyanidin phytoalexins especially luteolinidin and apigeninidin in red rot resistance. Recent studies using semiquantitative RT-PCR after pathogen inoculation from sugarcane cultivars varying in red rot resistance, revealed differential accumulation of transcripts of the flavanoid biosynthetic pathway like coumarate-4-hydroxylase, chalcone synthase, chalcone reductase, flavanoid 3-5 hydroxylase and flavanoid glycosyl transferase and this transcript analysis, further confirmed the role of sugarcane phytoalexins in red rot resistance. Similarly the role of PR- proteins like chitinase and -1,3-glucanase was established at the transcript level. Detailed molecular studies using differential display (DD)-RT-PCR identified expression of more number of differentially expressed transcripts during the host-pathogen interaction. Full length sequences of many potential transcripts were identified and are being characterized. Also to identify specific proteins involved in host resistance, proteomic approach has been attempted by optimizing sample preparation from stalk tissues, 2-D electrophoresis (2-DE), down stream processing of identified spots and bioinformatics. Several resistance associated proteins spots were identified and they are being analyzed critically. Overall, application of molecular techniques was found to be useful in identifying

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transcripts/proteins involved in host defence. Further studies are in progress to validate their specific involvement in red rot resistance. Durairaj Mohanraj, Purushotham Padmanaban, Rasappa Viswanathan (India) Screening for Red Rot Resistance in Sugarcane ABSTRACT Invited Review: Red rot of sugarcane caused by Colletotrichum falcatum Went is a serious fungal disease affecting sugarcane stalks, the economical part of sugarcane in different countries. In India, the disease has been a menace for sugarcane cultivation and many elite varieties were removed from cultivation due to their susceptibility to the disease. The commercial varieties are recommended for cultivation in India if they possess red rot resistance. Hence screening for red rot resistance became an integral part of varietal development in the country. Plug method of pathogen inoculation is the most common method used for red rot screening in sugarcane and here, severity score is being assigned based on the red rot symptoms inside the canes. The key parameters of the lesion such as lesion width, nodal transgression and nature and severity of white spots determine the disease severity. Since a bore-hole injury is caused on the stalk to inoculate the pathogen, there is an apprehension that natural barrier is breached to introduce the pathogen in this method. So many other methods were developed to facilitate the pathogen to enter the cane tissues in a natural way. However the disease reaction in these methods is influenced by environmental factors which lead to disease escapes under field conditions. Recently a controlled condition testing was developed to screen large number of progenies using cane tops in a short time. Overall, the plug method is used to assess resistance potential of a genotype in a foolproof way in sugarcane varieties proposed for commercial cultivation and the controlled condition testing is followed initially to screen large population. Amalraj Ramesh Sundar, Nagarathinam Selvaraj, Muthumeena Muthiah, Ashwin Ramadass, Palaniyandi Malathi, Rasappa Viswanathan (India) Induced Resistance A Potential Supplementary Strategy for the Management of Red Rot in Sugarcane ABSTRACT Invited Review: Induced resistance (IR) is the phenomenon by which a plant exhibits an increased level of resistance without changes of its basic genetic constitution. IR utilizes the plants own defense mechanisms to restrict pathogen development. Resistance can be induced locally on the pretreated plants of the plant as well as systemically on untreated parts and is then called systemic acquired resistance (SAR). Several chemicals such as salicyclic acid, acibenzolar-S-methyl, isonicotinic acid, jasmonic acid and others are known to induce resistance in certain plants, but only a few of them fulfil the criteria for being real plant activating compounds. The introduction of novel synthetic signal molecules is thought to be the first step for the integration of the principle of IR in a modern environmentally friendly plant protection concept. Plant diseases account for a considerable proportion of crop loss worldwide. In sugarcane, red rot disease caused by the fungus Colletotrichum falcatum is one of the important pathogens causing significant loss to the growers and the sugar industry. Many of the commercially superior varieties are out of cultivation because of either high levels of red rot susceptibility or breakdown of resistance, which is attributed to the emergence of new virulent pathotypes. In addition to breeding for durable red rot resistance, other viable options need to be explored for successful management of the disease. In this background, exploitation of the inherent SAR potential of highly successful commercial varieties, otherwise susceptible to red rot is a viable option to manage the disease. With the advent of tools of biotechnology, it has now become possible to understand better the molecular basis of IR and to regulate defense genes for augmenting disease resistance in crop plants. This review attempts to comprehensively resent all the available information on inducing disease resistance in sugarcane, with a special focus on red rot disease. Porntip Wongkaew (Thailand) Sugarcane White Leaf Disease Characterization, Diagnosis Development, and Control Strategies ABSTRACT Invited Mini-Review: Sugarcane white leaf (SCWL) is the most destructive disease of sugarcane in Thailand where major cane and sugar production of the world are situated. The severity degree and symptom variance are depended on soil fertility, temperature, cane sette quality, cultural practice and the phloem-colonized phytoplasma amount. The leafhopper Matsumuratettix hiriglyphicus is the first known vector that transmits SCWL phytoplasma. Recently at least other six leafhoppers have been found containing SCWL phytoplasma DNA. However only Yamatotettix flavovitatus has been closely

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investigate and found its transmissibility but with lesser extent. Outbreak of SCWL disease is mainly activated by transportation and propagation of endemic cane setts. This SCWL phytoplasma is closely related to Thailand sugarcane green grassy shoot (SCGGS) and India sugarcane grassy shoot (SCGS) with 96-98% similarity in their 16S-23S rDNA sequences. Some diversity among SCWL phytoplasma isolates from different sources and growing locations has also been indicated within the range of 89-98% similarity by the 16S-23S rDNA sequence analyses. In addition to existed conventional diagnosis methods, the electrochemical DNA sensors have been proposed as a new tool for reliable routine practice. At present, control of the SCWL disease is difficult due to is systemic nature and the lack of desirable resistant variety. Application of insecticide is ineffective either. The disease control measures thus are mostly emphasized on the other strategies such as physical and chemical treatment of the cane setts, disease free plant production via tissue cultures, sanitation and crop rotations, and regulatory quarantine. Cooperative assistance among growers and involving association is strongly needed to accomplish effective control. Mara Blanch, Mara-Estrella Legaz, Carlos Vicente (Spain) Structure and Biosynthesis of a Xanthan-like Polysaccharide Produced by Xanthomonas albilineans ABSTRACT Invited Mini-Review: Leaf scald is a vascular disease of sugarcane plants caused by Xanthomonas albilineans. Scalded leaves show white-yellowish streaks alternating with green zones in parallel to the main veins. The white-yellowish streaks show both phloem and xylem completely occluded by a gum and the overall mesophyll appears to be full of this bacterial secretion. The gum in conducting tissues has been purified from juices obtained from scalded stalks and it was identified as a xanthan-like polysaccharide composed by repeated tetrameric units containing two rests of fructose, one of mannose and one of glucuronic acid. Hydrolysis of xanthan with selective mannosidases and -1,4-glucanases reveals that the macromolecule consists of a linear, -1,4-backbone of -glucose units to which mannose in -1,3 bonds is linked. Since xanthans contain glucuronic acid, the ability of Xanthomonas to produce an active UDP glucose dehydrogenase is often seen as a virulence factor. X. albilineans produces a UDP-glucose dehydrogenase growing on sucrose. The enzyme oxidizes UDP-glucose to UDP-glucuronic acid by using molecular oxygen and NADPH. The value of pI of the purified enzyme is 8.98 and its molecular mass has been estimated as about 14 kDa. The enzyme is inhibited by UDP-glucose concentrations higher than 1.3 mM X. albilineans cultured axenically does not secrete xanthan to liquid media but it is produced by bacteria growing in sugarcane tissues. This host-dependence can be explained on the basis of the action of bacterial proteases upon the dehydrogenase. Glycoproteins from sugarcane, the natural host of the bacterium, produced as a response to the bacterial invasion assure the production of the active enzyme by inhibiting bacterial proteases. Jean Heinrich Daugrois, Rosiane Boisne-Noc (Guadeloupe F.W.I.), Patrice Champoiseau (Guadeloupe F.W.I./USA), Philippe Rott (France) The Revisited Infection Cycle of Xanthomonas albilineans, the Causal Agent of Leaf Scald of Sugarcane ABSTRACT Invited Mini-Review: Leaf scald, a sugarcane disease caused by Xanthomonas albilineans, has been largely studied since its discovery in 1911. Numerous data were obtained and reported, from isolation, transmission and diagnosis of the pathogen to the complete sequence of its genome. Although X. albilineans was thought to be exclusively transmitted by infected cuttings and contaminated cutting implements for more than 80 years, the existence of an epiphytic life of this pathogen was reported in the late 1990s. We highlight herein the role of the epiphytic populations of X. albilineans in plant and field contamination and the impact of environmental conditions, especially rainfall, on these populations of the pathogen. Data obtained from experimental research in Guadeloupe showed the capacity of X. albilineans, when rainfall is sufficient, to spread by aerial means within the sugarcane canopy in a field or between fields. This new epidemiological trait helps X. albilineans in contaminating rapidly healthy sugarcane plants and, in certain geographical locations, it can be much more important than disease spread by cutting implements. These new epidemiological data of sugarcane leaf scald lead us to propose a revisited infection cycle for X. albilineans. Mara Francisca Perera, Mara Paula Filippone, Aldo Sergio Noguera, Mara Ins Cuenya, Atilio Pedro Castagnaro (Argentina) An Overview of the Sugarcane Mosaic Disease in South America ABSTRACT

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Invited Review: Sugarcane mosaic, one of the most important viral diseases of sugarcane, is widely distributed in the world and its economic significance varies among regions. Economic losses depend on varietal susceptibility, virus strains, its interaction with other diseases, vector population and environmental conditions. Although not a major problem in some countries, sugarcane mosaic has caused substantial yield losses in other countries (Argentina, Brazil) due to severe outbreaks. Numerous strains of Sugarcane mosaic virus (SCMV) and Sorghum mosaic virus (SrMV) are commonly associated with mosaic symptoms. Both viruses are members of the SCMV subgroup in the genus Potyvirus of the family Potyviridae and their genetic variability could be effectively assessed only through DNA sequence comparisons. The greater genetic variability of viruses associated with sugarcane mosaic needs to be taken into consideration in breeding and biotechnology programmes for resistance to mosaic. The most effective way to control sugarcane mosaic has been through the use of resistant cultivars, which requires a complete understanding of the genetic diversity of the pathogens as well as their interaction with cultivars; resistance breakdown can occur when new strains or viruses appear. However, the production of healthy and genetically pure seed cane could be an available tool to reduce the pathogenic load in sugar cane growing areas. This could be achieved through hydro-heat-treatment followed by apical meristem in vitro culture and micropropagation. It is also relevant to implement extreme quarantine measures to prevent the entry of new pathogens or variants of the established ones through germplasm exchange. Marcos Cesar Gonalves, Luciana Rossini Pinto, Silvana Creste Souza, Marcos Guimares A. Landell (Brazil) Virus Diseases of Sugarcane. A Constant Challenge to Sugarcane Breeding in Brazil ABSTRACT Invited Review: Sugarcane mosaic virus (SCMV) and Sugarcane yellow leaf virus (SCYLV) are the two main viruses infecting sugarcane in Brazil. The authors analyze the emergence and current status of these pathogens and what has being done to minimize losses and prevent new outbreaks in sugarcane. It was shown that SCMV is the only causal agent of mosaic in sugarcane in the country. Most sugarcane cultivars are believed to be tolerant or intermediate resistant to mosaic, although new cases have been reported in the field. One recently characterized SCMV isolate comprises a novel severe strain capable of infecting cultivars previously considered to be resistant. SCYLV, the causal agent of sugarcane yellow leaf, is widely distributed in Brazil and other sugarcane producing countries causing significant yield losses. The virus became widespread in the field and in parental clones used in sugarcane breeding programmes. Sensitive and reliable detection methods for SCYLV were developed and have been routinely applied for diagnosis while screening for resistance, virus elimination for germplasm exchange, and production of virus-free seed cane. Screening for resistance to sugarcane mosaic and sugarcane yellow leaf is considered a primary and essential step in sugarcane breeding programmes in Brazil. Trinh Xuan Hoat, Dang Vu Thi Thanh (Viet Nam), Matthew Dickinson (UK), Ngo Gia Bon, Mai Van Quan, Vu Duy Hien, Nguyen Duc Thanh, Le Thanh Thuy, Ngo Vinh Vien (Viet Nam) Disease Problems of Sugarcane in Vietnam, with Special Reference to Phytoplasma ABSTRACT Invited Mini-Review: Seventeen diseases of sugarcane have been found in Vietnam including 13 fungal, 2 bacterial and 2 phytoplasmal diseases. Among them, the most important diseases are smut, pineapple, red rot, sugarcane white leaf and sugarcane grassy shoot diseases. We applied nested PCR using universal primers for detection and characterization of phytoplasmas from infected tissues. PCR products of the expected size (1200bp) were obtained from the 16S rRNA of the phytoplasma. The RFLP profiles indicated that all the samples were infected by the same pathogen. Nucleotide sequence analysis of the 16S rRNA genes revealed that the phytoplasma causing sugarcane grassy shoot disease in Vietnam is very similar to the phytoplasmas causing sugarcane grassy shoot disease in India sharing a sequence similarity of 99%. Phylogenetic trees showed that SCGS and SCWL strains collected from Vietnam and other reported SCGSs belong to the 16SrXI subgroup RYD (Rice Yellow Dwarf) 16S rRNA Group. Lilik Koesmihartono Putra, Tri Asmira Damayanti (Indonesia) Major Diseases Affecting Sugarcane Production in Indonesia ABSTRACT Invited Mini-Review: Although a large number of sugarcane pathogens have been recorded in Indonesia, the current major sugarcane diseases affecting sugar production in Indonesia are ratoon stunting disease, smut, mosaic and leaf scorch. A new type of mosaic disease caused by Sugarcane streak mosaic virus is now widely distributed in most commercial sugarcane areas in Java (Indonesia). An integrated management strategy is used for controlling the disease and minimizing their impact

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on sugar production. Heather McCafferty, Yun J. Zhu (USA) Transgenic Approach to Improve the Resistance of Sugarcane to Sugarcane Yellow Leaf Virus (SCYLV) ABSTRACT Invited Mini-Review: Genetic transformation of sugarcane using various methods has been reported. In this review, an overview of the sugarcane yellow leaf virus (SCYLV) will be presented. The technical approaches used to create transgenic sugarcane through transformation, selection and regeneration will be reviewed. The genetic engineering methods for sugarcane include Agrobacterium-mediated transformation, electroporation and particle bombardment. In particular, work in Hawaii and Florida with constructs conferring resistance to sugarcane yellow leaf virus will be highlighted. Finally, thoughts about the future of sugarcane transformation will be discussed. Ana Carolina Wanderley-Nogueira, Ederson Akio Kido, Nina da Mota Soares-Cavalcanti, Luis Carlos Belarmino, Joo Pacfico Bezerra-Neto, William Lee Burnquist, Sabrina Moutinho Chabregas, Jos Ivo Baldani, Ana Maria Benko-Iseppon (Brazil) Insight on Pathogen Defense Mechanisms in the Sugarcane Transcriptome ABSTRACT Original Research Paper: Recognition of pathogen and activation of defense mechanisms is a common feature known from all multicellular organisms. Among higher plants, systemic acquired resistance (SAR) is known to activate pathogenesis-related (PR) genes after recognition of the pathogen mediated by a resistance (R) gene. Both gene classes (R and PR) represent the main mechanism against biotic and sometimes also abiotic stresses. Therefore, the identification of SAR metabolism components is an important concern regarding plant breeding. Sugarcane (Saccharum officinarum) is a major tropical and subtropical crop, grown in more than 120 countries, being especially important due to its value for the production of bioethanol, constituting a renewable energy source, besides the sugar production. The present work brings an overview on sugarcane resistance and pathogenesis-related genes, regarding their structure, abundance and role in the plant-pathogen metabolic pathway and also regarding their distribution, as compared with rice. For this purpose a collection of 282,818 expressed sequences tags have been evaluated to identify R and PR genes as well as important factors identified in the classic plant-pathogen metabolic KEGG pathway using rice full length cDNA as seed-sequences. The identified sugarcane genes have been also used to screen four SuperSAGE libraries with 8,787,313 tags, allowing the identification of the main activated and repressed genes under abiotic stress (drought/salinity) conditions. The 1,460 identified genes have been plotted on a rice virtual karyotype inferring about their distribution, considering a putative synteny as a measure to infer about their relative position within rice and sugarcane chromosomes. The results revealed interesting insights on the variability and complexity of defense genes in sugarcane. SPECIAL ISSUE 2: Protease inhibitors. Guest Editor: Raj Deepika (Forestry and Agricultural Biotechnology Institute (FABI), Department of Genetics, Faculty of Natural and Agricultural Sciences, University of Pretoria, South Africa) ~ 2012 Tzi Bun Ng, Randy Chi Fai Cheung, Jack Ho Wong, Xiu Juan Ye (China) Protease Inhibitors of Botanical Origin ABSTRACT Invited Review: The intent of this article is to review recent literature on plant protease inhibitors which can be divided into different types comprising Kunitz, Bowman-Birk, squash and potato inhibitor types. Some of them specifically inhibit trypsin. Others also inhibit chymotrypsin and other proteases. The mode of inhibition can be competitive or noncompetitive. Some of them are characterized by remarkable thermostability and pH stability. In addition to protease inhibitory activity, antifungal, anti-insect, anticancer and /or HIV-1 reverse transcriptase inhibitory activity are present in some plant protease inhibitors. Vishnu Menon, Mala Rao (India) Protease Inhibitors: Emphasizing Functional Aspects of Aspartic Protease Inhibitors ABSTRACT Invited Review: Aspartic proteases are relatively a small group of proteolytic enzymes. Over the last decade, they have received tremendous research interest as potential targets for pharmaceutical intervention as many have been shown to play

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significant roles in physiological and pathological processes. Despite numerous efforts, however, the only inhibitors for aspartic proteases currently in the market are directed against the HIV protease of viral origin. Nevertheless, several inhibitors including those targeting renin-angiotensin system and -secretase are in clinical or preclinical developments and few other aspartic proteases are discussed as potential drug targets. Currently the research strategies are focusing on the need for improved comprehension of protease-regulated cascades, along with precise selection of targets and improved inhibitor specificity. There is plethora of synthetic inhibitory compounds targeting aspartic proteases; however there are few reports documented in literature on biologic inhibitors from microorganisms. Protease inhibitors (PIs) are widely distributed in the plant kingdom. One of the important defense strategies that are found in plants to combat predators involves PIs which are particularly effective against phytophagous insects and microorganisms. In plants, these PIs act as anti-metabolic proteins, which interfere with the digestive process of insects. The defensive capabilities of PIs rely on inhibition of proteases present in insect guts or secreted by microorganisms, causing a reduction in the availability of amino acids necessary for their growth and development. The present chapter is a comprehensive state-of-the-art review describing the aspartic protease inhibitors from microbial and plant origin. In addition, the chapter highlights the therapeutic perspectives of aspartic protease inhibitors and biocontrol aspects of protease inhibitors with special emphasis on aspartic protease inhibitors. Anussorn Wisessing, Kiattawee Choowongkomon (Thailand) Amylase Inhibitors in Plants: Structures, Functions and Applications ABSTRACT Invited Review: The -amylase inhibitors have been purified and characterized from several plant species. Six classes of -amylase inhibitors have been classified by their structures. They play important roles in the control of endogenous amylases as well as the protection against pathogens and pests. The purification methods, classification, function, inhibition mechanism together with their applications in agricultural, clinical and industrial processes have been discussed in this review. Vaijayanti A. Tamhane, Manasi Mishra, Neha S. Mahajan, Vidya S. Gupta, Ashok P. Giri (India) Plant Pin-II Family Proteinase Inhibitors: Structural and Functional Diversity ABSTRACT Invited Review: Potato inhibitor II (Pin-II) proteins are plant serine proteinase inhibitors (PIs) that occur in various plant species. Pin-II PIs are characterized by inhibitory repeat domains (IRDs) which form their functional units and show extensive sequence variation. Various studies have been conducted to gauge the occurrence (spatial and temporal) of Pin-II PIs and their appearance in response to biotic stresses mainly herbivore attack. Many hypotheses have been proposed to justify the mechanism which has led to the evolution of Pin-II PIs as well as the selection pressure in force. In spite of ample diversity, these molecules are highlighted by their conserved features with respect to their gene and protein sequence and structure. The structural studies highlight the crucial role of conserved residues in stabilizing the reactive loops and overall the three-dimensional conformation of Pin-II PIs. The remarkable flexibility of reactive loops allows their binding to a wide range of proteinases (either endogenous? or from the pest). Apart from defense, Pin-II PIs have also been speculated to have a significant role in endogenous functions, namely regulation of proteolysis, macromolecular trafficking, programmed cell death and consequently aid the plant growth and development in respective tissue. There have been attempts to test these candidates for their potential in insect control. In vitro experiments and insect bioassays at laboratory scale have given encouraging results and led to the field experiments in order to develop transgenic plants fortified with Pin-II PIs. This has also helped judging the fitness costs that usually transgenic palnts have to pay in return of incorporating the foreign trait. Tantravahi Srinivasan, P. B. Kirti (India) Protease Inhibitors and Stress Tolerance ABSTRACT Invited Review: Plant protease inhibitors (PIs) are extensively studied for their role in defense against pests and pathogens due to their ability to inhibit specific proteases of the intruder. All classes of PIs were found to play important roles in innate host defense mechanisms and are used to develop transgenic plants resistant to pests and pathogens. Recent reports suggest that they have a role in modulating abiotic stress tolerance also. The induction of PIs was observed in response to not only biotic but also abiotic cues suggesting their possible role in abiotic stress responses and in modulating tolerance to various stresses. During biotic stress, pathogen invasion or damage due to herbivory, induces SA/JA- mediated signalling pathways to combat the attack. Abiotic stress induces injury and different conditions like osmotic stress, oxidative stress etc. as long as the

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causative factor exists in the milieu. Unlike in the biotic stresses, the causative agent in abiotic stress cannot be countered by the plant. Hence, the abiotic stress has to be tolerated by the plant nullifying all its related damages mostly by adjustment through ABA-dependent, and sometimes through an independent pathway. The overexpression of PIs has been shown to enhance abiotic stress tolerance in the transgenic plants suggesting that their role is multidimensional. In this review, we focus on the possible role of PIs in plant growth, and biotic and abiotic stress tolerance. Yaxi Liu (China), Jirui Wang (China/USA) Molecular Evolution of Exogenous Alpha-amylase Inhibitors in Triticeae - An Update ABSTRACT Invited Review: Exogenous -amylase inhibitors in seeds and vegetative organs are attractive candidates for the control of seed weevils as these insects are highly dependent on starch as an energy source. In cereal seeds, -amylase inhibitors proteins were known as one great family on the basis of the homology of their sequences. The overall information on the monomeric, dimeric and tetrameric -amylase inhibitors strongly support the view that these inhibitors have evolved from a common ancestor gene through duplication and mutation. All -amylase inhibitors from Triticeae (21 different genomes) have 10 conserved cysteine (5 disulfide bonds) sharing common cysteine skeleton. In the past years the -amylase inhibitors have been well studied, including large amount of attention that directed towards their evolutionary relationship. Adaptive evolution of monomeric and dimeric -amylase inhibitor genes and its ecological association has been investigated in wild emmer wheat. They were contributed by both natural selection and co-evolution, which ensures the conserved function as well as the inhibition of a variety of insect amylases. The known aspects of -amylase inhibitors and their molecular evolution in Triticeae have been discussed in this review. Tzi Bun Ng, Randy Chi Fai Cheung, Jack Ho Wong, Xiu Juan Ye (China) Isolation and Characterization of Protease Inhibitors from Animal Tissues ABSTRACT Invited Mini-Review: The intent of this article is to review protease inhibitors produced from a diversity of animals including cnidarians, annelids, insects, crustaceans, mollusks, fish, amphibians, reptiles and mammals. They can be isolated from various tissues including pancreas, ova, liver, seminal plasma, serum, venom, skin secretion, hemolymph and salivary glands. Animal protease inhibitors display a variety of molecular masses ranging from several to over 50 kilodaltons. Some of them possess different protease specificity and inhibitory constants toward proteases. They play a role in various physiological processes such as reproduction, protection from viral infection, regulation of endogenous and exogenous (pathogen) proteases and anticoagulation activity. Marri Swathi, Soundappan S. Mohanraj, Chilka Srinivas, Elaprolu R. Prasad, Vantaku Venkatrao, Nalini Mallikarjuna, Aparna Dutta-Gupta, Kollipara Padmasree (India) Cajanus cajan (cv. C11): An Important Non-Host Resource for Proteinase Inhibitors Active against Larval Midgut Proteinases of Lepidopteran Insect Pest Achaea janata ABSTRACT Original Research Paper: Our previous studies indicated that seeds of various cultivars and wild relatives of pigeonpea possessed proteinase inhibitors (PIs) which were active against midgut trypsin-like proteinases of lepidopteran insects. In the present study a detailed analysis of PIs distribution in mature seeds of five different cultivars of Cajanus cajan and thirteen different accessions of Cajanus platycarpus was carried out. Further the inhibitory potential of these PIs against larval midgut trypsin-like proteinases of Achaea janata was also evaluated. Among the chosen cultivars and C. platycarpus accessions, the cultivar ICP 7118 also called as C11 showed highest inhibitory potential against A. janata larval midgut trypsin-like proteinases (AJPs). The inhibitory potential of PIs from C11 was 25 and 106 fold higher against AJPs, compared to bovine pancreatic trypsin and chymotrypsin, respectively. Further, the activity profile of PIs of C11 on gelatin-polyacrylamide gel demonstrated the presence of higher number of trypsin inhibitor isoforms as well as stronger inhibitory potential against AJPs, when compared with C. platycarpus accessions. Thus, the results from the present investigation confirmed that the cultivar C11 of C. cajan is the best non-host resource for PIs active against AJPs and it can be readily exploited further as ecofriendly pesticide in the management of this non-host lepidopteran insect pest A. janata using transgenic technology. Amarjit K. Nath, Kanika Chadha, Pratima Sharma (India), Raj Deepika (South Africa) Purification and Characterization of a

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Novel Inhibitor from Poinciana pulcherrima Seeds with Activity towards Pest Digestive Enzymes ABSTRACT Original Research Paper: Six tree legumes were analysed for trypsin inhibitor (TI) activity and inhibition of trypsin enzyme extracted from the Pieris brassicae larvae midgut. TI extracted from Poinciana pulcherrima seeds contained maximum TI activity (trypsin units inhibited (TUI) = 17.24) and it also inhibited gut TI of P. brassicae larvae to the maximum extent (TUI = 16.61); minimum inhibition of gut TI was observed in seeds of Poinciana regia (TUI = 2.46). The inhibitor was purified to 50.98-fold with 66.46% recovery by ammonium sulphate precipitation and gel filtration chromatography through a Sephadex G-100 column: a single band was observed in native-PAGE. The molecular weight of the purified protein was 17,782 Da as determined by SDS-PAGE and it was a monomer. The purified TI was heat stable over a wide range of temperatures (20-60C) and had a pH optimum of 7.5. The purified TI hence appears to be of the Bowman-Birk type that lost its activity when treated with different concentrations of 2-mercaptoethanol, indicating the role of disulphide linkages in maintaining its three-dimensional structure. The inhibition was non-competitive and had a Ki value of 0.6 M, indicating the high affinity of the inhibitor towards its substrate, BApNA. The purified inhibitor suppressed the growth of P. brassicae larvae, judged solely on the basis of the reduction in size and on the weight of excreta. The inhibitor also inhibited trypsin enzyme extracted from the mid-gut of Spodoptera littoralis larvae (36.3 TUI/30 g of purified inhibitor). The plant genes expressing such TIs can be isolated, cloned and introduced in vegetable crops, thereby conferring resistance and minimizing the devastating crop yield losses caused by various insect pests and pathogens. Huma Habib, Mohammad Afzal Zargar, Khalid Majid Fazili (India) Cloning and Expression of Trypsin Inhibitor Gene ti from Pea (Pisum sativum L.) cv. Arkel in Escherichia coli DH5 Cells ABSTRACT Original Research Paper: Protease inhibitors (PIs) play key regulatory roles in many biological processes. The single gene advantage associated with these inhibitors make them ideal candidates for gene transfer to produce pest resistant recombinant plants. The aqueous extracts from pea (Pisum sativum) displayed prominent trypsin inhibitory activity. For cloning of trypsin inhibitor gene (ti) into bacterial hosts, cDNA was first prepared from the RNA isolated from pea seedlings. Amplification of the Ti gene was carried out using two sets of primers, the 5-primer contained EcoRI restriction sequence, and the 3- primer contained HindIII restriction sites. The purified amplicons were cloned into pet 27b+ expression vector using EcoRI and HindIII restriction enzymes. The constructed vectors were transformed into Escherichia coli DH5 cells. The recombinant cells were grown in LB medium containing kanamycin and treated with IPTG to induce the expression of the cloned Ti gene. The expression profile of the cells revealed highly intense 12.6 kDa band in the induced samples. The gel print technique and Dot blot assay revealed that the protein showed significant inhibitory activity towards trypsin and enzyme assay with synthetic substrates showed that the protein caused 90 3% inhibition of trypsin. Taqi Ahmed Khan, Mohd Mazid (India), Jaime A. Teixeira da Silva (Japan), Firoz Mohammad (India), Mohd Nasir Khan (Kingdom of Saudi Arabia) Role of NO-Mediated H2O2 Signaling under Abiotic Stress (Heavy Metal)-Induced Oxidative Stress in Plants: An Overview ABSTRACT Review: Environmental stress, like heavy metal, constitutes the most significant factor leading to a substantial and unpredictable decrease in crop yield in agriculture. Hydrogen peroxide (H2O2) and nitrogen oxide (NO) are produced as primary signals in a stress signal cascade. H2O2 accumulation can lead to either enhanced expression of antioxidant enzymes or increased expression of other defense proteins. Alternatively, it can initiate programmed cell death, particularly when NO is also produced, depending on the intensity of the oxidative signal or oxidative load exerted by heavy metal toxicity in tissues. In this review, we examine the regulatory role of H2O2 and NO signaling in oxidative stress induced by heavy metals in plants, exemplified by a number of research studies. Finally, H2O2 and NO signaling in response to heavy metals may provide some clues to improving crop productivity.

Genes, Genomes and Genomics

Shibin Mohanan Nellikunnumal, Arun Chandrashekar (India) Computational Identification of Conserved MicroRNA and their Targets in Coffea canephora by EST Analysis

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ABSTRACT Original Research Paper: Comparative genomic tools have been successfully used to predict new miRNAs in different plant and animal species using expressed sequence tag (EST) and genome survey sequence (GSS) analysis. In the present study we identified conserved microRNAs in Coffea canephora identified by EST analysis using a new modified comparative genomics method. Use of this method eliminates false positives to a greater extent. Conserved microRNA belonging to 12 families was identified. All identified miRNA were used to search their potential target genes from the SGN (Solanaceace Genome Network) EST database and a total of 42 potential targets were identified for miRNA families from C. canephora. Most of the miRNA targets were transcription factors which appeared to be involved in plant growth, development and stress responses according to the mRNA target information provided by NCBI. The newly identified microRNA can help in better understanding of growth and development Coffea. The improved method can be used for more accurate prediction of microRNA from other plant species with sufficient EST database.

International Journal of Biomedical and Pharmaceutical Sciences

Arumugam Madeswaran, Muthuswamy Umamaheswari, Kuppusamy Asokkumar, Thirumalaisamy Sivashanmugam, Varadharajan Subhadradevi, Puliyath Jagannath (India) Docking Studies of Aldose Reductase Inhibitory Activity of Commercially Available Flavonoids ABSTRACT Original Research Paper: Molecular docking is a frequently used tool in computer-aided structure-based rational drug design. Flavonoids are a group of natural products which exhibits various biological and pharmacological activities. The primary objective of this study was to investigate the aldose reductase inhibitory activity of flavonoids using in silico docking studies. In this perspective, flavonoids like aromadedrin, eriodictyol, homoeriodictyol, isorhamnetin, okanin, pachypodol, peonidin, robinetin, tangeritin were selected. Epalrestat, a known aldose reductase inhibitor was used as the standard. In silico docking studies were carried out using AutoDock 4.2, based on the Lamarckian genetic algorithm principle. The interacting residues within the complex model and their contact types were identified. In the docking studies, three important parameters like binding energy, inhibition constant and intermolecular energy were determined. The results showed that all the selected flavonoids showed binding energy ranging between -9.20 kcal/mol to -8.02 kcal/mol when compared with that of the standard (-8.73 kcal/mol). Inhibition constant (181.13 nM to 1.32 M) and intermolecular energy (-10.99 kcal/mol to -9.81 kcal/mol) of the flavonoids also coincide with the binding energy. All the selected flavonoids contributed aldose reductase inhibitory activity because of its structural properties. These molecular docking analyses could lead to the further development of potent aldose reductase inhibitors for the treatment of diabetes. Further investigations on the above compounds and in vivo studies are necessary to develop potential chemical entities for the prevention and treatment of diabetes. Nitin Dubey, Ghanshyam Patil, Dinesh Kumar Jain, Subhash Chaturvedi (India) Simultaneous Estimation of Torsemide and Spironolactone in Combined Dosage Form Using Reverse Phase Liquid Chromatography ABSTRACT Techniques Paper: Simple, accurate, precise, and sensitive reversed-phase high-performance liquid chromatographic (RP-HPLC) method for simultaneous estimation of Spironolactone (SPR) and Torsemide (TOR) in combined tablet dosage form has been developed and validated. Beers law was obeyed in the concentration range of 5-25 g/mL for both drugs in methanol. The RP-HPLC method uses a Shimadzu LC 10 ATVP system with a Luna C18 column and methanol: acetonitrile: phosphate buffer, pH 3.5(60:20:20 %v/v) as the mobile phase. The detection was carried out using a diode array detector set at 238 nm. The recoveries were found to be in the range of 99.64 0.04 to 100.75 0.15 and 99.56 0.35 to 100.33 0.56 for TOR and SPR, respectively. Developed method was found to be simple, precise, sensitive and may be used for routine analysis of TOR and SPR in a pharmaceutical formulation. Results of analysis were validated statistically per ICH guidelines. Harpreet Walia, Saroj Arora (India) Delineation of Antioxidant Activity of Acetone Extract/Fractions of Fruits of Terminalia chebula Using TA 102 Strain of Salmonella typhimurium ABSTRACT Research Note: Antimutagenic or antioxidant properties elicited by plant species have many prospective applications in human

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healthcare. The fruits of Terminalia chebula are a rich source of tannins and other bioactive constituents. In Ayurveda, the plant is known as the wonder healer due to its extraordinary power of healing. In the present study, the Ames Salmonella histidine reversion assay was used to assess the antioxidant activity of the acetone extract of T. chebula fruits in the TA 102 strain of Salmonella typhimurium using hydrogen peroxide (H2O2) as a direct acting oxidant. The acetone extract was prepared by maceration and further fractionated with ethyl acetate and water. The antioxidant activity of ethyl acetate and water fractions was comparatively higher than that of the crude extract. The ethyl acetate fraction inhibited reduction of H2O2 by 72.48 and 72.83% in co and pre-incubation modes, respectively although there was no difference in co- and pre-incubation modes of experimentation.

International Journal of Plant Breeding

Sunita Singh, Srinivasulu K. Reddy, Narendra Jawali (India) Genetic Diversity Analyses of Mungbean (Vigna radiata [L]. Wilczek) by ISSR ABSTRACT Original Research Paper: In the present study 3 anchored ISSR primers were designed, and after optimizing each primer annealing temperature for obtaining a higher number of markers per primer, they were used to analyze genetic diversity among 87 mungbean genotypes from India and neighboring countries. Following screening the 24 ISSR primers, 8 primers that yielded a clear banding pattern were chosen. Out of a total of 145 bands, 135 were polymorphic. An average of 18.12 bands and 16.87 polymorphic bands per primer was obtained. A dendogram revealed clustering of genotypes into 5 clusters. The first two clusters individually included the genotypes developed at two of the agricultural research institutes in India. Cluster three included genotypes developed in and around India, and cluster 4 included the genotypes that were local collections of mungbean from India and the fifth cluster included exotic and old selections of mungbean Analyses showed that both resolving power of primer and number of polymorphic bands showed a positive correlation with the number of genotypes identified. Mukesh Singh, Rajat Kanti Chawdhari, Ila Chawdhari (India) Assessment of Genetic Diversity among Elite Cultivars of Ricinus communis L. Using RAPD Markers for Plant Breeding ABSTRACT Original Research Paper: The present investigation screened the diversity among 19 elite cultivars of castor bean (Ricinus communis L.) genotypes based on RAPD markers together with morphological traits in an approach to mine genes governing commercially important traits for future crop improvement programmes. A total of six primers were selected from an initial screening of 64 RAPD primers. The frequency of polymorphic amplicons observed among cultivars varied greatly from 0.05 to 0.95 in all 19 cultivars. A dendrogram showing genetic similarities among the cultivars was constructed based on the polymorphic bands using UPGMA cluster analysis. The UPGMA clustering method revealed two major groups. The first one was made up of two cultivars, DCH 190 and RG 90 while the remaining cultivars were clustered into the second group. Cultivars belonging to the second group were grouped into five sub-clusters. Genetic differences ranging from 0 to 0.86 among the cultivars indicate a diverse gene pool in the castor bean genotypes studied. Morphological and RAPD tools to determine genetic diversity among R. communis cultivars were established. Mejda Cherif, Dhekra Chaouachi, Wissal Friani, Salah Rezgui (Tunisia) Resistance of Wild Barley (Hordeum spontaneum, H. marinum and H. murinum) to Pyrenophora teres and Rhynchosporium secalis causing Net Blotch and Scald Diseases in Tunisia ABSTRACT Original Research Paper: Resistance to net blotch and scald in the seedling and in the adult growth stages were evaluated in 56 accessions of wild barley (Hordeum spontaneum, H. marinum and H. murinum) and seven varieties of H. vulgare. Generally, the screened barley genotypes were more resistant than the four checks for net blotch and scald diseases. Results of net blotch evaluation indicated that 88 and 51% of the barley genotypes were significantly more resistant than the most resistant check in the seedling and the adult growth stages, respectively. The H. marinum genotypes collected from Algeria and Egypt (accessions 4, 5 and 7) and the H. spontaneum genotypes collected from Afghanistan, Iraq and Syria (accessions 14, 25 and 43) were the most resistant in both growing stages. For both diseases (net blotch + scald), 36% of the evaluated barley genotypes were significantly more resistant than the most resistant check to a mixture of isolates. According to their reaction to

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both diseases, the accessions of H. murinum from Arminea, of H. marinum from Algeria, Egypt and Kazakhstan, and of H. spontaneum from Afghanistan, Azerbaijan, China, Egypt, Iraq, Lebanon, Libya, Syria, Turkmenistan, Turkey and Uzbekistan showed the highest level of resistance in the field. For scald, 45 and 15% of the evaluated barley genotypes exhibited more resistance than the mid check at seedling and adult growth stages, respectively. Among the evaluated barley genotypes, three accessions of H. marinum collected from Algeria and Egypt (accessions 4, 5 and 7) and two accessions of H. spontaneum collected from Afghanistan and Egypt (accessions 14 and 22) were resistant in both stages. Ramzi Chaabane, Sahari Khoufi, Khalil Khamassi (Tunisia), Jaime A. Teixeira da Silva (Japan), Emeni Ben Naceur, Houcine Bchini, Elyes Babay, Houda Ouji, Mbarek Ben Naceur (Tunisia) Molecular and Agro-physiological Approaches for Parental Selection before Intercrossing in Salt Tolerance Breeding Programs of Durum Wheat ABSTRACT Original Research Paper: A set of six Tunisian durum wheat varieties (Triticum turgidum L. subsp. durum Desf.) were used for developing and validating an approach to select parental genotypes before intercrossing them for salt tolerance breeding programs. This research was designed to study both molecular genetic diversity and differences in patterns of agro-physiological responses to salt stress. Different agro-physiological measurements were conducted for both the saline treatment (10 g/l NaCl) and the control. The data were converted to salt tolerance indices to allow comparisons among genotypes for salt sensibility. Genetic diversity among these varieties was determined by comparing their molecular traits. A dendrogram of genetic similarity was established using 46 polymorphic markers from eight SSRs and four RAPD primers. The results revealed that the most affected agro-physiological parameters were tiller number, chlorophyll content at 110 and 120 days, shoot dry weight, number of spikes/plant and total grain yield. For these parameters the most affected genotypes were Karim and Nasr and the least affected were Maali and Razzek. The dendrogram discriminated Karim from the remaining analysed varieties. Except for Karim, the remaining varieties were tightly clustered in the dendrogram, which reflects a narrow genetic base and low genetic variability among those genotypes. The genetic diversity of Tunisian durum wheat varieties could be enlarged by combining desired traits from different local and wild populations of different geographical origins into the breeding lines. Finally, we conclude that both molecular and agro-physiological approaches are helpful for current and future breeding programs in order to select genetically distinct parents with different salt tolerance components. Ezatollah Farshadfar, Reza Mohammadi, Zahra Vaisi (Iran) Screening and Repeatability of Quantitative Indicators of Drought Tolerance in Wheat-Barley Disomic Addition Lines ABSTRACT Original Research Paper: To locate the genes controlling drought tolerance in barley (Hordeum vulgare cv. Betzes), a set of complete wheat-barley disomic addition lines (DALs) along with two donor (barley) and recipient (wheat) parents were evaluated under drought stress and non-stress conditions for three cropping seasons. Several drought tolerance indices based on the grain yield of DALs in the stress and non-stress conditions were used to study the genetic variation in barley and to investigate relationship between the indices within and over the years. Differences in ranking of genotypes based on each index were observed from year to year, indicating that the drought tolerance of genotypes is influenced by year effect. The relationships among the indices and their association with mean yield based on Spearmans rank correlation were determined in each of the three cropping seasons. Principal component analysis (PCA) based on the Spearmans rank correlation matrix revealed that the screening methods were significantly inter-correlated with each other indicating that several of the statistics probably measure similar aspects of drought tolerance. The stress tolerance index (STI), geometric mean productivity (GMP), harmonic mean (HM) and mean productivity (MP) were consistently and highly correlated with each other over there cropping seasons, and, therefore, could be used to select drought tolerant genotypes with high yield performance in both stress and non-stress conditions. The stress susceptibility index (SSI), yield stability index (YSI), tolerance (TOL) and sensitivity drought index (SDI) showed consistent relationships with each other over years and can be used to screen the drought resistant and stable genotypes. According to multiple year data, most of the genes controlling drought tolerance are located on chromosome 7H in barley. Ezatollah Farshadfar, Hojjat Hasheminasab (Iran) Investigating the Combining Ability and Genetic Constitution of Physiological Indicators of Drought Tolerance in Bread Wheat (Triticum aestivum L.) Using GGE Biplot Methods ABSTRACT

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Original Research Paper: To study the combining ability and genetic constitution of physiological indicators of drought tolerance in bread wheat using GGE biplot techniques, an eight-parental diallel cross, excluding reciprocals, was grown in a randomized complete block design with three replications under two different water regimes (irrigated and rainfed) in the Agricultural Research Institute of Sararood, Kermanshah, Iran. Significant differences were found for yield potential (Yp), stress yield (Ys), relative water content (RWC) and leaf water potential (LWP). The result of biplot analysis showed that the Parent A, F, B, F and E were The best general combiners with positive effects, for improvement of Y, RWC, LWP, RWL and chlorophyll fluorescence (CHF) under drought conditions respectively, also parent C exhibited positive GCA for all traits. The crosses (A, C and H) (B, F and G), (F and D) (A, C and G), (F and E) (A, H, C and D), (F, B and E) (C, G and H) and (F and E) (A and H) for Y, RWC, LWP, RWL and CHF were heterotic groups with different dominant tolerance genes (D1 and D2), respectively. The polygon view of the biplot indicated combining A G and F E produced the best drought tolerance hybrids for all the traits through integrated the four tolerance genes (A1, A2, D1, and D2). G was the best tester in all, as it was very close to the ideal tester. The results showed additive gene effects mainly that control the physiological indicators of drought tolerance. Thus, genetic gain in developing tolerance in bread wheat can be achieved through selection. Md. Moniruzzaman, Md. Mahmudul Hasan (Bangladesh), Jaime A. Teixeira da Silva (Japan), Md. Jashim Uddin, Md. Mainul Hasan, Monjila Begum, Md. Abu Syed (Bangladesh) Variability Study in Inter-varietal Crosses of Aubergine (Solanum melongena L.) ABSTRACT Original Research Paper: A field experiment was carried out to assess the genetic variability of yield-attributing traits in 12 aubergine genotypes. The evaluated parameters were number of primary and secondary branches/plant, days to flowering, number of fruits/plant and fruit length, all of which differed significantly in different genotypes. Small differences in genotypic and phenotypic variance and coefficient of variation, high heritability as well as high genetic advance were observed for all traits, indicating additive gene effects. Line-23Line-24F2 performed best in terms of number of primary branches/plant (8.5), early flowering (48.18 days after transplanting), and maximum number of fruits/plant (37.29). Line-27 required more days to flowering (58.16) while BARI-2 produced most secondary branches/plant (20.59) and longest fruit (13.70 cm). Hence, a cross between Line-23Line-24F2 and BARI-2 is suggested for increased value-added traits. Solomon Ukaan, Peter Ogbonna (Nigeria) Genetic Variability and Character Association in Sesame (Sesamum indicum L.) Accessions ABSTRACT Original Research Paper: This study was conducted to determine the genetic variability and character association in 13 sesame (Sesamum indicum) accessions. The accessions were evaluated in replicated field experiments at the Department of Crop Science Teaching and Research Field, University of Nigeria, Nsukka under two planting seasons. The result showed significant differences among the accessions in most of the traits studied. The genotypic and heritability estimates were high for all the growth and yield attributes, indicating that these traits have high transmitting ability to subsequent generations. The highest genetic and phenotypic variance were observed in number of seeds/plant (5389.3 and 5391.3) in 2009 with number of capsules/plant (223.64 and 226.03) in 2010. However, for genetic and phenotypic coefficients of variation, the highest were observed in seed yield/ha and seed yield/plant (37.80 and 40.01) in 2009 while seed yield/plant (40.54 and 40.56) gave the highest in 2010. Apart from number of days to emergence in 2009 planting season, high heritability values were obtained on all the traits during the two planting seasons indicating that selection for those traits would be very efficient. The correlation matrix showed that seed yield/plant correlated positively and significantly with number of leaves, plant height, number of branches, stem girth, number of flowers/plant, number of capsules/plant, number of seeds/capsules, capsule length, capsule weight/plant, 1000-seed weight and number of seeds/plant. This indicates that increase in these traits will ultimately increase seed yield. Mohammad A. K. Azad (Bangladesh), Jaime A. Teixeira da Silva (Japan), Bhabendra K. Biswas (Bangladesh) Genetic Correlation among Various Quantitative Characters in Maize (Zea mays L.) Inbred Lines ABSTRACT Short Communication: Thirty maize (Zea mays L.) inbred lines were grown in a randomized complete block design with three replications to assess the genetic correlation among 13 quantitative traits. A positive correlation with grain yield/plant was recorded for plant height (r = 0.41), ear length (r = 0.54), ear diameter (r = 0.74), number of grains/row (r = 0.47), number of

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grains/ear (r = 0.45) and 1000-grain weight (r = 0.41). Days to 50% tasselling and days to 50% silking were negatively correlated with yield/plant while days to 50% maturity, ear height, number of ears/plant and number of kernel rows/ear were positively but insignificantly correlated with yield/plant. Ear diameter was the character that most contributed to high yield/plant (r = 0.74) followed by ear length (r = 0.54). A strong positive correlation was observed for grain yield/plant with plant height (r = 0.41), ear length (r = 0.54), ear diameter (r = 0.74), number of grains/row (r = 0.47), and number of grains/ear (r = 0.45) indicating that selection for these characters could help improve grain yield in maize inbred lines. Ravi V. Mural, Ravindra Sasalawad, Shailaja Hittalmani (India) Evaluation of Rice (Oryza sativa L.) Accessions for Growth and Yield Attributes in Aerobic Condition ABSTRACT Short Communication: The objective of this study was to evaluate 300 rice accessions and four check varieties for growth and yield component traits under aerobic condition. The analysis of variance for the ten characters studies revealed high significant difference among the genotypes both at five and one per cent level of significance except for test weight, seed length and single panicle weight thus, wide range of variability was observed for all the characters. In general, the phenotypic co-efficient of variation (PCV) was greater than genotypic coefficient of variation (GCV) for all the traits studied, indicating significant influence of environment on expression of the traits. The estimate of broad sense heritability (h2) was highest for biomass, genetic advance as per cent mean was high for total seed weight, single panicle weight and number of tiller per plant. High values of PCV, GCV, h2 and genetic advance as per cent mean were observed for grain yield per plant. Total seed weight was positively associated with above ground biomass, single panicle weight and number of tillers. A significant negative correlation between plant heights at 45 DAS with number of tillers was observed. The genotypes viz., MAS-IL858, MAS-IL869, MAS-IL748, MAS-IL835 and MAS-IL707 were found high yielders among introgression lines as compared with checks.

International Journal of Plant Developmental Biology

Aya Hatano-Iwasaki, Kenichi Ogawa (Japan) Biomass Production Is Promoted by Increasing an Aldolase Undergoing Glutathionylation in Arabidopsis thaliana ABSTRACT Original Research Paper: We previously identified a putative fructose-1,6-bisphosphate aldolase (FBA), which was designated as FBA1 (At2g01140.1), as a protein undergoing glutathionylation in Arabidopsis. Here we show that increasing FBA1 activity can improve biomass production by enhancing leaf area-based CO2 assimilation. Transgenic Arabidopsis plants with increased accumulation of FBA1 protein (35S-FBA1 plants) exhibited enhanced photosynthetic CO2 assimilation rate with increased FBA activity, and yielded more seeds and aerial biomass than wild-type plants under nitrogen nutrient conditions suitable for biomass production. FBA activity and photosynthetic CO2 assimilation rate decreased in glutathione-deficient mutants, cad2-1 and pad2-1, with decreased glutathione levels, while FBA protein levels were little affected in the mutant plants. An in vitro enzyme assay for FBA from mutant and wild-type plants revealed that FBA activity can be increased and recovered by 2.5 mM glutathione, but not by a strong reducing agent, 20 mM dithiothreitol. We found a positive correlation between photosynthetic CO2 assimilation rate and FBA activity among plants with various levels of FBA protein and of glutathione, and this correlation was strengthened (r2=0.955) with increasing light intensity from 25 to 1000 E m-2 s-1. On the other hand, there was much less correlation between photosynthetic CO2 assimilation rate and Rubisco protein (r2=0.013) or total activity (r2=0.005), which have been generally considered to be a major limiting factor of photosynthesis. These results indicate that FBA activity is a limiting factor for photosynthetic CO2 assimilation and biomass production, even at the light intensity at which Rubisco is fully activated, and that it is likely to be regulated by glutathione. Hussein Alzubi (Syria), Luz Marcela Yepes, Marc Fuchs (USA) Enhanced Micropropagation and Establishment of Grapevine Rootstock Genotypes ABSTRACT Original Research Paper: The effect of medium composition, antioxidant and adsorbent of polyphenolic compounds on micropropagation of grapevine rootstocks 3309 Couderc, 110 Richter, 101-14 Millardet et De Grasset, Vitis riparia Gloire de Montpellier and Teleki 5C was determined. Shoot length, leaf number and size, as well as root number, weight and length were consistently higher on woody plant medium (WPM) (Lloyd and McCown 1981) relative to full strength or half-strength Murashige

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and Skoog (MS) medium (Murashige and Skoog 1962) and Martin et al. medium (MM) (Martin et al. 1987). Supplementing WPM with 37 mg l-1 cysteine as antioxidant promoted better rooting and enhanced plant development for most rootstock genotypes. The presence of cysteine in WPM also provided homogeneous growth, regardless of the position of nodal segments on the initial subcultured plants, likely by breaking the apical meristem dominance inhibitory effect on axillary bud proliferation. Our micropropagation protocol facilitated a fast and uniform multiplication of grapevine rootstocks in tissue culture and a successful transfer and growth of micropropagated plants in the greenhouse. Vinay Kumar, Sudesh Kumar Yadav (India) Developmental Effect on Transcript Expression of Genes Encoding Enzymes for Flavan-3-ols Synthesis and its Content in Leaves and Flowers of Tea (Camellia sinensis (L.) O. Kuntze) ABSTRACT Original Research Paper: The production of monomeric flavan-3-ols mainly comprises of catechin and epicatechin, epigallocatechin and epicatechingallate, and their polymeric form as proanthocyanidins is highly controlled during the developmental stages of leaf and flower. We studied the accumulation of monomeric flavan-3-ols, anthocyanins and proanthocyanidins during the developmental stages of leaf and flower in relation to the expression of flavan-3-ols-specific genes of tea (Camellia sinensis). The expression analyses of flavan-3-ols-specific genes were also studied in the presence of exogenous monomeric flavan-3-ols such as catechin and epicatechin during different development stages of both organs. We selected two developmental stages of leaf (leaf bud (LB) and old leaf (OL)) and three stages of flower development (flower bud (FB), semi-mature flower (SMF) and fully mature flower (FMF)) to assess various activities. The monomeric flavan-3-ols were highest in LB and FB and lowest in OL and FMF. The pattern of accumulation of anthocyanins was opposite to monomeric flavan-3-ols content during different development stages of the leaf and flower. In leaves, proanthocyanidins, oligomeric and/or polymeric forms of flavan-3-ols were higher in OL than in LB while in flowers, proanthocyanidins were lowest in FMF. Expression of the DFR gene encoding dihyroflavonol 4-reductase, the LAR gene encoding leucoanthocyanidin reductase and the ANR gene encoding anthocyanidin reductase was correlated with the accumulation of monomeric flavan-3-ols during leaf and flower development. Expression of the ANS gene, encoding anthocyanidin synthase, was only detected in OL, which also contained the highest anthocyanin content. Exogenous exposure of flavan-3-ols (catechin and epicatechin) modulated the expression of DFR, LAR and ANR in OL and LB tissues but only modulated the expression of LAR during all stages of flower development. This study provides an overview of new insight for monomeric flavan-3-ols biosynthesis, and their transcript regulation during leaf and flower development of C. sinensis. Soumya V. Menon, T. V. Ramana Rao (India) Enzyme Activities during the Development and Ripening of Watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) Fruit ABSTRACT Original Research Paper: Watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai; Cucurbitaceae), is an excellent source of sugars and antioxidants, mainly lycopene, which makes it a nutritionally good fruit. The present study was carried out with the purpose of evaluating the activities of various enzymes in relation to the biochemical processes occurring at five sequential stages of watermelon fruit: young, pre-mature, mature, pre-ripened and ripened. Maximum activity (0.010 mg maltose released/mg protein) of -amylase (free), one of the starch hydrolyzing enzymes, was found in the pre-mature stage, while -amylase (bound) exhibited highest activity (0.063 mg maltose released/mg protein) during the pre-ripened stage. Sugar metabolizing enzymes in the relevant metabolic pathways of sugars were also quantified. Sucrose phosphate synthase and sucrose synthase exhibited maximum activities in the young and pre-mature stages, respectively. These activities were significantly higher (P < 0.05) than in other development and ripening stages. The sucrose-cleaving enzyme, acid invertase, showed remarkably high activity (0.014 mol/h/mg protein) in the ripe stage, as did neutral invertase in the mature stage (0.029 mol/h/mg protein). -Galactosidase activity (0.035 mol pnp/mg protein) was maximum in the mature stage, while activities of polygalacturonase (0.025 mg glucose released/mg protein) and cellulase (0.01 mg glucose released/mg protein) were maximum in the young stage. The specific activity of peroxidase was significantly high (0.0034 units/mg protein) in the pre-mature stage whereas polyphenol oxidase exhibited inconsistent activity, but with a maximum in the ripened stage (3.49 units/mg protein). The overall results indicate a positive relation between enzymes (sucrose phosphate synthase, sucrose synthase, -galactosidase, amylases and polyphenol oxidase) and the development and ripening of watermelon fruit. Tukaram Dayaram Nikam, Jitendra Gopichand Patil, Mahendra Laxman Ahire, Savaliram Goga Ghane, Kirti Manik Nitnaware, Vikas Bandu Naikawadi (India) Axillary Multiplication of Ceropegia mahabalei Hemadri & Ansari and Ceropegia

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media (Huber) Ansari: Critically Endangered Ethnomedicinal Herbs of Western Ghats, Maharashtra State of India ABSTRACT Original Research Paper: A rapid micropropagation system was developed for critically endangered ethnomedicinal herbs Ceropegia mahabalei Hemadri & Ansari and C. media (Huber) Ansari. For shoot multiplication nodal, internodal and leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinins [(6-Benzyladenine: BA or Kinetin: Kin (0.0-10.0 M)] alone and in combination with auxins [(-naphthaleneacetic acid: NAA; indole-3-acetic acid: IAA; 2,4-dichlorophenoxyacetic acid: 2,4-D (0.5-1.5 M)]. Maximum number of shoots were produced on subculture of nodal explants of C. mahabalei and C. media on MS medium supplemented with 5.00 M BA. The type and age of explant and addition of auxins (NAA, IAA or 2,4-D: 0.5-1.5 M) in the medium influenced shoot multiplication. The best medium for rooting of shoots of both the species was liquid MS medium containing 1.0 M NAA and 4% sucrose. Well rooted plantlets were transplanted into soil and about 88% of the plantlets survived well upon transferred to natural conditions. The plantlets were morphologically identical to the parental plants. This work may be helpful for in vitro propagation, ex situ conservation and genetic manipulation of these species. Mahendra Laxman Ahire, Pradip Pandit Patil, Polavarappu B. Kavi Kishor, Tukaram Dayaram Nikam (India) Micropropagation and Assessment of Antibiotic Selection in Vitro of Bacopa monnieri (L.) Pennell ABSTRACT Original Research Paper: Bacopa monnieri (L.) Pennell (Brahmi), Scrophulariaceae, is one of the sources of medhya rasayan drugs (that counteract stress and improves intelligence and memory) of Ayurveda. The aim of the present study was to assess tissue culture conditions and antibiotic selection which could be useful for genetic transformation of this important medicinal plant. An efficient, simple and reproducible system for plant regeneration through leaf and nodal explants was devised. Sensitivity of the nodal explants against cefotaxime (Cef), kanamycin (Kan) and hygromycin (Hyg) was established. Leaf and nodal explants produced about 90 shoots on Murashige and Skoog (MS) medium fortified with 1.0 mg/L 6-benzyladenine (BA) after 28 days of incubation. Subculture to fresh MS medium resulted in shoot elongation; the shoots obtained grew well and were healthy. The transfer of shoots to liquid MS medium supplemented with 0.2 mg/L -naphthaleneacetic acid (NAA) resulted in 100% rooting. The rooted shoots, on transfer to plastic containers containing a mixture of garden soil + sand (1:1), could acclimatize within 14 days under glasshouse conditions. The acclimatized in vitro-grown plants showed 100% survival after transfer to soil in earthen pots. A threshold limit of survival of nodal explants was observed at 500 mg/L Cef, 100 mg/L Kan and 15 mg/L Hyg. Shiwali Sharma, Anwar Shahzad (India), Mohd. Shahid (Kingdom of Bahrain/India), Noor Jahan (India) An Efficient in Vitro Production of Shoots from Shoot Tips and Antifungal Activity of Spilanthes acmella (L.) Murr. ABSTRACT Original Research Paper: An efficient method for the propagation of Spilanthes acmella (L.) Murr. through shoot tips (collected from 3 week-old axenic seedlings) has been successfully developed. This protocol can be employed on a commercial scale for the production of spilanthol. Among the cytokinins, 6-benzyladenine (BA), kinetin (Kn) and 2-isopentenylaminopurine (2-iP) were tested. Murashige and Skoog (MS) medium supplemented with 1.0 M BA was optimum for inducing bud break. When auxin was augmented with an optimal cytokinin concentration, the regeneration efficiency of explants was enhanced. The maximum response (96%) with highest number of shoots per explant (i.e., 33.0) was possible on MS medium containing 1.0 M BA and 0.1 M -naphthaleneacetic acid (NAA). Small shoots (3.0-4.0 cm) were rooted in vitro with half-strength MS medium containing 2.5 M NAA, forming a maximum of 32.2 roots/shoot. The well developed micropropagated plants were successfully acclimatized within 4 weeks in SoilriteTM and planted ex vitro in garden soil, farmyard soil and sand (2: 1: 1), where they grew well without any apparent morphological variation from the parent plant. Moreover, the phytomedicinal effect of various plant tissues was also evaluated against human pathogenic fungi. The alcoholic extracts of in vitro plant were more effective than in vivo plant materials. Maximum inhibition zone (MIZ) was noticed against Candida krusei followed by Candida albicans, Aspergillus fumigatus and Aspergillus niger. Among different explants, flower heads showed best response against C. krusei as highest of 12.3 and 12.0 cm MIZs were noticed for in vitro and in vivo source, respectively. Manohar Singh Rao, Anshul Goyal (India) High Frequency Rapid Clonal Propagation of Andrographis paniculata Using Cotyledonary Node Explants

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ABSTRACT Original Research Paper: Multiple shoot induction is reported from cotyledonary node (CN) explants cultured on MS (Murashige and Skoog 1962) medium containing different concentrations and combinations of 6-benzyl adenine (BA) and kinetin (Kn). However, maximum number of shoots/explant (9.6 shoots/explant) were proliferated on MS medium supplemented with a combination of 2.22 M BA and 2.32 M Kn. On this medium all the explants responded positively. Further, shoots were multiplied on the same medium at a rate of 1.4-fold but they remained stunted. The physical consistency of the medium and the concentration and combinations of BA and Kn evoked varied responses in terms of shoot multiplication and elongation. A liquid culture system supplied with 2.22 M BA significantly promoted shoot multiplication and overcame the problem of shoot elongation. On this medium, shoots multiplied at a rate of 3.88-fold where shoots attained an average height of 4.75 cm. Elongated shoots 2.5 cm in height were most suitable for in vitro root induction. The maximum rooting frequency (92%) and most roots (13.0 roots/shoot) were produced on 1/4-strength MS medium, 0.6% agar, 2.0% sucrose supplemented with 4.90 M indole-3-butyric acid. The rooted shoots transferred to sterilized jute compost moistened with 1/4-strength MS medium in 400-ml volume glass bottles (covered with polypropylene caps) hardened successfully under misthouse conditions. Hardened plantlets grew well and showed over 82% survival upon transfer to pots containing a mixture of garden soil: sand: farmyard manure (1: 1: 1) during acclimatization in a nursery. XiaoNan Yu, HongJuan Wu, Tong Pan (China), Jaime A. Teixeira da Silva (Japan) Multiple Shoot and Callus Induction of Herbaceous Peony (Paeonia lactiflora Pall.) ABSTRACT Original Research Paper: Underground buds of herbaceous peony (Paeonia lactiflora Pall.) Zhong Sheng Fen were used as explants for axillary shoot induction while stems, petioles and leaves of Da Fu Gui and Tao Hua Fei Xue were used as explants for callus induction. The effects of different basal media and concentrations of plant growth regulators (PGRs) on induction were investigated to establish an aseptic regeneration system. The best medium to induce and proliferate shoots was modified half-strength Murashige and Skoog (MS) medium with double the concentration of Ca2+ supplemented with 1 mg L-1 gibberellic acid (GA3) plus 1 mg l-1 6-benzylaminopurine (6-BA). Two successive steps were adopted for rooting shoots. Shoots were first cultured on Woody Plant Medium (WPM) plus 0.5 mg l-1 1-naphthyleneacetic acid (NAA) for 10-15 days in the dark, then shoots were transferred to PGR-free WPM medium containing 0.2% activated charcoal; in this case, rooting could reach 50%. The best explants for callus induction were young stems, and the best basal medium for callus induction was WPM for Da Fu Gui and Tao Hua Fei Xue. Syed Arshad Hussain (India), Jaime A. Teixeira da Silva (Japan), Azra Nahaid Kamili, Ali Mohammad Shah (India) In Vitro Propagation of Parrotiopsis jacquemontiana (Decne) Rehd. Using Mature Tree Explants ABSTRACT Original Research Paper: The vegetative propagation of Parrotiopsis jacquemontiana (Decne) Rehd., an endemic species of the Hamamelidaceae family growing wild in Kashmir, is still considered to be difficult owing to its difficult-to-root stem cuttings. The present study reports, for the first time, an in vitro propagation technique for this species using shoot apex and nodal stem segments of a mature tree as explants. The initial shoot cultures were established in agarified Murashige and Skoog (1962) basal medium supplemented with a cytokinin (6-benzylaminopurine (BA) or kinetin at 1-10 M). The explants secreted a large amount of phenolic substances which frequently led to tissue browning; this problem could be overcome by washing the explants for at least 30 min before inoculation and frequently transferring them to fresh medium during the culture establishment phase. The initial shoots were subcultured in a multiplication medium having a combination of an auxin (1-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA) or indole-3-acetic acid (IAA) at 2 M) and a cytokinin (BA) at 2.5-10 M. Shoot multiplication was best (5-6 shoots/culture) on MS medium having BA and IAA at 5 and 2 M, respectively. In vitro shoots 2.5 cm or longer were successfully rooted in auxin-supplemented -MS medium within 8 weeks. IAA, NAA or IBA at 2.5 and 5 M were the most effective concentrations for inducing rooting. The plantlets were acclimatized with 40% survival. Budi Winarto (Indonesia), Jaime A. Teixeira da Silva (Japan) Sterilization Procedure for in Vitro Culture of Leatherleaf Fern (Rumohra adiantiformis) ABSTRACT

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Original Research Paper: When explants are introduced to the in vitro environment, sterilization is a fundamental process allowing contamination to be eliminated. A new sterilization procedure for a novel in vitro propagation of leatherleaf fern (Rumohra adiantiformis) was successfully devised using rhizomes which are typically heavily contaminated as the donor explant. The most effective sterilization procedure for in vitro micropropagation involved pretreatment of rhizomes in 80% alcohol for 3 min, washing them under tap water for 3 h then immersing them in 0.05% mercuric chloride for 10 min, followed by 96% alcohol for 1 min. Finally explants were rinsed 6 times (5 min each rinse) using sterile distilled water. This combination of treatments reduced the percentage of contamination to 33% and stimulated the percentage of rhizome regeneration to 73% with 2.3 rhizomes regenerated/replication (= 5 ex vitro rhizomes). This in vitro sterilization procedure will have an effective impact on the establishment of explants in vitro and on the micropropagation of leatherleaf fern. XiaoNan Yu, LiHui Wang (China), Jaime A. Teixeira da Silva (Japan) Change of Endogenous Hormones Inside Paeonia lactiflora Buds during Winter Dormancy ABSTRACT Original Research Paper: Variation in and the ratios of endogenous hormones in the terminal buds of 4-year old traditional Chinese peony cv. Da Fu Gui throughout the dormant period were studied by HPLC to attempt to unravel the mechanism of winter bud dormancy in this woody ornamental. Abscisic acid (ABA) content began to increase on November 20 and peaked on December 10 while gibberellic acid (GA3) content started to increase on December 10 and dropped off by December 30 while the trend for cytokinin (CK) was similar to that of GA3. The level of indole-3-acetic acid changed little throughout the period of dormancy. GA3/ABA and CK/ABA values dropped as dormancy progressed and rose as dormancy was released. Vishnu Sukumari Nath, Muthukrishnan Senthil alias Sankar, Vinayaka Mahabaleswar Hegde, Muthulekshmi Lajapathy Jeeva, Raj Shekar Misra, Syamala Swayamvaran Veena (India) A Simple and Efficient Protocol for Rapid Regeneration and Propagation of Taro (Colocasia esculenta (L.) Schott.) in Vitro from Apical Meristems ABSTRACT Short Communication: An efficient and simple protocol was developed for the in vitro regeneration and propagation of taro (Colocasia esculenta) cv. Muktakeshi. Apical meristems (~1 cm) excised from leaf blight-resistant taro cultivar cv. Muktakeshi grown in a net house were used as explants. Most multiple shoots (i.e., 3.6/explant), which formed on Murashige and Skoog (MS) medium supplemented with 5 mg L-1 6-benzyladenine and 1 mg L-1 -naphthaleneacetic acid, could be rooted by transferring the 4-week-old shoots to MS basal medium without plant growth regulators. After 2 weeks, well developed plantlets were hardened in plastic cups in potting mixture (vermiculite + sand, 1:1, v/v). Acclimatized plants were transferred to 30-cm plastic pots containing top soil and vermicompost (3:1, v/v) where they grew well. Anu Bhatt, Shubhi Kansal, Rajinder Singh Chauhan, Hemant Sood (India) Modified Tissue Culture Procedures for Micropropagation of Apple Root Stocks ABSTRACT Research Note: The low yield of apples (Malus spp.) can be amounted to the uncertainties of the monsoon, dependence over the old cultivars and prone to pathogen infestation. This warrants the development of a cost-effective micropropagation technology for the rapid multiplication of commercially important root stocks which can be utilized for re-plantation in apple orchards. Axillary shoot tips of M9 rootstock formed most multiple shoots on Murashige and Skoog (MS) medium supplemented with 3 mg/l 6-benzyladenine (BA), 2 mg/l kinetin (KN), 3% (w/v) sucrose and 0.8% (w/v) agar-agar, with 85.7% of shoot apices forming multiple shoots. To reduce the cost of media components for commercial production of the planting material from root stocks, sucrose was replaced with table sugar and agar-agar was completely omitted. The low-cost medium combination of MS liquid medium supplemented with 2 mg/l BA, 3 mg/l KN and 3% (w/v) table sugar was best, resulting in 22 shoots/explants. There were no significant differences in multiple shoot formation/explant, relative growth and vigor of shoots and frequency of root formation in shoots independent of whether medium contained sucrose or table sugar. In vitro regenerated plantlets were successfully hardened and transferred to the field. The substitution of sucrose for table sugar and the omission of agar-agar from the medium reduced the cost/liter medium by approximately 80-fold when other cost-effective alternatives such as tap water and autoclavable polybags were used, assisting in easier and more effective commercialization.

Medicinal and Aromatic Plant Science and Biotechnology

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Volume 6 (Number 1) Animesh K. Datta, Aninda Mandal (India), Jaime A. Teixeira da Silva (Japan), Aditi Saha, Rita Paul, Sonali Sengupta, Priyanka Kumari Dubey, Sandip Halder (India) An Updated Overview on Aloe vera (L.) Burm. F ABSTRACT Review: Aloe vera (L.) Burm. f is a succulent shrubby perennial of the family Asphodelaceae (commonly known as Natural healer, Lily of the desert, Plant of immortality, Miracle plant, The Wand of Heaven, etc.) with immense therapeutic uses not withstanding its potential significance in cosmetic and food industries. The plant is the source of two products, gel and latex (commercially aloe products are pills, jellies and creams, drinks, liquid, sprays, ointments and lotions) obtained from its fleshy leaves. This unique plant also belongs to a larger plant family, the Xeroids. Considering the pharmacological and other potential uses of A. vera, an updated overview is being conducted on the species involving all essential aspects to provide necessary information to researchers for effective utilization of the species in human welfare. Khalid A. Khalid (Egypt), Jaime A. Teixeira da Silva (Japan) Biology of Calendula officinalis Linn.: Focus on Pharmacology, Biological Activities and Agronomic Practices ABSTRACT Invited Review: Calendula officinalis Linn. (Asteraceae), a medicinal plant, is used in traditional medicine around the world and is the subject of several chemical and pharmacological studies. Chemical studies have detected various classes of compounds in its organs, primarily volatile oil, carotenoids, flavonoids, terpenoids, coumarins, quinones, carbohydrates, lipids, amino acids, as well as other minor constituents. C. officinalis extract and pure compounds isolated from different organs possess multiple pharmacological activity, including anti-inflammatory, antioedematous, antioxidant, immunostimulant, anticancer, lymphocyte and wound healing, hepatoprotective, antibacterial and antifungal, anti-HIV, spasmolytic and spasmogenic, genotoxic and antigenotoxic, inhibition of heart rate, antiviral, inter alia. In this review, we explore the phytochemical and pharmacological activity of C. officinalis while also covering aspects of its culture and cultivation that would increase the production of its pharmacologically important compounds. Maria Rosario Alonso, Ignacio N. Peralta, Rita Lemos, Roberto Davicino, Claudia Anesini (Argentina) Influence of Environment Conditions on the Chemical Composition and Antioxidant Activity of Larrea divaricata Aqueous Extract ABSTRACT Original Research Paper: Natural antioxidants have gained considerable interest for their role in the prevention of autooxidation of fats and oils. Larrea divaricata Cav. is a South American plant widely distributed in Argentina with reported antioxidant, antitumoral and antimicrobial activities. The aqueous extract of the plant exerts antioxidant activity preventing the deleterious effect of temperature and oxygen on the antioxidant activity of vitamin C (Vit C). The aim of this work was: to assess the stability of the aqueous extract of L. divaricata (AE) under different environment conditions, such as different temperatures and air oxygen contents. The stability parameters evaluated included the percentage of majority compounds (flavonoid expressed as quercitrin (F) and nordihydroguaiaretic acid (NDGA)) and the antioxidant activity of the extract (AC). Results indicated that both compounds were affected by temperature and air oxygen, with F being more stable to high temperatures than NDGA, finding a decrease of 56% for F and a decrease 66% for NDGA. Nevertheless, NDGA was more sensitive to air oxygen, as an early decrease of 62% and a maximum decrease of 100% were found. The AC of AE decreased with high temperatures; however, air oxygen seemed to have a more important effect, for a decrease of about 42% was observed after 7 days of incubation. Even though, the environment conditions could affect the chemical composition of the whole extract, it was observed that not all the compounds were equally affected by temperature and air oxygen. Reaching a balance between these two parameters could allow a modulation of the antioxidant activity of this extract. Niraj Tripathi, Navinder Saini, Vandana Mehto, Sunil Kumar, Sharad Tiwari (India) Assessment of Genetic Diversity among Withania somnifera Collected from Central India using RAPD and ISSR Analysis ABSTRACT Original Research Paper: Withania somnifera is a multipurpose plant of immense therapeutic value and wide geographic

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distribution exhibiting extensive phenotypic and chemical variability. Characterization of plants using molecular markers is an ideal approach for improvement and conservation of plant genetic resources. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) molecular fingerprinting markers were employed as genetic markers to assay the genetic relationship of 16 accessions of W. somnifera collected from different locations of India. Randomly selected 25 decamer primers amplified 204 RAPD marker loci out of which 152 bands (74.5%) were polymorphic. Twenty eight ISSR primers were used to generate fingerprints and a total of 186 alleles were amplified, out of which 151 were polymorphic (81.1%). ISSR markers were more informative than the RAPD markers. Several ISSR markers amplified genotype-specific alleles that can be used for authentication and detection of adulteration in plant material. Similarity matrices were generated from RAPD and ISSR marker data separately using Jaccards coefficient and dendrograms were constructed based on UPGMA clustering. Genotypes were clustered into two groups and the grouping was similar for both marker systems. Mahmoud Ottai (Egypt), Jaime A. Teixeira da Silva (Japan), Sherin Mahfouze (Egypt) Phenotypic and Chemotypic Variation of Four Nigella (Nigella sativa) Varieties ABSTRACT Original Research Paper: The assessment of four Nigella sativa varieties from Egypt, Iran, Turkey and Syria indicated highly significant differences in phenotypic and chemotypic characters. Seed yield was the most important character to differentiate all Nigella varieties based on high genetic advance and heritability values. In addition, the relative percentage of all detected fatty acids differed (from trace amounts to 63.691%) among varieties although the major saturated and unsaturated fatty acids were palmitic, arachnidic and linoleic acids in all varieties. The relative percentage of 8 essential and 8 nonessential amino acids different in all varieties although the Turkish variety had only 7 nonessential amino acids. Arginine and proline (ranging from 4.94 and 46.07% to 6.40 and 53.70%, respectively) were the major essential and nonessential amino acids, respectively for all varieties. SDS-PAGE electrophoresis revealed a different number of protein bands ranging from 9 bands in the Syrian variety to 18 bands in the Egyptian variety. Khalid A. Khalid (Egypt) Influence of Hydro-Distillation Time on the Yield and Quality of Dill Volatile Constituents ABSTRACT Original Research Paper: The effects of hydro-distillation time (1, 2, 3, 4, 5 and 6 h) on the essential oil (EO) extracted from dill (Anethum graveolens L.) fruits were investigated by GC and GC/MS. EO content (ml g-2) increased with several treatments of hydro-distillation time compared with the first hour of hydro-distillation. After 5 h of hydro-distillation the yield of EO remained 2.5 ml g-2, then it stabilized. The main constituents of dill EO were apiol (34.6-48.7%), carvone (19.2-27.1%), (R)-limonene (7.7-15.0%) and -pinene (7.4-16.3%). Apiol and carvone gradually increased with an increase in hydro-distillation time but (R)-limonene and -pinene decreased. The monoterpene hydrocarbons presented a gradual decrease (39.3% in the first hour to 19.9% after 6 h), the total amount of oxygenated monoterpenes increased from 60.0% in the first hour to 79.5% after 6 h, the sesquiterpene hydrocarbons appeared to increase in the 3rd hour (1.1%) compared with other distillation times while oxygenated sesquiterpene appeared to increase after 4 h (0.4%). Mouna Ben Tarit Rayouf, Kamel Msaada, Karim Hosni, Brahim Marzouk (Tunisia) Essential Oil Constituents of Salvia argentea L. from Tunisia: Phenological Variations ABSTRACT Original Research Paper: The essential oils (EOs) from the aerial parts of Salvia argentea L. were analyzed at three developmental stages (vegetative, flowering and fruiting stages). The highest content of oil (0.15%, w/w) was obtained at full flowering. The current study showed consistent compositional variations among the three studied stages. In fact, manool and manoyl oxide characterised the vegetative stage while viridiflorol, camphor, methyl eugenol and 1,8-cineole prevailed during flowering and the fruiting phase was marked by the prevalence of viridiflorol, -humulene, -ionone and methyl eugenol. Additionally, a wide array of bioactive terpenic compounds was commonly found at different stages, making S. argentea an advocated herb in pharmaceutical science. Venugopal Rao Kokkirala, Srinivas Kota, Rajesh Yarra, Mallesham Bulle, Mahender Aileni, Kranthi Kumar Gadidasu (India), Jaime A. Teixeira da Silva (Japan), Sadanandam Abbagani (India) Micropropagation via Nodal Explants of Woodfordia fruticosa (L.) Kurz.

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ABSTRACT Original Research Paper: An efficient protocol for the in vitro clonal propagation of Woodfordia fruticosa (L.) Kurz using node explants from mature plants was established. Axillary bud proliferation was initiated on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA; 4.40, 8.90, 13.30, 17.70, 22.20 and 26.50 M) or kinetin (Kn, 4.60, 9.30, 13.90, 18.50, 23.20 and 27.20 M) alone, or with combinations of BA (2.22, 4.40, 6.62 and 8.90 M) and Kn (2.32, 4.60, 6.92 and 9.30 M). MS medium supplemented with BA (17.70 M) showed the highest regeneration efficiency with 35 1.65 multiple shoots/explant. The shoots that regenerated in vitro attained a maximum average length of 4.5 0.25 cm within 5 weeks of culture, 90% of which could root on medium containing half-strength MS salts fortified with 4.90 M IBA. The regenerated plantlets were established in a greenhouse with 85% survival. ISSR-PCR revealed genetic fidelity among micropropagated plants and the mother plant. Rumisa R. Quadri, Azra N. Kamili (India), Jaime A. Teixeira da Silva (Japan), A. M. Shah (India) In Vitro Multiplication of Arnebia benthamii Wall., a Critically Endangered Medicinal Herb of the Western Himalayas ABSTRACT Original Research Paper: Arnebia benthamii (Wall. Ex G. Don) Johnston [Syn Macrotomia benthamii (Wall.) DC.] (family Boraginaceae), locally known as Kahzaban, is a highly valued and critically endangered Himalayan medicinal plant, and ranks second in the list of medicinal plants prioritized for Western Himalaya and also figures among the 59 medicinal plants prioritized for conservation due to its high threat of extinction. An efficient in vitro multiplication and propagation system was developed for A. benthamii. Half-strength Murashige and Skoog (MS) medium augmented with different concentrations of 6-benzyladenine (BA) were used for shoot multiplication from shoot tip explants. The best response, i.e., multiple shoot formation, was with 5 M BA. In another experiment, the combined effect of BA with 1 M indole-3-butyric acid (IBA) was tested. The maximum number of multiple shoots was obtained on half-strength MS medium supplemented with 4 M BA and 1 M IBA. Different concentrations of IBA, indole-3-acetic acid (IAA) and -naphthaleneacetic acid (NAA) were used to induce roots from shoots. Roots formed best on half-strength MS medium supplemented with 4 M IBA, and 80% of plantlets transferred to field conditions survived. Smitha P. Venkata, Mohan Ch. Murali (India), Jaime A. Teixeira da Silva (Japan), B. Akondi Raju, R. Sravani (India) Screening the Antimicrobial and Antioxidant Potential of Ventilago denticulata, Scolopia crenata and Rivea hypocrateriformis from Maredumilli Forest, India ABSTRACT Original Research Paper: Despite the numerous advances in medicine, the prevalence of infectious diseases continues to rise due to the emergence of antibiotic-resistant pathogens, which is attributed to the widespread use of antibiotics. Antioxidants help to deal with oxidative stress, which is caused by free radical damage. Thus, the search for new antibacterial and antioxidant agents from plants has gained increasing importance. The methanolic extracts of Ventilago denticulata bark, Scolopia crenata bark and Rivea hypocrateriformis roots were screened for their antimicrobial potential against seven bacterial species (Bacillus cereus, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella sp., Staphylococcus aureus) using minimum inhibitory concentration (MIC) and against three fungal species (Aspergillus niger, Penicillium sp. and Trichoderma viride) using the disc diffusion method. The extracts were also evaluated for their in vitro antioxidant activity using the FRAP method. R. hypocrateriformis showed the highest zone of inhibition (8-11 mm) followed by V. denticulata (7-9 mm) and S. crenata (6-10 mm). Phytochemical analysis showed the presence of saponins, alkaloids, steroids, cardiac glycosides, tannins and phenolics. This is the first report on the antioxidant property of S. crenata, which has excellent antioxidant potential (73.56 1.34 mg GAE/g) among the three extracts tested. Results obtained in this study show that these plants have a broad spectrum antibacterial activity and constitute a potential source of new classes of antibiotics and antioxidants. Maryam Haghighi (Iran), Jaime A. Teixeira da Silva (Japan), Maryam Mozafariyan, Fatemeh Roustaii (Iran) Humic Acid Affects the Germination of Basil and Cumin and Alleviates the Negative Impacts of Salinity and Drought Stress ABSTRACT Original Research Paper: An experiment was conducted to investigate the effect of humic acid (HA; 0, 1000, 2000 mg/L) on

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germination and on the length of stems and roots of cumin (Cuminum cyminum) and basil (Ocimum basilicum) under salinity (0, 3, 6, 9 mmhos/cm with NaCl) and drought (0, -2, -4, -8 Pa by PEG-6000) stress. HA primarily at 1000 mg/L had a significant effect on the germination of both medicinal plants. The effect of the interaction of HA with salinity and drought was significant, proving that HA could improve germination and stem and root length in the presence of salinity stress and drought, particularly at 1000 mg/L. Ibironke A. Ajayi, Rotimi A. Oderinde, Joseph I. Uponi, Adewuyi Adewuyi (Nigeria) Evaluation of Selected Heavy Metals and Macronutrients Status of 10 Medicinal Plants from Nigeria ABSTRACT Original Research Paper: Ten important herbal plants from the South-west of Nigeria were analyzed for their heavy metal (K, Na, Ca, Mn, Mg, Cu, Fe, Zn, Pb and P) and macro-nutrient status using an atomic absorption spectrophotometer. The most prevalent heavy metals were K and Ca followed by Mg and Na. The highest level of K and Ca were found in Senna alata and Senna podocarpa respectively. Out of all the plants, Dissotia rotundifolia had the highest concentrations of Mg and Na. The results showed that the level of Cu and Zn found in the herbal plants were much lower than the reported range of the elements in agricultural products. Esther A. Adesegun, Oyeboade S. Adebayo, Aderonke K. Akintokun (Nigeria) Effect of Ocimum gratissimum and Cymbopogon citratus Extracts on Sclerotium rolfsii Sacc of Tomato (Lycopersicon esculentum Mill.) ABSTRACT Original Research Paper: Tomato (Lycopersicon esculentum Mill.) cultivation is severely affected by Sclerotium wilt caused by Sclerotium rolfsii Sacc. Chemical control has been a major strategy for its control. However, hazards associated with the use of chemicals have necessitated the search for alternatives, particularly among botanicals. An alternative method of control was examined by evaluating the inhibitory effect of two spices (Ocimum gratissimum and Cymbopogon citratus) on the growth of S. rolfsii in vitro and its subsequent development on tomato plants in vivo. FunguForce (Mancozeb 63% + Carbendazim 12.5 WP), a synthetic pesticide and sterile distilled water served as controls. The spices were extracted in water and ethanol and tested against the growth of S. rolfsii at six concentrations (0, 1, 2, 3, 4 and 5%, v/v). The highest inhibition of mycelial growth was recorded with 3% of the ethanolic extract of O. gratissimum in which no growth was recorded. The lowest inhibition (3.7 mm) was recorded with the control (0%). Generally better inhibition was possible with the ethanolic extract than with the aqueous extract. In pot experiments, the two plants reduced disease severity to 1.7 and 2.0, respectively at 5% and these values were comparable to 2.3 obtained with FunguForce and significantly lower to what was recorded in untreated plants. However, plants treated with O. gratissimum extract at 5% and FunguForce at 2.5 kg/ha gave significantly comparable plant height, number of leaves and fruit weight per plant. The plants in the control pot recorded the highest disease severity (4.7) and sclerotial rhizosphere population (82 Kg1 soil). The two spice extracts are potential options for the management of S. rolfsii on tomato. Sasan Mohsenzadeh, Javad Zaboli (Iran), Jaime A. Teixeira da Silva (Japan) Allelopathic Potential of Ajwain (Trachyspermum copticum) ABSTRACT Short Communication: The ethanolic extract obtained from ajwain (Trachyspermum copticum (L.) Link) seeds was evaluated in vitro to examine its potential allelopathic effects. The inhibitory effect of the extract at 0, 2.5 and 5% (i.e., g amounts of original extract in 100 ml of distilled water) on germination and seedling growth of corn (Zea mays), cowpea (Vigna unguiculata), redroot amaranth (Amaranthus retroflexus) and dandelion (Taraxicum officinalis) were tested. The effects of ajwain seeds extract on cowpea and corn as a cultivated crop were obviously different from the weeds (redroot amaranth and dandelion). At 2.5 and 5% of ajwain seed extract, seed germination and seedling length of both weeds were completely inhibited but in corn and cowpea there was no change or only a slight decrease. The inhibitory effects may be related to the presence of allelochemicals, including thymol, -terpinene and p-cymene of ajwain seed. Optimum concentrations of ajwain seed extracts could be used as herbicides in the control of weeds. Sasan Mohsenzadeh, Neamat Sayyadnia Tayyebi (Iran), Jaime A. Teixeira da Silva (Japan) Allelopathic Potential of Tecomella undulata (Roxb.) Seem

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ABSTRACT Short Communication: The aquatic extracts obtained from the dried roots and stem inner bark, leaves and flowers of Tecomella undulata (Bignoniaceae) were evaluated in vitro to examine their potential allelopathic effects. The inhibitory effects of the extracts at 0, 2.5, 5 and 10% (v/v) on germination and seedling growth of bread wheat (Triticum aestivum) and mung bean (Vigna radiata) were tested. The extract at all concentrations completely suppressed the germination of wheat. Root and stem inner bark and flower extracts reduced significantly (P 0.05) the length of wheat shoots and roots. The seed germination of mung bean was affected only by root and stem inner bark. Root and stem inner bark, leaf and flower extracts at 2.5% reduced the length of mung bean shoots and roots. The effect of root and stem inner bark extracts on seed germination and the length of shoots and roots were higher than by leaf and flower extracts, especially in wheat. Root and stem inner bark extracts could be used in herbicides against Gramineae weeds. Masoomeh Alipoor, Sasan Mohsenzadeh (Iran), Jaime A. Teixeira da Silva (Japan), Mehrdad Niakousari (Iran) Allelopathic Potential of Aloe vera ABSTRACT Short Communication: The ethanolic extract obtained from dried Aloe vera leaves and flowers was evaluated in vitro to examine its potential allelopathic effects. The inhibitory effect of the extract at 0, 2.5, 5 and 10% (i.e., g amounts of original extract in 100 ml of distilled water) on germination and seedling growth of wheat (Triticum aestivum), cereal rye (Secale cereale), garden cress (Lepidium sativum), redroot amaranth (Amaranthus retroflexus) and dandelion (Taraxicum officinalis) plants were tested. All concentrations of A. vera leaf and flower extracts suppressed the germination and growth of all tested plants seedlings significantly (P 0.05), except for wheat germination when 2.5% flower extract and epicotyl length when 2.5% leaf or flower extracts were used. The effects of leaf extract on epicotyl and radicle length were greater than when flower extract was used. Garden cress, redroot amaranth and dandelion seed germination and seedling growth were almost completely inhibited in the presence of 2.5% leaf or flower extract. Rumisa R. Quadri (India), Jaime A. Teixeira da Silva (Japan), Azra N. Kamili, A. M. Shah (India) Effect of 6-Benzyladenine, Kinetin and Thidiazuron on in Vitro Shoot Proliferation of Hyoscyamus niger L. ABSTRACT Research Note: Hyoscyamus niger is an important medicinal plant belonging to the Solanaceae family, commonly known as henbane and locally as Bazir bangh. A study was undertaken to investigate the effects of different plant growth regulators (PGRs) at various concentrations on shoot proliferation from shoot tip explants. Full-strength Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) were tested. Amongst the PGRs tested, BA at 12.5 M showed the best shoot proliferation and highest number of indirect multiple shoots while TDZ at 10 M formed intense disorganized friable light-greenish callus only; there was no response with Kn. In vitro shoots were separated and rooted under varying concentrations of different auxins viz. indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and -naphthaleneacetic acid (NAA). Best rooting response was recorded with 4.0 M IBA and 2.5 M NAA. However, no response was noted with any concentration of IAA. Plantlets transferred to field conditions showed 50% survival. Samuel O. Owolade, Gbenga S. Akinwumi (Nigeria) Estimation of Proximate and Phytochemical Constituents of Lactuca taraxacifolia (Wild Lettuce) ABSTRACT Research Note: An experiment was carried out to assess the proximate and phytochemical constituents of Lactuca taraxacifolia commonly referred to as wild-lettuce. It is one of the indigenous leafy vegetable in South Western Nigeria, which is being claimed in folk medicine to be used for the prevention of hepatic toxicity (hepatoprotective agent). The study was undertaken to evaluate its proximate, minerals, and phytochemical constituents which will provide biochemical information for further studies on its possible hepatoprotective activity as being claimed in folk medicine. Data on the bioactive component in food plants are needed so that research may be accurately conducted to assess their dietary intake, investigate their physiological functions and determine their relationship to health and diseases. The results revealed the presence of the basic bioactive constituents consisting of saponin (2.75 mg/100 g), tannin (2.67 mg/100 g), total phenol (2.23 mg/100 g), alkaloid (0.23 mg/100 g) and flavonoids (0.1 mg/100 g). Also, the result shows that this indigenous vegetable would likely be a good

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source of minerals such as K, Ca, Mg, and Fe. Hence, the presence of one or combination of some bioactive components present in the vegetable may be a factor responsible for its hepatoprotective activity as being claimed in folk medicine in Nigeria. SPECIAL ISSUE: Mineral Nutrition of Medicinal and Aromatic Plants. Guest Editors: M. Naeem, M. Masroor A. Khan, Moinuddin (Aligarh Muslim University) 2012 Brij K. Mishra, Anu Rastogi, Sudhir Shukla (India) Regulatory Role of Mineral Elements in the Metabolism of Medicinal Plants ABSTRACT Invited Review: Minerals have a diversified role in medicinal plant metabolism. Severity or scarcity of these causes multifarious effects in plant metabolism. Each and every aspect of plant biochemistry, physiology, anatomy, etc. is affected due to mineral nutrient composition of soils. Medicinal plants inherit resistance due to biosynthesis of bioactive substances (secondary metabolites) against the various types of diseases caused due to fungus, bacteria, viruses, mycoplasmas, insects and pests. The concentration of these minerals of both group i.e. activators or inhibitors present in the soil play a vital role in secondary plant metabolism. Minerals also play a major role in the reproduction of these medicinally important plants. Bioactive molecules of medical relevancy such as alkaloids, flavonoids, lignans, lipids, carbohydrates, resins, glycosides, phenolic compounds, volatile oils, vitamins, tannins etc. produced through various biosynthetic pathways of plants are a boon to urban, hilly and remote population of each nation. However, soils with different compositions of mineral elements adversely influence the metabolic activities of such valuable medicinal plants. Various physiological activities are governed through important mineral elements present in soils from where these are transferred to area where their need arises. So, accumulation and biosynthesis of these bioactive molecules in plant system are widely dependent on the availability of mineral elements in the soil. Different developmental stages of the medicinal plants need supplementation of different macro- and micro- elements during its various growth and biosynthesis steps. This review deals with regulatory role of various mineral elements in each biological activities of different medicinal plants viz. Papaver, Catharanthus, Withania etc. M. Naeem, M. Masroor A. Khan, Moinuddin (India) Role of Mineral Nutrients in Cultivation of Medicinal Legumes ABSTRACT Invited Review: Legumes (Fabaceae family) produce primary and secondary metabolites and other phytochemicals such as nutraceuticals, pharmaceuticals, pesticides and other industrial products. The medicinal legumes are potential sources of glycosides (aloe-emodin, chrysophenol, emodin, rhein, etc.), antibiotics, flavonoids, alkaloids and phytochemicals, which are used in drug manufacturing by the pharmaceutical industries. Thus, escalation of yield and quality of the medicinal legumes is of paramount importance. The various ways to improve yield and quality of medicinal legumes essentially include the supply of mineral nutrients as per the soil demand. In fact, balanced nutrition of crop plants plays a vital role in sustaining the yield and quality of medicinal plants together with maintaining the fertility status of soils on long-term basis. Among the macro-nutrients, phosphorus (P) is a major component of metabolic molecules involved in storage and utilization mechanism of energy that affects the growth and metabolism of plants significantly. Another element of macro-importance is calcium (Ca) that plays important structural and physiological roles in plants. It is essential for maintaining the stability of the membranes and walls of the cells and maintains the cell integrity. Above all, Ca is a second messenger and, thereby, controls the growth and differentiation of plants. In this review article, we have gathered important information about the individual effects of P and Ca on selected medicinal legumes. This review also covers the general description and therapeutic uses of medicinal legumes. The authors have themselves carried out a considerable work to evaluate the effect of sophera (Cassia sophera L.) and Cassia tora L. (Cassia obtusifolia L.). Remigius Chizzola (Austria) Metallic Mineral Elements and Heavy Metals in Medicinal Plants ABSTRACT Invited Review: A large proportion of the worlds population relies on medicines from the herbal sources. In this review, data about the mineral element content of medicinal and aromatic plants, including major elements, trace elements and toxic heavy metals, will be discussed and compared. Plants and drugs with remarkable elemental compositions will be pointed out. The P and Ca on selected medicinally important leguminous plants including hyacinth bean (Lablab purpureus L.), coffee senna (Senna occidentalis L.), senna

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main focus will be given on plants used in and originating from Europe and the Mediterranean region. The main metallic mineral macronutrients accumulate usually in the order K > Ca > Mg. The contents of beneficial trace elements decrease commonly from Fe > Mn > Zn > Cu > Ni > Mo. Higher concentrations occur in the leaves compared to other plant parts. Amongst the toxic heavy metals Cd, Pb and Hg are the best investigated. Due to its high mobility in the soil and good availability to plants, monitoring Cd merits special attention. A range of plants shows the tendency to accumulate higher concentrations of this element. For these plants, which may display problematic Cd levels, production measures should be taken to minimize the Cd accumulation, taking into consideration the growing site, planting material and fertilization regime. Interactions in the uptake between various elements have been described. The most prominent one occurs between Cd and Zn. Furthermore, it will be discussed that during the preparation of herbal tea using boiling water, only a proportion of the minerals is extracted. Finally regulations to set limits for toxic heavy metals at the national or international level are considered. Dragoja Radanovi, Svetlana Anti-Mladenovi (Serbia) Uptake, Accumulation and Distribution of Potentially Toxic Trace Elements in Medicinal and Aromatic Plants ABSTRACT Invited Review: This paper reviews contents and distribution of essential and potentially toxic trace elements (Fe, Zn, Cu, Mn, Co, Cd, Pb, Cr, Ni) in medicinal and aromatic plants (MAPs). A large difference in metal uptake between plant species has been demonstrated in many studies. Concentration of trace elements in the MAPs is largely determined by soil type, growing site conditions and soil chemical processes, particularly soil pH. Additionally, anthropogenic pollution of soil and air affects metal contents in the MAPs. Furthermore, the species and genotypes of a species differ greatly in their ability for metal uptake. Recent studies have shown capability of some of the MAPs for metal hyperaccumulation; particularly Hypericum perforatum accumulates more Cd than other plants under the same growing conditions. Uptake, accumulation and translocation of metals, and consequently, a good quality of MAPs and final products that should be free from potentially harmful constituents, can be controlled by combining: (i) adequate choice of growing site (especially regarding: soil pH, organic matter content, clay content, redox potential, distance from point pollution sources, etc.), (ii) choice of suitable genotypes and (iii) appropriate management of soil and crops. Additionally, the MAPs, and in particular the plant parts to be utilized (e.g. roots, leaves, flowers, herb) should be regularly tested for contaminant load before processing for pharmaceutical drugs. The present review also highlights the potential risks of human intoxication with trace elements after the consumption of pharmaceutical drugs (essential oils, extracts, and teas). Moinuddin, M. Masroor A. Khan, M. Naeem (India) Drought Stress Effects on Medicinal and Aromatic Plants and the Possible Stress Amelioration by Mineral Nutrition ABSTRACT Invited Review: Limited water supply is one of the major abiotic factors that adversely affects agricultural crop production worldwide. Drought stress disturbs the balance between the production of reactive oxygen species (ROS) and the antioxidant defense, causing the accumulation of ROS that induce oxidative stress to proteins, membrane lipids and other cellular components. Drought stress hampers photosynthesis harshly owing to interference with stomatal functioning and causing serious damage to photosynthetic apparatus through ROS production. Hence, water-stressed plants exhibit disturbance in metabolism, particularly that of carbon, nitrogen and oxygen. As a result of water stress, growth parameters associated with root and above-ground parts, e.g. leaf area, height, and fresh and dry matter of plants, are severely reduced, leading to drastic reductions in growth, yield and quality of plants. Mineral elements have numerous functions in plants including charge balance, electron carriers, structural components, and enzyme activation. Besides, these elements behave as osmotica for maintenance of turgor and growth under drought stress. However, their uptake is reduced to a great extent under water deficit. Hence, their additional supplementation under water deficit results in improvement in their uptake and mitigation of deleterious effects of drought stress. A wide range of studies on the effect of drought stress on growth, yield and quality of field crops have been conducted; however, such studies related to medicinal and aromatic plants (MAPs) are few. In this review, in addition to describing the adverse effects of water stress on the growth, yield and quality attributes of MAPs, the ameliorative role of some of the macronutrients (nitrogen, phosphorus, potassium, and calcium) is discussed. M. Masroor A. Khan, M. Naeem, Firoz Mohammad, Moinuddin (India) Establishment of Mineral Nutritional Requirement for Some Important Medicinal and Aromatic Plants in Climatic Conditions of Northern India

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ABSTRACT Invited Review: Exploiting the healing power of plants has been a human practice since prehistoric times. To meet the ever-increasing demand of medicinal plants, cultivation of these plants on scientific principles is the only solution. Mineral nutrients are indispensable for the growth and development of plants as their deficiency limits crop production significantly. Like other plants, medicinal plants also require mineral elements for their growth and development. However, all species do not require the same complement of minerals in the same amounts; in fact, the differences between species are much less variable than those between the amounts of specific minerals within a single species. With a view of improving yield and quality (active constituents) of medicinal and aromatic plants (MAPs), the mineral nutritional requirement of several MAPs in an Indian climate was calculated. The work was conducted in pots as well as in the field, supplying graded levels of the nutrients to the soil along with proper controls. This review provides information regarding the optimum requirement of N, P and K in Indian climatic conditions for the following MAPs: Anethum sowa, Cichorium intybus, Curcuma Longa, Cymbopogon flexuosus, Datura innoxia, Foeniculum vulgare, Lallemantia royleana, Linum usitatissimum, Mentha arvensis, Nigella sativa, Plantago ovata, Solanum nigrum, Tigonella foenum-graecum, Withania somnifera and Zingiber officinale. Jaime A. Teixeira da Silva (Japan), M. Naeem, Mohd. Idrees (India) Beneficial and Toxic Effects of Nickel in Relation to Medicinal and Aromatic Plants ABSTRACT Invited Review: Nickel (Ni) is a constituent of urease, and small quantities of Ni (0.01 to 5 g/g dry weight) are essential for some plant species. Ni has been considered as an essential micronutrient, which is required by urease for hydrolysing urea. Ni has a role in plant growth and N uptake in crops supplied with urea in calcareous soils. Ni deficiency affects plant growth, plant senescence, nitrogen metabolism, and iron uptake and it may play a role in disease resistance. Ni is absorbed and redistributed in plants via cation and/or metal-ligand complex transport systems. However, excessive Ni inhibits growth and development of plants, induces leaf chlorosis and wilting, and reduces total plant yields. Several investigators have performed several studies on the residual levels of heavy metals (HMs) in medicinal herbs. According to a study, the level of essential elements in MAPs is conditional, being affected by the geochemical characteristics of the soil and by the ability of plants to selectively accumulate some of these elements. The use of hyperaccumulators or wetland plants to remove and/or sequester Ni from soil and water has been proposed by various workers. The accumulation of HMs such as Ni in some desert plants may open a new perspective for the application of these species as accumulators of HMs to clean-up contaminated soils in dry environments. The researchers still debate over the causes and mechanisms of hyperaccumulation, which may depend on the morphological and physiological characteristics of particular plant species and the capacity of Ni to overcome the physiological barriers. We have reviewed the information regarding occurrence, sources, essentiality, toxicity and tolerance of Ni regarding medicinal and aromatic plants (MAPs), supporting our view discussing the effects of Ni on other plants. Azaizeh Hassan (Israel) Effects of Mineral Nutrients on Physiological and Biochemical Processes Related to Secondary Metabolites Production in Medicinal Herbs ABSTRACT Invited Mini-Review: Although biosynthesis of plant metabolites (e.g. alkaloids, terpenoids, phenolic compounds, etc.) is primarily controlled genetically, environmental factors such as stresses and nutrient elements affect the production of metabolites in plants. Many biological activities have been ascribed to these metabolites. In addition to their therapeutic effects, they play a role as chemical defense agents against microorganisms and herbivores. Therapeutic effects of medicinal plants are associated with their chemical peculiarities. Chemical features of these plants serve as an integral determinant of their species specificity and pharmacological properties and facilitate their wide use in medical practice and other uses. The relationship between the synthesis of physiologically active substances and accumulation of elements is mediated by several levels of molecular regulation. There is general agreement that various mineral nutrients increase the growth of individual plants and, consequently, enhance the total plant biomass yield. However, the effects of certain macro- and micro-nutrients on the production of secondary metabolites are also modulated by environmental conditions and depend on plant species. Incorporation of one or more of the trace elements could increase or decrease the production of secondary metabolites depending on the plant species as well as on the concentrations of these elements. The effect of the interaction of certain nutrients on the production of secondary metabolites in herbs is discussed in the present review as well. Katarzyna Seidler-oykowska, Anna Golcz (Poland) The Effect of Organic Cultivation on the Contents of Macro- and

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Micro-elements Studied in Polish Cultivars of three Medicinal Plants ABSTRACT Original Research Paper: Experiments were conducted to evaluate the contents of macro-elements [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), sodium (Na), and magnesium (Mg)] as well as micro-elements [iron (Fe), manganese (Mn), copper (Cu), and Zinc (Zn)] in organic herbs of sweet basil (Ocimum basilicum L.), savory (Satureja hortensis L.) and marjoram (Origanum majorana L.). For the chemical analysis, conducted on dry weight basis, the herbs were obtained from the experiments conducted in four certified organic farms in Poland. Among the herbs of basil, savory and marjoram, basil herb contained largest amounts of all macro- and micro-elements except that of Fe, which was maximal in marjoram. The average content of N, P, K, Mg Mn, Cu and Zn was higher in the organic herb compared with the conventional one. Enhanced content of macro- and micro-elements in the tested herbs had significant effect on the herb-quality. Kamel Msaada, Karim Hosni, Mouna Ben Taarit, Mohamed Hammami, Brahim Marzouk (Tunisia) Effects of Crop Season and Maturity Stage on the Yield and Composition of Essential Oil of Coriander (Coriandrum sativum L.) Fruit ABSTRACT Original Research Paper: The aim of this study was to determine the yield and chemical composition of the essential oil (EO) extracted from fruits of coriander (Coriandrum sativum L. cv. Menzel Temime with high essential oil yield: 0.35%, w/w) as affected by two successive crop seasons and different stages of maturity using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The maximal oil yields (0.17 and 0.32%) reached at the final stage of maturity in the season 2003 and 2004, respectively. Oil yields were significantly (P < 0.001) affected by the crop season, stage of maturity and their interaction. EO composition varied significantly (P < 0.05) with the stages of maturity. The compound linalool was the main compound in the season 2003 (79.86 8.16) as well as in 2004 (80.04 9.12). The strong effect of crop season, maturity stage and their interaction was found on 36 EO compounds. Several compounds (-terpinene, -terpineol, terpinene-4-ol, carvone and p-cymen-8-ol) showed a different response to crop season, stage of maturity and their interaction. Oyeboade Adebayo, Rukkayah Aminu-Taiwo, Esther Adesegun (Nigeria) Growth, Essential Oil Content and Antimicrobial Activity of Basil Influenced by Nitrogen Fertiliser and Plant Density ABSTRACT Original Research Paper: A field experiment was conducted to investigate the effect of different nitrogen (N) rates and plant densities on vegetative growth, yield and essential oil content of basil (Ocimum basilicum L). Three N rates (0, 60 or 120 kg N ha1) and three plant densities (6,250, 12,500 and 25,000 plants/ha) were assigned to main plots and sub-plots respectively in a split plot design with three replications. Basil essential oil was further evaluated for its in vitro effect on Meloidogyne incognita and Fusarium oxysporum. N fertilization up to 120 kg ha1 significantly (P > 0.05) increased yield of above-ground (40.1 t/ha) and leaf fresh biomass (122.6 t/ha), leaf essential oil yield (9.3%), but it did not affect plant height and the number of branches per plant. The increase in essential oil yield induced by N fertilization was related to leaf biomass. Plant density significantly increased the above-ground biomass yield and the plant N content but did not affect plant height and essential oil content. The highest above-ground biomass yield (47.6 t/ha) and plant nitrogen content (2.7%) were recorded with a density of 25,000 plants/ha. Basil essential oil completely prevented egg hatching in M. incognita at all concentrations tested while the mycelium growth of F. oxysporum was significantly reduced as oil concentration increased. The lowest mycelium growth of 1.5 mm was recorded on plates with 40% essential oil. Khalid A. Khalid (Egypt) Effect of Phosphorous Fertilization on Anise, Coriander and Sweet Fennel Plants Growing under Arid Region Conditions ABSTRACT Original Research Paper: Arid regions in Egypt are characterized by poor nutrients such as phosphorous (P) and unfavorable environmental conditions which negatively affect growth and productivity of medicinal and aromatic plants including anise (Pimpinella anisum L.), coriander (Coriandrum sativum L.) and sweet fennel (Foeniculum vulgare var. Dolce) plants. Thus, the main objective of the present investigation was to study the effect of different levels of P fertilizer as calcium superphosphate (15% P2O5) at 0 (control), 37.5, 56.3 and 75 kg ha-1 P2O5 on the morphological and biochemical contents of these three plants under arid conditions over two successive seasons. The most effective rate was 56.3 kg ha-1, resulting in a positive increase in

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vegetative growth characters [plant height (cm), leaf number (plant ), branch number (plant ), umbel number (plant ), herb fresh weight (plant ), herb dry weight (plant ) and fruit yield (plant )]. The highest values of vegetative growth characters were 44.5, 35.6, 10.9, 31.2, 20.9, 6.8 and 6.9, respectively for anise; 83.0, 66.1, 7.8, 24.7 19.9, 8.7 and 6.9, respectively for coriander; 98.9, 32.8, 6.9, 16.8, 113.4, 85.8 and 24.7, respectively for sweet fennel. 75kg ha-1 led to higher biochemical contents than the control. The increases were 1.2, 0.4 and 0.6% in essential oil; 3.7, 6.0 and 2.1% in fixed oil, 6.6, 11.0 and 9.0% in total carbohydrates; 0.6, 0.3 and 0.4% in soluble sugars; 3.7, 4.9 and 3.9% in crude protein; 1.4, 1.3 and 1.2% in nitrogen; 1.9, 0.8 and 1.5% in phosphorous; 1.5, 1.1 and 0.7% in potassium for anise, coriander and sweet fennel, respectively. Reham Sabry, Adel Salama, Mahmoud Sharaf-Eldin (Egypt) Effect of Plant Spacing and Nitrogen Levels on Growth, Yield and Essential Oil Content of Marrubium vulgare Plant ABSTRACT Short Communication: Marrubium vulgare L. (Lamiaceae) is a perennial herb commonly known as "horehound" and grows wild in the Egyptian desert. We report the first systematic cultivation of this plant in Egypt for the estimation of optimal plant spacing and nitrogen dose. Three plant spacings were studied (25, 35 and 50 cm apart) and three nitrogen doses were applied (N1 (80), N2 (120) or N3 (160) kg N ha-1). The maximum mean values of plant height were obtained with a 35-cm plant spacing. The lowest number of branches and fresh and dry weights of plants were observed with the 25-cm plant spacing in harvest. Nitrogen fertilization had a significant effect on most of agronomic parameters studied. N2 exhibited the best growth attributes, although the differences between N2 and N3 were not significant in most harvests. On the other hand, oil content was not influenced by plant spacing or nitrogen fertilization in the three harvests. A plant spacing of 35 cm and an N2 rate of nitrogen application resulted in the best horehound plant growth. SPECIAL ISSUE: Medicinal Plants and Cancer Research. Guest Editor: Dilfuza Egamberdieva (National University of Uzbekistan, Faculty of Soil Science and Biology, Tashkent, Uzbekistan), Antonio Tiezzi (Laboratory of Plant Cytology and Biotechnology, Department of Environmental Sciences, Tuscia University, Italy). 2012/2013 Mohamed L. Ashour, Sherif S. Ebada (Egypt) Plant Natural Products as Potential Modulators of the Transcription Factor NF-B ABSTRACT Invited Review: Nuclear factor-B (NF-B) is an inducible transcription factor which plays important role in the regulation of the immune, inflammatory and carcinogenic responses. Partial NF-B activation is necessary for normal cell survival and immunity; however the deregulated NF-B expression is associated with cancer development, metastasis, and in several inflammatory disorders with a resistant phenotype and poor prognosis. Therefore, NF-B has become an interesting target for drug discovery and several natural and synthetic products have been screened for their ability to inhibit NF-B pathway. This review surveys the plant natural products with significant NF-B inhibitory activity focusing on their potential mechanism of action and their implications for cancer therapy. Donatella Ceccarelli, Elisa Ovidi, Doriana Triggiani, Carlo F. Morelli, Giovanna Speranza, AnnaRita Taddei, Antonio Tiezzi (Italy) Antiproliferative Activity of Aloe arborescens Leaf Skin Extracts Tested on Murine Myeloma Cells: Cytological Studies and Chemical Investigations ABSTRACT Original Research Paper: MTT assays showed an intense antiproliferative activity (80%) of Aloe arborescens leaf skin extracts tested on murine myeloma cells. Bioassay-guided fractionation carried out by TLC allowed the identification of a spot showing antiproliferative activity; HPLC and NMR investigations showed that the TLC spot consisted of aloenin A and aloins A and B. The effects of the leaf extract and of the TLC spot were evaluated both by immunofluorescence techniques in order to test the microtubular array and at the morphological level by SEM and TEM observations. Doriana Triggiani, Donatella Ceccarelli, Maria Grazia Cusi, Alessandro Paffetti, Daniela Braconi, Lia Millucci, Giulia Bernardini, Annalisa Santucci (Italy) Rubus ulmifolius Leaf Extract Inhibits Proliferation of Murine Myeloma Cells ABSTRACT
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Original Research Paper: In the present paper we report preliminary results on the antiproliferative activity of Rubus ulmifolius leaf extract against murine myeloma cells (P3X63-Ag8.653). Cytotoxic effects of plant extract (range 3-30 L/mL of cell culture) were evaluated by microscopy analyses and viability assays, revealing a clear dose-response relationship for cytotoxicity in treated cells vs. controls. The number of viable cells was reduced to nearly 30% when plant extract was tested at 3 L/mL, and to nearly 3% when plant extract was tested at 5 L/mL. Higher concentrations of plant extract allowed the detection of a very limited number of viable cells. Furthermore, the effects of R. ulmifolius leaf extract at the molecular level were investigated through a comparative proteomic approach, which allowed us to highlight how it might modulate protein expression in murine myeloma cells. In particular, only one protein spot was found under-expressed, whereas 44 protein spots showed significantly higher levels in plant extract-treated cells when compared to control cells. Our results may thus lay the basis and open new perspectives for future investigations of the effects of plant extracts in mammalian cells.

Pest Technology
SPECIAL ISSUE: Potato pathology. Guest Editor: Mejda Daami-Remadi (Centre Rgional des Recherches en Horticulture et Agriculture Biologique, Chott-Mariem, Tunisia), 2012 Dennis Halterman, Amy Charkowski, Jeanmarie Verchot (USA) Potato, Viruses, and Seed Certification in the USA to Provide Healthy Propagated Tubers ABSTRACT Invited Review: Potatoes are vegetatively propagated and this can result in the dissemination of pathogens, and viruses in particular, in the tubers. Viruses infecting potato can be categorized by their mechanisms of transmission: aphid transmitted, mechanically transmitted, and soil-borne viruses. The most important viruses in North America include Potato leafroll virus, Potato virus Y, X, A, S, M, Tobacco rattle virus, and Potato mop top virus. The methods for chemical control of virus disease are greatly influenced by their mechanism of spread in the field. However, tubers play an important role in the spread of virus disease and this has led many regions to develop seed certification programs. The use of certified virus-free tubers by growers has been vital for control of disease worldwide. In addition, breeders have identified genetic resistance that can be introgressed into popular cultivated varieties and provides a method of control that is less costly than chemical application. In recent years there has been an emergence of viruses and recombinant virus strains that have posed new challenges to pathologists for seed certification and for breeders. Here we discuss the latest issues and challenges that viruses pose to potato production. Mejda Daami-Remadi (Tunisia) Potato Fusarium Dry Rot in Tunisia: Current Status and Future Prospects ABSTRACT Invited Review: Fusarium dry rot (FDR) is a postharvest disease of potato tubers of economic importance worldwide. In Tunisia, losses attributed to Fusarium spp. infections may be aggravated in the presence of other tuber rot pathogens. A review of published data indicates that five Fusarium species (F. sambucinum, F. graminearum, F. culmorum, F. oxysporum and F. solani) are involved in a disease complex in Tunisia. These species present a great range of inter- and intra-specific variability in growth, sporulation and aggressiveness. Their level of aggressiveness is variable, depending on storage conditions (mainly temperature) and potato cultivar. Although a large number of cultivars have been assessed, no cultivar was resistant to FDR although potato cultivars exhibited different levels of susceptibility to the disease. The ranking of cultivars with regard to their susceptibility to FDR changed depending on the Fusarium species involved in disease development and on the temperature used for tuber storage. Chemical-based treatments using old and new generations of fungicides tested against Fusarium species have successfully limited dry rot severity and revealed the appearance of benzimidazole-resistant isolates of F. sambucinum. Biological control studies undertaken based on various and complementary experiments in vitro, in vivo and under natural conditions permitted the selection of potentially important microbial agents (fungi and Gram-positive bacteria) for the control of potato FDR. Their mechanisms of action were also elucidated. Other alternatives of disease control which may be a part of an integrated management strategy are also cited. This review highlights the specificity of problems related to FDR in Tunisia, regarding mainly pathogen aggressiveness, genetic resistance and control alternatives and their limits which may be useful for the development of an integrated strategy for FDR management and for better guiding future research on the influence of the Fusarium species complex, potato cultivar, storage conditions and their interactions on the effectiveness of disease control.

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Kalthoum Harbaoui (Tunisia), Theo van der Lee (The Netherlands), Walid Hamada (Tunisia) Characterization of the Potato Late Blight Pathogen Phytophthora infestans in Tunisia ABSTRACT Invited Mini-Review: This mini-review focus on summarizing different phenotypic and genotypic analysis recorded on Phytophthora infestans population collected from the Northern area of Tunisia. Here, we demonstrated that mating type distribution plays a primordial role in genetic structure of the pathogen between sampling regions. Also, a probably strong correlation between mating type and metalaxyl resistance could be mentioned. Next to that, we found that the aggressiveness and the virulence patterns were highly effective parameters to outline phenotypic diversity in Tunisian population that showed specific characteristics comparing with Algeria and others countries in the world. Consistently, genotypic diversity based on SSR markers seems to be a very interesting key in the genetic understanding of the pathogen. In fact, we found that P. infestans populations in Tunisia were divided to two major phylogenic groups: a clonal lineage group shared between all sampling regions and a diverse group detected in potato population and specific to two sub-regions. Thus, we could conclude that genotypic diversity confirmed phenotypic diversity and both analysis leaded us to conclude that population structure of P. infestans in Tunisia is very specific either in North African area or all over the world. Pablo L. Cicore, Patricia A. Suarez, Adriana B. Andreu (Argentina) Phosphites Effect on Late Blight Control and Physiological Parameters in Commercial Potato (Solanum tuberosum L.) in Argentina ABSTRACT Original Research Paper: Phosphite (Phi) has been recommended to enhance plant resistance against Phytophthora spp. The purpose of this study was to evaluate the effect of calcium phosphite (PhiCa) and potassium phosphite (PhiK) under field conditions, taking into account physiological parameters, performance and protection against late blight in the foliage of two potato cultivars, and comparing these results to controlled in vitro experiments. Experimental design was a split plot in randomized complete blocks with four replicates. The main plot comprised two potato cultivars (Kennebec and Spunta) and the sub-plots three late blight control treatments: control, PhiCa and PhiK. The interaction between cultivar and treatment on the area under disease progress curve (AUDPC) was determined. In Kennebec, the PhiK treatment showed a significantly lower AUDPC than control and PhiCa treatments. No differences were detected between treatments on Spunta. In the detached-leaf tests, there was a significant treatment effect (P < 0.01). In both cultivars, PhiK application resulted in a significantly lower damage growth rate (DGR) compared to the control and PhiCa treatments, although there was less difference in Spunta. Treatments tested had no significant effect on yield. However, PhiK application tended to increase yields by 20.5 and 14.8% in relation to control and PhiCa treatments, respectively. Under experiment conditions, Phi decreased disease severity and increased yields, with differences between cultivars and chemical compounds. Mejda Daami-Remadi, Ins Dkhili, Hayfa Jabnoun-Khiareddine, Mohamed El Mahjoub (Tunisia) Biological Control of Potato Leak with Antagonistic Fungi Isolated from Compost Teas and Solarized and Non-Solarized Soils ABSTRACT Original Research Paper: Several fungal agents, isolated from compost teas and soil, were tested for their antagonistic activity against Pythium aphanidermatum and P. ultimum causing potato leak in Tunisia depending on the timing of their application. In vitro dual culture experiments showed that Aspergillus spp., Penicillium sp. and Trichoderma sp. isolates had inhibited differently the two Pythium species and that their inhibitory effect was improved when plated 24 h prior pathogen. Tested in vivo for their ability to control leak development, pooled data of antagonistic treatments revealed that all microbial agents had decreased rot incidence and severity compared with the untreated control. Moreover, their preventive application i.e. 24 h prior to pathogen inoculation has contributed by the reduction of leak severity by about 77.28% as compared to simultaneous application. Both Pythium species were differently controlled by the tested fungi. In fact, leak caused by P. aphanidermatum was completely suppressed (100% of inhibition) with all microbial agents applied 24 h before inoculation while for P. ultimum, the percentage of reduction of leak severity ranged between 15.37% and 95.94%. Boutheina Mejdoub-Trabelsi, Hayfa Jabnoun-Khiareddine, Mejda Daami-Remadi (Tunisia) Effect of Fusarium Species and Temperature of Storage on the Susceptibility Ranking of Potato Cultivars to Tuber Dry Rot ABSTRACT

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Original Research Paper: The relative susceptibility of 11 potato cultivars to Fusarium solani, F. oxysporum f. sp. tuberosi, F. sambucinum, and F. graminearum was assessed under different temperatures of storage. Obtained results revealed that these Fusarium species showed variable aggressiveness upon the 11 tested potato cultivars. In fact, F. sambucinum was the most aggressive on the majority of cultivars. F. graminearum showed comparable aggressiveness as F. sambucinum only on some cultivars. F. oxysporum f. sp. tuberosi and F. solani caused a relatively less severe dry rot on the cultivars tested. After 21 days of incubation at 15, 20, 25 and 30C, none of the cultivars tested was completely resistant to all Fusarium species and only some of them showed lesser susceptibility to at the most one species. This is the case of Spunta, Mondial and Nicola, the most cultivated in Tunisia, which tolerated at least one species of Fusarium: F. oxysporum f. sp. tuberosi for the two first cultivars and F. solani for the second. The rank order of susceptibility levels to Fusarium dry rot varied depending on Fusarium species and temperature of storage. When tubers were inoculated with F. sambucinum (the most aggressive species), cultivars placed in the less susceptible group at 30C were classified as highly susceptible at 15C. However, when tubers were inoculated with F. oxysporum f. sp. tuberosi (the least aggressive), cultivars placed on category of less susceptible were almost the same at all tested temperatures. Mejda Daami-Remadi, Rafik Bouallgue, Hayfa Jabnoun-Khiareddine, Mohamed El Mahjoub (Tunisia) Effect of the Level of Seed Tuber Infection by Colletotrichum coccodes at Planting on Potato Growth, Black Dot Severity and Subsequent Yield Reduction ABSTRACT Original Research Paper: A great deal of attention has been given to tuber infections of Colletotrichum coccodes as skin blemishes depreciate the tubers commercial value and serve as means of pathogen spread into new productions areas. This study was carried out on cv. Spunta potato plants to compare, quantitatively, the relative impact of different tuber-borne inoculum levels on black dot severity, plant growth and expected yield loss. Black dot severity recorded 60 (pot experiment) or 90 days post-planting (field experiment) on the below-ground plant parts varied significantly depending on inoculum levels tested. The severest symptoms were induced by the highest seed tubers infections corresponding to more than 50% of the seed tuber surface area covered with black dot lesions. The recorded disease severity was found to be highly and positively correlated with the level of seeds contamination. The aerial part fresh weight noted 60 DPP was reduced, compared to plants grown from the symptomless seed tubers, by 24 and 42% when heavily infested seeds were planted. This growth parameter was also negatively correlated with black dot severity on seed tubers and on the below-ground plant parts. In the pot experiment, tuber weight was reduced by about 14-21% on plants grown from extremely infected seeds as compared with the non infested control plants. Yield decrease, subsequent to the planting of diseased seeds, varied between 15 and 27% when C. coccodes inoculum levels ranged between 1 and 5. The field study revealed the existence of significant and negative correlations between the tuber yield, the level of seed-borne inoculum and disease severity on the below-ground stems, roots and stolons. Mejda Daami-Remadi, Hayfa Jabnoun-Khiareddine, Abir Sdiri, Mohamed El Mahjoub (Tunisia) Comparative Reaction of Potato Cultivars to Sclerotium rolfsii Assessed by Stem Rot and Tuber Decay Severity ABSTRACT Original Research Paper: Experiments were conducted to evaluate the relative susceptibility of 11 local potato cultivars to Sclerotium rolfsii by using several stem and tuber disease parameters. Based on stem rot severity, noted three weeks post-inoculation, pathogen penetration, and percentage of rotten tissue, recorded after 8 days of incubation at 30C, none of the cultivars tested was resistant to S. rolfsii. However, there was a variable degree of susceptibility in which cultivar Tango was found to be the most susceptible whilst Daisy was the most tolerant to the disease. The most severe stem rot attributed to pathogen inoculation was observed, three weeks post-inoculation, on Tango and Spunta plants. For all cultivars combined, the percentage of rotten tuber tissue was related to the lesion diameter formed on the tuber surface and to the pathogen penetration. Mohamed Ali Triki, Ines Hammami, Samira Krid Hadj-Taieb, Mejda Daami-Remadi, Aymen Mseddi, Mohamed El Mahjoub, Radhouane Gdoura, Nouri Khammassy (Tunisia) Biological Control of Atypical Pink Rot Disease of Potato in Tunisia ABSTRACT

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Original Research Paper: In order to biologically manage an atypical pink rot disease of potato, various indigenous antagonistic bacteria were tested against Pythium aphanidermatum. In vitro tests showed that Achromobacter xylosoxidans (B4), Pseudomonas putida (B7) and P. fluorescens (B10) isolates resulted in significant decreases in the mycelial growth of P. aphanidermatum ranging from 50 to 88%. Therefore, they were further evaluated for their ability to produce diffusible metabolites in culture filtrates. P. fluorescens was shown to be the most efficient by exhibiting the highest inhibitory activity in vitro. Total inhibition of P. aphanidermatum growth was achieved with the culture filtrates of this antagonistic agent applied at a 1/2 (v/v) ratio. The biological treatments of inoculated potato (Solanum tuberosum L.) tubers cv. Spunta by the bacterial culture filtrates were also efficient in reducing the rotting severity by more than 90% with P. fluorescens as compared to the untreated and inoculated tubers. Thus, the management of natural potato infection in storage and in the field by applying some antagonistic bacteria should be investigated. Heba Mahfouze, Khalid El-Dougdoug, Badawi Othman, Mostafa Gomaa (Egypt) Molecular Markers in Potato Cultivars Treated with Ribosome-Inactivating Proteins ABSTRACT Original Research Paper: Most plants produce ribosome-inactivating proteins (RIPs) that inhibit protein synthesis through modification of RNA. In this report we studied the inheritance of resistance to Potato virus Y (PVY) in five potato cultivars treated with RIPs using ISSR-PCR since genetic information on this topic is limited. Leaf extracts from pokeweed (Phytolacca americana and P. acinosa and the marvel of Peru (Mirabilis jalapa) were sprayed on five potato cultivars (Selan, Spunta, Cara, Diamond and Nicola) before virus inoculation, inhibiting infection by almost 100%, corroborated by DAS-ELISA. Changes in DNA (relative to PVYNTN-infected plants and the healthy control) in potato cultivars pretreated with 100 g/ml AVP resulted in genetic variations detected by ISSR-PCR analysis, which was performed using five primers. A total of 63 scorable amplified DNA fragments ranging from 90 to 1105 bp were observed using these primers, 43 of which were polymorphic while the others were monomorphic. The five primers showed a mean polymorphic percentage of 68.25%, the highest percentage derived from primer ISSR-1 (87.50%). The AVP-pretreated potato cultivars varied considerably in banding patterns using the five ISSR-PCR primers. Nicola had the highest number of unique markers (9), followed by Selan (8), Spunta (5), Dimond (1) and Cara (0). Leaf extracts from P. americana, P. acinosa and M. jalapa could be used in simple crop-protection agricultural systems by spraying these extracts on leaves of various crops to prevent or control viral infection. Lobna El Hajji, Najet Horrigue-Raouani (Tunisia) Efficiency of Solarization and Incorporation of Cattle Manure in Management of Meloidogyne javanica (Treub.) Chitwood and Globodera pallida (Stone) Behrens Associated with Potato ABSTRACT Original Research Paper: Potato is a strategic crop in Tunisia but is susceptible to many pests and diseases. Nematodes of the genera Meloidogyne and Globodera are important pests that cause yield losses and depreciation of tuber quality. A trial was carried out in a plot where the soil was naturally infested with M. javanica and G. pallida. Three soil treatments were tested and compared. Soil solarization (SS) and incorporation of cattle manure (70 T/ha), applied alone or in combination, were assessed for their impact on potato crop. SS alone improved potato growth by 13.56% and yield by 26.53%. Compared to the control, SS combined with cattle manure decreased the multiplication rate of M. javanica by 38.14% and enhanced plant height (29.47%), fresh root and shoot weight (47.89 and 59.53%, respectively) and yield (26.40%). V. Prathap Reddy, N. Narasimha Rao, P. S. Vimala Devi, M. Lakshmi Narasu, V. Dinesh Kumar (India) PCR-Based Detection of cry Genes in Local Bacillus thuringiensis DOR Bt-1 Isolate ABSTRACT Original Research Paper: A sporulating culture of Bacillus thuringiensis ssp. kurstaki strain DOR Bt-1 was isolated from castor semilooper (Achaea janata L.) cadavers from a castor bean (Ricinus communis L.) field at Kothakota mandal of Mahaboobnagar district, Andhra Pradesh, India. This strain has shown high toxicity against many insect pests and the wettable power formulation of this strain obtained through solid-state fermentation has been registered with the Central Insecticides Board (CIB), India under section 9 (3b). Based on the bioassay results and the efficacy of the formulation on different Lepidopteran insects, it was concluded that the strain must be carrying diverse cry genes. Therefore, to understand the cry gene profile of this strain, a PCR-based assay was carried out. Genomic PCR analysis of the DOR Bt-1 isolate with universal primers specific to cry gene classes indicated the presence of both cry1 and cry2 class-specific genes. Further, PCR analysis

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with cry gene-specific primers revealed the presence of cry1Aa, cry1Ab, cry1Ac, cry2Aa and cry2Ab genes. Thus, DOR Bt-1 isolate harbors a combination of different cry genes, such as Lepidoptera-active cry1Aa, cry1Ab and cry1Ac, and Lepidoptera-Diptera-active cry2Aa and cry2Ab genes, indicating that this isolate has the potential to be effective against both Lepidopteran and Dipteran insect pests.

Plant Stress
Mohd Mazid, Taqi Ahmed Khan, Firoz Mohammad (India), Jaime A. Teixeira da Silva (Japan), Mohd Nasir Khan (Saudi Arabia) Role of Hydrogen Peroxide in Tolerance to Abiotic Stress in Plants ABSTRACT Review: Plants have evolved a complex array of potential biochemical regulatory mechanisms to tolerate the effect of various abiotic environmental stresses. Hydrogen peroxide (H2O2) clearly functions as signaling molecule in plants and has gained a significantly critical position in plant stress biology, particularly due to its properties (evolutionary ancient molecule, long life span, not a free radical, potentially strong toxic oxidant, key systemic factor for programmed cell death, rapidly diffusing from cell to cell, transporting long distances from production sites) and multifunctional roles in plant growth and development. However, in the last decade, the potential of H2O2 as a signal of stress tolerance to abiotic stress in plants has positioned it for increasing attention. As highlighted in the present review, recent studies on H2O2s functionality in the induction of tolerance in plants to abiotic stresses have become unravelled and are very impressive. In plants, H2O2 possesses antioxidant properties, regulates the antioxidant defense system and might also act as a signal in the activation of a number of defense gene expression pathways through various sensing mechanisms under abiotic stress. In addition, H2O2 plays an important role in resistance to temperature, drought, heavy metal, UV-B and salt stress.

Seed Science and Biotechnology Terrestrial and Aquatic Environmental Toxicology

Yones Khaledian, Sohaila Ebrahimi, Hamed Bag Mohamadi (Iran) Effects of Urbanization and Changes Land Use on Water Geochemical Properties. A Case Study: Gharaso River in Golestan Province, North of Iran ABSTRACT Original Research Paper: Rapid growth of urban populations and the expansion of urban areas in the north of Iran have led to changes in land use and subsequently changes in water quality of the Gharaso River in Golestan Province. Thus, the aim of this research was to investigate the role of urbanization and land use change on water quality in seven stations (Siahab, Angirabad, Shastkalateh, Yasaqi, Naharkhoran, Abgir and Poleordogah). In this paper, principal component analysis and hierarchical cluster analysis were used to investigate the water quality of the Gharaso River. Water samples collected every month over a ten-year period from 7 sampling stations, along a 50 km section of Caspian Sea that is under the influence of anthropogenic and natural changes were analyzed. The highest pollutions were equal with, electrical conductivity 1448.6 and 1127.5 (mmho/cm), total dissolved solids 911.16 and 722.3 (mg/l), bicarbonate 293.37 and 331.83 (meq/l), chloride 221.7 and 139.56 (meq/l), calcium 81.74 and 93.44, sodium 150.7 and 86.49 (meq/l), potassium 6.9 and 14.31 (meq/l), sodium adsorption ratio 2.83 and 1.9 (meq/l), temporary hardness 230.6 and 262.19 (mg/l), total hardness 404.59 and 370.6, in the Siahabad and Naharkhoran, respectively, sulfate 195.6 and 153.9 (meq/l), magnesium 52.8 and 40.34 (meq/l), in the Siahabad and Abgir, respectively. The highest pH was 7.8 in the Naharkhoran station. The results indicated that water quality in Siahab and Naharkhoran stations was in the poorest quality among other stations because of anthropogenic effects. The best water quality was in Shast Kalateh station because there were no changes in the land uses. The Gharaso River joins the Caspian Sea creating sediment problems and an increasing threat to human and marine health. Sonia Sharma, Adarsh Pal Vig (India) Genotoxicity of Atrazine, Avenoxan, Diuron and Quizalofop-P-ethyl Herbicides using the Allium cepa Root Chromosomal Aberration Assay ABSTRACT Original Research Paper: Cytogenetic effects of different herbicides viz. Atrazine, Avenoxan, Diuron and Quizalofop-P-ethyl (QPE), were evaluated in the root tip meristem cells of Allium cepa. In the Allium root growth test, the effective concentration

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(EC50) value was determined as approximately 0.5 ppm in the case of Atrazine and Avenoxan and 1.0 ppm in the case of Diuron and QPE herbicides with a control for each combination. Mitotic index decreased with increasing herbicide concentration at each exposure time. In anaphase-telophase cells, the total percentages of different chromosomal aberrations like stickiness, bridges, break(s), ring chromosomes, vagrant chromosomes, c-mitosis, delayed anaphase, laggard(s) and micronuclei at high concentration (1 ppm) were calculated as 31.85% (Atrazine), 29.94% (Avenoxan), 36.66% (Diuron) and 41.04% (QPE). The total number of chromosome aberrations increased as herbicide concentration increased. Micronucleated cells were observed at different stages of the cell cycle. The frequency of the micronucleus was markedly higher at 1 ppm than at other test concentrations. Raj Kumar Salar, Jitender Kumar, Suresh Kumar (India) Isolation and Evaluation of Fungal Strains from Textile Effluent Disposal Sites for Decolorization of Various Azo Dyes ABSTRACT Original Research Paper: The potential of 10 indigenous fungi isolated from soil samples of dye disposal sites was evaluated to decolorize textile azo dyes viz., Reactive Blue MR, Orange M2R, Yellow M4G, Black HFGR and Red M8B. In pure culture, it was observed that Humicola insolens, H. brevis, Aspergillus terrus, A. flavus, A. niger and Rhizopus sp. were efficient in decolorizing textile dyes. The study also depicted that Rhizopus sp. was highly efficient in decolorizing (81.01%) a mixture of 5 dyes used in the present investigation followed by a fungal consortium consisting of all the ten fungal strains (78.73%). Recalcitrant dye yellow M4G was also efficiently degraded by fungal consortium (55.76%) compared to pure cultures. This study reinforces the potential of indigenous adapted fungal consortia for the decolorization of textile effluents. Natarajan Hemalatha, Othumbam Kat Remadevi, Chandrasekaran Ramanigopal (India) Influence of Tannin in Soil Adjacent to Acacia nilotica (L.) Willd. ex Del. on Microbial Population ABSTRACT Original Research Paper: The tannin polyphenol in soil influence microbial growth. Soil fertility is mainly due to the microbial load existing in a place which is decided by the physical and chemical nature of the soil. Hence in the present study, we attempted to estimate the different microbial populations in a tannin-rich soil adjacent to Acacia nilotica (L.) Willd. ex Del. trees and compared the results with microbial populations present in an area where there was no tannin. The present study revealed shifts from normal counts of predominant soil microbial groups. In non-tannin soil, bacterial counts (720 CFU 104/g) outnumbered fungal and actinomycete counts. Compared to other groups, fungi were the least abundant in non-tannin soils (5.8 CFU 104/g). The sequence of dominance in tannin soil followed the order: fungi < actinomycetes < bacteria. A comparatively higher total microbial count was responsible for high aggregate stability and reduced bulk stability of non-tannin soil. Nitrifiers in tannin soil are inhibited by tannins to account for lower soil nitrate content (4 g g-1 dry soil) compared to non tannin soil (33 g g-1 dry soil). The ecological consequences of elevated tannin levels of 5% may include allelopathic responses, changes in soil quality and reduced ecosystem productivity. Sanjeev K. Singh, Asha A. Juwarkar (India), Ackmez Mudhoo (Mauritius), Kirti V. Dubey (India) Remediation of Chromium-Contaminated Soils Using Pseudomonas aeruginosa Strain BS2 ABSTRACT Original Research Paper: The bioremediation of chromium-contaminated soil by a rhamnolipid biosurfactant produced by Pseudomonas aeruginosa strain BS2 has been explored through column studies using uncontaminated soil spiked with toxic concentrations of heavy metals i.e. 1000 mg/kg chromium. Results on removal of chromium from the spiked soil by using di-rhamnolipid and tap water have shown a high potential of di-rhamnolipid in mobilization and decontamination of contaminated soil. Within 36 h of the leaching study, di-rhamnolipid facilitated the removal of chromium 10-13-fold compared to tap water. These results indicated that the specificity of this biosurfactant towards chromium was very high and 92% removal of chromium was observed. Hence, di-rhamnolipid selectively favours mobilization of chromium from contaminated soil. Biosurfactant specificity observed towards a specific metal will help the preferential elution of a specific contaminant using di-rhamnolipid. Leachates collected from chromium-spiked soil column treated with di-rhamnolipid solution had a lower pH (6.60-6.78) than leachates from heavy metal-spiked soil column treated with tap water (pH 6.90-7.25), which showed high dissolution of metal species from the spiked soil, and effective leaching of metals.

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Neha Sharma, Gurjinder Singh Hundal, Indu Sharma, Renu Bhardwaj (India) Effect of 24-Epibrassinolide on Protein Content and Activities of Glutathione-S-Transferase and Polyphenol Oxidase in Raphanus sativus L. Plants under Cadmium and Mercury Metal Stress ABSTRACT Original Research Paper: Heavy metal toxicity results in oxidative stress in plants. Cadmium and mercury are non-essential elements for plants and are thus toxic even at low concentrations. Brassinosteroids, an important group of plant hormones have been reported to ameliorate abiotic stress in plants. The present study was undertaken to evaluate the role of 24-epibrassinolide in ameliorating the stress caused by Cd and Hg metals in raddish plants. The seeds of Raphanus sativus L. were soaked in 24-epibrassinolide (0, 10-7, 10-9, 10-11 M) and were sown in soil medium containing Cd and Hg (0, 0.5, 1.0, 1.5 mM). Oxidative stress caused by heavy metals was assessed by studying the protein content and activities of Glutathione-S-transferase (GST) and Polyphenol oxidase (PPO) enzyme activities in 60 and 90 days old raddish plants. Results revealed that presence of metals in the soil medium lead to decrease in protein content which was improved with the treatment of 24-epibrassinolide. The treatment of metals enhanced the antioxidative enzymes activities. These activities were further enhanced by 24-epibrassinolide treatments. Mukesh Kumar Kanwar, Renu Bhardwaj, Sikandar Pal Chowdhary, Priya Arora, Subodh Kumar (India) Nickel-induced Synthesis of Castasterone and 28-Homocastasterone in Brassica juncea L. ABSTRACT Original Research Paper: Heavy metals in the environment are increasing at an alarming rate. Metals like Cu, Zn and Ni are essential for the growth of plants, although elevated levels of these metals result in growth inhibition and toxicity symptoms. Plants possess a range of cellular mechanisms that may be involved in the detoxification of heavy metals and thus tolerance to metal stress. The concentration of these heavy metals in the environment has increased drastically, posing problems to the health of organisms. Plant growth regulators provide protection from various stresses. Brassinosteroids (BRs) are an important group of steroids hormones which are involved in the amelioration of several stresses. Aiming to understand the role of BRs under heavy metal stress, plants of Brassica juncea L. were grown in pots at the Botanical Garden of Guru Nanak Dev University, Amritsar. The plants were subjected to various concentrations of Nickel (0.0, 0.2, 0.4 and 0.6 mM) and harvested on the 45th day to observe the synthesis of these hormones. The GC-MS characterization of the compounds after physio-chemical analysis revealed the presence of castasterone (0.018 and 0.022 ug/Kg FW) and 28-homocastasterore (0.014 to 0.029 ug/Kg FW) in the leaves of B. juncea L. plants. Cyprian Y. Abasi, Augustine A. Abia, Jude C. Igwe (Nigeria) Sorption Kinetics and Intraparticle Diffusivities of Pb (II), Fe (III) and Cd (II) ions on Unmodified Raphia Palm Fruit (Raphia hookeri) Endocarp ABSTRACT Original Research Paper: Sorption kinetics of Pb (II), Fe (III) and Cd (II) ions from aqueous solution by unmodified raphia palm (Raphia hookeri) fruit endocarp was carried out through time dependent studies. The kinetic behaviour of the metal ions towards the adsorbent was studied with the pseudo-second order rate model. The rate constants (g/mg-min) were determined to be 7.02 10-3, 1.02 10-2 and 2.62 10-3 for Pb (II), Fe (III) and Cd (II) respectively. Sorption capacities based on the pseudo-second order model were found to be 196.08 mg/g, 169.5 mg/g and 188.68 mg/g for Pb (II), Fe (III) and Cd (II) respectively. Intraparticle diffusion models such as fractional attainment to equilibrium (FATE), McKay and Poots, Weber and Morris and Elovich equations were also used to investigate the predominant sorption mechanism. The poor fit of these intraparticle diffusion models as seen from their correlation coefficient values which ranged from 0.718 to 0.872 for Elovich equation, 0.730 to 0.837 for external film diffusion, 0.825 to 0.819 for particle diffusion, and 0.719 to 0.872 for percent adsorbed, showed that the sorption mechanism was not intraparticle diffusion controlled but film diffusion controlled. Akhil Pandey, Debdulal Banerjee (India) Biosorption of Cadmium (II) using Discarded Biomass of Aspergillus aculeatus DBF9 ABSTRACT Original Research Paper: The cadmium (Cd) biosorption potential of Aspergillus aculeatus DBF9 biomass was investigated. Among different forms of biomass, air-dried one was found most suitable in Cd removal. Maximum Cd (II) biosorption takes

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place at initial solution pH 4.5 after 90 min. Air-dried 150 mg of cell mass of A. aculeatus can remove about 78% of Cd (II) from 10 ml of 300 mg/L Cd (II) solution. The adsorption kinetics of Cd was modeled with a pseudo-second order equation to correlate the experimental data. The equilibrium data fitted very well to a Langmuir isotherm model more than to the Freundlich isotherm model. Metal accumulation was confirmed with FTIR, EDAX, and SEM analysis. This indicates that biosorption of Cd in A. aculeatus mainly occurs through ion exchange. Metal absorption properties of A. aculeatus can be used in Cd removal from industrial effluents. Etchie Ayotunde Titilayo, Adewuyi Gregory Olufemi, Etchie Tunde Ogbemi (Nigeria) Chronic Exposure to Heavy Metals in Public Water Supply and Human Health Risk Assessment ABSTRACT Original Research Paper: Historic disasters resulting from heavy metal contamination of drinking water emphasize the need to assess the quality of public water supplies. Currently, millions of people in and around Bangladesh are facing a chronic health risk of arsenic poisoning via drinking water. Therefore, to prevent the re-occurrence of such tragic episodes, heavy metal levels in public water supplies at Agodi-residential area of Ibadan, Nigeria were determined and human health risk assessment was done for metals whose levels were higher than the US EPA maximum contaminant levels for drinking purposes. Sampling was spread from January to July, 2010 to account for variations in rainy and dry seasons. 10 public taps were sampled at 5 different times, making a total of 50 water samples collected. Metals concentrations were determined by atomic absorption spectrophotometry. The mean concentrations (mg L-1) of Cd, Co, Cr+6, Mn and Pb in the sampled water were 0.05 0.02, 0.14 0.02, 0.21 0.07, 0.17 0.02 and 0.05 0.04, respectively. These values are significantly higher than the US EPA maximum contaminant levels in drinking water. Health risk assessment conducted for adult and child residents, via oral and dermal routes of exposure, gave overall hazard index values of 19 and 45, respectively, while the estimated cancer risks values were 2E-03 and 9E-04, respectively. These values significantly exceed the target non-cancer hazard index of 1 and target cancer risk of 1E-06. Hence, the concentrations of the investigating metals in Nigerian public water supplies are high enough to cause carcinogenic and non-carcinogenic systemic health effects to adults and children. Ayotunde Titilayo Etchie, Gregory Olufemi Adewuyi, Tunde Ogbemi Etchie (Nigeria) Source Apportionment of Heavy Metals in Utility Water Sources ABSTRACT Original Research Paper: Heavy metals cause cancer and non-cancer health effects in humans when consumed via drinking water. Therefore, the importance of assessing the levels and sources of heavy metals in utility water sources, with a view to proffering mitigation measures, cannot be overemphasized as not all toxic metals can be removed by conventional water treatment processes. A total of 40 composite water samples were randomly collected from utility dams and their feeding rivers from January, 2010 to December, 2011. The concentrations of metals were determined by atomic absorption spectrometry and were found to vary with season. The concentrations of Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn (mg L-1) in the water sources ranged from 0.02 to 0.49, 0.01 to 0.17, 0.03 to 0.38, 0.09 to 0.48, 0.17 to 3.25, 0.05 to 0.34, 0.02 to 0.33, 0.13 to 0.61 and 0.09 to 1.07, respectively, in the dry season, and from 0.08 to 0.68, 0.02 to 0.58, 0.13 to 0.87, 0.02 to 0.69, 0.28 to 5.62, 0.31 to 1.28, 0.09 to 1.08, 0.23 to 1.98 and 0.26 to 1.59, respectively, in the wet season, indicating that the concentrations of Cd, Cr, Fe, Ni, and Pb exceed regulatory limits in both seasons. Furthermore, the result of principal component analysis on absolute principal component scores (PCA/APCS) used to apportion sources to the metals in the dams and rivers revealed two major anthropogenic sources accounting for over 90% of the metals. Source 1 represents anthropogenic release from small-scale entrepreneurial enterprises, such as automobile-mechanic, panel beaters, vehicle sprayers and battery recharger workshops generating and spilling wastes such as lubricating oil, grease, petrol, diesel, battery electrolytes, contributing 96.2, 99.2, 95.9, 100, 99.4, 99.9, 100 and 95.2%, of Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn, respectively, while source 2 represents the release from agricultural activities, such as from fertilizers and fungicides and contributes 63.2, 100 and 51.6% of Cd, Co and Cr, respectively. Luke N. Ukiwe, Jude I. Alinnor, Chinonye I. Iwu (Nigeria) The Influence of Sludge Concentration and Sulphuric Acid on Bioleaching Efficiency of Thiobacillus thiooxidans on Sewage Sludge ABSTRACT Original Research Paper: Microbial leaching using Thiobacillus thiooxidans was investigated to solubilize heavy metals (HMs)

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from domestic sludge at different concentrations. Results obtained showed that 100% Cr was solubilized with 50 ml sludge when the bioleached sludge was digested with 50% (v/v) H2SO4, while 90% Cr was also solubilized with the same amount of sludge when the bioleached sludge could not be digested with H2SO4. However, 3.0% Cd was the metal least solubilized overall with 100 ml sludge when the bioleached sludge could not be digested with H2SO4. For high HM extraction efficiency, digestion using H2SO4 must be performed after bioleaching of sludge. Luke N. Ukiwe, Gerald O. Onyedika, Vivien I. Uche, Chinonye I. Iwu (Nigeria) Physicochemical Water Quality Indicators of Groundwater in Ishiagu, Ebonyi State, Nigeria ABSTRACT Original Research Paper: Physicochemical properties of groundwater in Ishiagu, Ebonyi State, Nigeria were investigated. Results obtained from analysis performed on samples of groundwater collected from boreholes of six sample areas (Amata, Ihie, Amaokwe, Ihetutu, Amaeze, Ngwo-Ngwo) during the raining season of June, July and August, respectively, showed that chloride (Cl-) concentration ranged from 26.9-49.7 mg/l, while sulphate (SO42-) and phosphate (PO43-) concentration ranged from 27.8-92.1 mg/l and 0.32-1.02 mg/l respectively. However, trace amounts of the metals Fe, Zn, Pb and Cd ranged from 0.01-0.05 mg/l. These values as well as those of the anions (Cl-, SO42-, PO43-) were well below WHO permissible standards. Luke N. Ukiwe, Jude I. Alinnor (Nigeria) Assessment of Polyacrylamide and Aluminum Sulphate Coagulants in Turbidity Removal in Wastewater ABSTRACT Original Research Paper: Turbidity adds an objectionable appearance to wastewaters. The present research investigates the application of a polymer (non-ionic polyacrylamide) and aluminum sulphate (AI2(SO4)3) in coagulation-flocculation to improve the quality of domestic wastewater. Results of the jar test conducted indicated that by adding 150 mg L-1 of AI2(SO4)3 at a contact time of 20 min, turbidity of the solution was reduced to about 90%. However, applying 200 mg L-1 of the polyacrylamide at a contact time of 35 min resulted in 89% turbidity removal. Evaluating the efficiency turbidity removal of both coagulants, no significant differences were noted in the ability of both coagulants to clarify wastewater. Jude C. Igwe, Emmanuel O. Odo, Solomon E. Okereke (Nigeria), Edidiong E. Asuqou (UK), Innocent C. Nnorom, Obinna C. Okpareke (Nigeria) Levels of Polycyclic Aromatic Hydrocarbons (PAHs) in Some Fish Samples from Mushin Area of Lagos, Nigeria: Effects of Smoking ABSTRACT Original Research Paper: Five different fish samples (mackerel (Scomber scombrus), tilapia (Oreochromis niloticus), croaker (Micropogonias unduletus), herring (Clupea harengus), horse mackerel (Trachurus trachurus) and blue whitting (Micromesistus pontassou)), fresh and smoked, bought from Mushin market in Lagos State, Nigeria, were sampled for polycyclic aromatic hydrocarbon (PAH) contaminants. The levels of PAHs ranged from 0.004 g/kg for acenaphthene in dry M. pontassou to 2.275 g/kg for phenathrene in dry S. scombrus. Florene, phenathrene, anthracene and fluoranthene were detected in all the fish samples. Indeno (1,2,3) perylene, dibenzo(a, h)anthracene and benzo(g, h, i)perylene were not found in any of the fish samples. Smoking increased the levels of PAHs in the fish samples. These results show that PAH levels in the fish samples are higher than the permissible limit set by the World Health Organization (WHO) and the Agency for Toxic Substances and Disease Registry (ATSDR). The implications of these contaminants in the environment together with their associated health hazards are also examined. Foluso O. Agunbiade, Bamidele I. Olu-Owolabi, Kayode O. Adebowale (Nigeria) Comparative Assessment of Coastal Water Usage Supports Using Water Quality Indices and Fuzzy Synthetic Evaluation Methods: A Case Study of Ondo State Estuary, Nigeria ABSTRACT Original Research Paper: The ability of water quality indices and fuzzy synthetic evaluation methods to combine large and technical water quality data into a single value that could help understand and communicate the status of water bodies and their usage supports for policy formulation is revealed by this study. Three water quality indices and two fuzzy synthetic evaluation methods were used to assess the usage support of Ondo estuarine water and the results compared. The indices revealed that

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the sites distant to the estuarine discharge point improved in quality and support uses than the ones nearer. Desalination of such will further improve is usage support. Anthropogenic activities are negatively affecting the quality of the environment and need to be managed if good quality water will be made available for economic uses and the vast aquatic life resources in the estuary and the neighbouring Atlantic Ocean will not be adversely affected. Jude C. Igwe, Felix C. Nwadire, Augustine A. Abia (Nigeria) Kinetics and Equilibrium Isotherms of Pesticides Adsorption onto Boiler Fly Ash ABSTRACT Original Research Paper: The adsorption of two herbicides paraquat dichloride (N-N-dimethyl-4,4-bipyridinium dichloride) and glyphosate (isopropyl amine glycine) from aqueous solution onto activated boiler fly ash (BFA) was investigated. Kinetics and equilibrium adsorption was followed by UV-spectroscopy in a batch system and the data were treated according to various rate and isotherm models. The results show that the adsorption capacities is dependent on both concentration and time and the adsorption rate increases rapidly with each specified concentration at initial time of one to ten min and attains equilibrium at about 10 min. Glyphosate was adsorbed more than paraquat dichloride. The adsorption dynamics of the pesticides were well fitted to the pseudo-second order kinetic model, with coefficient of determination of 0.9900 for glyphosate and 0.9714 for paraquat dichloride. The equilibrium adsorption was modeled by five isotherms namely; Langmiur, Freundlich, Dubinin-Radushkevich, Temkin and Harkins-Jura. From the coefficient of determination (R2), the Freundlich isotherm gave a better fit for paraquat dichloride (0.9899) while the Harkins-Jura isotherm gave a better fit for glyphosate (0.9719). The apparent energy of sorption or mean biosorption energy was found to be 50.0 and 48.83 KJ/mol for glyphosate and paraquat dichloride, respectively, indicating chemisorptions mechanism. Also, a two way ANOVA without replication shows that the differences in the results of the fitness of the isotherms were not significant. Hence, the use of BFA an agricultural waste in the adsorption of these toxic pesticides from aqueous solution is feasible and could be developed into a cheap, cost effective energy requirement and affordable technology for pesticide clean up from wastewater. Adewuyi Gregory Olufemi, Etchie Ayotunde Titilayo, Etchie Ogbemi Tunde (Nigeria) Evaluation of Total Petroleum Hydrocarbons (TPH) and some related Heavy Metals in Soil and Groundwater of Ubeji Settlement, Warri Metropolis, Nigeria ABSTRACT Original Research Paper: Soil and groundwater samples in areas of oil spill in Ubeji settlement, Warri metropolis, Nigeria, were collected in order to determine the oil and grease, total petroleum hydrocarbons (TPH) and some related heavy metals in the samples. Control samples were taken from a low density residential area in Delta State (Ekrejeta in Abraka) which was relatively free from petroleum activities. The parameters were evaluated in order to determine the quality of the soil and groundwater of Ubeji settlement, as a follow up on an earlier research conducted on the surface water and sediment of the settlements river. This research is so important because the local population uses the soil and water resources for agricultural purposes and domestic activities. Measurements of oil and grease and TPH in samples were done gravimetrically, while atomic absorption spectrophotometry was used for determination of heavy metals. For oil and grease and TPH, the soil had a mean of 1064.90 7.70 and 579.10 9.30 mg kg-1, respectively, while groundwater had 315.15 11.60 and 28.70 1.30 mg L-1, respectively. Mean heavy metal concentration of the soil and groundwater varied between 0.30 0.01 to 215.49 25.33 mg kg-1 and 0.03 0.01 to 3.05 0.07 mg L-1, respectively. These values indicate that the investigated samples had higher levels of oil and grease, TPH and heavy metals than control samples and regulatory standard limits. This result is an indicator of the high level of petroleum hydrocarbons pollution of soil and groundwater of the settlement. Princewill C. Ogbonna, Timothy O. Anigor (Nigeria), Jaime A. Teixeira da Silva (Japan) Bioaccumulation of Nutrients and Heavy Metals in Plants at a Coal Mine ABSTRACT Original Research Paper: This study assessed the bioaccumulation of nutrients (N, P, and Ca) and heavy metals (Ni, Pb and Cd) in plants in Onyeama coal mine, Nigeria. The highest concentrations of Ni (9.30 mg/kg) and Pb (7.90 mg/kg) in soil were observed 1 m from the coal mine site while Cd (0.08 mg/kg) was highest 100 m from the site. Ni in soil ranged from 2.16 to 9.30 mg/kg, Pb from 4.01 to 7.90 mg/kg, and Cd from 0.01 to 0.08 mg/kg. The highest concentrations of Ni (0.001-2.01 mg/kg) and Cd (0.001-2.41 mg/kg) were detected in Landolphia owariensis and Canarium schweinfurthii, respectively. Similarly, the highest concentrations of N (0.14-1.38 cmol/kg) and P (0.11-0.40 cmol/kg) were detected in L. owariensis. The level of Cd in soil

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reflected significant pollution compared to average global concentrations in soils. Ayodhya D. Kshirsagar, Mahendra L. Ahire, Venkat R. Gunale (India) Phytoplankton Diversity Related to Pollution from Mula River at Pune City ABSTRACT Research Note: Water samples were collected monthly (during October 2007 to September 2008) at three selected sampling stations (station I: Wakad; station II: Aundh; station III: Dapodi) from Mula River, Pune City. Collected water samples were analyzed qualitatively and quantitatively for their algal population. A total of 162 algal species belonging to 75 genera were recorded at selected sampling stations at Mula River throughout the study period. Among the different groups, Chlorophyceae was the most abundant followed by Bacillariophyceae, Cyanophyceae and Euglenophyceae. The greatest algal population was recorded at station III followed by stations II and I. Maximum abundance of Scenedesmus quadricauda, Chlorella vulgaris, Oscillatoria limosa and Melosira granulata at stations II and III throughout the year showed that these algal species could be considered as bioindicators of organic pollution. Algal monitors showed that water at stations II and III are highly polluted with organic pollutants in the Mula River. Highest algal populations were observed in April, May and June, i.e., summer, and fewest in winter and the monsoon season. Maimuna Waziri, Abdulrahman A. Audu (Nigeria) Consequences of River Obstruction: High Incidence of Pollutants in Kumadugu-Yobe Basin of Nigeria ABSTRACT Research Note: The obstruction of the flow of Kumadugu-Yobe River in the north eastern region of Nigeria has affected the entire river system. This study was undertaken to determine the levels of metal pollutants and organic pollution indicators in the water body in order to assess the impact of the obstruction and possible environmental threats posed by pollutants. Water and sediment samples were collected from five selected locations which consisted of three sampling sites each and subjected to standard analytical tests to determine the levels of Fe, Cu, Zn, Co, Cr, Ni and Pb as well as the DO, BOD, COD and TOC of the water body. Results showed some level of pollution by organic pollutants and heavy metals when compared with WHO and NAFDAC specifications for drinking water and aquatic ecosystems. The correlation coefficient (r) values of the organic pollution indicators with the metals revealed positive correlations. The obstruction of the natural flow of water into the river also resulted in the invasion and spread of aquatic weeds into the surroundings. It is postulated that the pollution in the river came from upstream as well as washings from the surrounding lands due to flooding at the peak of the rains during the months of August to September. It is recommended that the management of the Kumadugu-Yobe basin should take proper steps to constantly monitor the flow of metal and organic pollutants in to the river.

The African Journal of Plant Science and Biotechnology

Special Issue: Plant Science and Biotechnology in Ethiopia ~2012 Solomon Abate Mekonnen, Fikiremariam Haile (Ethiopia) Essential Oil Content Response of Lemongrass, Palmarosa and Citronella to Post Harvest Wilting and Chopping ABSTRACT Original Research Paper: An experiment was set to determine effects of wilting period and chopping on essential oil content of three aromatic grasses vis. Cymbopogon winterianus Jowitt, C. citratus Stapf and C. martinii. A completely randomized design with three replications was used in the experiment. Wilting the harvested part of C. citratus, C. winterianus and C. martinii for 96 hr yielded significantly higher essential oil (av. 1.31, 2.44 and 1.45%, respectively) than other treatment levels, except for 72 hr. Chopping to any level generally reduced essential oil content by 20.9% for C. citratus, 8.9% for C. winterianus and 9.24% for C. martinii. Thus, commercial production of essential oils from these plants needs to consider these two factors in order to optimize the quality and yield. Beemnet Mengesha Kassahun, Solomon Abate Mekonnen (Ethiopia) Effect of Cutting Position and Rooting Hormone on Propagation Ability of Stevia (Stevia rebaudiana Bertoni)

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ABSTRACT Original Research Paper: To fill in existing information and knowledge gaps on asexual propagation of stevia (Stevia rebaudiana Bertoni), this experiment was conducted at the Wondo Genet Agricultural Research Center nursery site in a plastic chamber. The experiment consisted of two levels of cutting positions (top and middle) and three levels of rooting hormones (zero hormone, 0.2% indole-3-butyric acid (IBA) and 0.4% 1-naphthalene acetic acid (NAA)). The experiment was laid out in a 32 factorial arrangement in a randomized complete block design with three replications. Data on leaf number/plant, survival count and rate were recorded and analyzed. Mean squares from analysis of variance revealed the existence of a very highly significant influence (P < 0.001) of cutting position on leaf number and survival rate. The application of rooting hormone exerted a significant influence (P < 0.05) on survival rate only. Interaction effect of cutting position and hormone application did not exert an influence (P > 0.05) on the parameters considered. Higher but statistically similar survival rate were recorded for the control and the 0.2% IBA treatment whereas the lowest values were recorded for the 0.4% NAA treatment. Top cuttings demonstrated significantly more leaves (eight) and survival rates (80.18%) and showed a 46.78 and 28.49% increase in leaf number and survival rate compared to middle cuttings. Therefore, it is recommended that the top part be used for the propagation of stevia. The application of rooting hormone might not necessarily be an economically sound option for vegetative propagation. Zewdinesh Damtew Zigene, Beemnet Mengesha Kassahun, Tsion Tesema Ketaw (Ethiopia) Effects of Harvesting Age and Spacing on Leaf Yield and Essential Oil Yield of Rosemary (Rosmarinus officinalis L.) ABSTRACT Original Research Paper: A study was conducted to evaluate the effect of harvesting age and spacing on agronomic and chemical traits of rosemary (Rosmarinus officinalis L.). Data on plant height, branch number/plant, fresh and dry leaf weight/plant, fresh and dry leaf yield/ha, essential oil (EO) content and EO yield/ha were collected and analyzed. Harvesting age exerted a very highly significant (P < 0.001) influence on plant height and dry leaf weight/plant, and a highly significant (P < 0.01) influence on dry leaf to stem ratio. Spacing affected fresh and dry leaf weight/plant very highly significantly (P < 0.001) and highly significantly (P < 0.01) plant height. The interaction effect of harvesting age and spacing was significant (P < 0.05) on dry leaf yield/ha, EO content and yield, and highly significant (P < 0.01) on fresh leaf yield/ha. Maximum plant height and dry leaf weight/plant were obtained 11 months after transplanting (MAT) and maximum dry leaf to stem ratio at 9 MAT. Higher plant height was attained at 60 60 cm while higher fresh and dry leaf weight/plant at 90 90 cm. Higher fresh leaf yield/ha (38.43 t) and dry leaf yield/ha (10.83 t) were obtained at 60 60 cm at 9 MAT. Maximum EO content (2.77 %) and EO yield (271.43 kg) were recorded at 10 MAT at a spacing of 90 120 cm and 60 60 cm, respectively. Solomon Abate Mekonnen (Ethiopia), T. P. S. Katiyar, H. Ravishankar (India) Yield Components and Yield of Haricot Bean (Phaseolus vulgaris L.) Under Different Irrigation Frequency and Planting Density Treatments ABSTRACT Original Research Paper: An experiment was undertaken to determine the yield component and yield of haricot bean (Phaseolus vulgaris L.) varieties in the semi-arid region of Dire Dawa, Ethiopia. The experiment was a split plot with three irrigation frequencies as main plots and a combination of three planting densities and two haricot bean varieties as subplots, all of which replicated three times. Irrigation frequencies consisted of 1, 0.67, and 0.5 irrigation water to cumulative pan evaporation (IW/CPE ratio) thereby fixing the depth of IW to 60 mm, planting densities of 15.6, 25, and 35.7 (plants m2) and two haricot bean varieties, Roba-1 and Mexican-142 were considered. Different planting densities were employed by changing planting distances within cultivation rows. Plant data on dry biomass/plant, number of branches bearing pods, number of pods/primary branch, number of pods/plant, pod length and width, number of beans/pod, number of beans/plant, 100-bean weight and harvest index at harvest were recorded. Increasing irrigation frequency from 0.5 IW/CPE to 0.67 IW/CPE and to 1 IW/CPE significantly increased all parameters. On the contrary, increasing population densities caused a significant reduction in the parameters. Averaged over planting density and variety, yield/ha and soil water depletion increased significantly with the increase in irrigation frequency. These results showed that irrigation given at 1 IW/CPE can give maximum yield of both haricot bean varieties in the semi-arid condition of Dire Dawa. Yared Semahegn Belete, Sentayehu Alamerew Kebede, Adugna Wakjira Gemelal (Ethiopia) Genetic Associations of Seed Oil Quality Traits and Selection Criteria in Ethiopian Mustard (Brassica carinata A. Brun) ABSTRACT

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Original Research Paper: Ethiopian mustard, as an oil seed crop, needs genetic improvement which integrates both quality and agronomic traits. In this study, the correlations of eight quality traits of the 36 genotypes were investigated using a 6 6 simple lattice design at Holetta Research Center, Ethiopia. Heritability and genetic advance (as percent of mean) of the studied traits ranged from 35.4-69.2 and 5-23.8%, respectively. A strong positive correlation (0.752) was observed between palmitic and erucic acid. Similarly, a strong but negative correlation (-0.866) was observed between oleic and erucic acid. In relation to agronomic traits, positive and significant correlation of stearic with number of primary branches (0.607), number of secondary branches (0.608) and number of pods (0.387) was observed. Palmitic showed negative correlation with seed yield/plot (-0.354), oil yield/plot (-0.393) and 1000-seed weight (-0.404). Oil content positively correlated with seed yield/plot (0.343) and oil yield/plot (0.446), while negative correlation was found with days to flowering (-0.373) and days to maturity (-0.394). Path analyses showed that oleic acid and oil content were found the most important components to be considered as selection criteria in the improvement of edibility of seed oil of Ethiopian mustard genotypes. This investigation also indicated that improvement in the oil content of the seeds of the genotypes would be possible through selecting early flowering genotypes. Beemnet Mengesha Kassahun, Hassen Nurhusain, Hailesilasie Gebremeskel, Solomon Abate Mekonnen, Zewdinesh Damtew, Muluken Philipos, Bekri Melka, Terutaka Niide (Ethiopia) Participatory Development of Quality Seedlings in Lemon Verbena (Alloysia triphylla L.) Using Stem Cuttings ABSTRACT Original Research Paper: This experiment fills in existing information and knowledge gaps on asexual propagation of lemon verbena by involving two farmers research groups (FRGs) at Sembero Rogicha and Dawile Kebeles during 2011 starting from the beginning of August to the end of October. Each FRG has 10 member farmers. The experiment consisted of three levels of the part used (top, middle and bottom) and four levels of node number (three, five, seven and nine) in a factorial combination. The nursery experiments were laid out in a randomized complete block design with three replications. Data on survival count, survival percentage, number of branches/seedling, number of leaves/branch and number of leaves/seedling were recorded in a participatory approach. Mean squares from analysis of variance revealed the existence of a very highly significant influence (P < 0.001) of cutting position, node number and the interaction effect of cutting position with node number on all parameters considered in the study. Bottom cuttings with 3 and 5 nodes demonstrated significantly higher respective values of survival rate (81.67 and 78.33%), number of branches/seedling (6), number of leaves/branch (25) and number of leaves/seedling (104 and 137) and lowest values of these parameters were recorded for top cuttings. Similar results were obtained with the FRG member farmers evaluation criteria. Therefore, bottom cutting position with 3 and 5 nodes could be recommended for the development of quality lemon verbena seedlings using stem cuttings under good nursery management. Solomon Abate Mekonnen (Ethiopia) Irrigation Frequency and Plant Density Affect Phenology and Crop Growth of Haricot Bean (Phaseolus vulgaris L.) ABSTRACT Original Research Paper: A field study was conducted involving a factorial combination of two haricot bean (Phaseolus vulgaris L.) varieties Roba-1 and Mexican-142 with three irrigation frequencies at 60, 90 and 120 mm cumulative pan evaporation (CPE) using an IW/CPE ratio thereby fixing the depth of irrigation water (IW) to 60 mm and three planting density levels (150000, 250000, 350000 plants/ha) to investigate their effects on phenology and crop growth at Tony farm, Dire Dawa during the winter (Bega) season of 2002. A split plot design was laid out with irrigation frequency as main plots while a combinations of planting density and variety as sub-plots with three replications each. Increasing irrigation frequency from 120 to 90 and to 60 mm CPE significantly increased days to maturity and crop growth parameters of haricot bean. Variety significantly influenced phenological stage, specifically days to flowering and days to maturity. Plant height and leaf number were significantly higher for Mexican-142, with a 17.7 and 29.0% increase, respectively. Increasing plant density significantly (P < 0.05) decreased leaf area and leaf number, and increased leaf area index of beans. The highest seed yield per plant was recorded with the minimum plant density, 150000 plant/ha and lowest seed yield per plant with the highest plant density, 350000 plants/ha. The increase in irrigation frequency from 120 to 60 mm CPE significantly increased seed yield per plant from 38.22 to 59.94 g. Mihiret Mekonnen Moges, Tanta val Selvaraj, Mitiku Tesso Jebessa (Ethiopia) Influence of Some Antagonistic Bacteria against Early Blight (Alternaria solani (Ell. & Mart.) Jones & Grout.) of Tomato (Lycopersicon esculentum Mill.)

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ABSTRACT Original Research Paper: Early blight caused by Alternaria solani (Ell. & Mart.) Jones & Grout., is amongst the most common foliar disease in tomato (Lycopersicon esculentum Mill.) farms of West Shewa sub-regions of Ethiopia that reduce yield and occasionally cause complete crop loss. To satisfy the contemporary market driven demand and supply for tomato products, the research need to focus on management options that are environmentally friendly. The present study was undertaken to evaluate the antagonistic effect of some rhizospheric bacteria (biocontrol agent) against A. solani and to study their influence on growth and development of tomato leaf of a farming cultivar, Romans VS. Ten local antagonistic bacteria were screened in vivo for suppressing the pathogen. Five promising antagonists exhibiting higher zone of inhibition (ZOI) (38 mm and above) and percent disease control (ranging from 38.16 to 43.79%) were selected. These are Pseudomonas fluorescens TK-1, P. fluorescens TK-3, Bacillus subtilis TK-4, P. fluorescens TK-8 and P. fluorescens TK-10. The greenhouse experiment revealed P. fluorescens TK-3 as a best biocontrol agent which increased plant height by 35.20% and biomass by 52.28%. The efficacy test results of antagonistic bacterial isolates have clearly indicated that the indigenous strain, P. fluorescens TK-3 followed by Bacillus subtilis Tk-4 is an efficient biocontrol agent against A. solani with good in vitro and in vivo antagonistic activity. Tewabech Tillahun (Ethiopia), Jaime A. Teixeira da Silva (Japan), Solomon Admassu (Ethiopia) Assessment of Yield Loss and Determination of Optimum Planting Date for the Control of Gray Leaf Spot on Maize (Zea mays L.) in South Ethiopia ABSTRACT Original Research Paper: Maize (Zea mays L.) is one of the most important strategic crops selected for food security mainly due to its high productivity and wider adaptability in Ethiopia. In recent years, gray leaf spot (GLS) has become a serious disease in different parts of the country. However, no research activity has been carried out to determine the role of planting date on the control of GLS. Therefore, the objectives of this study were to assess the damage and yield loss due to GLS and to identify and determine the optimum plating date of maize for the control of GLS. The experiment was carried out at Areka and Billito, Southern Ethiopia which are GLS hotspots during the 2004-2006 main cropping seasons. Fungicide treatment as the main plot and planting dates as the subplot treatment were arranged in a split plot design and replicated four times. The result of this experiment shows that there were significant differences among planting dates, fungicide spray, years and locations. Analysis of variance carried out across years at Areka and Billito indicated that there were statistically significant differences among planting dates for severity, upper ear leaf infestation, number of cobs harvested, number of diseased cobs, ear length, ear diameter, grain yield and 1000-kernel weight. The highest grain yield (8.12 and 9.09 t ha-1) was recorded from plantings on March 17 and April 18 at Areka and Billito, respectively. Planting on March 27 and April 18 increased yield by 55.5 and 43.0% more than maize planted on April 17 and May 18 at Areka and Billito, respectively. The highest grain yield (8.61 t ha-1) was recorded in plots sprayed with fungicide at Billito while the least was recorded in unsprayed plots (6.3 t ha-1) at Areka. The yield loss due to late planting was 29.5% more than early planting. Zewdinesh Damtew Zigene, Beemnet Mengesha Kassahun (Ethiopia) Agronomic Characteristics and Essential Oil Yield of Palmarosa (Cymbopogon martinii (Roxb.) Wats) as Affected by Population Density and Harvesting Age at Wondo Genet, Southern Ethiopia ABSTRACT Short Communication: A study was conducted to evaluate the effect of harvesting age and population density on agronomic characteristics and essential oil yield of palmarosa (Cymbopogon martinii (Roxb.) Wats). Data on number of tiller/bunch, number of leaf/bunch, fresh weight/bunch, fresh herbage yield/ha, dry weight/bunch, dry herbage yield/ha, essential oil (EO) content and EO yield/ha were collected and analyzed. Population density exerted a very highly significant (P < 0.001) influence on fresh herbage yield/ha, dry herbage yield/ha and essential oil yield/ha of palmarosa. A very highly significant (P < 0.001) influence of harvesting age was observed on number of leaves/bunch, fresh weight/bunch and fresh herbage yield/ha while a highly significant (P < 0.01) effect was observed on EO yield/ha. Dry weight/bunch, dry herbage yield/ha and EO content was affected significantly (P < 0.05) by harvesting age. Maximum fresh herbage yield/ha (20.86 t), dry herbage yield/ha (6.85 t) and EO yield/ha (100.65 kg) were obtained at 60 60 cm spacing for harvests made at every 3 months after plating and the preceding harvests. Higher and statistically similar EO content were obtained for harvests made at every 3, 4 and 5 months after planting regardless of spacing. Zewdinesh Damtew, Bizuayehu Tesfaye, Daniel Bisrat (Ethiopia) Growth and Essential Oil Yield of Artemisia (Artemisia annua L.) as Affected by Growth Stage

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ABSTRACT Short Communication: A study was conducted to observe the growth trend and essential oil yield of Artemisia annua L. under varying growth ages at the Wondo Genet Agricultural Research Center. The experiment consisted of six growth stages (2, 3, 4, 5, 6 and 7 months after transplanting). The treatments were arranged in randomized complete block design with three replications. Data on plant height, branch number/plant, fresh leaf weight/plant, dry leaf weight/plant, fresh leaf/stem ratio, dry leaf/stem ratio essential oil content (EOC) and essential oil yield (EOY) were collected and analyzed. As mean squares from analysis of variance indicated, different growth ages affected plant height, fresh leaf/stem ratio and dry leaf/stem ratio very highly significantly (P < 0.001) and affected EOC and EOY highly significantly (P < 0.01). Fresh leaf weight/plant and dry leaf weight/plant were affected significantly (P < 0.05) by different growth ages. Branch number/plant did not vary significantly (P > 0.05) at different age of growth. In this study, the highest values were recorded at 5 months after transplanting (MAT) for plant height (194 cm), fresh leaf weight/plant (382.47 g) and dry leaf weight/plant (98.36 g) and at 2 MAT for fresh leaf/stem ratio (0.69) and dry leaf/stem ratio (0.64). The highest EOC (1.08%) and EOY (21.78 kg) values were obtained at 6 MAT. Zenebe Mekonnen Gebretsadik (Ethiopia) The Effect of Slope on Diameter and Height Growth of Grevillea robusta at Wondo Genet, Southern Ethiopia ABSTRACT Short Communication: Grevillea robusta (A. Cunn. ex R.Br.) is widely used in Africa and grows on fairly well drained and neutral to acidic soils in altitude ranging 0-3,000 m a.s.l. but does not tolerate water logging or heavy clays. In Ethiopia, it does well in agro-climatic zones ranging 1,500-2,700 m a.s.l. In this study, three south facing slope categories with mean slope in degree and mean slope length in meters {(15, 50); (8, 70); (4, 100)} are selected coincidently in ten-year old G. robusta plantation perpendicular to the contour. Ten trees at each slope range are selected randomly. The objectives of the study were i) to identify the slope factor on height and Diameter at Breast Height (DBH i.e.1.3 m above the ground) growth of G. robusta trees and ii) to compare responses of apical and lateral growths of trees on three slope categories to give reasonable assurance of detecting meaningful differences. The statistical analysis from one-way ANOVA at = 5% has shown that there is highly significant difference (P < 0.0001) in height growth of G. robusta planted on different slope ranges. Similarly, it was also recognized that there is significant difference (P < 0.04) in lateral or diameter growth of G. robusta trees planted at hillsides of Abaro Mountain at Wondo Genet. Volume 6(1) Samuel Adelabu, Onisimo Mutanga, Moses Azong Cho (South Africa) A Review of Remote Sensing of Insect Defoliation and its Implications for the Detection and Mapping of Imbrasia belina Defoliation of Mopane Woodland ABSTRACT Review: Forest health, especially insect defoliation monitoring in forest using direct sampling and visual estimation has been only moderately successful due to its cost, time required for sampling, and most importantly the need to collect data immediately before and after an extreme event. However, remote sensing techniques offer timely, up-to-date, and relatively accurate information for sustainable and effective management of forest health. In this paper, we discuss the different approaches including the remote sensing platforms and techniques that have been used for assessing insect defoliation and its implications for detecting and monitoring mopane worm defoliation of mopane woodland, highlighting their strengths and weakness. Research gaps in the detection of insect defoliation with remote sensing are highlighted and future directions of research are also proposed. Stanley Mukanganyama (Zimbabwe), Simbiso C. Dumbura, Leseilane Mampuru (South Africa) Anti-Proliferative Effects of Plant Extracts from Zimbabwean Medicinal Plants against Human Leukaemia Cell Lines ABSTRACT Original Research Paper: A selected group of 14 medicinal plants was screened for antiproliferative activity against two human leukaemia cell lines Jurkat T and Wil 2 in vitro. The Trypan Blue assay was used to assess antiproliferative and cytotoxic activity. The five most potent medicinal plants showed the following order of potency against Wil 2 cell line: Parinari curatellifolia > Aloe barbadensis > Croton gratissimus > Syzigium guineense > Vernonia adoensis with IG50s of 93, 115, 148, 149.8 and 130

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g/ml, respectively. The plants had comparable proliferation inhibition to the cancer drug doxorubicin. From these species, high levels of cytotoxicity were detected in extracts from Parinari curatellifolia and Aloe barbadensis to Wil 2 cell line at concentrations of 500 and 1 000 g/ml. Croton gratissimus, Syzigium guineense and Vernonia adoensis extracts were found to be antiproliferative and not cytotoxic at the same concentrations. P. curatellifolia extract at a concentration of 10 g/ml reduced cell proliferation of Jurkat T cells by 70% after 48 h of incubation. Studies were also carried out where the extract from P. curatellifolia combined with doxorubicin at concentrations 10 to 0.50 g/ml were tested for antiproliferative activity against a Jurkat T cell line. The top five plant extracts had IG50s of less than 150 g/ml. The results show that plants used traditionally for treatment of diseases such as tuberculosis, mycosis and diarrhea can be used to inhibit cell proliferation in leukaemia cell lines. The extract from P. curatellifolia could be a potential source of lead compounds that may be used as anticancer drugs. The plant extracts that showed cytotoxicity and inhibition of cell growth will be further characterized to identify the active fractions and compounds. Sofiane Abdelhamid (Tunisia) Genetic Variation and Characterization of Swiss Chestnut Cultivars (Castanea sativa Mill.) Using RAPD and AFLP Markers ABSTRACT Original Research Paper: In order to study the genetic variation and to provide specific markers for genetic characterisation and identification of Swiss chestnut cultivars, two sets of molecular markers were explored: Random Amplified Polymorphic DNAs (RAPDs) and Amplified Fragment Length Polymorphic DNAs (AFLPs). 98 RAPD and 222 AFLP polymorphic markers were amplified using 12 and 4 primer combinations, respectively. Clustering analysis performed with the two sets of markers to group cultivars according to their similarity coefficients separated the genotypes into clear groups with different grades of success. The genetic identification of cultivars was more correct by RAPD than by AFLP. AFLP generated the highest number of polymorphic bands and clustered fairly closely-related chestnut cultivars. These results allowed problems generated by synonyms and homonyms in different chestnut accessions to be resolved. Molecular marker results were comparable. The similarity matrices based on the two sets of data give a highly significant positive correlation between RAPD and AFLP (r = 0.78). Michael I. Uguru (Nigeria), Benedict C. Oyiga (Germany), Elias A. Jandong (Nigeria) Responses of Some Soybean Genotypes to Different Soil pH Regimes in two Planting Seasons ABSTRACT Original Research Paper: Seven genotypes of soybean (Glycine max (L.) Merrill) were screened in 2004 and 2005 planting seasons at the experimental research farm of the University of Nigeria, Nsukka to identify the acid tolerant genotypes using some agronomic and yield traits. These genotypes were screened under varying soil pH conditions. The results revealed that the genotypes varied considerably in the agronomic and yield traits at the different pH values. The soil pH, genotype and their interactions had significant effects on most of the traits evaluated in both planting seasons. The traits were greatly reduced at soil pH < 5.5 and, it increased progresively with increase in the soil pH up to 6.0. The principal component analysis (PCA) indicated that the first three principal components contributed 71.12 and 69.28% of the total variability among the genotypes in the 2004 and 2005 plantings, respectively. Thus, under soil acid conditions traits such as root length, fresh root weight and number of nodules are discriminating and can serve as selection criteria to distinguish between acid tolerant and acid-sensitive genotypes. In the 2004 planting, Digil, Garikida and Sunkani were identified as tolerant; Kyado and Sebore as moderately tolerant; Gembu is moderately susceptible while TGX1448-2E is suceptible to soil acidity. In 2005, Sebore and Digil were identified as acid-tolerant; Gembu and Sunkani as moderately tolerant; Garikida and TGX1448-2E as moderately susceptible and Kyado as suceptible to low soil pH. The tolerant genotypes had normal root growth and higher seed yield at pH < 5.5 in both planting seasons indicating their tolerance to low soil pH. The results suggest that Kyado, Sebore and Digil can therefore be included in breeding programs to develop new genotypes that can withstand low soil pH conditions in the derived savanna region of south eastern Nigeria. Ibrahim Y. Dugje, Adam L. Ngala, Haliru Yakubu (Nigeria) Effects of Cowpea Planting Date and Phosphorus Level on Striga Infestation, Growth and Yield of Cowpea Intercropped with Pearl Millet in a Nigerian Sudan Savanna ABSTRACT Original Research Paper: Grain yield of cowpea (Vigna unguiculata (L.) Walp is usually reduced when intercropped with pearl

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millet (Pennisetum glaucum (L.) R. Br.)) in the low fertility soils in the Sudan savanna of Nigeria. Field experiments were conducted to determine the influence of cowpea planting date and phosphorus (P) levels on Striga gesenerioides infestation and grain yield of cowpea grown in mixture with pearl millet during 2005 and 2006 rainy seasons. Five cowpea planting dates: 0, 10, 20, 30 and 40 days after millet planting (DAMP) and three P levels: 0, 30 and 60 kg P2O5/ha, were evaluated in randomized complete block design in a factorial arrangement. Each treatment was replicated three times. The results showed that cowpea growth, yield and yield component parameters and Striga infestation of cowpea were significantly reduced with delay in planting cowpea. Increase in P levels significantly increased vegetative and yield parameters of cowpea and reduced Striga infestation. There were significant positive linear relationships among cowpea agronomic parameters (r = 0.25 to 0.96). Seed yield/ha was negatively correlated with Striga count (r = -0.51) and 100-seed weight (r = -0.38). The simultaneous planting of cowpea and pearl millet or planting cowpea within 10 days after pearl millet and each combined with 30-60 kg P2O5/ha improved cowpea grain yield and reduced Striga infestation in cowpea when intercropped with pearl millet in the Sudan savanna. Ibrahim Y. Dugje, Adam L. Ngala (Nigeria) Influence of Cowpea Planting Date and Phosphorus Level on Striga Infestation and Performance of Pearl Millet in Mixture with Cowpea in a Nigerian Sudan Savanna ABSTRACT Original Research Paper: Pearl millet is traditionally grown in mixture with cowpea in the savanna region of Nigeria. Field experiments were conducted to determine the response of pearl millet to varying dates of planting cowpea and phosphorus levels during the 2005 and 2006 rainy seasons. A randomized complete block design in factorial arrangement was used to evaluate five cowpea planting dates (0, 10, 20, 30 and 40 days after planting millet) and three phosphorus levels (0, 30 and 60 kg P2O5/ha), each replicated three times. The results showed that Striga counts and pearl millet plant height significantly increased with delayed in planting cowpea, while number of grains/panicle, seed weight and grain yield/ha significantly decreased with delay in planting date. Increase in P levels significantly reduced Striga counts, but had no significant effect on grain yield/ha. Grain yield/ha was inversely associated with Striga counts (r = -0.28 to -0.42), but positively correlated with number of grains/panicle (r = 0.52 to 0.89) and seed weight (r = 0.27 to 0.35). The competitive effects of cowpea on pearl millet were balanced by substantial gain in cowpea grain yield for simultaneous or planting cowpea within 10 days after pearl millet. Therefore, both pearl millet and cowpea should be planted simultaneously or within 10 days after planting pearl millet as each proved beneficial to pearl millet and enhanced the productivity of the system in the Nigerian savanna. Oladayo O. Idowu-Agida, Dotun J. Ogunniyan, Emmanuel O. Ajayi (Nigeria) Screening Long Cayenne Pepper (Capsicum frutescens L.) Accessions Collected from Southwest Nigeria for Agronomic Performance ABSTRACT Original Research Paper: Germplasm collections and selection for agronomic traits are essential for genetic enhancement. Capsicum has great genetic diversity; greater than 200 landraces are grown in Nigeria, but there are no recommended cultivars. An experiment was conducted at the National Horticultural Research Institute of Nigeria, Ibadan to evaluate and select long cayenne pepper accessions, collected from 31 locations in Southwest Nigeria, for agronomic performance in rainy and dry seasons of 2008. Six-week-old seedlings of the pepper accessions were transplanted using 0.6 0.6 m spacing (28 plants/plot) into 3.6 2.4 m beds, separated by 1 m. Data were collected on eight middle plants on growth, flowering and fruit traits in both seasons. Data collected were subjected to analysis of variance; least significant difference was used to separate means, and least means square used to explain results of interactions. Different accessions were adapted to each season. Season had no effect on seedling vigour, plant height, number of seed/fruit and 1000-seed weight; but dryness hastened flowering and fruiting. Fruit yield/plant and number of seed/fruit were higher in the dry than rainy season. Accessions IA108-9, IA108-10, IA108-12, IA108-13, IA108-14, IA108-20 and IA108-28 performed best under rain-fed; accessions IA108-2, IA108-5, IA108-8, IA108-19, IA108-23 and IA108-31 did best in the dry season; and accessions IA108-3, IA108-4, IA108-7 and IA108-29 performed well in both seasons. Fruit yield/plant in IA108-28 (>100 g) was stable across seasons; the accession could be selected for further studies in Ibadan. Gbenga Akinwumi, Vincent Umeh, Isaac Olabode, Semeton Amosu, Babashola Adelaja (Nigeria) The Performance of Two Guava (Psidium guajava L.) Varieties Supplied with Organic and Inorganic Fertilizer under Tropical Conditions ABSTRACT Original Research Paper: Mineral nutrition is a factor that can easily be modified for plant growth, and consequently soil can

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be amended to alleviate limitations of nutrient availability and/or absorption in many horticultural practices. Generally, the amendments provide soil and biological factors necessary for improved plant growth by affecting changes in plant growth and development, and particularly soil chemical composition. A trial was conducted to evaluate the performances of two varieties of guava supplied with poultry manure and inorganic fertilizers applied alone or in combination for over a period of two years. Two varieties of guava seedlings namely Allhabad and White delicious were transplanted to the field. Cured poultry manure and NPK15:15:15 were used as organic and inorganic fertilizer respectively at the rates of 0.28 t/ha organic fertilizer + 75 kg/ha NPK inorganic fertilizer at four application per year (F1), inorganic fertilizer at 150 kg/ha NPK at four application per year (F2), 0.56 t/ha organic fertilizer only (F3) and control (no fertilizer) F0. Result showed that a combination of organic + inorganic fertilizers performed better in terms of growth parameters assessed among the treatments in both varieties than plants supplied with full organic fertilizer (0.56t/ha poultry manure). However, Allahabad had the highest stem girth and canopy cover while White delicious had the tallest plants. The combined application of poultry manure and NPK fertilizer gave the best growth performance. Oyebamiji O. Oyegoke, Samuel A. Babarinde, Akinola J. Akintola, Zainab B. Olatunji (Nigeria) Bioactivity of Ocimum sanctum Linn. Leaf Powder and Extracts against Tribolium castaneum Herbst ABSTRACT Original Research Paper: Two formulations of Ocimum sanctum L. (powder and extract) were used to test for its toxic and repellent properties against Tribolium castaneum in the laboratory. A toxicity test consisted of exposing T. castaneum larvae to millet (Pennisetum glaucum (L.) R. Br.) seeds treated with three dosages (0, 0.15 and 0.3 ml/5 g seeds) and millet flour admixed with three dosages (0, 0.25 and 0.5 g/5 g flour). In the case of the leaf powder, mortality increased with increase in concentration, with highest percentage mortality (63.5%) observed with 0.5 g O. sanctum. However, when two extracting solvents were used for extraction, acetone evoked greater mortality than petroleum spirit. Although petroleum spirit had greater repellent effects, it was not dependent on dose. Percentage repellency followed a progressive increase when acetone was used for extraction with the significantly (P < 0.05) highest percentage repellency (86%) observed with 0.3 ml/30 cm2. Sherin Mahfouze, Eman Khattab (Egypt), Nour Gadalla (Egypt/Kingdom of Saudi Arabia) Resistance of Faba Bean Accessions to Bean Yellow Mosaic Virus and Broad Bean Stain Virus ABSTRACT Original Research Paper: The primary objective of this study was to find new faba bean (Vicia faba) accessions resistant of Bean yellow mosaic virus (BYMV) and Broad bean stain virus (BBSV). In addition, since little genetic information is available on the resistance of faba bean to these viruses, this study aimed to investigate the changes in peroxidase activity and protein composition in faba bean leaves resistant to both BYMV and BBSV. The 15 faba bean accessions had different resistance to BYMV and BBSV based on disease index. Accessions No. 3, 4, 8 and 12 were immune while accessions No. 1, 5, 7, 9, 10, 11, 13 and 14 were highly resistant; accessions No. 6 and 15 were moderately resistant and accession No. 2 was tolerant. SDS-PAGE and POD-isozymes patterns were used to studying genetic variability among the immune, tolerant, or susceptible 15 faba bean accessions under mixed infection with both BYMV and BBSV. 12 protein markers with molecular sizes ranging from 145.2 to 6 KDa were observed in immune, tolerant and resistant plants but which were not present in the control. The highest number of markers appeared in accession No. 3 (six markers). Most pathogen-related proteins were observed in diseased plants with both BYMV and BBSV, e.g., accession No. 14 in which four bands appeared (92, 86.5, 21 and 14.8 KDa). Peroxidase activity increased in all faba bean accessions except for accessions No. 2, 11 and 15. Increasing peroxidase activity was related with host resistance to both viruses. Jetro Nkengafac Njukeng, Eugene Ejolle Ehabe, George Elambo Nkeng (Cameroon) Estimating Organic Carbon from Loss-on-Ignition and pH in Some Cameroon Hevea brasiliensis Plantation Soils ABSTRACT Short Communication: Recent studies on carbon sequestration and climate change require accurate estimates of soil organic carbon (SOC) following its importance in carbon storage studies. This study evaluates the use of loss-on-ignition (LOI) as a rapid, less costly and accurate method for estimating SOC considering the errors associated with estimation of the latter. LOI-SOC-pH regression equations were obtained using data from analyses of topsoils (0-15 cm deep) in mature rubber plantations in the humid forests of south west Cameroon. Indeed, significant linear equations characterized SOC-LOI and

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SOC-pH (KCl) relations. The rather weak LOI-SOC relations (r 0.38) would indicate that this parameter may serve as an accurate estimate for SOC upon its modification following a mastery of factors responsible for its variability. Olagorite Adetula, Gbenga Akinwumi (Nigeria) Assessment of Varietal Growth of Plantain and Banana in South-western Nigeria ABSTRACT Short Communication: Southwestern parts of Nigeria are located within savanna and forest agro-ecological zones in which the economy is agrarian in nature. The major crops intercropped with plantain and banana are cocoa (when young) and cocoyam. This research was therefore geared toward the collection of various types of plantains and bananas commonly grown in the south west in order to increase genetic lines in the gene bank of the National Horticultural Research Institute, Ibadan. Information was obtained from six states (Lagos, Ogun, Osun, Ondo, Ekiti and Edo). Data were collected from farmers in 50 farms from each state who had large and small farms using direct questioning, measuring tapes for plant height, stem girth, leaves length and breadth. Soil observations on each farm were carried out and soil characteristics such as texture, stoniness and colour were determined. The results of the survey indicate that farmers had given more attention to the cultivation of the crops either intercropped with cocoyam or cocoa (when young) or when planted as a sole crop. The majority of farmers had their plantain and banana farms in their backyards. Most of their cultural practices were rain-fed with uniform population variability. Saro banana variety showed significant difference (P < 0.05) among the mean value in plant height and stem girth with Owode Owena banana recorded highest plant height and stem girth when compared with Owan and Aramoko. The major constraints to production were nematodes, weevil infestation and the lack of knowledge of cultural and maintenance practices. Abioye Olabode, Olagorite Adetula, Gbenga Akinwumi (Nigeria) Prospects and Constraints of Marketing Musa spp. in Southwest Nigeria ABSTRACT Research Note: The bulk of Musa spp. output come from large numbers of very small scattered farms making the task of collection and distribution more complicated. A survey was undertaken to study the prospects and constraints of marketing of banana and plantain in south west Nigeria, during the period of January to May 2008. 150 marketers were selected in six states at random for the interview using structured questionnaires. Data were analyzed using descriptive statistics, marketing margin and t-test. The result of the analysis showed that 80% of respondents were female and 20% male. Majority of respondents had ten years experience. Above (68%) were middle aged people. Analysis also indicated that 40, 35 and 53% sourced their fund through contribution (Esusu), family and personal savings respectively. The majority of respondents (72%) had the problem of collection since farms are small and scattered with little access. According to respondents the average purchasing price of big and small bunch of plantain were 251 and 95.60 respectively while, big and small banana bunch were found to be 104.60 and 62.80 respectively. Also average selling price of big and small bunch of plantain were 573 and 188.60 respectively. While big and small bunch of banana were sold for 250.80 and 125.00 respectively. The result of t-test analysis showed that the cost price was significant (P<0.05) different from the selling price of plantain and banana. This indicated that marketing of the crops is highly profitable. Finally, these surveys revealed high level of profitability associated with the marketing of plantain and banana, if government assists in the provision of credit facilities to the sellers and accessible roads to the producers. Next issue Sofiane Abdelhamid, Naziha Grati-Kamoun (Tunisia), Francesco P. Marra, Tiziano Caruso (Italy) Genetic Diversity of Major Olive Varieties from Southern Tunisia ABSTRACT Short Communication: Considering the importance of olive-growing in Tunisia, microsatellite (SSR) analysis was used to study the genetic variation among twenty olive accessions from southern Tunisia. This set of olive microsatellites showed potential utility for genetic studies and it could contribute to the development of strategies for Tunisian germplasm conservation and breeding. Unweighted pair group method cluster analysis was performed and cultivars separated in three main groups. Five polymorphic simple sequence repeats (SSR) loci were employed and they revealed 38 alleles with a mean number of 7.6 alleles per locus. Genetic variability was wide as indicated by high values of both observed heterozygosity (mean value = 0.79) and PIC values (average value = 0.60). Cultivars formed 3 distinct and clear groups. Var. Chemlali was grouped with the others

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cultivars and showed low genetic diversity. We hypothesize that this variety is a population of cultivated varieties, with the presence of different clones of the same cultivar.

The Americas Journal of Plant Science and Biotechnology

SPECIAL ISSUE: Rusts. Xianming Chen (USA) High-Temperature Adult-Plant Resistance, Key for Sustainable Control of Stripe Rust ABSTRACT Invited Review: High-temperature adult-plant (HTAP) resistance expresses when plants grow old and the weather becomes warm. This non-race specific and durable type of resistance has been used successfully in control of wheat stripe rust in the U.S. since early 1960s. This article describes practical procedures for identification and characterization of HTAP resistance and reviews recent studies on discovery of genes conferring HTAP resistance. Recent studies providing insights to the molecular basis for the durability of HTAP resistance will be presented. Strategies for improving levels of HTAP resistance and improving control of stripe rust through combining HTAP resistance with effective all-stage resistance will be discussed. Elsa Ballini (France/USA), Nick Lauter, Roger Wise (USA) Prospects for Advancing Rust Disease Defense through Genetical Genomics ABSTRACT Invited Review: Rusts are one of the most severe threats for cereals because new races emerge regularly, typically resulting in infestations that lead to large yield losses. In 1999, a new race of stem rust, Puccinia graminis f. sp. tritici (Pgt TTKSK or Ug99), was discovered in Uganda. Most of the wheat and barley cultivars grown currently worldwide are susceptible to this new race. Pgt TTKSK has already spread northward into Iran and will likely spread eastward throughout the Indian subcontinent in the near future. This scenario is not unique to stem rust; new races of leaf rust (Puccinia triticina) and stripe rust (Puccinia striiformis) have also emerged recently. One strategy for countering the persistent adaptability of these pathogens is to stack complete- and partial-resistance genes, which requires significant breeding efforts in order to reduce deleterious effects of linkage drag. These varied resistance combinations are typically more difficult for the pathogen to defeat, since they would be predicted to apply lower selection pressure. Recent technological developments allow the fusion of quantitative genetic and molecular methods, often referred to as genetical genomics or expression Quantitative Trait Locus (eQTL) analysis, to efficiently identify regulatory loci that control the expression of many to hundreds of genes. Integrated deployment of these technologies coupled with efficient phenotyping offers significant potential to elucidate the regulatory nodes in genetic networks that orchestrate host defense responses. The focus of this mini-review will be to present advances in genetical genomic experimental designs and analysis, particularly as they apply to the prospects for discovering partial disease resistance alleles in cereal crops. Thiago Lvio P. O. Souza, Ana L. Alzate-Marin, Fbio G. Faleiro, Suelen N. Dessaune, Trazilbo J. de Paula-Junior, Maurilio A. Moreira, Everaldo G. de Barros (Brazil) Breeding for Common Bean Rust Resistance in Brazil ABSTRACT Invited Review: Common bean (Phaseolus vulgaris L.) is an economically, nutritionally, and socially important crop. It is grown in distinct regions and different seasons around the world by subsistence level farmers with low-technology input as well as by farmers that use high input technologies. One important factor that can limit the bean growing and drastically affect grain yields is the high number of destructive pathogens that attack P. vulgaris and cause serious damage to the crop. Among them is bean rust, incited by the highly variable fungus Uromyces appendiculatus. This disease is distributed throughout the world, but it effectively causes major production problems in humid tropical and subtropical regions. In Brazil, rust causes major losses in south, southeast, and central regions of the country. Bean rust control by resistant cultivars is an easy and economical strategy to be used in association to other rust management practices. The pyramiding of different race-specific resistance genes in association with other genes conferring adult plant resistance, slow rusting, and reduced pustule size can prolong the lifespan of a common bean cultivar by creating a more durable resistance complex against the rust pathogen. This review manuscript presents an overview on common bean rust and reports some breeding efforts aiming to develop rust resistant cultivars in Brazil.

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Lus Antnio Siqueira de Azevedo, Fernando Cezar Juliatti (Brazil) Strategies of Chemical Protection for Controlling Soybean Rust ABSTRACT Invited Review: Asian soybean rust (Phakopsora pachyrhizi) is a very destructive disease that undermines the current soybean production system in Brazil, and in the 23 million ha available for production, the disease is present throughout the entire cultivated area. The disease control has required a combination of several practices, in order to avoid losses. The chemical control of the Asian soybean rust is the most widely used method for the control of the disease. In the last two harvests, the application of fungicides has been shown as an effective alternative for the producer in the control of this aggressive disease. Under the technical and economic point of view, the best results have been obtained with two to three applications of triazole or triazole + strobilurin fungicides. Since the first fungicides emergency recommended for the 2002/03 harvest (azoxystrobin, difenoconazole, fluconazole, pyraclostrobin + epoxyconazole and tebuconazole), a large number of new formulations was added to the arsenal to control rust. There are today recorded in MAPA about 40 active ingredients (alone or in combination), trademarks and formulations for the rational use against rust. Among fungicides, there are differences in efficacy, residual period, metabolic stability and translocation rate, requiring care from producer and technical assistance in the choice of product to be used in each situation. In the present review, the chemical control of rust is analyzed in Brazil from 2001/02 to 2006/10; its economic importance, strategic variables for the rational use of fungicides, factors that complicate the control chemical and the risk of resistance to the main chemical groups. Arianne Tremblay, Benjamin Matthews (USA) Understanding the Soybean Rust Interaction with Soybean using Biotechnology ABSTRACT Invited Review: United States, Brazil and Argentina are the three biggest producers of soybean in the world at a combined 180 million metric tons each year. All soybean fields growing in these countries are susceptible to soybean rust; caused by Phakopsora pachyrhizi Sydow. With new and emerging biotechnology techniques we may develop new strategies for broadening resistance of soybean to soybean rust. Using these new techniques, scientists can identify and study genes expressed on both sides of a plant-pathogen interaction to better understand the infection process. The recent development of techniques, such as microarray analysis, high-throughput DNA sequencing and laser capture microdissection, has dramatically improved our ability to identify new genes and to examine gene expression. Microarrays can be used to study changes in expression of thousands of genes per experiment in infected leaves. The complete soybean genome sequence now available provides a template for analyzing gene expression results from deep sequencing of transcripts during the infection process. Deep sequencing provides unprecedented amounts of data for analysis of the genome and gene expression. Laser capture microdissection allows the isolation of specific cells from infected leaf cross-sections that can be analyzed for pathogen gene expression using diverse DNA sequencing approaches. There is still a great lack of knowledge of the function of soybean and fungal genes and their spatial and temporal expression. However, these new techniques will provide new information about pathogen development in its host. Some of these plant and fungal genes may be useful to broadening soybean resistance to soybean rust. David Henry Moon, Thiago Falda Leite, Juliano Bragatto, Luis Felipe Boaretto, Carlos A. Labate (Brazil) Eucalyptus Rust (Puccinia psidii): Strictly a Brazilian Problem? ABSTRACT Invited Review: Eucalyptus is one of the most important arboreal cultures with more than 19 million hectares planted worldwide, of which 6.4 million are planted in the Americas and it is used as the raw material for several industrial purposes. In 2010 the Brazilian pulp and paper industry produced 14.1 million tonnes of cellulose (almost exclusively from Eucalyptus spp.) and 9.8 million tonnes of paper. Currently the greatest single threat to Eucalyptus monoculture is tropical rust caused by Puccinia psidii, which attacks trees up to 2 years-old. This biotrophic fungus primarily infects the younger actively growing leaves, quickly covering these organs with the characteristic yellow pustules containing large numbers of spores in susceptible plants. If the infected individuals are able to recover they exhibit reduced height when compared to resistant individuals resulting in reduced yield at harvesting age. We will try to put into perspective potential the economic losses due to tropical rust by reviewing the current threat of P. psidii rust to Eucalyptus plantations in and outside Brazil. This review also contains a brief introduction into

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the life cycle of the fungus and evidence for genetic variation within and between populations of this pathogen. The interaction between Eucalyptus grandis-Puccinia psidii will be discussed with respect to differential gene and protein expression during the exposure of resistant and susceptible individuals to infection, indicating probable mechanisms of resistance and potential molecular markers. Finally, these results will be discussed in light of the chemical analysis carried out on the on leaves from a susceptible and a resistant clone in the presence and absence of the fungus. Eleonora Barilli (Spain), Amero A. Emeran (Egypt), Josefina C. Sillero, Ana M. Torres, Elena Prats, Diego Rubiales (Spain) The Pea Rusts, Uromyces pisi and U. viciae-fabae: Pathogen Identification and Sources of Resistance ABSTRACT Invited Review: Pea is the second most important feed legume crop in the world. Pea rust has become an important pea pathogen being widely distributed, particularly in regions with warm, humid weather. Most reports refer to Uromyces viciae-fabae as the causal agent of pea rust, however, pea rust can also be incited by U. pisi. Similarity of their macroscopical features makes difficult to distinguish among them, being necessary to resort to specific details of substomatal vesicle or to molecular identification. It seems that U. vicia-fabae on pea prevails in tropical and subtropical regions as India and China, while U. pisi might be prevalent in temperate regions. In this review we describe and discuss methods of identification, the occurrence and incidence of both pathogens on peas and the availability of sources of resistance. Maria Carlota Vaz Patto (Portugal), Diego Rubiales (Spain) Rust Resistance in Lathyrus cicera L. and Lathyrus sativus L. ABSTRACT Invited Review: Several Lathyrus species have a high potential both as food and fodder crops. Rust fungi are among the most important pathogens of major grain legumes including Lathyrus species. L. sativus and L. cicera are known for its tolerance to abiotic factors such as drought, heavy and poor soils, and to biotic factors such as rust fungi. We review and critically discuss available knowledge on existence of resistance and the underlying resistance mechanisms against rust fungi in the L. sativus and L. cicera. Cheng-ming Tian (China), Peng Zhao (Japan), Ying-mei Liang, Zhi-min Cao (China) Research Progress on Poplar Leaf Rust in China ABSTRACT Invited Review: Poplar leaf rust disease is a worldwide disease that causes damage in short rotation coppice (SRC) poplar plantations for renewable energy. The situation is especially severe in China as poplars are grown as a major SRC crop for energy for their yield potential and coppicing ability. From disease investigation to disease control, fungi taxonomy, physiological specialization and host resistance breeding, the present paper introduced the progress in researches on poplar leaf rust. Comments and the prospective of the researches on poplar leaf rust were discussed in terms of the research progress and current research situation in China. Suvendu Mondal, A. M. Badigannavar (India) Peanut Rust (Puccinia arachidis Speg.) Disease: Background and Recent Advances ABSTRACT Invited Review: The peanut rust disease caused by Puccinia arachidis Speg. is an important biotic stress that reduces pod and fodder yield and oil quality significantly. The fungus belongs to the class urediniomycetes like other rust fungi but occur to a lesser extent in teliospore form. P. arachidis is predominantly spread by the repeated cycle of urediniospores. The disease is prevalent in most countries where it is cultivated and favored by warm and humid climatic conditions. Despite its economic importance, limited work has been carried out on host-fungus interaction, fungal diversity and physiological specialization. The present mini-review describes different aspects of P. arachidis especially symtomatology, histopathology, epidemiology as well as developments on host resistance, genomics, breeding and control measures. Mitchell L. Wise (USA) Relationship between Crown Rust and Avenanthramide Biosynthesis in Oat (Avena sativa) ABSTRACT

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Invited Mini-Review: Avenanthramides are phenolic alkaloids produced in oat. These secondary metabolites are found in the grain as well as leaf and other tissues in the plant. A growing body of evidence indicates that these compounds impart beneficial health effects in animals thus may be of interest as phytonutrients. Originally termed avenalumins and described as phytoalexins by Mayama in 1981, the chemical structures of the avenanthramides were correctly determined in 1989. There is abundant evidence that avenanthramides are elicited in oat leaf and other tissue by infection with crown rust (Puccinia coronata) or molecular mimics of fungal infection. Correlations between crown rust infection and levels of avenanthramides in the grain have also been observed. This manuscript will review the current literature relating to the biosynthesis of avenanthramides in response to crown rust and other fungal elicitors, the efficacy of these phytoalexins in combating crown rust and implications for breeding oat cultivars for enhanced avenanthramide production. J. C. Comstock (USA), W. Ovalle (Guatemala), E. Chavarra (Costa Rica), N. C. Glynn, L. A. Castlebury, R. N. Raid (USA), Hector Orozco (Guatemala) Sugarcane Orange Rust, an Emerging Disease in North and Central America: Its Impact and Comparison to Sugarcane Brown Rust ABSTRACT Invited Mini-Review: Sugarcane orange rust, caused by Puccinia kuehnii, was detected in June 2007 in Florida and subsequently in several Caribbean, Central American and South American countries, as well as, Cameroon and the Ivory Coast in Africa. It has reduced production in several locations, primarily on CP 80-1743 in Florida; on CP 72-2086, SP 71-5574 and SP 79-2233 in Costa Rica and CP 72-2086 in Guatemala. The disease has also impacted variety development programs and has necessitated a rapid transition to more resistant varieties. Orange rust is characterized by the light orange coloration of pustules and poses a legitimate threat to all sugarcane producing countries in the Western Hemisphere. Orange rust develops in warmer weather conditions than brown rust and is active for a longer duration on the crop than brown rust, caused by P. melanocephala. Orange rust may be differentiated from brown rust by its light orange urediniospores, which have a prominent apical thickening. Urediniospores of P. melanocephala are darker brown and have uniformly thickened walls. Resistance ratings are given for a number of sugarcane varieties. Mohamed Bechir Allagui (Tunisia) Recent Progress in Oat Crown Rust in Tunisia ABSTRACT Invited Mini-Review: Crown rust caused by Puccinia coronata Cda. f. sp. avenae Eriks, is a major fungal disease of cultivated oats (Avena sativa and A. byzantina) in Tunisia. In a pure stand or in association with vetches, oats are the dominant fodder crops grown mainly for hay and silage for livestock. Surveys conducted during different cropping seasons showed that crown rust severity in farmers fields varied with the year; the humid year favourable to oat culture was also favourable to crown rust epidemics and vice versa mainly when the year is too dry. Trials on sowing dates showed that, for early-seeded plantings, early development of crown rust occurred on susceptible varieties. The attack was delayed when sowing date was late and the oat variety had efficacious resistance gene. The Pc-gene differential oat lines, used by oat researchers to study the virulence pattern in oat crown rust populations showed the virulence phenotypes locally recorded were TJFR, TDFL and TBLM; indicating that the Pc-genes still with resistance to oat crown rust were Pc38, Pc39, Pc52 and Pc68. Results on alternate host indicate that Rhamnus lycioides L., a common and endemic part of the vegetation in Tunisia, is a new aecial host of oat crown rust that the aeciospores released in April and May constitute the source of the fungus virulence diversity. Landraces of oats recently collected from different regions were evaluated for leaf rusting and for agronomic traits. The results suggest that vast areas in Tunisia are still rich with oat crown rust resistance as many landraces displayed important agronomic traits and good resistance to crown rust making it useful in breeding programs. Stephen J. Molnar, Winson Orr, David L. De Koeyer, James Chong, Nicholas A. Tinker (Canada) Development and Validation of PCR-based Markers Linked to Pc68/Pg3/Pg9 and Pg4/Pg13 Rust Resistance Gene Clusters in Oat ABSTRACT Original Research Paper: Genetic resistance to pathogens causing crown and stem rust is essential for oat varieties developed worldwide. We report the development, testing, and deployment of five new PCR-based markers linked to the complex gene cluster containing the Pc68 crown rust resistance gene and the Pg3 and Pg9 stem rust resistance genes. These markers are based on sequence characterized amplified regions (SCAR), on cleaved amplified polymorphic sequences (CAPS), and on an amplified microsatellite or simple sequence repeat (SSR). Marker-assisted selection (MAS) with these markers was

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used to identify oat lines containing Pc68 resistance in four oat populations. The SCAR and CAPS markers can be used as either dominant or co-dominant markers depending on the alleles present in the population of interest, while the SSR marker is co-dominant. At a second complex gene cluster, the Pg4 stem rust locus was mapped close to markers previously shown to be linked with the Pg13 stem rust resistance locus, validating the map location of both rust loci. For this cluster, a new SCAR marker was developed that has potential for MAS for Pg4 and Pg13. Md. Shahidul Haque, Robert F. Park, Colin R. Wellings (Australia) Postulation of Seedling Resistance Genes and Assessment of Adult Plant Response to Stem Rust in Two International Oat Nurseries ABSTRACT Original Research Paper: Multipathotype greenhouse tests were conducted to postulate seedling resistance to stem rust in entries in the 1998 (71 lines) and 1999 (95 lines) North American Quaker Oat Nurseries. The seedling tests were conducted with seven Australian races of the oat stem rust pathogen (Puccinia graminis f. sp. avenae; P. g. avenae), one of which was also used in field nurseries to investigate the adult plant rust responses of the oat lines. The 166 lines carried eight oat stem rust seedling resistance genes, namely, Pg-a (41 lines), Pg-1 (17), Pg-2 or Pg-4 (34), Pg-2 + Pg-4 (24), Pg-8 (1), Pg-10 (4), Pg-13 (2) and Pg-Sa (2), none of which were effective to all seven P. g. avenae races used. A total of 26 lines had unknown resistance to P. g. avenae and 15 lines were susceptible to all P. g. avenae isolates used. All 166 lines were rated as susceptible at adult plant growth stages, however, several had lower disease severities and may therefore carry slow rusting against P. g. avenae. Further field tests are however needed to examine this in more detail. Evsey Kosman, Pnina Ben-Yehuda, Jacob Manisterski (Israel) Virulence Dynamics of Puccinia triticina on Wheat in Israel ABSTRACT Original Research Paper: Pathotype and virulence structure of four annual collections (1994, 1998, 2002 and 2006) of wheat leaf rust (P. triticina) in Israel were analyzed in order to reveal major trends of the pathogen evolution in the course of time. Virulence phenotypes of 60 to 84 isolates were annually determined on 17 Thatcher isogenic lines that possess Lr1, Lr2a, Lr2c, Lr3, Lr3ka, Lr9, Lr10, Lr11, Lr15, Lr16, Lr17, Lr18, Lr21, Lr23, Lr24, Lr26 and Lr30 resistance genes. The number of detected pathotypes varied from 18 to 38 in 1998 and 1994, respectively, which were years of the least and highest richness of the pathogen annual collections. Some population parameters were also compared with the corresponding ones in the collection of P. triticina on wheat in 1993. Changes in the Israeli population of wheat leaf rust may be attributed to the possible massive migration of leaf rust urediniospores from the neighboring regions in 1994 and selection pressure of new wheat cultivars since 1997. The pathogen collections of 1994 and 1998 were significantly different from those of 2002 and 2006. Many new pathotypes that emerged in 1994 belonged to the M-race with virulences on Lr1 and Lr3, and the T-race with virulences on Lr1, Lr2a, Lr2c and Lr3. Isolates of the T-race were prevalent in 1994 and 1998, but were nearly not detected in 2002 and 2006. On the other hand, the frequency of the M-race drastically increased in 2002 and 2006. These two pairs of annual collections can also be distinguished on the basis of virulence dynamics on Lr2a, Lr17 and Lr15 resistance genes. The frequency of virulence on Lr2a was moderate in 1994 and 1998, but nearly nil in 2002 and 2006. In contrast, the frequency of virulence on Lr15 and Lr17 resistance increased considerably in 2002 and 2006. Patterns of genotypic and genetic diversity within the annual collections of wheat leaf rust were not congruent. Mary Frances Mahalovich (USA) Genetic Differentiation of Pinus albicaulis Engelm. Populations from the Northern Rocky Mountains ABSTRACT Original Research Paper: Survival, height, cold hardiness and blister rust resistance of 107 individual-tree and one bulk lot were used to summarize patterns of genetic differentiation for whitebark pine across 3,600 km2 of forested lands in Idaho, Montana, and eastern Washington, U.S.A. The artificial inoculation trial demonstrates the efficacy of screening for resistance to the recently introduced blister rust fungus. Significant proportions of genetic variance are associated with seed source (1-44%) and families within seed sources (7-59%). Family heritabilities for survival, height, cold hardiness, and blister rust resistance are moderate to high in the inoculated seedlings (0.68-0.99) and low to high in the control seedlings (0.33 to 0.92). Rust resistance as described by seven traits is weakly correlated with 6th year height, where seed sources from northwestern locations are taller and have more rust resistance (r=0.14). Rust resistance is weakly correlated (r=0.27) to freezing injury, where populations from lower elevations or geographically milder sites are more resistant but have low freezing tolerance. Survival is moderately

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correlated to freezing injury (r=0.45) and 6 year height (r=0.36). Seed sources from lower elevations or geographically milder sites are taller but have low freezing tolerance. Regression models explaining 15-68% of the variability suggest that seed transfer can be geographically broad, 1.85 latitude, 2.15 longitude, and 245 m elevation. Moderate seed source differentiation, low QST index values, and variation explained by random events not included in the regression models characterize whitebark pine as having a generalist adaptive strategy in this portion of the species distribution. Omar Perdomo Snchez (Dominican Republic), Meike Piepenbring (Germany) Species of Uromyces (Pucciniales, Basidiomycota) on Loranthaceae ABSTRACT Original Research Paper: Two new species of Uromyces with reticulated teliospores are compared with nine species known from Loranthaceae. The new species Uromyces brasiliensis has smaller spores than all known species of Uromyces with reticulate teliospores. Uromyces struthanthi is characterized by a large pedicel of the teliospores and spinose-echinulate aeciospores. In addition new details of ornamentation of aecidio- and teliospores of known species are presented. Francisco das Chagas Oliveira Freire (Brazil), Reinhard Berndt (Switzerland) An Updated List of Rust Species on Plants from Cear State (Brazil) ABSTRACT Research Note: A survey of rust fungi (Basidiomycota, Uredinales) from plants in Cear State (Brazil) is presented. Puccinia is the most common genus, followed by Uromyces, Achrotelium, Chaconia, Dasyspora, Esalque, Hemileia, Kimuromyces, Maravalia, Melampsora, Phragmidium, Physopella, Porotenus, Prospodium, Pucciniosira, Sphaerophragmium, Sphenospora and Ypsilospora. At least five new species have recently been described. The species Coleosporium solidaginis, Phakopsora euvitis, Puccinia horiana and Uromyces transversalis were introduced into Cear State through infected seedlings from the southeast region of Brazil. The importance of studying the fungi biodiversity on different types of vegetation of Cear State is discussed.

th

The Asian and Australasian Journal of Plant Science and Biotechnology

Mahendra L. Ahire, Vinayak H. Lokhande, Polavarappu B. Kavi Kishor, Tukaram D. Nikam (India) Brinjal (Solanum melongena Linn.) Varieties Accumulate both Na+ and K+ under Low NaCl Stress, but Excludes Na+ and Accumulate K+ under High Salt Levels ABSTRACT Original Research Paper: The effect of NaCl stress (0 200 mM) was investigated on the accumulation of mineral nutrient and antioxidant enzyme activities in two brinjal varieties MEBH 10 (salt sensitive) and MHBJ 112 (salt tolerant) that differ in their salt sensitivity. NaCl stress resulted in an increase in sodium (2.55- and 3.10-fold), potassium (1.95- and 2.65-fold) and calcium (1.55- and 1.48-fold more) content in the seedlings of MEBH 10 and MHBJ 112 up to 100 mM NaCl level over control, respectively. The magnitude of accumulation of Na+ and K+ ions was more in salt tolerant variety MHBJ 112 as compared to salt sensitive variety MEBH 10. Both the lines maintained significantly lower Na+/K+ but not Na+/Ca2+ ratios. Under high salt stress, brinjal varieties excluded Na+ and accumulated K+. Catalase (CAT) activity increased with increasing NaCl level in both varieties. About 225% more CAT activity in MEBH 10 and 249% more in MHBJ 112 was recorded at 200 mM NaCl as compared with the respective control. The activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and guaiacol peroxidase (GPX) increased up to 100 mM NaCl but decreased at higher concentrations (150 200 mM) of NaCl. An about 124% and 291% increase in SOD activity in MEBH 10 and MHBJ 112, respectively were recorded at 100 mM NaCl. Similarly, a 124% increase in AXP activity in MEBH 10, 118% in MHBJ 112 and 175% and 168% increase in GPX activity was recorded in MEBH 10 and MHBJ 112, respectively at 100 mM NaCl. Thus the mechanism of high salt tolerance in brinjal appears to be reduced Na+, increased K+ and by maintaining higher activity of antioxidant enzymes. Hameedunnisa Begum, Medagam Thirupathi Reddy, Surapaneni Malathi, Boreddy Purushotham Reddy, Sunil Arcahk, Javaregowda Nagaraju, Ebrahimali Abubaker Siddiq (India) Molecular Analysis for Genetic Distinctiveness and Relationships of Indigenous Landraces with Popular Cultivars of Mango (Mangifera indica L.) in Andhra Pradesh, India

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ABSTRACT Original Research Paper: Mango (Mangifera indica L.) is mostly distributed in the two eco-geographical regions (Coastal and Rayalaseema) of an Indian state of Andhra Pradesh, where large morphological and genetic diversity has been detected due free pollination among plants. A juicy landrace-specific exploration survey covering the two eco-geographical regions was undertaken during 2008 to identify new juicy landraces of mango with high fruit quality. Following a selective sampling strategy, 20 indigenous landraces of mango were selected according to fruit morphology and their fruit and leaf samples were collected for morpho-physiological and molecular characterization, respectively. Morpho-physiological characterization and evaluation based on fruit characteristics revealed that six landraces viz., DM Acc-3, 4, 7, 15, 17 and 18 were elite with respect to fruit characteristics, which were further characterized for their genetic distinctiveness and relationships with the choicest juicy cultivars of mango in Andhra Pradesh (Peddarasam, Chinnarasam, Cherukurasam, Panchadarakalasa and Suvarnarekha) at the molecular level, using 109 mango-specific microsatellite markers (SSRs). Jaccards similarity coefficient ranged from 0.40 to 0.69 signifying wide variability in the landraces of mango to an extent of 31-60%, offering ample scope for selection. Of the 109 SSR markers validated, 57 were polymorphic, of which 10 were highly polymorphic. Diversity was high among 11 genotypes with a total number of 198 alleles displayed. Microsatellite SSR-84 was able to differentiate and/or identify all of the 11 genotypes under study. UPGMA cluster analysis revealed that the six elite landraces viz., DM Acc-3, 4, 7, 15, 17 and 18 clustered according to their geographical origin. Being horticulturally superior and genetically distinct, these six land races can be clonally selected for further testing and multiplication for commercial exploitation. Koona Subramanyam, Kondeti Subramanyam, Pinnamaneni Rajasekhar, Chirra Srinivasa Reddy (India) Assessment of Genetic Relationships among South Indian Chilli (Capsicum annum L.) Cultivars Using RAPD and ISSR Markers ABSTRACT Original Research Paper: The present investigation was carried out with the objective of evaluating genetic diversity in chilli (Capsicum annum. L). A total of 24 south Indian chilli cultivars including five commercial hybrids were characterized using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses. Out of 50 primers employed to generate RAPD profiles, reproducible bands were obtained with 16, 15 (93.75%) of which could detect polymorphism. A total of 121 bands were scored, out of which 59 bands (48.76%) were polymorphic. RAPD primer SKURAPD18 exhibited the highest level of polymorphism (81.81%). ISSR analysis was carried out by using 13 ISSR primers, 10 of which produced reproducible amplified fragments, and 9 of which (i.e., 90%) showed polymorphic bands. From 9 ISSR primers 97 fragments were amplified, 61 (62.88%) of which were polymorphic. Primer SKUISSR8 showed maximum polymorphism (83.83%). A dendrogram was developed using Jaccards coefficient of similarity and UPGMA with RAPD and ISSR data. The constructed dendrograms revealed that the commercial hybrid cultivars formed separate clusters. All 15 RAPD primers and 9 ISSR primers could distinguish all chilli cultivars. Only 9 ISSR primers were needed to generate sufficient information about genetic diversity, whereas 15 RAPD primers were required. Thirupathi Reddy Medagam, Hari Babu Kadiyala, Ganesh Mutyala, Hameedunnisa Begum (India) Diallel Analysis for Yield and its Components in Okra (Abelmoschus esculentus (L.) Moench) ABSTRACT Original Research Paper: Gene action and combining ability effects of yield and its components of okra (Abelmoschus esculentus (L.) Moench) were studied through 10 10 half-diallel analysis. Forty five single crosses were developed by crossing 10 lines of okra viz., P1(IC282248), P2(IC27826-A), P3(IC29119-B), P4(IC31398-A), P5(IC45732), P6(IC89819), P7(IC89976), P8(IC90107), P9(IC99716) and P10(IC111443) in a half-diallel manner during summer. All 45 F1s along with their 10 counterpart parental lines were evaluated in a randomized block design with three replications during early kharif (June to September) of the same year at the Vegetable Research Station, Rajendranagar, Andhra Pradesh, India. Combining ability analysis following Method-II and Model-I of Griffings approach revealed the preponderance of non-additive gene action for plant height, internode length, days to 50% flowering, first flowering node, first fruiting node, fruit length, fruit weight, total number of fruits/plant, number of marketable fruits/plant, total yield/plant, marketable yield/plant, fruit and shoot borer infestation on fruits and shoots and Yellow vein mosaic virus infestation on fruits and plants and additive gene action for number of branches/plant and equal importance of both additive and non-additive gene action for fruit width. The parental lines P5(IC45732), P6(IC89819) and P7(IC89976) were high general combiners for total and marketable yield per plant and other yield associated traits. The crosses C23(IC29119-B IC99716), C17(IC27826-A IC111443), C42(IC89976 IC111443) and C43(IC90107 IC111443) were superior specific combiners for total and marketable yield/plant and other yield-related traits

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could be exploited for the production of F1 hybrids. The cross C42(IC89976 IC111443), having one of the parents with significantly positive general combining ability effect for total and marketable yield/plant, could be utilized in recombination breeding. Jaime A. Teixeira da Silva (Japan), Judit Dobrnszki (Hungary), Pham Thanh Van (Japan/Vietnam), Nafees Khan (India), William A. Payne (USA/Burkina Faso) Corresponding Authors: Rules, Responsibilities and Risks ABSTRACT Research Note: It is generally understood that the corresponding author (CA) is responsible for all communications related to the submission of a manuscript to a journal. However, it is quite common that the CA be a student or inexperienced scientist, which can lead to often very damaging results arising from the allocation of responsibility to that person. Errors most commonly made by these CAs include (despite signed declarations to the publisher or journal): submission of a manuscript without knowledge of the co-authors; falsification of data or double submissions; inclusion of false authors or those who should not be authors. Most of these errors could be eliminated if: 1) there were full, open and transparent communication between the CA and the other co-authors and between the CA and the publisher; 2) the CA selected were a senior member of the research group; 3) all key points during the publishing process were shared with all co-authors, including submission, main revisions and acceptance. The choice of the CA should not lie with the journal or publisher, but the choice should be made smartly in line with the guidelines and advice held within this paper.

The European Journal of Plant Science and Biotechnology

SPECIAL ISSUE: Buckwheat 3, 2012 Tatsuro Suzuki, Yuji Mukasa, Tosikazu Morishita (Japan), Sun-Ju Kim, Sun-hee Woo (Korea), Takahiro Noda, Shigenobu Takigawa, Hiroaki Yamauchi (Japan) Possible Roles of Rutin in Buckwheat Plant ABSTRACT Invited Mini-Review: Buckwheat contains rutin, a kind of flavonol, not only in the seeds but also in the cotyledon, leaf, stem and flower. To date, several reports have been published on the physiological roles of rutin. In this review, mainly based on our recent results, we summarize the main aspects of rutin related to its possible physiological roles. The enzymes, which catalyzed a part of rutin synthesis (quercetin glycosyl transferase) and decomposition (rutinosidase), have lower Km especially for quercetin/UDP-glucose and rutin respectively compared to other similar compounds. This indicates that buckwheat developed glycosyl transferase and rutinosidase suit for rutin metabolism. The time course studies for rutin accumulation at seed ripening and leaf/cotyledon expansion show that rutin accumulation pattern is different for each organ. In seeds, rutin content per seed increases along with seed development, and mature seed contains the largest rutin concentration. In leaves, rutin content also increased along with development whereas senescent leaves contain little amount of rutin. In the cotyledon and mature leaves, rutin is distributed in the epidermis, and large amount of rutinosidase activity is present on surface of cotyledon during cotyledon expansion. In addition, rutin concentration and rutinosidase activity was increased by some stresses (UV-B radiation, cold and moisture stress) in leaves. Quercetin, the hydrolyzed moiety of rutin, possesses high antioxidant activity and the ability to be a precursor of a kind of anti fungal agents. Therefore, rutin and rutinosidase are suggested to be related to the enhancement of the defense system against stress conditions in buckwheat. Tatsuro Suzuki, Yuji Mukasa, Tosikazu Morishita (Japan), Sun-hee Woo, Sun-Ju Kim (Korea), Takahiro Noda, Shigenobu Takigawa, Hiroaki Yamauchi (Japan) Possible Roles of Lipase, Lipoxygenase and Peroxidase in Buckwheat Flour and Noodles ABSTRACT Invited Mini-Review: The freshness of buckwheat flour and its unique flavor is important for the quality of buckwheat products. Several reports have shown that lipid degradation and oxidation in buckwheat flour are the main causes of measurable quality deterioration during storage. On the other hand, some flavor compounds are produced by lipid degradation and oxidation pathway. Therefore, understanding lipid degradation and oxidation pathways is important in the quality control of buckwheat flour and products. In some crops, lipoxygenase pathway is important for lipid degradation and oxidation. The pathway includes some enzymes such as lipase (triacylglycerol lipase EC 3.1.1.3) (LIP), lipoxygenase (EC 1.13.11.12) (LOX) and peroxidase (EC

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1.11.1.7) (POX). This review, mainly based on our recent results, summarizes the main aspects of the possible relation between these enzymes, their substrates and flour deterioration/flavor generation as well as purification and characterization of related enzymes. LIP and POX activity in buckwheat flour apparently plays a role in the lipid degradation and quality deterioration whereas LOX does not have significant influences. LIP and POX activity in buckwheat flour also plays an important role for flavor generation of boiled buckwheat noodles whereas LOX does not have. This indicates that the mechanism of quality deterioration and flavor generation in buckwheat flour is different from that of rice and soybean. Svetlana Radovi, Jovanka Milju-Djuki, Jelena Samardi, Bojana Banovi, Dragana Nikoli, Mira Milisavljevi, Gordana Timotijevi (Serbia) Buckwheat as a Model Plant in Molecular Biology ABSTRACT Invited Mini-Review: Buckwheat (Fagopyrum esculentum Moench) is a pseudocereal crop, mostly grown in the Northern Hemisphere. It is desirable for human consumption because buckwheat seeds have a high content of proteins (with high concentrations of essential amino acids) and minerals (e.g. iron, zinc and selenium). Concerning their high nutritive value, buckwheat seed storage proteins (SSPs), and genes that code for them, are of importance to study. Our research focus is the structure and the expression profile of selected buckwheat genes coding for proteins of known functions (such as SSPs), as well as proteins of undefined functions possibly involved in protein degradation/processing, and/or in the stress response (e.g. aspartic proteinases and metallothionein). These genes, their promoters and translational products are important, not only from the aspect of fundamental research, but also in regard to their potential biotechnological application in agriculture and land preservation. In particular, we are interested in the processes taking place during the last stage of buckwheat embryogenesis, especially in the analyses of specific gene expression regulation under normal physiological and/or stress conditions, which is the subject of our present research. Asha Panwar, Nidhi Gupta, Rajinder S. Chauhan (India) Biosynthesis and Accumulation of Flavonoids in Fagopyrum spp. ABSTRACT Invited Review: Buckwheat is a multipurpose crop used for both grains and greens and known to have several medicinal and nutritional properties. Buckwheat contains flavonoids such as rutin, anthocyanins, catechins, chlorogenic acid, 4-hydroxy-3-methoxy benzoic acid, caffeic acid, epicatechins, p-courmaric acid, ferulic acid etc. Fagopyrum esculentum and Fagopyrum tataricum are the major source of flavonoid called rutin. Seeds of F. tataricum contain higher rutin content in comparison to F. esculentum. This review discusses the physiological and molecular basis of flavonoid biosynthesis and accumulation in plants in general and rutin and anthocyanin content in Fagopyrum species, its correlation with the expression of flavonoid pathway genes and the effect of different environmental factors on flavonoid biosynthesis. The understanding of rutin biosynthesis in buckwheat is expected to supplement for genetic improvement of buckwheat for higher nutritional value. Jai C. Rana, Rajinder C. Chauhan, Tilak R. Sharma, Nidhi Gupta (India) Analyzing Problems and Prospects of Buckwheat Cultivation in India ABSTRACT Invited Mini-Review: Buckwheat is a multi-purpose crop grown widely at higher elevations in the Indian Himalayan region. Buckwheat contains high amounts of antioxidant compounds, mainly rutin (a flavonol glycoside) with several medicinal properties. The physiological and biological properties of rutin include anti-oxidation, anti-inflammation, anti-hypertension, vasoconstrictive, spasmolitic and positive inotropic effect. Out of 20 species reported world-wide only two, Fagopyrum tataricum and F. esculentum are cultivated in India and elsewhere in the world. The majority of the varieties grown are farmers own selections, however, five varieties have been released through the All India Coordinated Project on underutilized crops. Besides, around 911 germplasm accessions have been conserved in medium term storage at Shimla; 837 out of these have also been maintained as base collection in the National Gene Bank. Although buckwheat has diverse uses as food, medicinal and industrial plant, the area under its cultivation is decreasing at an alarming rate mainly due to changing cropping patterns, migration, low productivity, changing food habits, and lack of alternative uses/products. Screening of germplasm for biotic and abiotic stresses, restructuring of research and development activities towards value addition by exploiting its potential as fast food, medicinal and beverage plant and awareness of its food and medicinal value at grass root level are some of the new thrust areas for expanding buckwheat cultivation.

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Saswati Sen, S. K. Dutta (India) Evaluation of Anti-cancer Potential of Ragi Bifunctional Inhibitor (RBI) from Eleusine coracana on Human Chronic Myeloid Leukemia Cell Line ABSTRACT Original Research Paper: Finger millet (ragi), a staple food crop, is well known as a super cereal for its excellent nutritional value, long-term storage potential and ethno-medicinal characteristics. Ragi bifunctional inhibitor (RBI) from Eleusine coracana Gaertn. (ragi/finger millet) belonging to cereal alpha-amylase/protease inhibitor family, inhibits alpha-amylase and trypsin simultaneously. Several seed protease inhibitors are known for their protective as well as curative role against many types of human cancers. Recently, the anti-cancer activity of ragi seed extract on K562 chronic myeloid leukemia cells was explored. In the present study, RBI was purified from finger millet seeds by affinity chromatography followed by FPLC (Fast Protein Liquid Chromatography) size exclusion chromatography. Purified RBI showed cytotoxicity against K562 chronic myeloid leukemia cells (IC50 = 20 g/ml) but, not against normal human peripheral blood mononuclear cells. Reduction of cellular proliferation and induction of apoptosis of K562 cells by purified RBI was determined by MTT (3-4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assay and flow cytometry analysis. This investigation being the first report on the anti-proliferative potential of RBI from edible ragi seeds, might provide a future preventive as well as curative natural solution for chronic myeloid leukemia. Bojana Filipev, Olivera imurina, Marija Bodroa-Solarov (Serbia) Enrichment of Ginger Nut Biscuits with Wholegrain Buckwheat and Rye Flour ABSTRACT Original Research Paper: To broaden the use of common buckwheat in human nutrition, an attempt was made to use wholegrain buckwheat flour in ginger nut biscuit formulations. The aim of the study was to develop a ginger nut biscuit formulation in which wheat flour is partially substituted with buckwheat flour in such doses to obtain nutritionally improved and acceptable product. Buckwheat flour was tested at doses 30, 40, 50% (flour weight basis). Proximate composition (basic chemical composition, fibers, micro elements), physical (height, width, spread, density, color) and textural attributes (hardness, fracturability) of buckwheat enriched ginger nut biscuits were analyzed and compared to those made with wheat flour solely. Since buckwheat tends to increase hardness of the products due to the peculiar gelling properties of buckwheat starch, inclusion of rye flour at 10 and 20% dose was also investigated. The dimension of ginger nut biscuits decreased but since the biscuit height was more affected by the rising doses of buckwheat, the spread increased in the composite biscuits (significant difference was noted at 40 and 50% levels of replacement). This could be attributed to the coarse granularity of buckwheat flour. Hardness and fracturability of ginger nut biscuits increased significantly with the addition of 50% buckwheat. The enriched biscuits were nutritionally improved as they contained more fibers, minerals and proteins (percent increase ranges depending on the replacement level were: for fibers 41.8-141.4%, for Fe 132.3-189.2%, for Zn 115.4-223.1%, for Mn 43.9-72.7%, for Cu 212.5-262.5%, for proteins 6.6-17.1%). Yellow tone decreased in all buckwheat containing biscuits at all substitution levels. All composite biscuits over 40% buckwheat substitution level showed an increased red tone. The majority of composite biscuits did not differ significantly from the control regarding darkness. Danuta Zieliska, Jacek Kwiatkowski, Henryk Zieliski, Dorota Szawara-Nowak, Magorzata Przygodzka, Grzegorz Lamparski (Poland), Fridrich Zeller (Germany) Antioxidant Properties and Flavonoid Composition as Quality Index of the Hulls and Groats from Common, Tartary and an Interspecific Hybrid of Buckwheat ABSTRACT Original Research Paper: The interest in buckwheat flavonoids as therapeutic agents against diseases involving free radical damage and possible allelopathic compounds is growing. This paper presents a field program addressed to buckwheat genotypes with the aim to develop cultivars of high antioxidant capacity and high flavonoid concentration in the hulls and groats. Therefore, both parameters were considered as new quality index of buckwheat cultivars. The antioxidant capacity of groat and hull separated from buckwheat seed was evaluated against
+

stable,

non-biological

radicals

such

as

2,2-azinobis-(3-ethylbenzothiazoline-6-sulphonate) radical cation (ABTS ) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), and against the key reactive oxygen intermediate superoxide anion radical (O2-) while the reducing capacity was measured directly by cyclic voltammetry (CV) method. The extent of variation in the content of rutin, quercetin and flavone C-glucosides in hull and groat was determined by high-performance liquid chromatography with diode array detection (HPLC-DAD). The content of flavonoids determined in the hulls and dehulled seeds of common, tartary and hybrid buckwheat varieties was

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correlated with antioxidant capacity determined by updated analytical strategies. The applied analytical strategy for the determination of the antioxidant capacity, based on the four analytical assays offering different chemical principles, created a useful quality parameter for buckwheat. The applied methods provided the same rank of the antioxidant capacity among hulls and groats however the real antioxidant capacity values differed according to the methods. The rank of values of the antioxidant capacity vs. applied methods was as follows: ABTS assay PCL assay > DPPH RSA CV assay. The rutin and flavone C-glucosides content in the groats from common buckwheat cultivars and the interspecific hybrid were lower than those noted in the hulls. In contrast, groat and hull from tartary buckwheat accession contained comparable high level of flavonoids. The extent of variation in the content of rutin, quercetin and flavone C-glucosides in the hull may be useful for researchers working on the resistance of buckwheat lines against pathogens and diseases whilst extent of variation in the flavonoid composition in groats should be important from the nutritional point of view. Therefore, flavonoid composition of the hulls and groats from different buckwheat genotypes may serve as a quality index. Andrea Brunori, Gerardo Baviello (Italy), Ferenc Kajdi (Hungary), Jaime A. Teixeira da Silva (Japan), Tibor Gyri, Gyrgy Vgvri (Hungary) Grain Yield and Rutin Content of Common and Tartary Buckwheat Varieties Grown in North-Western Hungary ABSTRACT Original Research Paper: Grain yield and rutin content was assessed in 31 common buckwheat (Fagopyrum esculentum Moench) and three tartary buckwheat (Fagopyrum tataricum Gaertn.) varieties grown in the summer of 2008 at the Research and Experimental Farm of the Faculty of Agricultural and Food Sciences, University of West Hungary, Mosonmagyarvr, Hungary. Grain yield of common buckwheat varied from a high of 0.98 t/ha in Vlada, 0.94 t/ha in Koto and 0.92 t/ha in Anita Belorusskaya to lows of 0.31 t/ha in Arakawa Village, 0.47 t/ha in Kora and 0.48 t/ha in Springfield. Much lower grain yield was observed in the three tartary buckwheat varieties: 0.12 t/ha in Ishisoba, 0.37 t/ha in Donan and 0.38 t/ha in Golden. As expected, and unlike grain yield performance, the rutin content observed in F. tataricum was as much as two orders of magnitude higher than that of F. esculentum, ranging between 974 mg/100 g DW in Golden and 1196 mg/100 g DW in Ishisoba. In common buckwheat, grain rutin content ranged from 8 mg/100 g DW in Darja and Kitawasesoba to 24 mg/100 g DW in La Harpe. The best compromise between grain yield and rutin content in common buckwheat was observed in var. Vlada, which had the top yield and ranked third in rutin content. Tartary buckwheat grain was a valuable source of rutin. In general, grain yield and rutin content were largely not correlated, almost as if grain development and rutin accumulation were not competing processes. Rather, rutin accumulation appeared to be the function of a varietys aptitude likely related to origin. European varieties, regardless of their yield potential, expressed quite clearly a somewhat higher rutin content than varieties originating from Pacific areas, namely Japan and Canada. SPECIAL ISSUE: Millets, 2012 Arun Gupta, Salej Sood, Pawan Kumar Agrawal, Jagdish Chandra Bhatt (India) Floral Biology and Pollination System in Small Millets ABSTRACT Invited Mini-Review: The flowers and flowering of small millets are poorly understood taxonomically. The knowledge of floral structure, floral biology and pollination behaviour are pre-requisite for understanding the system and its manipulation for developing a proper crossing technique, which is largely lacking in small millets. The present manuscript deals with floral morphology of small millets in particular in the light of the Poaceae family. Shantharaj Deepak, Sathyanarayana Niranjan-Raj (India), Axel Mithfer (Germany), Shekar H. Shetty (India) Nutritional Bio-fortification in Pearl Millet ABSTRACT Invited Mini-Review: Research on millets and their food value is in its infancy and its potential is vastly untapped. This review focuses on nutritional enhancement and improvement in pearl millet (Pennisetum glaucum (L.) R. Br.), a staple food crop of the semi-arid tropics largely grown for food and fodder in sub-Saharan Africa and parts of India. Climate change will greatly affect what crops we grow and where we grow it. Two of the most comprehensive models of climate change suggest that pearl millet is among the winner crops which are likely to be most suited and widely cultivated in future. Pearl millet is mainly grown for grain

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but it is also valued for its stove and/or forage. Apart from being used as food for human consumption and feed for livestock and poultry, pearl millet grain is also gaining importance as a cheap source of starch for fine quality brewing and in other diversified food uses. However, pearl millet being a crop grown and consumed by the poorest farmers needs nutritional bio-fortification by conventional and transgenic approaches to counteract the present nutritional deficiency in sub-Saharan Africa and parts of India. Kajal Srivastava, A. K. Sharma (India) Nutraceutical Importance of Finger Millet (Eleusine coracana) for Improved Human Health ABSTRACT Invited Mini-Review: Diet is a major focus of public health strategy aimed at maintaining optimum health throughout life thus preventing early onset of chronic diseases as well as promoting healthier ageing. Studies on the different properties of foods have shown that consumption of certain foods may provide greater health benefits. One such group of healthy food is the nutraceutical, which can be any substance that is a food or component of a food that provides medical, health benefits, including the prevention and treatment of disease. They are often used in nutrient premixes or nutrient systems in food and pharmaceutical industries. Such foods items or food components that help in prevention or treatment of diseases are made from herbal/botanical raw materials. Finger millet (Eleusine coracana) is one of the oldest cereal grains in the Indian sub-continent having high nutraceutical value. It grows well in harsh environments and on poorly fertilized and dry soils where other crops give poor yield. It also grows well in hot climates with short rainfall periods and cool climates with warm summers. A multitude of small farmers grow finger millet with limited water resources and in many countries this crop is often referred as poor people's crop. Finger millet as compared to the other crops is a very rich source of calcium; the calcium content is thirty times more than that of rice and wheat. It is not only a rich source of calcium but contains also other micronutrients essential for good health. It can replace or complement traditional crops and, with vertical integration into agriculture and manufacturing, it will also have impact on rural economic development. Gbor Gyulai, Lszl Holly, Richrd Lgler, Lajos Horvth (Hungary) The Hungarian Gene Bank Collections of Common Millet (Panicum miliaceum) and the Application for Conservation Genetics ABSTRACT Original Research Paper: Over the past decades gene bank of RCAT (Research Center of Agrobiodiversity, Tpiszele, Hungary), specialized for field and vegetable crops, has collected 250 accessions of heirlooms, old varieties, landraces and breeding materials of common millet (Panicum milaceum). The accessions have been characterized by descriptor list and compiled according to the international gene bank standards (IPGRI). In the study presented, short history of the RCAT, and a case study of common millet accession (P. miliaceum) are presented. Molecular analysis of DNA samples of twenty current millets and two archaeological seed samples excavated from the 4th and 15th centuries are compared at loci of gln4 (nuSSR), ISSRs and 18S mtDNA. The in silico analyses at loci of ITS1-5.8S-ITS2 (nuDNA), rbcL (cpDNA) and protein RuBisCO are also presented to trace molecular events occurred during the evolution and domestication of common millet. Shashi Kumar Gupta, Thirunavukkarasu Nepolean, Kedar Nath Rai, Charles Thomas Hash (India), Ranjana Bhattacharjee (India/Nigeria), Abhishek Rathore (India) Within-line Genetic Variation for Quantitative Characters and SSRs in Long-time Maintained Inbreds in Pearl Millet [Pennisetum glaucum (L). R. Br.] ABSTRACT Original Research Paper: Six maintainer (B-) and restorer (R-) lines each from ICRISATs pearl millet hybrid parental line breeding program were investigated for within-line genetic variation for quantitative characters and simple sequence repeats (SSRs). Thirty two progenies of each inbred line were evaluated under two contrasting seasons and observed for 5 quantitative characters, and for 20 SSRs. Some B- and R- lines had small but significant within-line genetic variation for certain traits. Higher number of significant differences for within-line variation observed among progenies for B-lines (48%) than for R-lines (33%) indicated that ear-to-row procedure of line-maintenance was more effective than bulking the phenotypically similar plants, for maintaining the genetic uniformity in inbreds. Amongst B-lines, ICMB 89111 had the maximum within-line variation for both quantitative characters and SSR markers. Amongst R-lines, IPC 802 and IPC 909 had within-line variation for maximum of 4 quantitative characters. Wide range of variability was observed for all the characters in both B- and R- lines, but genotypic coefficient of variation (GCV) was very low. SSRs were able to detect low level of residual heterozygosity in some of the

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inbreds. Mahalingam Govindaraj, Kedar N. Rai, Ponnusamy Shanmugasundaram, Aluri S. Rao (India) Efficiency of Single Plant Selection for Grain Iron and Zinc Density in Pearl Millet ABSTRACT Original Research Paper: Single plant selection, if effective, can make significant contributions to enhance breeding efficiency. This hypothesis was tested for grain iron (Fe) and zinc (Zn) density in four populations of pearl millet (Pennisetum glaucum (L.) R. Br.). Inbreeding and selection in advancing generations is normally practiced by evaluating progenies in unreplicated nurseries or at most in 2-replication trials. In each population in this study, grain samples of 40 random individual plants (hereafter referred to as S0 plants) and their S1 progenies grown in 2-replication trials for two seasons (called as environments) were analyzed for Fe and Zn density using ICP analytical method. In each population, correlation coefficients between S0 plants and their respective S1 progenies (whether individual environment or the mean of both environments) both for Fe and Zn density were positive, highly significant, and of the similar order as the correlation coefficients between the two environments for the S1 progeny performance. Also, the patterns of correlation coefficients between the S0 plants and either of the two replications of the S1 progenies in each environment were similar to those between the two replications for S1 progeny performance in both environments and in all four populations. While the Fe and Zn density were positively and highly significantly correlated, these were not correlated with grain mass. The patterns of these associations were similar both at the S0 plant level as well as at the S1 progeny level in each population. These results suggest that individual plant selection can be effectively used for simultaneous genetic improvement of both grain Fe and Zn density without compromising on grain size. Kedar N. Rai, Michael Blmmel, Akhilesh K. Singh, Aluri S. Rao (India) Variability and Relationships among Forage Yield and Quality Traits in Pearl Millet ABSTRACT Original Research Paper: Pearl millet [Pennisetum glaucum (L.) R. Br.], owing to its high photosynthetic efficiency and biomass production ability, fewer disease and insect pest problems, and tolerance to multiple environmental stresses, is a valuable forage crop, especially in view of climate change consequences. Nine open-pollinated varieties (OPVs) and 27 top-cross hybrids made on three male-sterile lines (A-lines) were evaluated in Alfisols at ICRISAT, Patancheru in the rainy season for two years. When harvested at 50 days after sowing, top-cross hybrids out-yielded OPVs, on an average, by about 30%, most likely due to relatively earlier flowering and higher biomass accumulation. At 80 d harvest, the dry forage yield of OPVs was similar to those of the hybrids. Forage nitrogen (N), in vitro digestibility and metabolizable energy content were used as laboratory fodder quality traits. Significant differences among the OPVs and among the hybrids were observed for these three quality traits, both at 50 d and 80 d harvest. While forage N declined by 49% at 80 d harvest, in vitro digestibility and metabolizable energy declined by 16-18%. At 50 d harvest, forage N content, in vitro digestibility and metabolizable energy were all significantly negatively correlated with forage yield both in OPVs and hybrids. At 80 d harvest, forage yield was not associated with any of the three quality traits in OPVs. In hybrids, forage yield was significantly negatively correlated with forage N content, while it was significantly positively correlated with the other two quality traits. These results indicate better prospects of combining high forage yield with high levels of in vitro digestibility and metabolizable energy in hybrids than in OPVs of pearl millet. Lszl Radics, Izra Gl, Imre Kdr (Hungary) The Effect of Rising Doses of NPK Fertilizers on Weeds of Proso Millet (Panicum miliaceum) on a Calcareous Loamy Chernozem Soil ABSTRACT Original Research Paper: The effect of different NPK levels (poor, satisfactory, excessive and toxic) and their combinations on the soil cover percentage of the natural weed flora of proso millet (Panicum miliaceum) was studied on a loamy chernozem soil with lime deposit. The N levels were 0 (poor), 100 (satisfactory), 200 (excessive) and 300 (toxic) kg ha-1 year-1; P and K fertilizing was done with 0 (poor), 500 (satisfactory), 1000 (excessive), 1500 (toxic) kg ha-1 P2O5 and K2O refilling doses. Later, only the sustaining of PK levels was targeted, refilling was repeated every 5-10 years. The P and K fertilizers and half of the N fertilizer was spread in autumn before ploughing the plots, the other half of N was spread in spring in the form of 25-28% calcium ammonium nitrate, 18% superphosphate and 40-60% potassium chloride. The series of mineral fertilization experiments was established in the autumn of 1973. Different crops were produced on these plots every year. Proso millet was

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sown in 1996. This long-term series of experiments consisted of 4N 4P 4K = 64 treatments, or nutritional levels, in two replications, giving a total of 128 plots. The plot size was 6 6 = 36 m2 in mixed factorial design. Weed surveys were done in 4 m2 quadrats in two replications and contained not only weed flora but crop cover too. The number of weed species, the soil cover of weeds and the crop were highly dependent upon the doses and ratio of different fertilizers. Sunita Choudhary, Fran R. Bidinger, C. Tom Hash, Vincent Vadez, Michael Blmmel (India) Gene Action Governing Pearl Millet Stover Nitrogen and in Vitro Digestibility and Opportunities for Improvement ABSTRACT Short Communication: Two pearl millet [Pennisetum glaucum (L.) R. Br.] stover fodder traits, nitrogen content (N) and in vitro digestibility, were targeted to investigate their modes of inheritance and to assess the scope for improving them relative to a base population. From a full-sib (FS) base population of pearl millet variety ICMV 221 three high and low nitrogen and three high and low in vitro digestibility FS were selected. Crosses were made for high high (H H), low low (L L) and high low (H L) FS trait contrasts and evaluated at Patancheru in the rainy seasons of 2007 and 2008. The high and low nitrogen FS parents contrasted 0.85 and 0.72% for nitrogen (N). In the crosses stover N contents were: HN HN = 0.85%, LN LN = 0.73% and HN LN = 0.80% (P < 0.05). The high and low digestibility FS parents contrasted 43.3 and 40.3% for in vitro digestibility. In the crosses stover in vitro digestibilities (D) were: HD HD = 43.7%, LD LD = 40.3% and HD LD = 42.2% (P < 0.05). The intermediate results of H L crosses strongly indicates the additive nature of the stover quality traits nitrogen and in vitro digestibility and suggest the application of cyclic breeding methods for increasing stover nitrogen content and in vitro digestibility in pearl millet. va B. brahm, Nra ri, Sndor Szab, Lszl Romhny, Lajos Blask, Gyrgy Zsigrai (Hungary) Quality of Grain of Different Proso Millet (Panicum miliaceum L.) Varieties ABSTRACT Short Communication: The quality of the grain of 5 millet varieties was examined during our research work. There were 3 varieties with white coloured grain and 2 ones with red coloured grain among the tested 5 varieties. Millet grains were husked with a laboratory rice mill, than we milled them. We determined the total scavenger capacity, the total phenolic content and different mineral element (N, P, K, Mg, Ca, Cu, Mn, Fe, Zn) contents of whole and dehulled millet grains. The tannin content was detectable only in the bran therefore we cannot take this parameter into account during the data analyses. The quality differences of the tested millet varieties were characterized. The total phenolic content of red coloured whole millet grains was significantly higher compared to that of light coloured millet grains, but there was not significant difference between the dehulled millet grains of different cultivars. The dehulling process resulted in a significant decrease in the Fe- and Mn-contents of each variety. SPECIAL ISSUE: Sorghum, 2012 Yanna Liang (USA) Sorghum: Genetic Improvement for Biofuel ABSTRACT Invited Review: During recent years, sorghum has drawn intensive attention from the scientific community and policy decision-makers. Three categories of sorghum, grain, forage, and sweet sorghum can be used differently for producing bioethanol and other kinds of biofuels. While grains from grain sorghum and juice from sweet sorghum are readily fermentable, the sorghum stalk which is a lignocellulosic material is recalcitrant. To utilize sorghum as a bioenergy crop, however, the biomass quality and yield must be enhanced. Highly fermentable and highly digestable lines are extremely desirable but research on this aspect is lagging behind those of other crops, such as maize. With numerous genes affecting sorghum quality being known, it is reasonable to expect that sorghum can be improved to serve as a biomass plant well through genetic and molecular breeding. Over the years, genetic modification of sorghum has been mainly conducted in three areas: 1) reducing lignin content by establishing brown midrib mutants and understanding the genetic mechanisms for lignin reduction; 2) developing cold tolerant progenies and identifying quantitative trait loci (QTL) related to cold tolerance; and 3) investigating sugar related traits with the aim of increasing contents of sugar and nonstructural carbohydrates. This review provides detailed information on how these studies are carried out. Future research directions are also recommended for transforming sorghum from a promising crop to a truly invaluable plant in the biofuel arena.

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Christopher R. Little, Ramasamy Perumal, Tesfaye Tesso, Louis K. Prom, Gary N. Odvody, Clint W. Magill (USA) Sorghum Pathology and Biotechnology - A Fungal Disease Perspective: Part I. Grain Mold, Head Smut, and Ergot ABSTRACT Invited Review: Three common sorghum diseases, grain mold, head smut and ergot, each of which is directly related to seed production and quality are covered in this review. Each is described with respect to the causal organism or organisms, infection process, global distribution, pathogen variability and effects on grain production. In addition, screening methods for identifying resistant cultivars and the genetic basis for host resistance including molecular tags for resistance genes are described where possible. Tesfaye Tesso, Ramasamy Perumal, Christopher R. Little, Adedayo Adeyanju, Ghada L. Radwan, Louis K. Prom, Clint W. Magill (USA) Sorghum Pathology and Biotechnology - A Fungal Disease Perspective: Part II. Anthracnose, Stalk Rot, and Downy Mildew ABSTRACT Invited Review: Foliar diseases and stalk rots are among the most damaging diseases of sorghum in terms of lost production potential, thus commanding considerable research time and expenditure. This review will focus on anthracnose, a fungal disease that causes both foliar symptoms and stalk rots along with the stalk rots caused by Fusarium spp. and Macrophomina phaseolina. Although the downy mildews are caused by oomycetes rather than true fungi, recent outbreaks have revealed resistance to previously effective chemical seed treatments and the evolution of new pathogenic races, once again pointing out the need for continuous vigilance. Sorghum diseases are described with respect to the causal organism or organisms, infection process, global distribution, pathogen variability and effects on grain production. In addition, screening methods for identifying resistant cultivars and the genetic basis for host resistance including molecular tags for resistance genes are described where possible along with prospects for future advances in more stable disease control. I. K. Das, S. Audilakshmi, J. V. Patil (India) Fusarium Grain Mold: The Major Component of Grain Mold Disease Complex in Sorghum (Sorghum bicolor L. Moench) ABSTRACT Invited Review: Sorghum grain mold, particularly Fusarium and Curvularia grain mold, are important on improved, short- and medium-duration sorghum cultivars worldwide. There is hardly any literature that deals with Fusarium grain mold as a component of sorghum grain mold disease complex in detail. This review summarizes many aspects related to Fusarium grain mold: typical symptoms, causal organisms in relation to mold development phases, colonization processes, nature of damage and its significance on yield and quality and management options. The review also attempts to shortlist probable mechanisms that might be useful for developing resistance against early infection events. There are three major and proven pathogenic species of Fusarium (F. andiyazi, F. proliferatum and F. thapsinum) that are capable of infecting sorghum flower. Others are predominantly saprophytes. Some strains of these species are highly toxigenic and responsible for Fusarial-toxicoses in human beings, animals and poultry birds. There is necessity to identify resistance against Fusarium grain mold in general and toxigenic strains in particular and incorporate the resistance in new varieties and hybrids of sorghum. Joshua H. Wong, Jeffrey F. Pedersen, Bob B. Buchanan, Peggy G. Lemaux (USA) Western Blot Analysis Uncovers Clues to Prolamin Digestibility in Uncooked and Cooked Sorghum and Corn Meal ABSTRACT Original Research Paper: Digestibility of cereal grain protein is conventionally expressed as percentage of protein digested by pepsin in a defined period. Sorghum has the lowest protein digestibility among cereals especially after wet cooking. If additional details were known as to which proteins are resistant to digestion, it should be possible to find means to improve this property. In this paper, two approaches are described to extract and identify proteins undigested by pepsin in uncooked and cooked meal from numerous sorghum and corn varieties. The first approach involves extracting residues undigested after 2 h with Borate-SDS-ME and separated by PAGE. Improved separation of undigested sorghum proteins was achieved using NuPAGE Bis-Tris gels. Western blots with antibodies against particular zeins and kafirins, used to monitor fates of different kafirins, revealed differential digestion rates. A second approach involved extracting undigested residues sequentially with 60%

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t-butanol, 60% t-butanol-ME and Borate-SDS-ME. The second approach, coupled with western analysis, revealed the following. (i) Existence of oligomeric forms of certain kafirins that differ in the degree of their susceptibility to pepsin digestion. (ii) Effect of cooking on the formation and digestion of the oligomers. (iii) Cross-linked forms of most -kafirins became more resistant to digestion after cooking. (iv) Most -kafirins are preferentially extracted in 60% t-butanol-ME while most -kafirins are extracted in Borate-SDS-ME buffer. (v) Monomeric -kafirins are resistant to pepsin digestion. (vi) -Kafirins form a series of oligomers that exhibit differential resistance to digestion. Our results suggest that the presently described systematic approach to analyzing the digestion by pepsin of sorghum prolamins should lead to greater insights into the digestion of specific types of sorghum grain proteins. A. Ashok Kumar, Belum V. S. Reddy, B. Ramaiah, K. L. Sahrawat (India), Wolfgang H. Pfeiffer (Colombia) Genetic Variability and Character Association for Grain Iron and Zinc Contents in Sorghum Germplasm Accessions and Commercial Cultivars ABSTRACT Original Research Paper: Sorghum a widely consumed cereal stapled in sub-tropical and semi-arid regions of Africa and Asia. Sorghum is the second cheapest source of energy and micronutrients (after pearl millet). Micronutrient malnutrition, primarily the result of diets poor in bio-available vitamins and minerals, causes blindness and anemia (even death) in more than half of the worlds population, especially among women and children. Biofortification wherever possible, is a cost effective and sustainable solution for tackling the micronutrient deficiencies as the intake of micronutrients is on a continuing basis with no additional costs to the consumer. ICRISAT is working on sorghum biofortification for enhancing Fe and Zn contents of the grain. Large variability for grain Fe and Zn contents was found from assessment of core germplasm collections (>2200), and from these, promising donors were identified for further improvement. Significant positive association observed between grain Fe and Zn contents indicated that it is feasible to develop high Fe and Zn containing cultivars with high yielding and different maturity backgrounds. The commercial sorghum cultivars (66) currently being cultivated by the farmers in India, were evaluated to identify high Fe and Zn cultivars in adapted backgrounds. Identification of commercial cultivars with high grain Fe and Zn content would help in wider dissemination of the cultivars to complement the on-going efforts for combating the micronutrient malnutrition. Pinnamaneni Srinivasa Rao, Santosh Deshpande, Michael Blmmel, Belum V. S. Reddy, Tom Hash (India) Characterization of Brown Midrib Mutants of Sorghum (Sorghum bicolor (L.) Moench) ABSTRACT Original Research Paper: Twenty brown midrib (bmr) mutants of sorghum (Sorghum bicolor (L.) Moench) were evaluated for agronomic traits, forage quality traits and the relationships between these traits. Potential fodder quality was assessed by laboratory analysis. Significant differences were observed among the bmr mutants (bmr1, 3, 6, 7, 8 and two new mutants) for stover yield, plant height, stover nitrogen (N) content, in vitro organic matter digestibility (IVOMD), metabolizable energy (ME) content and acid detergent lignin (ADL) content. The bmr mutants such as IS 23253, IS 21549, IS 23789 and IS 23787 had high stover yield coupled with better forage quality, hence these mutants can be exploited in future bmr hybrid breeding programs. All of these had reduced cinnamyl alcohol dehydrogenase (CAD) activity except for the new and uncharacterized bmr mutant, IS 23253, whose allelic relationship is yet to be determined. Pinnamaneni Srinivasa Rao, Michael Blmmel, Belum V. S. Reddy (India) Enhancement of In Vitro Digestibility of Sorghum (Sorghum bicolor (L) Moench) in Brown Midrib (bmr) Mutant Derivatives of bmr1 and bmr7 ABSTRACT Original Research Paper: Ten sorghum derivatives with brown midrib alleles (bmr1 and bmr7) were investigated for stover fodder quality traits, grain yield and the relationships between these productive traits. Potential fodder quality was assessed by laboratory analysis. Significant differences were observed among the tested derivatives of bmr1 allele for stover nitrogen (N) content, in vitro organic matter digestibility (IVOMD), and acid detergent lignin (ADL) content. In contrast, no significant genotypic differences were observed among bmr7 derivatives. In the bmr1 gene derivatives stover N content ranged from 0.63 to 0.81%, ADL content varied from 2.6 to 3.2% and IVOMD ranged from 52.9 to 55.4%. Two lines have recorded over 55% IVOMD and higher grain yield than the bmr1 source. In bmr7 derivatives IVOMD ranged between 52.9 and 54.4%. The enhanced IVOMD coupled with improved grain and biomass yield in bmr1 and bmr7 derivatives will increase overall benefits

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from sorghum in mixed crop livestock systems. This is the first study on utilization of bmr1 and bmr7 mutants in improving the forage quality of tropical sorghums with perceptible genetic enhancement of candidate biomass and grain yield traits. Dariush Minai-Tehrani, Aidin Tavakoli Tameh, Ali Rashidfarokhi, Alireza Noormohammadi, Amirabbas Khodakarami, Majid Talebi (Iran) The Effect of Light Crude Oil-Contaminated Soil on the Growth and Germination of Sorghum bicolor ABSTRACT Original Research Paper: Light crude oil has volatile components that can be toxic to living organisms when spilled to the soil and water. The use of plants for treatment of crude oil-contaminated soil has been of particular interest in environmental cleansing. Some plants have demonstrated to have a good capacity for biodegrading oil in soil. In this study the growth and germination of Sorghum bicolor was studied in the presence of different concentrations of light crude oil (1-10%) in the soil. Root and shoot biomass, leaf length and germination percentage decreased as crude oil concentration increased. Light crude oil delayed germination and affected the normal growth of S. bicolor, inducing chlorosis in plants. The measurement of total petroleum hydrocarbons (TPHs) of soil at the end of treatment (45 days) showed that TPH was more reduced in the 1% sample than in the 7 and 10% samples. S. bicolor can be a good plant for phytoremediation of oil-contaminated soil when the concentration of crude oil is lower than 3% in the soil. Lszl Radics, Izra Gl, Imre Kdr (Hungary) Effect of Rising Dose of NPK Fertilizers on Weeds of Grain Sorghum (Sorghum bicolor) on a Calcareous Loamy Chernozem Soil ABSTRACT Original Research Paper: The effect of different NPK levels (poor, satisfactory, excessive and toxic) and combinations of these on the soil cover percentage of natural weed flora of grain sorghum (Sorghum bicolor) was studied on a loamy chernozem soil with lime deposit. The N levels were 0 (poor), 100 (satisfactory), 200 (excessive) and 300 (toxic) kg ha-1 year-1; P and K fertilizing was done with 0 (poor), 500 (satisfactory), 1000 (excessive), 1500 (toxic) kg ha-1 P2O5 and K2O refilling doses. Later only sustaining of PK levels were targeted, reapplication was repeated every 5-10 years. P and K fertilizer and half of the N fertilizer was spread in autumn before ploughing the plots, the other half of N was spread in spring in the form of 25-28% calcium ammonium nitrate, 18% superphosphate and 40-60% potassium chloride. The mineral fertilization experiment series was initiated in autumn 1973. Different crops were produced on these plots annually. Grain sorghum was sown in 1992. This long-term experiment series consisted of 4N 4P 4K= 64 treatments, or nutritional levels, in two replications, giving a total of 128 plots. The plot size was 6 6 = 36 m2 in mixed factorial design. Weed surveys were done in 4 m2 quadrats in two replications and contained not only weed flora but crop cover, too. The number of weed species, the weed and crop soil cover were highly dependent upon the doses and ratio of different fertilizers. va B. brahm, Nra ri, Sndor Szab, Lajos Blask, Gyrgy Zsigrai (Hungary) Effects of Whitening Process on the Quality of Different Grain Sorghum Hybrids ABSTRACT Short Communication: The effect of whitening on the nutritional profile of grain sorghum was examined in this work. Three commercial hybrids, the white seeded Albita and the red seeded Zador along with another red-seeded hybrid check were examined. Grain samples drawn from the grain sorghum crop were whitened using the whitener part of a laboratory rice mill. Tannin content, total scavenger capacity, and total phenol content were determined in the whole grain sample, the whitened grains and the bran fraction. The concentration of essential mineral elements (N, P, K, Mg, Ca, Cu, Mn, Fe, Zn) was also measured in whole grains and whitened grains. Tannin content, scavenging capacity and phenol concentration were significantly different among the hybrids and between grain sample fractions; bran had the highest proportion of these compounds in all genotypes. Differences among hybrids with respect to mineral content were not significant in most of the examined elements. Larger differences between the hybrids were only found for N and Fe content. Whitening did not change the element content of the hybrids. SPECIAL ISSUE: Plant science and biotechnology in Turkey. Guest Editor: Dr. mr Baysal (West Mediterreanean Agricultural Research Institute (BATEM) Plant Pathology and Molecular Biology Departments, Antalya, Turkey), 2012 Ahmet Balkaya, Onur Karaaa (Turkey) General Status of Leguminous Vegetable Genetic Resources in Turkey

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ABSTRACT Invited Mini-Review: The legume family is of great significance because so many species are used throughout the world as sources of food and medicine. Turkey is described as a microcenter for bean (Phaseolus vulgaris L.), faba bean (Vicia faba L.), and cowpea (Vigna unguiculata) species. Wild, weedy and cultivated forms of Pisum are found in Turkey where the primary and secondary gene centres of origin overlap. The national collection of Leguminosae species at National Gene Bank of Turkey (AARI) consist of many accessions based on collection-related activities which systematically started since the early 1960s: 79.2% of these sources are beans (P. vulgaris L.), 12.4% is faba bean (V. faba L.) 5.8% is pea (Pisum sativum L.) and 2.6% is cowpea (V. unguiculata). In this review, the number of leguminous vegetable accessions of Turkish origin at different seed gene banks around the world is provided. The aim of this review is to outline some of the legume vegetable genetic resources of Turkey, their distribution, collection and characterization, and to describe cultivar breeding programmes. Hale Gunact, Ali Erklc, Hulya Ozgonen (Turkey) Status of Potato Wart Disease (Synchytrium endobioticum) in Turkey and Control Methods ABSTRACT Invited Mini-Review: Potato (Solanum tuberosum) is one of the most important global food sources and Turkey is one of the major seed and table potato-producing countries. There are many important pests and diseases preventing the production of potatoes. Potato wart disease caused by Synchytrium endobioticum was recently considered to be the most destructive fungal disease of potato. Typical symptoms of the disease occurring on tubers are cauliflower-like warts or tumours of different size. The disease can cause symptoms on the underground parts of potato plants including the crown, stolons and tubers, but not roots. Solanum tuberosum and other species of Solanum are the primary hosts of the disease. The disease prefers cool climates and is known to exist in 43 countries. Losses due to the disease range between 50 and 100% worldwide. Contaminated potato-growing areas ban seed potatoes because of the disease and there is zero tolerance in production. Recently, the disease has been well managed by strict quarantine measures and resistant varieties. However, it has still causing serious losses due to the existence of different races in different locations. In addition, some cultural methods are important to prevent the dispersal of this disease and, to date, there is no effective chemical application program against it. Despite control methods, the disease still remains economically significant. In this review, some informations about potato wart disease, including general characteristics of the disease, signs and symptoms, status in Turkey, epidemiology and disease management, are presented. Hulya Ozgonen, Ali Erkilic (Turkey) Reactions of 11 Potato Cultivars against Some Important Soil-Borne Pathogens ABSTRACT Original Research Paper: In this study, surveys and the reaction to diseases of some potato (Solanum tuberosum L.) cultivars (Satina, Vangogh, Marabel, Latona, Marfona, Vericus, Jearla, Cosmos, Granola, Hermes, Agria) against important soil-borne pathogens were conducted. In surveys, the most isolated genus was Rhizoctonia sp., Fusarium sp., Phytophthora sp. and Pythium sp. The isolation ratio of each genus varied between locations. Eleven commercial potato cultivars were evaluated for their reactions to Rhizoctonia solani (stem cancer and black scurf), Fusarium solani (Fusarium wilt), Phytophthora erythroseptica (pink rot) and Pythium deliense (root rot). Cultivars exhibited different levels of susceptibility to the pathogens. The most susceptible cultivars were Satina and Jearla. Satina, the most sensitive cultivar, had high disease severity ratios to R. solani, F. solani, P. erythroseptica and P. deliense (40, 100, 80 and 65%, respectively). Hermes and Agria were resistant to all diseases. Taner Yldz, Y. Benal Yurtlu, Elin Yeilolu (Turkey) Determination of Some Physical Properties of Some Pumpkin (Cucurbita moschata Duch.) and Winter Squash (Cucurbita maxima Duch.) Genotype Seeds ABSTRACT Original Research Paper: In this study, some physical properties of pumpkin line (14 BO 01) (Cucurbita moschata Duch.) and winter squash variety seeds (55 A 15 and Arican 97) (Cucurbita maxima Duch.) were evaluated at five different moisture content levels. Length, width and thickness increased linearly, as the moisture content increased from 8.62 to 25.49% dry weight basis (dwb) for Arican 97, from 9.60 to 25.60% for 55 A 15 and from 6.81 to 23.07% for 14 BO 01. All the diameters of seeds were significantly related with the seed moisture contents (P < 0.05). The sphericity varied from 0.548 to 0.565 for

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Arican 97, from 0.548 to 0.565 for 55 A 15 and from 0.482 to 0.504 for 14 BO 01 as the moisture content increased. With the increase in moisture content, the surface area varied from 290.08 to 330.86 mm2 for Arican 97, from 349.22 to 382.12 mm2 for 55 A 15 and from 333.29 to 398.35 mm for 14 BO 01. The bulk and true densities decreased from 411.86 to 366.03 kgm , from 390.35 to 347.37 kgm and from 375.54 to 316.55 kgm -3 for Arican 97, 55 A 15 and 14 BO 01, whereas the seed moisture content increased, respectively. The angle of repose increased from 17.11 to 22.87 for Arican 97 as the moisture content increased. These values increased from 15.25 to 23.87 for 55 A 15 and from 16.42 to 24.17 for 14 BO 01 with the increase in moisture content. Porosity values ranged between 41.3043.20%, 45.0847.36% and 45.4750.49% with the increasing moisture contents for Arican 97, 55 A 15 and 14 BO 01, respectively. Cneyt Civelek, Ahmet Balkaya (Turkey) The Nutrient Content of Some Wild Plant Species Used as Vegetables in Bafra Plain Located in the Black Sea Region of Turkey ABSTRACT Research Note: Some wild plant species have been used as vegetables in several parts of the world. These species are also used with respect of their aromatic and medicinal properties. Furthermore, these plants are evaluated as alternative food sources in periods where crop vegetables were not available. In this study, nineteen wild plant species, which are used as vegetable by local people in Bafra Plain, were collected and identified systematically. Dry matter, ash, protein, phosphorus, potassium, magnesium, calcium, iron, manganese and zinc contents were analyzed in order to determine the nutritional values of consumed parts of these wild species. Significant differences were found between species regarding their nutritional values. In fresh plant samples; the highest dry matter content was determined in Trachystemon orientale (14.73 g/100 g) and the highest ash content in Chenopodium album L. (24.73 g/100 g-dry sample). Within the species the highest protein content was observed from Taraxacum officinale Weber (32.03 g/100 g-dry sample) and Coronopus squamatus Forssk. Aschers. (31.73 g/100 g). Species displaying the highest phosphorus, potassium, magnesium, calcium, iron, manganese and zinc contents were respectively Chenopodium album L. (0.49 g/100 g), Trachystemon orientale L. (2.90 g/100 g), Chenopodium album L. and Rumex acetocella L. (0.47 g/100 g), Malva silvestris L. (1.54 g/100 g), Coronopus squamatus Forssk. Asch. (583.40 mg/kg), Ocimum basilicum L. (39.86 mg/kg) and Asparagus acutifolius L. (44.13 mg/kg). The results of this study showed that most of these examined nutritional values of these species are higher than cultivated vegetables such as spinach, lettuce and parsley.
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Transgenic Plant Journal Tree and Forestry Science and Biotechnology