Genet Resour Crop Evol DOI 10.

1007/s10722-012-9852-4

RESEARCH ARTICLE

Origin and cytology of a novel cytotype of Allium tuberosum Rottl. ex Spreng. (2n 5 48)
Geeta Sharma Ravinder N. Gohil

Received: 14 December 2011 / Accepted: 3 May 2012 Springer Science+Business Media Dordrecht 2012

Abstract Naturally occurring/spontaneously produced polyploids with six/more genomes are rarely found in Alliums. A hexaploid form of Allium tuberosum with 2n = 48 chromosomes has been isolated for the rst time amongst the open-pollinated seedlings of a hypotetraploid plant (2n = 4x = 31); latter being the seed-derived product of a normal tetraploid stock (2n = 4x = 32) growing in Jammu University Botanical Garden. Except for the guard cells and pollen grains that are of increased size, this form compared to its progenitor is dwarf, has smaller leaves and bears inorescences with few owers. This plant is also different from its progenitor in having nearly one-fourth (27.8 %) of its pollen mother cells (PMCs) with varying chromosome number viz. 2764, with the remaining cells having somatic or double the somatic number of chromosomes. To assess the nature of hexaploid form, its chromosomes were studied for morphological details, putative grouping and pairing properties during reduction division. Morphological similarity in the chromosomes of the present cytotype and its progenitor, arrangement of 48 chromosomes in eight groups of six chromosomes each and presence of
G. Sharma (&) University of Jammu, Jammu, Jammu and Kashmir, India e-mail: geetaji@yahoo.com R. N. Gohil Centre for Biodiversity Studies, School of Biosciences and Biotechnology, B.G.S.B. University, Rajouri 185131, Jammu and Kashmir, India

21.88 % euploid cells with eight hexavalents pointed towards the autopolyploid nature of the present strain. Regarding the origin of this strain, observation made on the meiosis in the two sex mother cells of the progenitor provides some clues. In the later plant, presence of most of embryo-sac mother cells with 62 chromosomes that showed 31:31 segregations and existence of majority of the PMCs with 31 chromosomes exhibiting erratic segregations indicate that the hexaploid strain has probably originated as a result of the fusion of reduced male (n = 17) and unreduced female gamete (2n = 31). Keywords Allium tuberosum Rottl. ex Spreng. Chromosome morphology Hexaploidy Karyotype analysis Multivalents

Introduction Allium tuberosum Rottl. ex Spreng., a crop species of genus Allium, is often confused with wild growing Allium ramosum because of resemblance in gross morphological features (rhizomatous cylindrical bulbs, linear leaves, hemispherical white inorescences, laments shorter than tepals). Despite these similarities which led Hanelt (1988) to purpose conspecic status for the two taxa, Stearn (1946) on the basis of presence of differences in A. tuberosum and A. ramosum with regard to oral traits (stellate/

123

Genet Resour Crop Evol

companulate owers; narrow-ovate tepals with green mid-line/lanceolate-oblong tepals with reddish midline; stamens 4/5th the length of the tepals/stamens 1/2 the length of the tepals) and owering period (August October/JuneJuly) advocated their maintenance as separate species. Previous report of present workers (Geeta 2005) regarding marked differentiation of oral and life history traits (owering period) of A. tuberosum (4x) growing in Jammu University Botanical Gardens from those reported for A. ramosum (Stearn 1946), further supported Stearns (1946) suggestion. Recent report of Blattner and Friesen (2006) highlighting clear genetic differentiation between two taxa corraborates that the species status assigned to A. tuberosum stands conrmed (Stearn 1946; Geeta 2005) and is being maintained. Commonly called Chinese Chive, A. tuberosum is cultivated on large scale, more extensively than A. cepa in China, Japan, Korea, Vietnam and Taiwan for seasoning dishes (Jones and Mann 1963). In India, leaves and bulbs of wild growing plants found in Diskit and Hunder in Nubra valley of Ladakh in Khasi hills (Gohil 1998; Pandey et al. 2008) are used by locals as a spice. Because of having therapeutic properties, whole plant extracts of Chinese chive are used as antimicrobial and carminative agent as well as for relieving hypertension (Mau et al. 2001; Pandey et al. 2008). Besides, extracts of its bulbs are potent, even more than A. sativum for reducing hyperlipidemia and arterosclerosis (Choudhary 2008). Except for the two reports describing the naturally occurring diploid (2n = 16) forms of the species (Ohno 1964; Yang et al. 1998), all other forms are known to be tetraploid having 2n = 4x = 32 chromosomes (Mathur and Tandon 1965; Gohil and Koul 1971, 1973a, b; Sen 1974; Pandita and Mehra 1981; Talukder and Sen 2000). Nevertheless, nding of some aneuploids (32 23 chromosomes) and an octaploid (2n = 6164 chromosomes) by the previous workers (Gohil and Kaul 1979; Kojima et al. 1991) amongst the seedlings of open-/cross-pollinated and manually-selfed tetraploids indicated that the tetraploid forms have tremendous capability of throwing up numerical variants in the seed progeny. To nd variants having traits of signicance in plant breeding, populations were raised from the seed obtained from the tetraploid population cultured in Jammu University Botanical Gardens and screened for chromosome count. Four aneuploids with 2933 chromosomes were

found. While most of these bred true to type, a single hexaploid was isolated from the progeny of a hypotetraploid (2n = 4x = 31 chromosomes). Detailed cytology of this hexaploid, recovered for the rst time till date, along with its nature and genesis form the basis of present communication.

Materials and methods Morphological details of the vegetative parts of the present cytotype (voucher specimen deposited in Jammu University Herbarium vide number 13930) were taken at the time of ower initiation and those of reproductive parts when plants were in full bloom. Data for each trait is based on fteen observations. For karyotypic details, root-tips of the present hexaploid were procured by uprooting bulbs. These were pretreated in 0.3 % aqueous colchicine solution (3 h), xed in acetic-alcohol (3:1), stained in Feulgen after hydrolyzing in 1 N HCl (60 C for 12 min) and squashed in 1 % acetocarmine. Fifteen root-tip cells with good chromosome spreads were analysed for chromosome count. Chromosome sizes and index numbers of all the chromosomes were taken as mean of the ve cells studied. Following Sharma and Gohil (2008), chromosomes with index number = 0.671, 0.660.33 and\0.33 have been classied as metacentric, submetacentric and subtelocentric respectively. For studying meiosis in the pollen mother cells (PMCs), young inorescences were xed in 1:3 aceticethanol for 24 h, washed in tap water and preserved in 70 % ethanol. Meiotic details were studied by squashing anthers in 1 % acetocarmine. Filled and well-stained pollen grains were counted as viable and shriveled and unstained grains were taken as non-viable.

Results Cytological analysis of forty seedlings obtained from the open-pollinated hypo-tetraploid (2n = 31 chromosomes) led to isolation of a single plant with fortyeight chromosomes; 39 other plants had 2n = 31 chromosomes. The hexaploid plant had short and narrow leaves (14.22 0.01 9 3.1 0.005 mm) and small sized inorescences (24.7 0.648 9 22.9 0.378 mm)

123

Genet Resour Crop Evol

with on an average 17 owers born on 14.3 0.448 mm long pedicels. These owers had variable number of tepals and stamens viz. 47. Tepals were 5.1 0.001 mm long and laments 2.4 0.076 mm long. Guard cells and pollen grains were 59.92 0.421 9 14.95 0.009 lm and 450.03 0.687 9 278.69 0.004 lm in size.

Karyotype In all, 15 root-tip cells with clear 48 chromosomes were analyzed for karyotypic details (Fig. 1A). Of these, 12 were median, 30 sub-median and 6 subtelocentric. Latter had secondary constrictions on their short arms. The total chromatin length of the complement was

Fig. 1 Metaphase chromosomes (A) and putative chromosome grouping of hexaploid cytotype of Chinese chive (B). The chromosome marked in A is of deviant type. (Bar = 10 lm)

123

Genet Resour Crop Evol

Fig. 2 PMCs at diakinesis with 26 (A), 27 (B), 36 (C) and 48 (D) chromosomes. A PMC at metaphase I with 96 chromosomes (E). Note that the two bivalents marked in E are about to separate. (Bar = 10 lm)

Table 1 Number and percentage of PMCs with varying number of chromosomes Chr. no No. of cells (%age) Fig.
a

26 4 (6.4) 2A

27 3 (4.8) 2B

32 2 (3.2)

36 1 (1.6)

37 1 (1.6) 2C

42 1 (1.6)

46 2 (3.2)

48 32 (51.6) 2D

49 1 (1.6)

64 2 (3.2)

96a 13 (20.9) 2E

Occurrence of cells with double the somatic number of chromosomes is a normal feature for A. tuberosum (Talukder & Sen 2000)

632.41 lm, the smallest and the longest chromosomes being 9.13 and 17.12 lm respectively. Meiosis Of 62 PMCs available at diakinesis/metaphase I, 32 (51.6 %) had somatic (2n = 48) and 13 (20.9 %) had double the somatic number of chromosomes (2n = 96). Besides, 17 (27.2 %) cells had deviant

number of chromosomes ranging from 26 to 64. Table 1 presents the frequency of cells with varying number of chromosomes. It is clear from Table 1 that frequency of deviant cells with\2n = 48 number is less for this plant; only three cells having higher numbers. Except for the cells with 36 and 37 chromosomes that had quadrivalents plus bivalents i.e. 8IV ? 2II and 6IV ? 6II ? 1I, most of the cells with variant number had univalents.

123

Genet Resour Crop Evol

Like the normal tetraploids, cells with double the somatic number of chromosomes consistently had 48 bivalents and those with normal count either had complex associations or 48 univalents. Of the 32 cells with normal count, 25 cells (78.12 %) had 48 univalents, seven cells (21.88 %) appeared to possess eight hexavalents. Anaphase I segregations were equal in cells with double the somatic number of chromosomes whereas those with normal count showed erratic segregation patterns (Table 2). At telophase I and II, 58 and 51 % cells had micronuclei with their number ranging from 1 to 6. Pollen stainability of this cytotype was found to be as low as 1.33 %. Only 23 seeds, formed in three inorescences, were empty and did not germinate.

Discussion Polyploidy is quite common in the genus Allium, with tetraploidy being the most common condition (Fedorov 1969). For A. tuberosum, naturally occurring diploid (Ohno 1964; Yang et al. 1998) and tetraploid forms (Mathur and Tandon 1965; Gohil and Koul 1971, 1973a, b; Sen 1974; Pandita and Mehra 1981; Talukder and Sen 2000) as well as an experimentally produced octaploid (manual selng of a tetraploid) form is earlier on record (Kojima et al. 1991). As such, the present nding of a hexaploid among the open-pollinated seedlings of a hypo-tetraploid (2n = 4x = 31) completes the polyploid series for this taxon.

Table 2 Distribution of chromosomes at anaphase I in 18 PMCs Distribution patterns No. of cells (% age) 20:28 4 (22.2) 20:3:25 2 (11.11) 23:25 4 (22.2) 18:1:29 2 (11.11) 20:5:23 2 (11.11) 48:48 4 (22.2)

Table 3 Morphology and fertility of the present hexaploid and its hypotetraploid progenitor (Geeta 2005)

Characteristics

Hexaploid (2n = 6x = 48) 14.22 0.01 3.10 0.005 August 24.5 0.956 24.7 0.648 22.9 0.378 16.9 0.378 Variable; 47 5.1 0.001 14.3 0.448 2.4 0.076 3.0 0.008 59.92 0.421 9 14.95 0.009 450.03 0.687 9 278.69 0.004 1.33 23 Nil

Hypo-tetraploid (2n = 4x = 31) 17.75 0.024 3.43 0.096 July 41.1 0.457 26.9 0.904 25 0.348 33.9 0.388 6 5.4 0.008 21.0 0.430 3.2 0.042 3.3 0.046 45.68 0.007 9 13.81 0.008 385.88 0.11 9 235.82 0.0114 76.47 45 68

Leaf length (cm) Leaf width (mm) Month of owering Scape length (cm) Inorescence length (mm) Inorescence width (mm) No. of owers per inorescence No. of tepals/stamens per ower Tepal length (mm) Pedicel length (mm) Filament length (mm) Style length (mm) Guard-cell size (lm) Pollen grain size (lm) Pollen stainability (%age) Seed set per inorescence Seed germination (%age)

123

Genet Resour Crop Evol

Comparison of the morphological traits of the hexaploid isolated and its progenitor (Table 3) revealed that except for increase in guard-cells and pollen grains, indicator of higher ploidy level, hexaploid compared to its parent is dwarf, has smaller leaves and bears inorescences with fewer owers. Overall decrease in vigor with increase in ploidy level is on record for plants like foxtail millet (Wang et al. 1999). Delayed owering as noticed in the present polyploid is in consonance with the earlier observations and is supposed to be a result of increased duration of mitotic cycle in higher polyploids (Stebbins 1971). Compared to the somatic cells of the present cytotype that consistently had 48 chromosomes, 27.2 % of its PMCs studied had deviant number of chromosomes (Range = 2664) whereas remaining meiotic cells like those in the normal tetraploids had somatic (48) or double the somatic number of chromosomes (96). Existence of cells with deviant numbers (Range = 833) is previously on record for a seed-derived tetraploid plant of the present basic stock growing in Jammu University Botanical Garden (Sharma and Gohil 2002), Coix gigantea Koenig (Sapre and Naik 1990) Astragalus subuliformis DC. (Ashraf and Gohil 1994). While in Coix gigantea, deviant PMCs had one/two chromosomes in addition or missing, present polyploid like the earlier reported tetraploid Chinese chive and Astragalus subuliformis exhibited wide variation in chromosome count. It is important to mention here that existence of wide variation in the chromosome count within the PMCs of an anther is generally attributed to cytomixis. In the present case too, this phenomenon appears to be the most plausible reason for the occurrence of chromosomal chimeras in the PMCs. However, since no cytoplasmic channels or connections between PMCs were observed, some other mechanism might also be operating to bring this anomaly. For determining the nature of this cytotype, somatic chromosomes were analysed for putative grouping and the conclusions thus found were complemented by pairing properties during meiosis. Ignoring minor differences, 48 chromosomes, on the basis of size, form and presence of secondary constrictions, got arranged in eight groups of six chromosomes, though one chromosome of the fourth group was of somewhat deviant morphology (6th chromosome of third group of Fig. 1B). In having a deviant type chromosome and

on account of overall morphology of somatic chromosomes, present complement more or less corresponds with that of its hypo-tetraploid progenitor (Geeta 2005). This feature in conjunction with arrangement of the 48 chromosomes in eight groups of six chromosomes each suggested the autoploid origin of present cytotype. Because all the PMCs with double the normal count exhibited autobivalent formation and most of the cells with deviant number showed desynapsis, euploid cells of present cytotype were considered for determining the frequency of various chromosomal associations present. As mentioned earlier, while 78.12 % cells had 48 univalents, 21.88 % appeared to have eight hexavalents. Presence of cells with eight hexavalents, though in low frequency support the autoploid origin of the neo-polyploid. Observations made on the meiosis in the two sex mother cells of the hypotetraploid progenitor (2n = 4x = 31) of present hexaploid provides some clues as to its origin. It has been observed that most of the embryo-sac mother cells of the parent plant had 62 chromosomes that formed 31 bivalents and exhibited 31:31 segregations whereas majority of PMCs had 31 chromosomes that formed multivalent and showed erratic chromosome segregations viz. 17:14, 18:13 and 16:15 at anaphase I (Geeta 2005). While former sex mother cells seem to have formed unreduced megaspores, latter must have developed into aneuploid reduced microspores with n = 1318, etc.; aneuploidy (n = 1019) in the male gametes is earlier on record for A. tuberosum (Gohil and Koul 1973b). Present polyploid probably originated as a result of fusion of a hyperploid male (n = 17) and unreduced female gamete (2n = 31). This process called unilateral sexual polyploidisation is considered to be dominant one for higher polyploid formation in angiosperms. In facultative apomicts as A. tuberosum, polyploids produced in this way are referred to as B-III (Rutishauser 1948)/2n ? n (Harlan and De Wet 1975)/U-hybrids (Asker 1977). Contrary to present view, Kojima et al. (1991) attributed the origin of octaploid Chinese chive (2n = 8x = 6164) to extrapremeiotic doubling of chromosomes in the EMCs. Latter assumption, however, lacked proof. Inter-/intraspecic hybridization leading to the formation of neo-polyploids confers these with greater pool of genes/alleles for selection and potential benets of increased heterosis/heterozygosity and

123

Genet Resour Crop Evol

evolutionary diversity (Adams and Wendel 2005; Doyle et al. 2008). Though polyploidy promotes the latter two processes in auto- and allopolyploids, these are expected to differ in their abilities to get beneted. Genomic merger and addition in allopolyploid systems like Arabidopsis suecica (Fries) Norrlin (Maldung et al. 2002), Gossypium (Adams et al. 2004), Spartina angelica C. E. Hubb. (Salmon et al. 2005) hexaploid wheat (Bottley et al. 2006) and Brassica napus L. (Gaeta et al. 2007) is known to initiate a large number of genetic and gene expression alterations as gene loss, homeologous recombination, gene silencing by methylation/histone modication and complementation of silenced orthologs by homeologs which may allow for rapid subfunctionalisation/ neo-functionalisation. These changes are supposed to underlie heterosis and origin of evolutionarily novel phenotypes in a wide range of organisms (Wittkop et al. 2004). In autopolyploids, the effect of genome doubling on gene expression is on record for a few species. While autoploids (4x and 6x) of some species as Brassica oleracea L. (Albertin et al. 2005) and Helianthus decapetalus L. (Church and Spaulding 2009) did not differed signicantly from the diploids, mono- to tetraploid lines of maize (Guo et al. 1996) and potato (Stupar et al. 2007) exhibited radical alterations in the expression of a few genes (18 in maize and 10 % of 9,000 analysed in potato). Using diploid and autotetraploid lines of six species of Cymbopogon, Lavania et al. (2012) revealed that the change from diploid to tetraploid result in accumulation of more secondary metabolites as well as enhanced DNA methylation. As autopolyploids lack homeologous alleles; dramatic alterations in the expression of some genes, if occurs, would be affecting the organism and not lead to evolutionary advantageous subfunctionalisation. Autopolyploidy which often result from with-in species mating may not lead to evolutionary innovations more frequently. It, however, provides an easy and safer path to the doubled genome. Rapid formation of such polyploids (Ramsey and Schemske 1998) in conjunction with increased vigorosity which is generally associated with autopolyploid condition (particularly heterozygous) seems to be responsible for common occurrence of autopolyploids at least in some families as Saxifragaceae (Soltis 2007) and Cactaceae (Hamrick et al. 2002). It is pertinent to

mention here that traditionally naturally occurring autopolyploids were reported to be extremely rare (Stebbins 1950; Grant 1971, 1981) because of the difculties associated with identifying autopolyploids in eld conditions (Hegarty and Hiscock 2007), relying of previous investigators on pairing properties of chromosomes which in some cases may not indicate the type of polypoidy (Qu et al. 1998) and nonavailability of genetic markers that are used these days to distinguish polyploids exhibiting disomic/polysomic inheritance (Soltis et al. 2007). Apart from providing short term tness to the individuals with doubled genomes, autopolyploidy can lead to lineage diversication by producing heterozygous progeny in high proportion/progeny with hetrozygosity at more loci than the diploid progenitor. As increased heterozygosity is associated with suppressed homozygosity, autoployploidy may have protective effect against deleterious recessive mutations that express in homozygous condition (Comai 2005). Nascent autopolyploids can exploit long-term advantages only if these out-cross, though with the individuals of same species, undergo few segregation losses and overcome fertility barriers, if any (Hegarty and Hiscock 2007, 2008; Jauhar 2010). Like autopolyploid Lathyrus palustris (Khawaja et al. 1995), present plant exhibited very low pollen stainability (1.33 %) and high sterility. This sterility seems to be on account of disturbed meiosis resulting in imbalance of chromosome number in gametes or failure of zygotic or endosperm development. As such, in A. tuberosum, mainly propagating via seed, with plant biomass constituting the economic product, chromosome addition seems to be hindrance rather than help. Despite sterility, present plant has survived because of its having efcient means of vegetative propagation and perennial habit. Since enhanced ploidy level is often accompanied by enhanced concentration of secondary metabolites (Lavania 2005; Lavania et al. 2012), the isolated genetic material, A. tuberosum with 2n = 48, is now being chemically analysed in order to determine its potential.
Acknowledgments The rst author is grateful to the Head, Department of Botany, University of Jammu, Jammu for providing the necessary facilities and to DST, Govt. of India, New Delhi for providing nancial assistance.

123

Genet Resour Crop Evol

References
Adams KL, Wendel JF (2005) Polyploidy and genome evolution in plants. Curr Opin Plant Biol 8:135141 Adams KL, Percield R, Wendel JF (2004) Organ-specic silencing of duplicated genes in a newly synthesized cotton allotetraploid. Genetics 168:22172226 Albertin W, Prabant P, Catrice O, Eber F, Jenczewski E et al. (2005) Autopolyploidy in cabbage (Brassica oleracea L.) does not alter signicantly the proteomes of green tissue. Proteomics 5:21312139 Ashraf M, Gohil RN (1994) Cytology of legumes of Kashmir Himalaya. V. Cytomixis and chromosome migration in Astragalus subuliformis D.C. Nucleus 37:119122 Asker S (1977) Pseudogamy, hybridization and evolution in Potentilla. Hereditas 87:179184 Blattner FR, Friesen N (2006) Relationship between Chinese Chive (Allium tuberosum) and its putative progenitor as assessed by random amplied polymorphic DNA. In: Zeder MA, Bradley DG, Emshwiller E, Smith BD (eds) Documenting domestication: new genetic and archeological paradigms. University of California Press, Berkley and Los Angeles, California Bottley A, Xia GM, Koebner RMD (2006) Homeologous gene silencing in hexaploid wheat. Plant J 47:897906 Choudhary R (2008) Benecial effect of Allium sativum and Allium tuberosum on experimental hyperlipidemia and arterosclerosis. Pak J Physiol 4(2):79 Church SA, Spaulding EJ (2009) Gene expression in a wild autopolyploid sunower series. J Hered 100:491495 Comai L (2005) The advantages and disadvantages of being polyploid. Nat Rev Genet 6:836846 Doyle JJ, Flagel LE, Paterson AH, Rapp RA, Soltis DE, Soltis PS, Wendel JF (2008) Evolutionary genetics of genome merger and doubling in plants. Annu Rev Genet 42:443 461 Fedorov AA (ed) (1969) Chromosome numbers of owering plants. Komarov Botanical Institute, Academy of Sciences of the USSR, Leningrad, USSR Gaeta RT, Pires JC, Iniguez-Luy F, Leon E, Osborn TC (2007) Genomic changes in resynthesized Brassica napus and their effect on gene expression and phenotype. Plant Cell 19:34033417 Geeta (2005) Cytogenetic studies in Allium tuberosum Rottl. ex Spreng. with emphasis on the phenomenon of apomixis. Unpublished PhD thesis, University of Jammu, Jammu, India Gohil RN (1998) Cytology of Indian Alliumsa brief overview. In: Kachroo P (ed) Progress in cytogenetics. Bishen Singh Mahindra Pal Singh, Dehra Dun, pp 95115 Gohil RN, Kaul R (1979) Seed progeny studies in Alliums. I. Numerical variants in the progeny of tetraploid Allium tuberosum Rottl. ex Spreng. Beitr Biol Panzen 54:305 309 Gohil RN, Koul AK (1971) Desynapsis in some diploid and polyploid species of Allium. Can J Genet Cytol 13: 723728 Gohil RN, Koul AK (1973a) Some adaptive genetic-evolutionary processes accompanying polyploidy in the Indian Alliums. Bot Notiser 126:426432

Gohil RN, Koul AK (1973b) Bearing of meiotic irregularities on karyotype alterations in some polyploid taxa of genus Allium. In: Kachroo P (ed) Advancing frontiers in cytogenetics. Hindustan Publishing Corporation, New Delhi, pp 136142 Grant V (1971) Plant speciation. Columbia University Press, New York Grant V (1981) Plant speciation, 2nd edn. Columbia University Press, New York Guo M, Davis D, Birchler JA (1996) Dosage effects on gene expression in a maize ploidy series. Genetics 142:1349 1355 Hamrick JL, Nason JD, Fleming TH, Nassar JM (2002) Genetic diversity in columnar cacti. In: Fleming H, Valiente-Banet A (eds) Columnar cacti and their mutualists: evolution, ecology, and conservation. University of Arizona Press, Tuscon, pp 122133 Hanelt P (1988) Taxonomy as a tool for studying plant genetic resources. Kulturpanze 36:169187 Harlan JR, De Wet JMJ (1975) On O. winge prayer: the origin of polyploids. Bot Rev 41:361390 Hegarty M, Hiscock S (2007) Polyploidy: doubling up for evolutionary success. Curr Biol 17:927929 Hegarty MJ, Hiscock SJ (2008) Genomic clues to the evolutionary success of polyploid plants. Curr Biol 18:435444 Jauhar PP (2010) Genetic control of chromosome behaviour: implications in evolution, crop improvement and human biology. Nucleus 53:312 Jones HA, Mann LK (1963) Onions and their allies. Botany, cultivation and utilization. Leonard Hill (Books) Limited, London Khawaja HIT, Ellis JR, Sybenga J (1995) Cytogenetics of Lathyrus palustris, a natural autohexaploid. Genome 38: 827831 Kojima A, Nagato Y, Hinata K (1991) Degree of apomixis in Chinese chive estimated by esterase isozyme analysis. Jpn J Breed 41:7383 Lavania UC (2005) Genomic and ploidy manipulation for enhanced production of pharmaceuticals. Plant Genet Resour 3:170177 Lavania UC, Srivastava S, Lavania S, Basu S, Misra NK, Mukai Y (2012) Autopolyploidy differentially inuences body size in plants, but facilitates enhanced accumulation of secondary metabolites, causing increased cytosine methylation. Plant J. doi:10.1111/j.1365-313X.2012. 05006.x Maldung A, Masuelli RW, Watson B, Reynolds SH, Davison J, Comai L (2002) Remodeling of DNA methylation and phenotypic and transcriptional changes in synthetic Arabidopsis allotetraploids. Plant Physiol 129:733746 Mathur M, Tandon SL (1965) Cytology of an autotetraploid Allium tuberosum. Indian J Hort 22:382384 Mau JL, Chen CP, Hsieh PC (2001) Antimicrobial effects of extracts from Chinese chive, and cinnamom and corni fructus. J Agric Food Chem 49:183188 Ohno R (1964) Chromosomes in Allium tuberosum. J Hokkaido Gakugei Univ II B 16:1718 Pandey A, Pandey R, Negi KS, Radhamani J (2008) Realizing value of genetic resources of Allium in India. Genet Resour Crop Evol 55:985994

123

Genet Resour Crop Evol Pandita TK, Mehra PN (1981) Cytology of Alliums of Kashmir Himalayas, II. Cultivated taxa. Nucleus 43:4656 Qu L, Hancock JF, Waylon JH (1998) Evolution in an autopolyploid group displaying predominately bivalent pairing at meiosis: genomic similarity of diploid Vaccinium darrowi and autotetraploid V. corymbosum (Ericaceae). Amer J Bot 85:698703 Ramsey JR, Schemske DW (1998) Pathways, mechanisms, and rates of polyploid formation in owering plants. Ann Rev Ecol Syst 29:467501 Rutishauser A (1948) Pseudogamie und Polymorphie in der Gattung Potentilla. Arch Julius Klaus Stiftung 23:267424 Salmon A, Ainouche ML, Wendel JF (2005) Genetic and epigenetic consequences of recent hybridisation and polyploidy in Spartina (Poaceae). Mol Ecol 14:11631175 Sapre AB, Naik AS (1990) Formation of aneuploid gametes through non-disjunction during pre-meiotic mitoses in Coix gigantea. Cytologia 55:7177 Sen S (1974) Floral biology, meiosis, pollen cytology and cause of seed setting in Allium tuberosum Rottl. Caryologia 27:716 Sharma G, Gohil RN (2002) Chromosomal chimeras in the male track of Allium tuberosum Rottl. ex Spreng. Caryologia 57:158162 Sharma G, Gohil RN (2008) Intrapopulation karyotypic variability in Allium roylei Stearna threatened species. Bot J Linn Soc 158:242248 Soltis DE (2007) Saxifragaceae: a taxonomic treatment. In: Kulbitzki K (ed) The families and genera of vascular plants, vol 9. Springer, New York Soltis DE, Soltis PS, Schemske DW, Hancock JF, Thompson JN, Husb BC, Judd WS (2007) Autopolyploidy in angiosperms: have we grossly underestimated the number of species? Taxon 56:1330 Stearn WT (1946) Nomenclature and synonymy of Allium odorum and A. tuberosum. Reprinted from Herbertia (1944) 11:226245 Stebbins GL (1950) Variation and evolution in plants. Columbia University Press, New York Stebbins GL (1971) Chromosomal evolution in higher plants. Edward Arnold, London Stupar RM, Bhaskar PB, Yandell BS, Rensink WA, Hart AL, Ouyang S, Veilleux RE, Busse JS, Erhardt RJ, Buell CR, Jiang J (2007) Phenotypic and transcriptomic changes associated with potato autopolyploidization. Genetics 176:20552067 Talukder K, Sen S (2000) Chromosome characteristics in some Allium species and assessment of their interrelationship. Nucleus 43:4657 Wang R, Gao J, Liang GH (1999) Identication of primary trisomics and other aneuploids in foxtail millet. Plant Breeding 118:5962 Wittkop PJ, Haerum BK, Clark AG (2004) Evolutionary changes in cis and trans gene regulation. Nature 430:8588 Yang L, Xu JM, Zhang XL, Wan HO (1998) Karyotypical studies of six species of the genus Allium. Acta Phytotaxonomica Sinica 36:3646

123