Transgenic Plants in Agriculture

Proceeding of the UGC Sponsored National Seminar
on
''THE ROLE OF BIOLOGY IN BRINGING SECOND
GREEN REVOLUTION"
Organised by
Department of Botany, M. S. College, Saharanpur,
(CCS University, Meerut, UP.)
on
11 & 12 October, 2015
Editor
Dr. Vijai Malik

M.Sc., M.Phil., Ph.D., F.A.P.S., F.A.P.T., F.B.S.
Assistant Professor, Department of Botany
M. S. College, Saharanpur (UP) India
2015
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TABLE OF CONTENT
S. Title Name of the Authors Page

No. No.
1 Green Revolution for Qualitative Traits in Plants: Rajni Shukla, Rashmi
Biofortification of Nutrients 1-8
Upadhyay and Yogesh Kumar
Sharma
2 Shoving Towards a New Green Revolution Vijai Malik 9-13

3 Biopesticide: An Ecofriendly Approach For Second Rashmi Nigam 14-18
Green Revolution in Agriculture
4 Rhododendron Diversity and their Conservation in Vishal Kaushik 19-25
Sikkim Himalayas
5 Changes in Micromorphology of Plant Sida veronicaefolia in Shiv Kumari and Ila Prakash 26-32
Response to Air Pollution Stress in Meerut City
6 In vitro study of an endangered Miracle plant: Shalini Sharma and Y. Vimala 33-40
Mandookparni (Centella asiatica (L.) Urb.)
7 Role of Nanotechnology in Pollution Control: Shalini Singh 41-45
Review
8 Identification of Geminivirus in Cowpea (Vigna unguiculata) Shail Pande 46-50

Through Amplification of Selective DNA Fragment Using
Degenerate Primers
9 Science and Technology in Rural India Richa Atreya 51-55
10 Effects of Distellery Effluent Irrigation on Mustard (Brassica Renu Choudhary, Naresh 56-66
campestris) Kumar and Harendra Malik
11 Role of Transgenic Plants in Agriculture Renu Rani 67-71
12 Depression in Serum Zinc Concentration and Elevation in Punam Yadav 72-79

Serum Potassium Concentration in Chronic Renal Failure
Patients
13 Enhancement of Fe and Zn in Cowpea grain by applying soil Namita Yadav & Y.K. Sharma 80-83
and foliar application
14 Adoption of Improved Technologies in Black Gram Crop Lokendra Kumar Singh 84-88
15 Study on SO2 Induced Effects and their Amelioration in Arhar Harendra Malik, Naresh Kumar 89-98
(Cajanus cajan)
and Renu Choudhary
'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION iii

16 Synthesis, Characterization and Biological Activities of Some Dinkar Malik 99-105

Metal Complexes with 2- Amino -4- (p- Ethoxy Phenyl)
Thiazoline Ligand
17 Nutritional sink formation in galls of Alstonia scholaris Linn. Deepak Kumar, Vijai Malik 106-114
(Apocynaceae) by the insect Pauropsylla tuberculata and S.C. Dhiman
(Homoptera: Psyllidae).
18 Perspective of Stem Cell Research in India Bindu Sharma 115-120
19 Study of Medicinal Angiosperms of Rampur District (UP), Beena Kumari 121-129
India With Special Reference to Their Sustainable Use
20 Water Quality Control of Industrial Effluents Using Anuja Agarwal & Vaishali 130-137
Crosslinked Chitosan Hydrogel Beads.
21 Clinical Application and Potential Role of Stem Cells Abhishek Gupta & Bindu 138-143
Sharma
22 Wild Life and Biodiversity in the Plays of Shakespeare Rashmi Rana 144-148
23 Conservation of Withania somnifera and Commiphora wightii Anshu Dhaka & Vinit Kumar 149-152
medicinal plants
24 The zero energy sewage treatment plant (ZESTP). A Shyam Singh 153-162
conventional or alternative method of sewage treatment to
enhance the livelihood of marginal formers in peri urban
region of Gorakhpur (U.P.)
25 Health Issues in Old Women: Reasons and Suggestion for Meenakshi 163-164
Improvement
26 Morphological Studies of Certain Plant Parasitic Nematodes Om Dutta 165-171
Associated with Vegetable Crops of District Bulnadshahr
27 Genetics of Hg++ -tolerance in Barley Ritu Aggarwal 172-179
28 Stem Cell Therapy Pragati 180-182
'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION iv

Themes to be Covered
Role of Biotechnology and Nanotechnology to revolutionize Agriculture and Food

Industry.
Means of enhanced productivity by employing high yielding varieties of crop
plants.
Pest and disease control.
Soil chemistry.
Conventional manures and recent thoughts.
Water management-water scarcity and water security.
Mechanization.
Due stress on forestry and ecological balance.
Conservation of plants and animals.
Agricultural management.
Human health.
Biodiesel and alternate energy sources.
Generating gainful self employment.
Government policies and recent initiatives.
Climate change and food security.
Food and nutrition security.
Stem cell therapy.
Science education in rural areas
'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION v

Green Revolution for Qualitative Traits in Plants: Biofortification of Nutrients
Rajni Shukla, Rashmi Upadhyay and Yogesh Kumar Sharma
Department of Botany, University of Lucknow, Lucknow (226 007)
The Agricultural Revolution was a period of technological improvement and increased crop productivity that
occurred during the 18th and early 19th centuries in Europe. The need of food is based on nutrient requirement.
More than 12% of soils are deficient in available iron. Zinc deficiency also occurs in crops and human on a world
scale and is now regarded as next to iron. Iron deficiency is growing a health concern in the developing world, and
responsible for diverse of health complications including anemia and impairments in immune system. Zinc is an
essential micronutrient for plant and human, and over 40% of children in the world are suffering from Zn
malnutrition. Iron and Zinc deficiency in human is mainly caused from low Fe or Zn concentration and their
bioavailability in foods or edible parts of crop plants. Cereal grains are the most important dietary source of
micronutrients in many developing countries. Micronutrient concentrations and bioavailability in cereal grain is
generally low. Biofortification appears to be a most sustainable and cost-effective approach to solve human
micronutrient malnutrition.
The genetic capacity of the newly developed (biofortified) cultivars to absorb sufficient amount of Fe and
Zn from soil and to accumulate it in the grain may not be expressed to the full extent. It is, therefore, essential to
have a short term approach to improve Fe and Zn concentration in cereal grains. Application of FeSO 4 as foliar
spray, Zn fertilizers or Zn-enriched NPK fertilizers (e.g., agronomic biofortification) offer a rapid solution to the
problem, and represents useful complementary approach to on-going breeding programs. There is increasing
evidence showing that foliar or combined soil+foliar application of Fe and Zn fertilizers under field conditions are
highly effective and very practical way to maximize uptake and accumulation of Fe and Zn in whole cereal grains,
raising concentration up to 20mg kg-1 Fe and 60 mg kg1 Zn. Increasing the micronutrient concentration of cereal
grains has been identified as a way of addressing human micronutrient deficiencies.
Revolution
The Agricultural Revolution was a period of technological improvement and increased crop productivity
that occurred during the 18th and early 19th centuries in Europe. In this lesson, learn the timeline, causes, effects
and major inventions that spurred this shift in production.
There have been so many revolutions in the system time to time as per the need of human kind-
* Black Revolution - Petroleum Production : 1970

* Blue Revolution - Fish Production : 1960 Father of Blue revolution Prof. Hiralal Chaudhuri.
* Brown Revolution - Leather/non-conventional/Cocoa production
* Golden Fiber Revolution - Jute Production
* Golden Revolution - Fruits/Overall Horticulture development/Honey Production
'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 1

* Green Revolution - Food grains : Father of Green Revolution M.S. Swaminathan.

* Grey Revolution - Fertilizer : 1964
* Pink Revolution - Onion production/Pharmaceutical/Prawn production
* Red Revolution - Meat & Tomato Production
* Round Revolution Potato
* Silver Fiber Revolution Cotton Production
* Silver Revolution - Egg/Poultry Production
* White Revolution (In India: Operation Flood) - Milk/Dairy production : 1970 Father of White revolution Dr.
Verghese Kurien
* Yellow Revolution - Oil Seeds production : Father of Yellow Revolution Sam Pit Roda
Orange Revolution:
After 20 years of persistent research, city-based National Research Centre for Citrus (NRCC) has sown the
seeds of a new orange revolution in near future. The Centre has succeeded in increasing the productivity and
tolerance to diseases of Nagpur orange or mandarin and acid lime manifold by simply replacing the conventionally
used root stock of Rangpur lime and rough lemon with an exotic root stock from USA called as Alemow (Citrus
macrophylla) during budding/grafting.
The process of bringing this change and actual scientific trials took over 18-20 years. "Alemow is being used
worldwide as a root stock for improving productivity of fruits, especially citrus family. Our Centre began work in
this direction as an in-house research way back in 1992. The root stock has much superior horticultural
characteristics like smaller canopy, higher productivity and better disease tolerance compared to the domestic root
stocks," said the NRCC director V J Shivankar.
The productivity of orange has increased from just 9-10 tonnes/ha to 21 tonnes/ha in orange and from five
tonnes/ha to 13 tonnes/ha in acid lime. Both orange and acid lime developed using Alemow root stock also have
increased tolerance towards main citrus diseases, especially soil borne diseases like phytophthora fungus, the biggest
threat to orange. Right now, the success of technology is also being tested at national level through an All India
Coordinated Research Project (AIRCP) on orange by the Indian Council of Agricultural Research (ICAR) in all the
citrus growing belts of the country.
Principal scientists in horticulture and the project leader of the research R K Sonkar, who was recently
honoured with Vidarbha Bhushan award for leading the work, told TOI that the centre was also planning to try the
technology on other orange varieties like the Kinno, Khasi and Kurg mandarins if these national level trials yielded
good results. "The technology is not new. World over, Alemow is being used as root stock for many citrus crops. It
is being tried in India for the first time. We started experimenting with 12 different root stocks but Alemow turned
out to be the best," said Sonkar.
To develop planting material for orange, nurseries use Rangpur lime and rough lemon as root stocks on
which the buds of Nagpur orange are put. Both these root stocks have much less yield and are susceptible to
diseases. A good root stock provides the grower a useful tool to manipulate the vigour and performance of the
orchard. Though even Alemow is not completely resistant to phytophthora, the fungus attack is delayed by many

years and is comparatively milder. Also, since the canopy size is reduced with Alemow use, sunlight penetrates the
lowest part of the tree and increases fruit bearing ability. This makes the new trees more suitable for 'high density
plantation' the technique being promoted for all horticultural crops now. Instead of conventional 6mX6m distance
these can be planted at a distance of 6mX4m. Sonkar explained that acid lime was generally grown through seedling
but in this technology, budding is used. The production through seedling is just 5 tonnes/ha whereas budding on
Alemow root stock increased this yield to 13 tonnes/ha due to vigorous growth that occurs. Some farmers who
regularly visit NRCC on their own decided to take up cultivation using Alemow root stock. Bandu Wasankar from
Paratwada has tried it successfully. Mohan Tambi has gone step ahead and imported the root stock from USA for
his nursery and is developing planting material from them. Some farmers in Solapur too are trying it. Centre in the
long run will try to work on drought tolerance capacity too.
Red Revolution:
The hill state of Uttarakhand is silently scripting a 'red revolution' of sorts on the Himalayan reaches. Growing a
lesser known red cereal crop, amaranth, is opening up new vistas for the farmers settled in the upper areas where
staple plantations of wheat and paddy are not possible because of extreme climate conditions and mountainous
topography. Amaranth is a traditional fibre-rich plant used in baby food products, breads, etc. for its high protein
content and other nutritional values. In the past two years, international demand for amaranth has risen to the extent
that the farmers in the state are unable to meet the supply orders. "Demand for amaranth is largely from the overseas
markets of South Africa and the Netherlands. The cereal is grown only in this state. Even as the area under amaranth
cultivation has increased over the years, the demand and supply is a mismatch, perhaps to the advantage of the
farmers," said Binita Shah, senior programme manager, Uttarakhand Organic Commodity Board (UOCB). Shah said
the state, which acts as a facilitator, has an estimated standing demand of over 1,000 tonnes from overseas buyers,
against an estimated production of close to 600 tonnes expected this year.
"A few years ago, our visit to higher areas in the state brought to the fore a sordid tale. Farmers used to barter
amaranth for 2 kg of wheat, salt and a little more. Today, their income has increased manifold," she said. To
augment the demand, both in terms of export orders and domestic supplies, the government is laying more emphasis
on plantation of organic amaranth to empower farmers, said Chief Minister Ramesh Pokhriyal "Nishank".
From insignificant gains till some years ago, farmers this year got a handsome Rs 3,500 per quintal of
amaranth, up nearly 30 per cent since last year. Close to 5,000 hectares of area in upper areas of Garhwal,
Uttarakashi, Chamoli, Rudraprag is under amaranth production and just about 500 hectares under certified organic
produce in Uttarakhand.
An increase in food production, especially in underdeveloped and developing nations, through the introduction
of high-yield crop varieties and application of modern agricultural techniques. The introduction of high-yielding
varieties of seeds and the increased use of chemical fertilizers and irrigation are known collectively as the Green
Revolution, which provided the increase in production needed to make India self-sufficient in food grains, thus
improving agriculture in India. High-yielding wheat was first introduced to India in 1968 by American agronomist
Norman Borlaug. Borlaug has been hailed as the Father of the Green Revolution but M.S. Swaminathan is known as
the "Father of the Green Revolution in India". The methods adopted included the use of high yielding varieties

(HYV) of seeds. The production of wheat has produced the best results in fueling self-sufficiency of India. Along
with high yielding seeds and irrigation facilities, the enthusiasm of farmers mobilized the idea of agricultural
revolution and is also credited to M. S. Swaminathan and his team had contributed towards the success of green
revolution. Due to the rise in use of chemical pesticides and fertilizers there were many negative effects on the soil
and the land such as
land degradation.
Blue Revolution
Blue Revolution means the adoption of a package programme to increase the production of fish and marine
products. The Blue Revolution in India was started in 1970 during the Five-Year Plan when the Central Government
sponsored the Fish Farmers Development Agency (FFDA). Subsequently, the Brakish Water Fish Farms
Development Agency were set up to develop aquaculture. The Blue Revolution has brought improvement in
aquaculture by adopting new techniques of fish breeding, fish rearing, fish marketing, and fish export. Under the
Blue Revolution programme, there had been a tremendous increase in the production of shrimp. Andhra Pradesh and
Tamil Nadu have developed shrimp in a big way. The Nellore District of Andhra Pradesh is known as the 'Shrimp
Capital of India'.
Pink Revolution
Pink Revolution is a term used to denote the technological revolutions in the meat and poultry processing
sector. India has already seen the green and white revolutions in its food industry related to agriculture and
milk respectively, now thrust is upon meat and poultry sector. India being a country of huge cattle and poultry
population, has high potential for growth if this sector is modernized.
Green Revolution:
The Green Revolution refers to a series of research and development and technology transfer initiatives,
occurring between the 1930s and the late 1960s (with prequels in the work of the agrarian genetist Nazareno
Strampelli in the 1920s and 1930s), that increased agricultural production worldwide, particularly in the developing
world, beginning most markedly in the late 1960s. The initiatives, led by Norman Borlaug, the "Father of the Green
Revolution," who won the Nobel Peace Prize in 1970, credited with saving over a billion people from starvation,
involved the development of high-yielding varieties of cereal grains, expansion of irrigation infrastructure,
modernization of management techniques, distribution of hybridized seeds, synthetic fertilizers, and pesticides to
farmers.
The term "Green Revolution" was first used in 1968 by former United States Agency for International
Development(USAID) directorWilliam Gaud, who noted the spread of the new technologies: "These and other
developments in the field of agriculture contain the makings of a new revolution. It is not a violent Red
Revolution like that of the Soviets, nor is it a White Revolution like that of the Shah of Iran. I call it the Green
Revolution."
Effect of Green Revolution on Crop Production:
Cereal production more than doubled in developing nations between the years 19611985. Yields of rice,
maize, and wheat increased steadily during that period. The production increases can be attributed roughly equally

to irrigation, fertilizer, and seed development, at least in the case of Asian rice. While agricultural output increased
as a result of the Green Revolution, the energy input to produce a crop has increased faster, so that the ratio of crops
produced to energy input has decreased over time. Green Revolution techniques also heavily rely on
chemical fertilizers, pesticides and herbicides and rely on machines, which as of 2014 rely on or are derived
from crude oil, making agriculture increasingly reliant on crude oil extraction. Proponents of the Peak Oil theory
fear that a future decline in oil and gas production would lead to a decline in food production or even a Malthusian
catastrophe .
Technologies used in Green Revolution:
Technology is the application of organized and scientific knowledge to solve practical problems. In
agriculture, the success of a technology can be measured only when it gets transferred for increased crop production.
The following technologies have helped in increasing crop production in the World:
1. Dwarfing gene
2. Hybrid Technology
3. Biotechnology particularly cry genes in cotton
4. Seed Technologies: Seed Vigor, seed coating
5. Agronomic Technologies: Row-Plant Spacing minimum tillage etc
Introduction:
The diets of over two-thirds of the worlds population lack one or more essential mineral elements. This
can be remedied through dietary diversification,mineral supplementation, food fortification, or increasing the
concentrations or bioavailability of mineral elements in produce (biofortification). Humans require at least 22
mineral elements for their wellbeing(Welch & Graham, 2004; White & Broadley, 2005a; Graham et al., 2007).
These can be supplied by an appropriate diet. However, it is estimated that over 60% of the worlds 6 billion people
are iron (Fe) deficient, over 30% are zinc (Zn) deficient, 30% are iodine (I) deficient and c. 15% are selenium (Se)
deficient (see Supporting Information References S1). In addition, calcium (Ca), magnesium (Mg) and copper (Cu)
deficiencies are common in many developed and developing countries (Frossard et al., 2000; Welch & Graham,
2002,2005; Rude & Gruber, 2004; Grusak & Cakmak, 2005;Thacher et al., 2006). Currently, mineral malnutrition is
considered to be among the most serious global challenges to humankind and is avoidable (Copenhagen Consensus
2004;http://www.copenhagenconsensus.com). Mineral malnutrition can be addressed through dietary diversification,
mineral supplementation, food fortification and/or increasing mineral concentrations in edible crops
(biofortification).
Because the seeds of many cereals are often consumed after milling or polishing, it is pertinent to consider
whether genetic variation in the distribution of mineral elements within the seed can be utilized in biofortification
strategies. Mineral elements are nonhomogenously distributed within the seed and the concentrations of many
mineral elements are highest in the husk and/or aleurone layers (see References S12). Milling or polishing cereal
seeds can, therefore, remove large quantities of mineral elements from the diet and the extent of these losses is
genotype dependent (Gregorio et al., 2000; Vasconcelos et al., 2003; Ma et al., 2004; Lyons et al., 2005; Prom-u-
thai et al., 2007).

Improving the crop yield and quality through Biofortification:

Regarding the nutritional traits, one of the most promising application of transgenic technology has been the
development of vitamin A enriched varieties, popularly known as Golden Rice due to the slightly yellow colour
conferred to the endosperm. First successful product of GM based biofortification with pro-vitamin A trait.
Researchers have found that in Japonica Golden Rice, an intake of only 120 g is required for meeting the Vit As
RDA. (Paine et al, 2005;Tang et al, 2009) Bio fortification of rice with iron and zinc are also being carried out by
several public and private sector organizations. Increasing the concentration of micronutrient (especially Zn) in food
crop plants is a growing global challenge, with potentially great implications for both crop production and human
health. It is believed thatZn deficiency is the most widespread micronutrient deficiency in crop plants and human
beings (Alloway, 2004; Hotz and Brown, 2004). Zn deficiency in humans causes a wide range of health
complications, including impairments in the immune system, learning ability and physical growth, and increases in
mortality and infections (Hotz and Brown, 2004; Cunnigham-Rundles et al., 2005). Zinc deficiency also induces
DNA damage and increases the risk of cancer occurrence (Ho, 2004).
Zinc and Fe deficiencies are a growing public health and socioeconomic issue, particularly in the developing
world (Welch and Graham 2004). Recent reports indicate that nearly 500,000 children under 5 years of age die
annually because of Zn and Fe deficiencies (Black et al.2008). Zinc and Fe deficiencies together with vitamin A
deficiency have been identified as the top priority global issue to be addressed to achieve a rapid and significant
return for humanity and global stability (www.copenhagenconsensus.com). Low dietary intake of Fe and Zn appears
to be the major reason for the widespread prevalence of Fe and Zn deficiencies in human populations. In countries
with a high incidence of micronutrient deficiencies, cereal-based foods represent the largest proportion of the daily
diet (Cakmak 2008). Cereal crops are inherently very low in grain Zn and Fe concentrations, and growing them on
potentially Zn- and Fe-deficient soils further reduces Fe and Zn concentrations in grain (Cakmak et al. 2010). Thus,
biofortification of cereal crops with Zn and Fe is a high-priority global issue. HarvestPlus (www.harvestplus.org) is
the major international consortium to develop new plant genotypes with high concentrations of micronutrients by
applying classical and modern breeding tools (i.e. genetic biofortification). Although plant breeding is the most
sustainable solution to the problem, developing new micronutrient-rich plant genotypes is a protracted process and
its effectiveness can be limited by the low amount of readily available pools of micronutrients in soil solution
(Cakmak 2008). Application of Zn- and Fe-containing fertilizers (i.e. agronomic biofortification) is a short-term
solution and represents a complementary approach to breeding.
Cereals (e.g.wheat, rice and maize) with inherently low Zn concentrations in the grain are the most
important source of calories in the developing world. In the present study, we used a range of wheat genotypes with
different concentrations of Zn and other micronutrients to determine whether the DTZ method could be useful as a
rapid screening method for seed Zn concentration.

GW-366T1SEED GW-366T2SEED GW-366T3SEED

(Control)
High Efficient Variety
DL-803(T3)SEED
DL-803(T1)SEED DL-803(T2)SEED
(Control)
Low Efficient Variety
Biofortification of Zinc in two different variety of Wheat through soil application (T2)and
(T3)Foliar application
Here we have taken two different variety of wheat where one is High efficient variety name GW-366 mean
that variety which perform better in sense of yield in case of adverse condition and the second variety is Low
efficient variety name DL-803 means that variety is not performing better in unfavourable condition. Now the
biofortification of zinc is provided in two ways the first one is through soil application (T1) and second is soil +
Foliar application (T3). Here through analytical study biofortification of zinc is occurs more in foliar
application.variety GW-366 is much better highly fortified in comparision to the variety DL-803. The localization
and accumulation of zinc is found in foliar application of mature seed of GW-366 variety.
References:
1. Black RE, Lindsay HA. Bhutta ZA, Caulfield LE, De Onnis M, Ezzati M, Mathers C, Rivera J (2008).
Maternal and child undernutrition: global and regional exposures and health consequences. Lancet
371,243-260.
2. Cakmak I (2008) Enrichment of cereal grains withzinc: Agronomic or genetic biofortification? Plant and
Soil 302,1-17.
3. Cakmak I, Pfeiffer WH, McClafferty B (2010) Biofortification of durum wheat with zinc and iron. Cereal
Chemistry 87,10-20.
4. Welch RM, Graham RD (2004) Breeding for micronutrients in staple food crops from a human nutrition
perspective. Journal of Experimental Botany 55,353364.
5. White PJ, Broadley MR. 2005. Biofortifying crops with essential mineral elements. Trends in Plant Science
10: 586593.

6. Hotz C, Brown KH (2004) Assessment of the risk of zinc deficiency in populations and options for its
control. Food Nutr Bull 25: 94204.
7. Alloway BJ (2004) Zinc in Soils and Crop Nutrition.International Zinc Association Communications.
IZAPublications, Brussel.
8. Graham RD, Welch RM, Saunders DA, Ortiz-Monasterio I, Bouis HE Bonierbale M, de Haan S, Burgos G,
Thiele G, Liria R et al. 2007. Nutritious subsistence food systems. Advances in Agronomy 92: 174.
9. Frossard E, Bucher M, Mchler F, Mozafar A, Hurrell R. (2000). Potential for increasing the content and
bioavailability of Fe, Zn and Ca in plants for human nutrition. Journal of the Science of Food and
Agriculture 80:861879.
10. Welch RM, Graham RD. 2002. Breeding crops for enhanced micronutrient content. Plant and Soil 245:
205214.
11. Welch RM, Graham RD. 2005. Agriculture: the real nexus for enhancing bioavailable micronutrients in
food crops. Journal of Trace Elements in Medicine and Biology 18: 299307.
12. Rude RK, Gruber HE. 2004. Magnesium deficiency and osteoporosis: animal and human observations.
Journal of Nutritional Biochemistry 15: 710716.
13. Grusak MA, Cakmak I. 2005. Methods to improve the crop-delivery of minerals to humans and livestock.
In: Broadley MR, White PJ, eds. Plant nutritional genomics. Oxford, UK: Blackwell, 265286.
14. Thacher TD, Fischer PR, Strand MA, Pettifor JM. (2006). Nutritional rickets around the world: causes and
future directions. Annals of Tropical Paediatrics 26: 116.

Shoving Towards a New Green Revolution
Vijai Malik
Department of Botany, M. S. College, Saharanpur
Abstract
India is an agricultural country due to its strategic geographical, climatic and topographical location on the
world map. Even with industrial revolution, the importance of agriculture has not reduced. Agricultural self-
sustenance has brought India pride during the first green revolution, as the country had sustained several famines
from British ruled period till just after independence. The seat of first green revolution was north- west India mainly,
with wheat being the focal staple crop. Second Green Revolution in fact should be focused to practice sustainable
agriculture at regional level and crop wise. It must aim at raising the productivity of other important food crops
such as Sorghum, Millets and Cassava- food produced and consumed mainly by the worlds poor. The present
article includes aim, calls, and some of the approaches that may push India towards Second Green Revolution.
Key Words: Second Green Revolution, Food Security, Nanotechnology
Introduction:
Research in applied Botany is playing and will play important role in self sufficiency towards food production.
Advance research in Genetics, Cytogenetics, Plant Breeding, Molecular biology, Biotechnology etc. are helping a lot
of in this direction. Not only Yoga and use of medicinal plants are helping in building good character and health of
human beings but also recent advances in Molecular Biology, Molecular Cytogenetics and Pant Biotechnology are
further enhancing food production and human health. We have seen a revolution in the past concerning with food
production, i.e. Green Revolution. Due to shrinkage of agricultural land by industrialization, urbanization and many
fold increase in population, this revolution has no means. We need big revolution at this juncture.
The term Green revolution is applied to the period 1967-1978. In 1968, William S. Gaud coined the term Green
Revolution to describe phenomenal growth in agriculture. The first green revolution relied heavily on the use of
large amounts of fertilizers, pesticides and other agricultural inputs. This coupled with continued expansion of
farming areas led to self-sufficiency in food production. A quantum jump in the productivity and production of
wheat and then rice transformed the image of India as a begging bowl to a bread basket. Second Green Revolution in
fact refers to practicing sustainable agriculture. That is, protecting natural resources from becoming increasingly
degraded and polluted and using production technologies that conserve and enhance the natural resource base of
crops, forests, inlands and marine fisheries [1].
Aim of Second Green Revolution:
Declining productivity due to unstable agriculture practices over the years and galloping rate of population rate
have both put a severe strain on the food supply situation in the country. To meet the food requirements of the

growing population a second green revolution has become imperative. Second Green Revolution must aim at raising
the productivity of other important food crops such as Sorghum, Millets and Cassava- food produced and consumed
mainly by the worlds poor.
Calls for a Second Green Revolution
The Second Green Revolution is a change in agricultural production widely thought necessary to feed and sustain
the growing population on Earth. These calls have precipitated in part, as a response to rising food commodity
prices, and fears of peak oil among other factors.
Calling for a second Green Revolution, Prime Minister Sh. Narendra Modi has asked farmers to adopt scientific
methods to enhance food grain production and reduce imports by using one-fifth of their farming land to cultivate
lentils.
"Unless we prepare a balanced and a comprehensive integrated plan, we will not be able to change the lives of
farmers," PM said, adding that Indian farmers are still lagging behind in terms of availability of good quality seeds,
adequate water, power, right price and market for their produce. Invoking former Prime Minister Lal Bahadur
Shastri's slogan "Jai Jawan, Jai Kisan," Modi asked farmers to try and grow pulses on part of their land.
"The production of pulses in the country is very low. I (PM) urge farmers that if they have five acres of farming
land, use four acres for other crops but cultivate pulses on at least one acre. This basic source of protein should be
available to the poor at affordable prices," he said. The Centre is willing to pay more than the announced minimum
support price to farmers for pulse production, he said.
"We have seen the first Green Revolution, but it happened several years ago. Now it is the demand of time that there
should be a second Green Revolution without any delay. And where is it possible? It is possible in eastern UP,
Bihar, West Bengal, Jharkhand, Assam, and Odisha", PM said. "The government is making every effort to make the
farmers aware of modern scientific advancements," he added. The PM said only one model of farm technology
cannot be adopted uniformly in the country as there are variations in soil and climatic conditions to consider.
Pitching for 'per drop, more crop', he stressed the need for research in the field of agriculture to determine the health
of soil and its needs in terms of seeds, water quantity, amount of fertilization, etc.
"We want private individuals to own laboratories, similar to pathological laboratories, to test soil and issue a health
card so that a farmer is aware of the deficiency, fertilizer requirements and type of crop suitable for his plot of land,"
he said, adding this will also lead to job creation. He also added that students enrolling in IARI from different parts
of the country would come for higher education and research and would then return to their respective states after
developing a model suitable for that geographical region [2].
Role of Nanotechnology to Revolutionize Agriculture

The technology applied in agriculture is referring as green technology. The green technology subjects are energy,
green buildings, environmentally preferred purchasing, green chemistry and green nanotechnology. Nanotechnology
in agriculture has gained momentum in the last decade with an abundance of public funding, but the pace of
development is modest, even though many disciplines come under the umbrella of agriculture. This could be
attributed to: a unique nature of farm production, which functions as an open system whereby energy and matter are
exchanged freely; the scale of demand of input materials always being gigantic in contrast with industrial
nanoproducts; an absence of control over the input nanomaterials in contrast with industrial nanoproducts (eg, the
cell phone) and because their fate has to be conceived on the ecosphere-biosphere-hydrosphere-atmosphere
continuum; the time lag of emerging technologies reaching the farmers field, especially given that many emerging
economies are unwilling to spend on innovation; and the lack of foresight resulting from agricultural education not
having attracted a sufficient number of brilliant minds the world over, while personnel from kindred disciplines
might lack an understanding of agricultural production systems. If these issues are taken care of, nanotechnologic
intervention in farming has bright prospects for improving the efficiency of nutrient use through nanoformulations
of fertilizers, breaking yield barriers through bionanotechnology, surveillance and control of pests and diseases,
understanding mechanisms of host-parasite interactions at the molecular level, development of new-generation
pesticides and their carriers, preservation and packaging of food and food additives, strengthening of natural fibers,
removal of contaminants from soil and water, improving the shelf-life of vegetables and flowers, clay-based
nanoresources for precision water management, reclamation of salt-affected soils, and stabilization of erosion-prone
surfaces, to name a few.
Time for a Second Green Revolution
After Green Revolution India had become self-sufficient in basic food grains (wheat and rice), but recently we are
facing recurrent spells of shortages in essential items like lentils, edible oil, sugar and onions. This has resulted in
their knee-jerk imports at exorbitant prices. According to experts if we do not focus immediately on increasing food
production by rationalizing priorities in the agriculture sector, we may be in for a rude shock of unmanageable food
scarcity. In the present scenario cultivable land and water availability are shrinking. We have almost dried our rivers
and are emptying groundwater reservoirs by reckless extraction. Extensive use of chemical fertilizers and pesticides
has polluted surface water and but groundwater resources.
Researchers should focus on alternative organic fertilizers and pesticides which will need less water. Irrigation
should aim at giving only the required amount of water in the root zone of plants for which drip and/or sprinkler
system must be applied in the entire cultivable areathrough subsidy, wherever necessary. Provide water to every
field. This has to be done by making field channels, wherever terrain permits. At all other places sprinklers with
minimal extraction of groundwater should be used to meet the target of total irrigation coverage to cultivated land.
We will have to collect and store each drop of water, check its misuse and wastage, and use it frugally yet
efficientlyespecially in agriculture, because that is the biggest user sector of water. This should be the target of the
second Green Revolution.

For Second Green Revolution we will have to develop techniques which would deliver more crops for each drop of
irrigation as well as more yield per field of cultivated land. Although land available for agriculture is declining there
is still scope to increase productivity from it, mainly because we are far behind most countries in per hectare yield.
With new technology, improved seeds, rationalized crop rotation and balancing soil chemistry etc., we can obtain
much more crop from the available land. Genetically improved seeds, which are being scoffed atdue to lack of
proper education and informationcan enhance productivity greatly.
Time has come to launch a second Green Revolution to provide long-term food security to the nation, and it is the
direction in which agriculture scientists and institutions need to focus. Agriculture scientists should start their
research by accepting that whatever water is available for irrigating crops is not only limited but also mostly
polluted. Their target should be to obtain maximum production from frugal use of this water in irrigation. Of course,
reuse and recycling of unclean water would be attempted after treating it to acceptable parameters for agricultural
application [3].
The Second Green Revolution through Biotechnology:
The worlds food grain production was tripled and saved millions of life from famine. This could happen because of
the Green Revolution during 1960s. Despite of that, feeding an estimated worldwide human population of over 9
billion by 2050 and alleviating the malnutrition in the poor nations are still the massive global challenges.
Conventional breeding strategies alone may not be able to increase the productivity without disturbing the
environment. The biotechnological interventions including tissue culture, genetic engineering and molecular
breeding have proved as a game changer.
In India, Bt-cotton the only transgenic crop is the live demonstration of the importance of genetic engineering that
has revolutionized cotton production by escalated yields, reduced insecticide applications, and improved socio-
economic livelihood of farmers. But misinformation, lack of public awareness and current national policies are the
major bottleneck in the adoption of this technology.
A second revolution has been made possible by the sequencing of the rice genome in 2005 (the first cereal crop to be
sequenced). This enabled breeders to discover the genes for flood resistance in one obscure variety from eastern
India and transfer them to varieties all round the world. Breeders will soon do the same for genes that provide other
valuable traits.
Conclusion:
70% of Indian population lives in rural areas and 60% of its workforce is agriculture. As we all know current state
of agriculture in India is result of green revolution which is in place since late 1960s, which was heavily backed by
government. It has delivered India food security and sufficiency which was critical at that time. This progress and
security had its own costs in terms of environment and economic viability. Green revolution rampantly used
fertilizers and other chemicals, which made food and water, toxic to some extent. Indias 80% of fresh water is

consumed by agriculture, more particularly by rice farming. Consequently, new agriculture policy of India aims at
sustainable agriculture, which is popularly called Second Green Revolution or Evergreen Revolution. Declining
Indian economy and growing food insecurity policy and environmental experts has called for a Second Green
Revolution. The only way to attain food and nutritional security is by improving marginal and dry land agriculture in
a sustainable manner by implementing Second Green Revolution.
References:
Kumar S. Towards a Second Green Revolution. Science Reporter (June) 9-15 (2008).
Deogharia J. PM calls for Second Green Revolution. The Times of India, June 29, (2015)
Narain Y. and Kumar S. K. Time for a Second Green Revolution. Indian Express, June 26, (2015).

Biopesticide: An Ecofriendly Approach For Second Green Revolution in Agriculture
Rashmi Nigam
Assistant Professor, Department of Plant Pathology, Janta Vedic College, Baraut, Baghpat(U.P.)
ABSTRACT
Agriculture plays a vital role in a developing country like India. Apart from fulfilling the food requirement of the
growing Indian population, it also plays a role in improving economy of the country. Emphasis of present day
agriculture is to produce more with lesser land, water and man power. The Green Revolution technology adoption
during 1960 to 2000 has increased wide varieties of agricultural crop yield per hectare which increased 12-13% food
supply in developing countries. Inputs like fertilizers, pesticides helped a lot in this regard. But in spite of this fact,
food insecurity and poverty still prevails prominently in our country. Use of chemical pesticides and fertilizers have
caused negative impact on environment by affecting soil fertility, water hardness, development of insect resistance,
genetic variation in plants, increase in toxic residue through food chain and animal feed thus increasing health
problems and many more which makes bio pesticides to come into picture. The present study deals with different
type of biopesticide used in agriculture. Bio-pesticides are ecofriendly pesticides which are obtained from naturally
occurring substances (biochemicals), microbes and plants. They are also biochemical pesticides that are naturally
occurring substances that control pests by nontoxic mechanisms. Biopesticides are living organisms (natural
enemies) or their products (phytochemicals, microbial products) or by products (semiochemicals) which can be used
for the management of pests that are injurious to plants. The most commonly used biopesticides are living
organisms, which are pathogenic for the pest of interest. These include biofungicides (Trichoderma), bioherbicides
(Phytopthora) and bioinsecticides (Bacillus thuringiensis). There are few plant products also which can now be used
as a major biopesticide source. The stress on organic farming and on residue free commodities would certainly
increased adoption of biopesticides by the farmers.
Keywords: Biopesticide, Ecofriendly, Biofungicide, Bacillus thuringensis.

Introduction
Agriculture plays a vital role in a developing country like India as it plays a important role in improving economy of
the country. Agriculture is adversely affected by destructive activities of numerous pests like bacteria, fungi, weeds
and insects, leading reduced yields. Since 1960s, the most common method for pest control has been based on the
intensive use of synthetic organic pesticides. The Green Revolution technology adoption during 1960 to 2000 has
increased wide varieties of agricultural crop yield per hectare which increased 12-13% food supply in developing
countries. Inputs like fertilizers, pesticides helped a lot in this regard. The indiscriminate use of chemical pesticides
in modern agriculture resulted in the development of several problems such as pesticide resistance insects,
resurgence of target and non target pests, destruction of beneficial organisms like honey bees, pollinators,
parasitoids and predators and pesticide residue in food, feed and fodder (Al-Zaidi et al., 2011). Hence, the todays
need is to produce maximum from the decreasing availability of natural resources without adversely affecting the
environment. Therefore, alternative, environmentally safe methods are needed for pest management. As a result,

bio-pesticides using eco-friendly, plant or microbial derived ingredients showing broad insecticidal effects with
minimal damage to the environment have been increasingly developed. Biopesticides are generally less toxic than
chemical pesticides, often target specific, cause minimal harm to birds, insects and mammals. In addition, even if
used in an open field, they decompose quickly, thereby minimizing the risk of environmental pollution or residual
toxicity. Bio-pesticides are ecofriendly pesticides which are obtained from naturally occurring substances
(biochemicals), microbes and plants (Dutta, 2015). Biopesticides are certain types of pesticides derived from such
natural materials as animals, plants, bacteria, and certain minerals. The most commonly used biopesticides are
livingorganisms, which are pathogenic for the pest of interest. These include biofungicides (Trichoderma),
bioherbicides (Phytopthora) and bioinsecticides (Bacillus thuringiensis).There are few plant products also which can
now be used as a major biopesticide source (Salma et al., 2011). Plant-incorporated protectants include substances
that are produced naturally on genetic modification of plants. Such examples are incorporation of BT gene, protease
inhibitor, lectines, chitinase etc into the plant genome so that the transgenic plant synthesizes its own substance that
destroys the targeted pest. In India, some of the biopesticides like Bt, NPV, neem based pesticides, etc. have already
been registered and are being practiced. (Gupta, 2010). In the present study, an attempt has been made to provide a
comprehensive study or review on biopesticide, its ecofriendly approach on the sustainable agriculture or for an
evergreen revolution in agriculture. The study deals with different type of biopesticide, used in agriculture, its
application and advantages. The study is based on secondary data which has been collected from the different
sources.
Types of Biopesticide
Biopesticides are an important ingredient of Integrated Pest Management (IPM) packages due to their capability in
maintaining the natural diversity without the use of any artificial or synthetic residues. The origin of Biopesticides
can be microbial (bacteria, fungi or virus), herbal (plant extracts) or genetically modified plants (GM). Beauveria
spp., Trichoderma spp., and Bacillus spp., are some of the microbial biopesticides. Biopesticides may be broadly
categorized into three major groups: 1) Biochemical biopesticides 2) Microbial pesticides and 3) Plant-Incorporated
Protectants (PIPs).
1. Biochemical pesticides: Biochemical pesticides are naturally occurring substances that control pests by non-toxic
mechanisms. Conventional pesticides are generally synthetic materials that directly kill or inactivate the pest.
Biochemical pesticides are herbal-based substances that are naturally produced by a plant or an organism. They are
non-toxic and biodegradable. They help the plant in counter-attacking its pests or producing chemicals that would
prevent pest attack on the plant. Biochemical pesticides include substances that interfere with growth or mating,
such as plant growth regulators, or substances that repel or attract pests, such as pheromones. Because it is
sometimes difficult to determine whether a substance meets the criteria for classification as a biochemical pesticide,
EPA has established a special committee to make such decisions. Plant biochemicals are collectively called
botanicals and the most important botanical is pyrethrum, followed by neem, rotenone and essential oils, typical
used as insecticides (e.g. pyrethrum, rotenone, rape seed oil, quassia extract, neem oil, nicotine), repellents (e.g.
citronella), fungicides (e.g. laminarine, fennel oil, lecithine), herbicides (e.g. pine oil), sprouting inhibitors (e.g.
caravay seed oil) (Isman, 2006).

2. Microbial pesticides: These pesticides originate from micro-organisms such as bacteria, fungi or other protozoan
groups. These are mostly target-specific organisms that are aimed at killing one or a group of pests. Microbial
pesticides consist of a microorganism (e.g., a bacterium, fungus, virus or protozoan) as the active ingredient. Desai,
(1997) concluded that microbial pesticides can control many different kinds of pests, although each separate active
ingredient is relatively specific for its target pest. Forexample, there are fungi that control certain weeds, and other
fungi that kill specific insects. Microbial pesticides can control many different kinds of pests these include
biofungicides (Trichoderma), bioherbicides (Phytopthora) and bioinsecticides (Bacillus thuringiensis and
Baculovirus). The most widely used microbial pesticides are subspecies and strains of Bacillus thuringiensis, or Bt.,
can control certain insects in cabbage, potatoes, and other crops Each strain of this bacterium produces a different
mix of proteins, and specifically kills one or a few related species of insect larvae. Some Bt's control moth larvae
found on plants, other Bt's are specific for larvae of flies and mosquitoes. The target insect species are determined
by whether the particular Bt produces a protein that can bind to a larval gut receptor, thereby causing the insect
larvae to starve. (Kalra, and Khanuja, 2007). The widely used microbial pesticides are Trichoderma viride, Bacillus
thuringenesis, Bacillus sphaericus, Pseudomonas fluorescence (Bacteria), Beauveria bassiana (Fungi), Baculo virus
and Nucleopolyhedrosis Virus.
3. Plant-Incorporated-Protectants (PIPs): These are genetically modified materials produced by modifying a
protein and introduced into the plant so that it produces its own pesticide. Plant-Incorporated-Protectants are
pesticidal substances that plants produce from genetic material that has been added to the plant. For example, the
gene for Bt pesticidal protein, was introduced into the genetic material of cotton plant and plant manufactures the
substance that destroys the pest. The protein and its genetic material, but not the plant itself, are regulated by EPA.
(Thakore, 2006). Transgenic plant produces biodegradable protein with no harmful effect on animals and human
beings, and thus minimizes the use of hazardous pesticides.
Applications of Biopesticide
Biopesticides are usually applied in a manner similar to chemical pesticides, but achieve pest management in
an environmentally friendly way. With all pest management products, but especially microbial agents, effective
control requires appropriate formulation (Burges,1998) and application (Matthews et al., 2014, Lacey and Kaya,
2007). Biopesticides for use against crop diseases have already established themselves on a variety of crops. For
example, biopesticides already play an important role in controlling downy mildew diseases. A major growth area
for biopesticides is in the area of seed treatments and soil amendments. Fungicidal and biofungicidal seed treatments
are used to control soil borne fungal pathogens that cause seed rots, damping-off, root rot and seedling blights. They
can also be used to control internal seedborne fungal pathogens as well as fungal pathogens that are on the surface
of the seed. Many biofungicidal products also show capacities to stimulate plant host defence and other
physiological processes that can make treated crops more resistant to a variety of biotic and abiotic stresses.
Biopesticide Technology in India

The Bio pesticide segment occupies a small portion of the large pesticide market in India. In 2005, it accounted for
just 2.89%, which was expected to increase by 2.3%. Globally, there are 175 registered biopesticide active-

ingredients and 700 products available in the market (Hajeck and Leger, 1994). In India, so far only 12 types of
biopesticides have been registered under the Insecticide Act, 1968 (Thakore, 2006). Neem based pesticides, Bacillus
thuringensis, NPV and Trichoderma are the major biopesticides produced and used in India. Last decade, has
witnessed a rapid growth in this segment, especially on standardization of production techniques of Trichoderma,
Gliocladium, Paecilomyces, Pseudomonas, Trichogramma, NPV and Bacillus to use them against many insect pests
and diseases. Today, these biological control agents have been successfully employed in India. Trichogramma,
which is a stingless wasp that feeds on the eggs of sugarcane borers, has been used against borers in the states of
Tamil Nadu, Rajasthan, UP, Bihar and Haryana. Similarly Trichogramma, Bracon, Chelonus and Chrysopaspp are
being used for the control of cotton bollworms. Trichogramma has also been used against rice stem borer and leaf
folder. The sugarcane scale insect has been controlled with the help of predatory Coccinellid beetles in UP, West
Bengal, Gujarat and Karnataka. Most of the biopesticides find use in public health, except a few that are used in
agriculture as transgenic plants and beneficial organisms called bio-agents: are used for pest management in India
(Kalra, and Khanuja, 2007). Important biopesticide registered in India are Bacillus thuringiensis var. israelensis,
Bacillus thuringiensis var. kurstaki, Bacillus thuringiensis var. galleriae, Bacillus sphaericus, Trichoderma
viride, Trichoderma harzianum and Pseudomonas fluoresens. Shia and Feng, (2004) reported some successful
utilization of biopesticides and bio-control agents in Indian agriculture like control of rots and wilts disease in
various crops by Trichoderma based products, control of mango hoppers and mealy bugs and coffee pod borer by
Beauveria, control of Helicoverpa on cotton, pigeon-pea, and tomato by Bacillus thuringiensis, control of white fly
on cotton by neem products, control of diamondback moths by Bacillus thuringiensis, control of sugarcane borers by
Trichogramma and control of Helicoverpa on gram by N.P.V.
Advantages of Biopesticide
Biopesticides are usually inherently less toxic than conventional pesticides. No harmful residues detected.
Biopesticides generally affect only the target pest and closely related organisms, in contrast to broad spectrum,
conventional pesticides that may affect organisms as different as birds, insects and mammals.
Biopesticides often are effective in very small quantities and often decompose quickly, resulting in lower
exposures and largely avoiding the pollution problems caused by conventional pesticides. It can be cheaper than
chemical pesticides when locally produced.
When used as a component of Integrated Pest Management (IPM) programs, biopesticides can greatly reduce
the use of conventional pesticides, while crop yields remain high. Biopesticides can be more effective than
chemical pesticides in the long-term
Conclusion
India has a vast potential for biopesticides. To develop eco-friendly pest control technologies are the need of day
and challenging tasks for developing countries to improve agricultural productivity in a sustainable manner.
Biopesticides are considered as one of the eco-safe alternatives due to their biodegradation in nature, multiple mode
of action on target pests and may not leave toxic residues,are used globally for controlling insect pests and diseases.
Thus bio-pesticides would achieve the target of evergreen revolution or second green revolution if these bio-

molecules can be characterized for their potency using available modern biotechnological tools, to develop low cost
technology for compound isolation, formulation and commercial scale establishment of plant resource and microbial
pesticides. The stress on organic farming and on residue free commodities would certainly warrant increased
adoption of biopesticides by the farmers.
References
1. Al-Zaidi, A. A., Elhag, E. A., Al-Otaibi, S. H. and Baig, M. B. (2011). Negative effects of pesticides on
the environment and the farmers awareness in Saudi Arabia: a case study. J. Anim. Plant Sci. 21(3): 605-
611.
2. Burges, H.D. (1998). Formulation of Microbial Biopesticides, beneficial microorganisms, nematodes and
seed treatments Publ. Kluwer Academic, Dordrecht, 412 pp.
3. Desai, S. T. (1997). Chemical industry in the post independence era: a finance analysis point of view.
Chemical Business, 11(1): 25 - 28.
4. Hajeck, A. E and Leger, St. (1994) Interactions between fungal pathogens and insect hosts, Annual Review
of Entomology, 39: 293 - 322.
5. Isman, M. B., (2006) Botanical insecticides, deterrents, and repellents in modern agriculture and an
increasingly regulated world, Annu. Rev. Entomol. 51 4566.
6. Kalra, A. and Khanuja, S. P. S. (2007). Research and Development priorities for biopesticide
andbiofertiliser products for sustainable agriculture in India. In. Business Potential for Agricultural
Biotechnology (Teng, P. S. ed.), Asian Productivity Organisation, 2007; 96-102.
7. Lacey, L. and Kaya, H. (2007). Field Manual of Techniques in Invertebrate Pathology2nd edition. Kluwer
Academic, Dordrecht, NL
8. Matthews, G. A., Bateman, R. P., Miller, P. C. H. (2014). Pesticide Application Methods (4th Edition),
Chapter 16. Wiley, UK.
9. Salma Mazid,Ratul Rajkhowa, and Jogen Kalita ( 2011). Article A Review on use of Biopesticides in
Insect Management, International Journal of Science and Advanced Technology: 1 (7).
10. Shia, W.B. and. Feng, M.G. (2004) Lethal effect of Beauveria bassiana, Metarhizium anisopliae, and
Paecilomyces fumosoroseus on the eggs of Tetranychus cinnabarinus (Acari:Tetranychidae) with a
description of a mite egg bioassay system, Biological Control; 30: 165 173.
11. Dutta, S. (2015) Biopesticide: An ecofriendly approach for pest control.World Journal of Pharmacy and
Pharmaceutical Sciences. 4,( 06),, 250-265
12. Gupta, S. (2010). Biopesticides: An eco-friendly approach for pest control, Journal of Biopesticides,; 3(1
Special Issue): 186 - 188.
13. Thakore, Y. (2006). The biopesticide market for global agricultural use. Industrial Biotechnology; 194-
208.

Rhododendron Diversity and their Conservation in Sikkim Himalayas
Vishal Kaushik, Department of Botany, N. R. E. C. College, Khurja.
vishalkaushik18@rediffmail.com
Abstract
The genus belongs to family Ericacae of order Ericales. In India the members of the family are confined to
higher altitudes and grow from 1,500 to 6,000 meters. But most of the species are found at altitudes between
2,500 to 4,500 meters. The Rhododendron are found along with Pines and Junipers Elnus, Popular, Betula and
Quercus in Eastern Himalayas. The plants sprout in February, March and profuse flowering continues till July.
Flowers with beautiful shapes and shades add to the beauty of the high altitudes. Forty three species of
Rhododendron are found in India out of which thirty six species are confined to Sikkim alone. The
Rhododendron is a keystone species in the higher altitudes of Himalayas. It is a sacred plant and has a lot of
social importance attached to it. It also has aesthetic, ethano - medicinal, medicinal, and commercial value. The
plant is also used as fuel wood. Due to anthropogenic activities the natural populations and diversity of
Rhododendron species in the entire Himalayas are gradually diminishing. The major threats to Rhododendron
population and diversity are deforestation, and unsustainable extraction for firewood and incense by local
people. Many species of Rhododendron which are classified as rare or endangered may soon become extinct
and will we lost forever if proper conservation measures are not taken. The present study deals with the study of
diversity of the plant, its conservation status and strategies being used to tackle the situation.
Key Words: Rhododendron, Diversity, Conservation, Sikkim, Himalayas.
Introduction:
The genus Rhododendron, belongs to the family Ericaceae and was founded by Linnaeus [1]. The
word Rhododendron itself is derived by two Greek words rhodon (rose) and dendron (tree) which mean, rose
tree. This genus is represented by 850 species in the world [2], which are mostly distributed at higher elevations
in the Sino-Himalayan region with maximum concentration in Western China [3]. In India, the genus is mostly
confined to the Himalayan region, especially in the Eastern Himalayas. A revision of the genus was carried out
by Cullen [4], Chamberlain [5], Philipson and Philipson [6],Chamberlain and Rae [7], Kron [8] and, Judd and
Kron [9]. Some inventories of the genus were also made by Pradhan [3,10], Ghosh and Samaddar [11],
Bhattacharyya and Sanjappa [12]. Sastry and Hajra [13] and Mao et al. [14] contributed towards the study of
rare and endemic Rhododendrons of India. Extensive study of Rhododendrons of the Sikkim-Himalayan region
were conducted by Pradhan and Lachungpa [15] and Singh et al. [16].
The Indian Himalayas are one of the most recent and fragile mountain regions of the world. They
house a large number of biological species, and harbour one of the biological hot spots of the world. The region
is quite rich in a variety of flora, fauna, human settlements, tribes and culture. Of the estimated 8,000 species of
vascular plants in the Himalayan region, around 3,160 are endemic and 450 species are endangered [17, 18].

This diversity is constantly and increasingly under pressure from anthropogenic activities. The indiscriminate
exploitation of the flora, fauna and other natural resources present in the region, destruction of natural habitats,
spread of non biodegradable organic chemicals, introduction of alien species, have taken a toll on the number of
a number of plant and animal species, which have disappeared completely while others await the same fate [19].
Due to this disappearance of the natural flora and fauna, an alarm has been raised by the enlightened gentry of
the world towards the conservation of the various species present in the world. In this regard, efforts are being
made to conserve the various species and genera endemic to the respective areas. One such plant species
endemic to the Himalayas is the Rhododendron. It is found at high altitudes in Himalayan regions of both the
Western and Eastern Himalayas at very high altitudes. Again in the North - Eastern states a very large number
of species of the plant ( Rhododendron ) are present. Due to human interference the natural populations of
Rhododendrons in the entire Himalaya are gradually diminishing. The major threats to Rhododendrons are
deforestation and unsustainable extraction for firewood and incense by local people. A set of Rhododendrons
which are classified as rare/endangered may be wiped out from the biota in the near future if proper
conservation measures are not made. [20] In the present paper we deal with the diversity and conservation status
of Rhododendron in the state of Sikkim.
Material And Methods:
The present work is based on extensive literature surveys made in the state of Sikkim., Published scientific
papers, monographs, red-list documents, IUCN list, etc. were consulted, for the threat categories. All the taxa
observed and studied, have been meticulously listed in table form with their vital information and their threat
categories have also been listed.
Results and Discussion:
A total of 38 species, 3 subspecies and 2 varieties of Rhododendrons were recorded in the state of Sikkim in the
study, the details are provided in the table. The distribution of species in relation to altitude is as follows: The
maximum numbers of Rhododenrons are present at an altitude of 3001-3500 mts. Minimum number of the
said plant are found at the lower altitudes of 500-1000 mts. and above 5000 mts. The genera was absent at an
altitude of less than 500 mts. Only one species viz., namely R. arboreum Sm. is only found at an altitude of less
than 1000 mts. (800 m onwards). R.nivale Hook. f. was the single species found above the altitude of 5000
mts. The three taxa found endemic to Sikkim were R. candelabrum Hook. f. , R. decipiens Lacait. and R.
sikkimense U. C. Pradhan & S. T. Lachungpa. The following four taxa were found to be endangered, these are
R. leptocarpum Nutt. , R. maddenii Hook. f., R. niveum Hook. f. and R. pumilum Hook. f.. of the observed taxa
Nine were found to be rare which are, R. baileyi Balf. f., R. campylocarpum Hook. f., R. edgeworthii Hook. f.,
R. keysii Nutt., R. maddenii Hook. f., R. papillatum Balf. f. & Copper, R. pendulum Hook. f., R. wightii Hook. f.
and R. xanthostephanum Merrill out of which R. maddenii Hook. f. is both endangered and rare.
The species of Rhododendrons exhibits significant diversity in habit and broad range of distribution
from the altitude of 800-6000 mts. A total of 38 species, 3 subspecies and 2 varieties of Rhododendrons were
recorded in the state of Sikkim in the study. Out of these species, 3 species were endemic to Sikkim; four

species were found to be endangered, while nine species were found to be rare and 23 were not yet evaluated till
the time of study[14,16,20]. On comparison with neighboring states the maximum concentration of species is
observed in Arunachal Pradesh , where 75of the total 87 species of Rhododendron, found in the eastern
Himalayas are observed, after that the states in the order of decreasing diversity of Rhododendron species are
Sikkim, Manipur, Mizoram, Nagaland and Meghalaya. Compared with other Rhododendron rich regions China
has a total of 571 species of Rhododendrons, out of which 409 species are endemic [21]. Countries like
Pakistan, Bhutan, Nepal, etc. are have very little species diversity of Rhododendrons. Considering the altitude
the maximum number of Rhododendron species are present in the altitudes of 3001-3500 mts. These altitudes
are considered as optimum suitable heights for the growth the Rhododendrons and for their conservation and
multiplication.
In the present times, the diversity as well as the number of species of Rhododendrons is adversely
affected due to threats posed by natural calamaties and anthropogenic activities. The rise in population with
demand on land for farming, increased animal husbandry practices, construction of roadways, hydel-power
stations and allied works, army personnel garrisoned at alpine locations and lately the tourist influx have
collectively resulted in the building up of considerable pressure on the availability of Rhododendron species
[20,21]. The major threats to Rhododendrons are deforestation and unsustainable extraction for firewood and
incense by local people. Due to the presence of polyphenols and flavonoids, Rhododendrons make excellent
firewood that burns even under wet conditions. Rhododendron firewood is also being used in the high-altitude
trekking corridor for the purpose of tourism. Some of the species have already become scarce, for example, R.
leptocarpum is endangered and reported to have only 16 surviving individuals at present in the Sikkim [16, 20].
The conservation of Rhododendron species can be done by, the in-situ and ex-situ modes of conservation. In-
situ conservation can be brought about by establishing Rhododendron sanctuaries, Parks, etc. which is being
looked into by the Sikkim government and the Government of India. Two Rhododendron sanctuaries have
already been established at Barsey in West Sikkim and at Shingba in North Sikkim[22]. Other than that various
governmental agencies and NGOs also take up programs from time to time to create awareness for the
environment. Some efforts by Sikkim forest department and Sikkim Rhododendron Society have been made by
fencing the Rhododendron rich sites and declaring them as Rhododendron Sanctuary between Lachung and
Yumthang in the State.
The ex-situ conservation can be made by cultivating Rhododendron species in the gardens and parks under
suitable climatic conditions or by using tissue culture techniques. The plants can be introduced in Botanical
Gardens and Parks also. The species of Rhododendron arboreum is being propagated through cuttings [23,24].
Tissue culture studies of Indian Rhododendrons are also being used to propagate the species of which
Rhododendron maddeni is a successful example [25]. In many countries, these techniques are already in use for
commercial cultivation of Rhododendrons [26-32]. The success of the conservation programs depends as much,
on Govermental agencies, NGOs, as much as on the awareness of local people. But an onus also lies with
common tourist also. It is imperative to educate the local inhabitants and the tourism industry operators and

tourists about the wealth of Rhododendrons and importance towards the conservation of biodiversity in the
region.
References
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nd
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Pogonanthum, Notes from the Royal Botanic Garden Edinburgh Vol. 39, pp. 1-207, (1980).
[5] Chamberlain, D. F. A Revision of Rhododendron II. Subgenus Hymenanthes, Notes from the Royal
Botanic Garden Edinburgh, Vol. 39, pp. 209-486, (1982).
[6] Philipson, W. R. and Philipson, M. N. A Revision of Rhododendron III. Subgenera Azaleastrum,
Mumeazalea, Candidastrum and Therorhodion, Notes from the Royal Botanic Garden Edinburgh, Vol. 44,
pp. 1-23, (1986).
[7] Chamberlain, D.F. and Rae, S. J. A revision of RhododendronIV. Subgenus Tsutsusi, Edinburgh Journal
of Botany, Vol. 47, pp. 89-200,(1990).
[8] Kron, K. A. A Revision of Rhododendron Section Pentanthera,Edinburgh Journal of Botany, Vol. 50, pp.
249-364, (1993).
[9] Judd, W. S. and Kron, K. A. A Revision of RhododendronVI. Subgenus Pentanthera (Sections
Sciadorhodion, Rhodora, and Viscidula), Edinburgh Journal of Botany,Vol. 52, pp. 1-54, (1995).
[10] Pradhan, U.C. A Preliminary Enumeration of Rhododendrons of the Indian regionPart 2, Himalayan
PlantJournal, Vol. 4, No. 11-12, pp. 73-76, (1986).
[11] Ghosh, R.B. and Samaddar, U.P. The Rhododendrons of the North-East India, Journal of Economic and
Taxonomic Botany, Vol. 13, No. 1, pp. 205-220, (1989).
[12] Bhattacharyya, D. and Sanjappa, M. Rhododendrons Habitats in India, Journal of American
Rhododendron Society Vol. 62, No. 1, pp. 14-18, (2008).
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Study, In: S. K. Jain and R. R. Rao, Eds., An Assessment of Threatened Plants of India, BSI, Calcutta,
pp. 222-231, (1983).
[14] Mao, A. A. Singh, K. P. and Hajra, P. K. Rhododendrons, In: N. P. Singh and D. K. Singh, Eds.,
Floristic Diversity and Conservation Strategies in India, BSI, Calcutta, pp. 2167-2202, (2002).
[15] Pradhan, U. C. and Lachungpa, S. T. Sikkim Himalayan Rhododendrons, Primulaceae Books,
Kalimpong, (1990).
[16] Singh, K. K., Kumar, S., Rai, L. K. and Krishna, A. P. Rhododendron Conservation in Sikkim Himalaya,
Current Science, Vol. 85, pp. 602-606,(2003)

[17] Singh, D. K. and Hajra, P. K. Floristic Diversity, In: G. S. Gujral and V. Sharma, Eds., Changing
Perspectives of Biodiversity Status in the Himalaya, British Council, New Delhi, (1996).
[18] Samant, S. S., Dhar, U. and Palni, L. M. S. Medicinal Plants of Indian Himalaya: Diversity Distribution
Potential Values, Gyanodaya Prakashan, Nainitial, (1998).
[19] Singh, N. P., Sharma, J. R., Singh, K. P. and V. Mudgal, Species Diversity in Angiosperms, In: N. P.
Singh and D. K. Singh, Eds., Floristic Diversity and Conservation Strategies in India, Botanical Survey
of India, Kolkata, Vol. 5, pp. 1631-1674.( 2002)
[20] Chandra Sekar, K. and Srivastava, S. K. Rhododendrons in Indian Himalayan Region: Diversity and
Conservation American Journal of Plant Sciences, , 1, 131-137, (2010).
[21]Paul, A., Khan, M.L., Arunachalam, A. and Arunachalam, K. Biodiversity and conservation of
rhododendrons in Arunachal Pradesh in the IndoBurma biodiversity hotspot. Current Science 89(4): 623-
634, (2005).
[22] http://nbaindia.in/uploaded/state-wise/sikkim/1.list_Protectedareas_SIKKIM.pdf
[23] Thakur, P., Sharma, Y. D., Kashyap B. and Thakur, A. Vegetative propagation of native ornamentals of
Himachal Pradesh in India, Abstract of XXVII International Horticultural CongressIHC2006, Himachal
Pradesh, (2006).
[24] Singh, K. K., Kumar ,S. and Shanti, R. Raising Planting Materials of Sikkim Himalayan Rhododendron
through Vegetative Propagation Using Air-Wet Technique, Journal of American Rhododendron Society,
Vol. 62, pp. 136-138, (2008).
[25] Singh, K. K. and Gurung, B. In vitro Propagation of R.maddeni Hook. f. an Endangered Rhododendron
Species of Sikkim Himalaya, Notulae Botanicae Horti Agrobotanici Cluj-Napoca, Vol. 37, No. 1, pp. 79-
83, (2009).
[26] Lloyd, G. and McCown, B. Community-Feasible Micropropagation of Mountain Laurel, Kalmia latifolia,
by Use of Shoot Tip Culture, Proceedings of International Plant Propagation Society, Vol. 30, pp. 421-
427, (1981).
[27] W. C. Anderson, A revised Medium for Shoot Proliferation of Rhododendron, Journal of American
Society & Horticultural Science, Vol. 109, pp. 343-347, (1984).
[28] Douglas, G. C. Propagation of Eight Cultivars of Rhododendron in vitro Using Agar-Solidified and
Liquid Media and Direct Rooting of Shoots in vivo, Scientia Horticulturae,Vol. 24, pp. 337-347, (1984).
[29] McCown, B.H. and Lloyd, G.B. A Survey of the Response of Rhododendrons to in vitro Cultures, Plant
cell tissue and organ culture, Vol. 2, pp. 77-85, (1985).
[30] Briggs, B. A., McCulloch, S. M. and Caton, L. A. In vitropropagation of Rhododendron, Acta
Horticulturae, Vol.364, pp. 21-26, (1994).
[31] Evers, P. W., Donkers, J., Prat, A. and Vermeer, E. Micropropagation of forest trees through tissue
culture, Centre for Agricultural Publishing and Documentation, Wageningen, (1988).

[32] R. Almeida, S. Gonalves and A. Romano, In vitro Micropropagation of Endangered Rhododendron

ponticum L.subsp. baeticum (Boissier & Reuter) Handel-Mazzetti, Biodiversity and Conservation Vol. 14,
No. 5, pp. 1059-1069, (2005).
Table 1: Rhododendrons of Sikkim
S. NAME OF THE TAXA ALTITUDE STAT

No. (MTS) US
1 R. anthopogon D. Don 3350-5000 N.E.
2 R. anthopogon D. Don subsp. hypenanthum (Balf. f.) J. Cullen 3350-5000 N.E.
3 R. arboreum Sm. 800-3000 N.E.
4 R. arboreum Sm. subsp. Cinnamomeum (Wall. ex G. Don) Tagg c. 2500 N.E.
5 R. arboreum Sm. var. roseum Lindl. 2500-3600 N.E.
6 R. baileyi Balf. f. 3000-4000 R.A.1
7 R. barbatum G. Don 2500-3700 N.E.
8 R. camelliaeflorum Hook. f. 2700-4000 N.E.
9 R. campanulatum D. Don 2500-4300 N.E.
10 R. campanulatum D. Don subsp. aeruginosum (Hook. f.) D. F. Chamb. 4500-5000 N.E.
11 R. campanulatum D. Don var. wallichii Hook. f. 4000 N.E.
12 R. campylocarpum Hook. f. 3300-4300 R.A.1
13 R. candelabrum Hook. f. 3600-4300 E.N.1
14 R. ciliatum Hook. f. 2700-3400 N.E.
15 R. dalhousiae Hook. f. 1800-2300 N.E.
16 R. decipiens Lacait. 2500-3000 E.N.1
17 R. edgeworthii Hook. f. 2100-3300 R.A.1
18 R. falconeri Hook. f. 2100-4000 N.E.
19 R. glaucophyllum Rehder 3080-3700 N.E.
20 R. grande Wight 2160-3385 N.E.
21 R. griffithianum Wight 2160-2770 N.E.
22 R. hodgsonii Hook. f. 3080-3690 N.E.
23 R. keysii Nutt. 2440-3650 R.A.1
24 R. lanatum Hook. f. 3080-4000 N.E.
25 R. lepidotum Wall. ex D. Don 2160-4620 N.E.
26 R. leptocarpum Nutt. 2300-4310 E.D.2
27 R. lindleyi T. Moore 1850-3080 N.E.
28 R. maddenii Hook. f. 2400-3650 RA1,
ED2
29 R. nivale Hook. f. 4000-6000 NE

30 R. niveum Hook. f. 3080-3700 ED2

31 R. papillatum Balf. f. & Copper 1800-3300 RA1
32 R. pendulum Hook. f. 2270-3650 RA1
33 R. pumilum Hook. f. 3500-4500 ED2
34 R. setosum D. Don 2160-4950 NE
35 R. sikkimense U. C. Pradhan & S. T. Lachungpa 3700 EN1
36 R. smithii Nutt. 2160-3700 NE
37 R. thomsonii Hook. f. 3390-4000 NE
38 R. triflorum Hook. f. 2160-2930 NE
39 R. vaccinioides Hook. f. 1850-3700 NE
40 R. virgatum Hook. f. 2160-2770 NE
41 R. wallichii Hook. f. 4000-4500 NE
42 R. wightii Hook. f. 3050-4310 RA2
43 R. xanthostephanum Merrill 1500-3000 RA1
1 - Mao et al., 2002; 2 Singh et al., 2003; ED - Endangered; EN Endemic; NE Not Evaluated; RA Rare;

Changes in Micromorphology of Plant Sida veronicaefolia in Response to Air Pollution

Stress in Meerut City
Shiv Kumari* and Ila Prakash

Department of Botany
D.N. College Meerut, (U.P.) India
drshivasharma85@gmail.com
ABSTRACT
The race for rapid development has resulted in unscrupulous exploitation of natural resources. This has disturbed
the delicate ecological balance between living and nonliving components of the biosphere. Of the various changes
disturbing the ecological balance most originate from industries. Emissions from various industrial units, thermal
power plants, etc. have immensely contributed to environmental pollution. Entire global biota (flora and fauna) has
become vulnerable to this ever increasing menace. Present study deals with the observation made on the plants on
roadside in comparison to plants in controlled area. On roadside plants having heavy auto-exhaust pollution load.
Present studies were made on Sida veronicaefolia taken from Garh road, Railway road, Delhi road and University
road. These changes have been worked out on the basis of percentage of reduction in various parameters in tested
plants. Due to high concentration of automobile pollution, the number of stomata, stomatal index and stomatal
density were reduced and Number of epidermal cells and epidermal density were increased. Variation in stomatal
index and stomatal density on the adaxial and abaxial surface has been observed in these plants.
Key Words: Stomatal Density, Stomatal index, epidermal density, Vehicular exhaust pollution.
INTRODUCTION:
Our environment is a complex mixture of a number of constituents like air, water, soil, plants and animals, all of
which maintain a dynamic inter-relationship and interdependence. The earth is the only planet known in the entire
universe capable of supporting life which is due to its unique environment. Any undesirable change in the
environment, which may be due to addition of unwanted substances results in atmospheric pollution and disturbs the
normal functioning of the ecosystem. Zielinska et al., 2004 reported that the composition of emissions from
automobiles highly depend on the fuel type, the state of vehicular maintenance and ambient conditions. Sarkar et al.
1986 observed the high effects of automobile exhaust pollution on Clerodendron incerme, Solanum torum and
Calotropis procera along a road carring dense traffic. And found visible injury, necrosis, chlorosis and reduction in
leaf area due to air pollution. Decrease in stomatal frequency and occurence of aborted stomata were reported in the
leaves of some woody perennials as a result of air pollution (Chattopadhyay, 1996 ). Similarly, decrease in stomatal
index, density and coverage area was observed in Dahlia and Tagetus (Dhaka, 1999; Prakash et al., 2008;
Helianthus annus (Goswami, 2002; Zea mays (Jeyakumar et al., 2003) and Mammillaria fragilis (Joshi et al.,2004);
in sunflower and napier grass (Marie and Romeo, 2008); and Miyazawa et al. 2006.
MATERIAL AND METHOD:

Leaves collected from different sites and studied. For the study of stomata parameters, the replica technique
(conservative facsimile technique) was adoptd (Prakash and Kumar, 1995). In this method, an adhesive such as
quick-fix was applied on the surface of the leaf. It was allowed to dry for a few seconds and then a cellotape was
placed on the leaf surface. The tape was then pulled off and the impression of leaf epidermis than was obtained
under the microscope. Stomatal index was determined as follows.
Number of stomata
Stomatal density

Field area mm 2
Number of epidermal cells
Epidermal density 100

Field area mm2
S
Stomatal index 100 (Salisbury, 1927)
E S
Where,
S = no. of stomata in microscope field area-1,
E = no. of epidermal cells microscopic field area-1
RESULTS & DISCUSSION:
The reduction in the number of stomata on adaxial and abaxial surface was Sida veronicaefolia 32.0% and 15.75 at
Delhi road (Table: 1, 2). The number of epidermal cells however recorded at increase. The number of epidermal
cells was higher at high polluted sites. In Sida veronicaefolia plant the stomatal index at Delhi road was 15.3783 on
adaxial surface and 20.6632 on abaxial surface (Table 1, 3).
Stomatal density also recorded a decrease at different polluted sites. In In Sida veronicaefolia the stomatal density
was 120.3966/ 180.594 on adaxial/ abaxial surface at Delhi road and 173.5127/ 254.957 on adaxial/ abaxial surface
at University road (Table 1, 3). Number of epidermal cells got increased in number due to reduction in number
stomata was observed with an increase in concentration of pollutants at polluted site. As a result of reduction in the
number of stomata, the epidermal density was increased. In Sida veronicaefolia the epidermal density was recorded
as 662.1813 mm-2 / 686.968 mm-2 on adaxial / abaxial surfaces at Delhi road. In Sida veronicaefolia, the stomatal
coverage area is 2521.2464/ 3271.9546 on adaxial/ abaxial surface at Delhi road and 5545.3257/ 7963.8810 on
adaxial/ abaxial surface at University road (Tables 2, 4). In Sida veronicaefolia the number of trichomes recorded an
increase of 3.25/1.5 on adaxial/ abaxial surface at Delhi road and 1.0/0.75 on adaxial/ abaxial at University road. T
he results were statistically analyzed and interpreted by three way ANOVA. All the data were subjected to statistical
analysis to find out Critical Difference at (CD) 5% and 1% level (Fisher, 1951) is superscripted with single star (*)
and double star (**)respectively.

Effect of pollution on various other parameters of plants growth growing under polluted condition might be
attributed to the disturbances caused in habitat due to pollutants emitted by diesel engines of locomotives, deposition
of smoke, carbon, dust and gaseous pollutant, etc. present study number of stomata decreases with increase in
pollution on roadside. As the number of stomata decreases, consequently the number of epidermal cells increases.
Decrease in stomata could be regarded as an adaptive feature developed by plants in order to cope up with the effect
of the gaseous pollutant which enters the leaf, injuries the tissue and causes death (Chattopadhyay, 1996); Marie et.
al., 2008; Prakash et al., 2008. Gaseous pollutant enters the leaves through stomata following the same diffusion
pathway as CO2. Stomatal opening is checked by the entry of gases in leaf. The pollutants after entering the leaf
dissolve in the apoplastic water to produce mainly sulphite and bisulphite ions as (SO 3-2, HSO3-) which are toxic at
high concentrations but at low concentrations are effectively detoxified by plants to sulphate ions which then work
as sulphur source for the plant. Kulshreshtha et. al., 2005); Salsbury 2006; Sunstar.com 2009.
Urban areas are characterized by higher concentration of SO2, hence the plants in these areas cannot be
detoxified rapidly and adapt themselves by following some line of defense against SO 2 stress. It may include
stomotal closure and reduction in number of stomata was observed at all sites, so stomatal index and stomatal
Density was also reduced accordingly. The number of trichomes was high at upper surface and low at lower surface.
Higher number of trichomes was found on the upper surface of the leaf in order to keep the pollutants away from the
leaf surface.The gradual increase in trichome number may be due to the tendency of plants to increase the resistance
to air pollutants as the trichomes may increase boundary layer resistance and hence reducing the gaseous diffusion
into leaves. The effect was more severe on adaxial surface than on abaxial surface. This might be due to direct
exposure of the adaxial surface to the pollutant. As the number of stomata decreases, consequently number of
epidermal cells increases. This resulted in increase in epidermal density.
CONCLUSION:
Present paper shows an overview that high concentration of pollutants was observed at Delhi road and low at
University and moderate at Garh road and Railway road in comparison to control site. The adverse effects of air
pollution on plants were greater in the area receiving higher pollution load and vice-versa. Due to higher level of
dust on upper surface of plant, stomatal process and food synthesis in leaves were highly affected. According to the
study it can be concluded that Delhi road was most polluted site followed by Garh road, Railway road, and
University road in comparison to control. It can be concluded that the high level of pollution in busy roadsites
hampers the plant growth and development to an extent that the plant is disturbed.
ACKNOWLEDGEMENT:
The author is thankful to centre for the study of D.N.College laboratory, Meerut, U.P. India to providing me all the
facilities for experimental work. author is also thankful to Dr. (Smt) Ila Prakash for her guidance and supervision for
this work.
REFERENCES:

1) Chattopadhyay, S.P. Leaf surface effects of air pollution on certain tree species in Calcutta, Adv Plant Sci.,
9(1), 1-4 (1996).
2) Dhaka, R. Impact of air pollution on some plants. A ph.D. Thesis, C.C.S. Uni. Meerut India, (1999).
3) Fisher, R.A. The Design of Experiments. Six Eds. Oliver and Boyd, London, (1951).
4) Goswami, R. Toxicity of air pollution to plants. A Ph.D Thesis. C.C.S.Univ., Meerut India, (2002).
5) Jeyakumar, M., Jayabalan, N. and Arockiasamy, D.I. Effect of sulphur dioxide on maize (Zea mays L.var.
Co-1) seedling at lethal dose 50. Physiol. Mol Biol Plants 9 (1), 147-151 (2003).
6) Joshi, R. Choudhary, M., Tyagi, K. and G. Prakash Experimental analysis on stomatal behaviour in cacti
exposed to sulphur dioxide pollution. Prog Agric 4(1), 22-24 (2004).
7) Kulshreshtha, K., K. Srivastava and K. J. Ahmed Effect of automobile exhaust pollution on leaf surface
structures of Calotropis procera L. and Nerium indicum L. Feddes repertorium, Environ. Sci. Hlth. Part A,
Environ. Sci and Egg. (105), 185- 189 (2005).
8) Marie C.G. Duldulao and Romeo, A. Gomez Effect of vehicular emission on morphological characteristics
of young and mature leaves of sunflower and Napier grass. Benguet State University, Research Journal,
Volume (xvi), 142-151 (2008).
9) Miyazawa, S., Livingston, N. J. and Turpin, D. H. Stomatal development in new leaves is related to the
stomatal conductance of mature leaves in Populas trichocarpa. J. of Experimental Botany, 57 (2), 373-380
(2006).
10) Prakash., Joshi, C. and Chauhan, A. Performance of locally grown rice plants (Oryza sativa) exposed to air
pollutants in a rapidly growing industrial area of Haridwar. Life Science Journal. 5(3), 57-61 (2008).
11) Prakash, G. and Kumar, V. Stomatal study on defense organs by conservative facsimile method. J. Ind. Bot.
Soc. (74), 263-268 (1995).
12) Prakash, G., Poonia, S., Sharma, S., Sangita and Dhaka, R. Stomatal response to sulphur dioxide exposure
in Dahlia variabilis L. and Tagetes patula L. Role of Biological Sciences in New Millennium 39-44 (2001).
13) Salisbury, E.J. On the causes and ecological significance of stomatal frequency with special reference to
woodland flora. Phill. Trans, R. Soc. B. Vol. (216), PP. 1-65 (1927, 2006 a).
14) Sarkar, R.K., Banerjee, A. and Mukherji, S. Acute ration of peroxidase and catalase activities in leaves of
wild dicotyledonous plants as an indication of automobile exhaust pollution. Environ. Pollut (42), 289-295
(1986).
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50 (2004).
TABLES:
Table. 1 Stomatal response in terms of no. of epidermal cells, no. of stomata, epidermal density (mm-2), stomatal
-2
density (mm ) and stomatal index on adaxial surface in Sida veronicaefolia plant.

Different Study Sites Statistical Value
Attribute
Railway University
Control Delhi Road Garh Road CD 5% CD 1%
Road Road
No. of epidermal 27.0 46.75 40.25 39.25 38.5
cells
+ 1.1547 + 0.9574 + 1.5 + 2.5 + 1.732
12.5 8.5 9.25 10.75 12.25

No. of stomata
+ 0.5773 + 0.5773 + 0.5 + 0.5 + 0.5
382.4362 662.1813** 570.1133** 555.9490** 545.3257**

Epidermal
47.756 113.830
density (mm-2) + 16.355 + 13.5612 + 21.246 + 35.4107 + 24.533
Stomatal density 177.0538 120.3966* 131.01983* 152.2662* 173.5127

23.878 56.916
(mm-2 ) + 8.1777 + 8.177 + 7.0821 + 7.0821 + 7.0821
31.6548 15.3783** 18.6826** 21.5339** 24.1515*

Stomatal index 3.976 9.478
+ 1.36190 + 0.8467 + 0.5084 + 1.3383 + 0.9969
Table 2. Stomatal response in terms of length and breadth of stomata and stomatal coverage area (mm -2) on adaxial
surface in Sida veronicaefolia plant.
Different Study Sites Statistical value
Attribute
Railway University
Road Road
Length of 210.0 157.5 180.0 195.0 202.5
stomata(
+ 24.494 + 15.0 + 24.494 + 17.320 + 15.0
Breadth of 150.0 120.0 127.5 135.0 142.5
+ 24.494 + 0.0 + 15.0 + 17.320 + 15.0

Stomatal
6157.932 2521.2464* 3349.8583* 4472.3796 5545.3257
coverage area
2755.393 6567.650
+ 943.628 + 187.9107 + 665.7223 + 849.730 + 561.0567
(mm-2 )
Table. 3 Stomatal response in terms of no. of epidermal cells, no. of stomata, epidermal density (mm-2), stomatal density
(mm-2) and stomatal index on abaxial surface in Sida veronicaefolia plant.
Different Study Sites Statistical Value
Attribute
Railway University
Road Road
No. of epidermal 40.25 48.5 46.0 44.0 40.75
cells
+ 1.258 + 0.5773 + 0.816 + 0.816 + 1.5
20.75 12.75 13.5 15.75 18.0

No. of stomata
+ 0.9575 + 2.2173 + 2.516 + 0.9574 + 1.4142
570.113 686.968** 651.558* 623.229* 577.195

Epidermal density
52.043 124.048
(mm-2 )
+ 17.823 + 8.1777 + 11.565 + 11.565 + 21.246
293.909 180.594** 191.218** 223.087* 254.957

Stomatal density
39.598 94.384
(mm-2 )
+ 13.561 + 31.407 + 35.646 + 13.561 + 20.031
34.008 20.6632** 22.6100** 26.346* 30.631

Stomatal index 1.483 3.535
+ 0.508 + 3.005 + 3.642 + 1.067 + 2.177

-2
Table. 4 Stomatal ) on abaxial surface
in Sida veronicaefolia plant.
Different Study Sites Statistical value
Attribute
Railway University
Road Road
Length of 202.5 127.5 142.5 165.0 187.5
+ 15.0 + 15.0 + 15.0 + 17.320 + 15.0
Breadth of 16.5 127.5 127.5 135.0 150.0
+ 17.320 + 15.0 + 28.722 + 17.320 + 24.494
Stomatal coverage
10949.008 3271.9546* 3788.951* 5541.784* 7963.8810
area
5138.590 12248.148
+ 1759.7914 + 814.991 + 906.035 + 1168.555 + 1550.862
(mm-2 )
32
In vitro study of an endangered Miracle plant: Mandookparni (Centella asiatica (L.)

Urb.)
Shalini Sharma* and Y. Vimala
Department of Botany, C. C. S. University, Meerut
[*shalini2976@gmail.com]
Abstract
Plants have been used as treatments for thousands of years, based on experience and folk remedies and continue
to draw wide attention for their role in the treatment of chronic diseases. Mandookparni (Centella asiatica (L.)
Urb.) or Indian pennywort medicinal herb belongs to family Apiaceae. It is valued in Indian systems of
medicines for improving memory, for treatment of nervine disorders, skin diseases and wound healing. The
bioactive compounds responsible for its medicinal properties are triterpene saponins as Asiaticoside, Asiatic
acid, Madecassocide etc. Rapid urbanization, degradation of plant habitat, ruthless collection of herb and other
anthropological activities have markedly depleted the wild stocks of plant. It has been listed as Threatened plant
species by IUCN and an endangered species. Present study reports the standardized protocol for callus induction
from Centella plant and comparative account of yield of asiaticoside from this callus by treating with various
growth regulators, salt and colchicine treatment. Present study reports significant amount of yield of asiaticoside
from callus with that of 60 days old leaves of plants, so that overexploitation of this herb can be avoided.
Key words: Mandookparni, Callus induction, Salt treatment, Colchicine treatment, Asiaticoside.
Introduction:
Plants have been enumerated as an efficient basis of medicine since immemorial past. Drugs based on
the plants are of prime importance for several remedies in traditional and conventional medicine throughout the
world and serve as a substitute for drug supply in modern medicine. Now a days world markets are turning
towards plants as the source of ingredients in manufacturing health care products. Secondary metabolites
obtained from the plants are found to be an important source of various phytochemicals that could be used
directly or as an intermediate for the production of pharmaceuticals. Approximately 80% of the population still
relies on the traditional medicine derived from the plants for health care needs 3-5. Thus the demand for herbal
medicines is continuously increasing day by day due to lesser side effects in comparison to the synthetic drugs.
Unfortunately, in the present day, precipitous economic development and suburbanization resulted in
overexploitation and loss of valuable natural resources, including the medicinally important plants. As a result,
many of the plant species are endangered or threatened with extinction leading to severe depletion of
biodiversity. C. asiatica L. (Fig. 1) is one of the threatened medicinal herbs, generally endemic to Western
Ghats of South India.
The medicinal properties of this plant are due to secondary metabolites triterpenoidsaponins, that can
be defensive substances such as phytoanticipins, antifeedant, attractants, phytoalexins and pheromones. The
triterpene of C. asiatica composed of several compounds which include asiatic acid, madecassic acid,
asiaticoside, madecassoside, brahmic acid, thankunoside, isothankunoside, centelloside, sceffoleoside, madsiatic
acid, centic acid and centillic acid and alkaloid hydrocotylin. In addition to these bioactive components it also
contains high phenolic content which is contributed by the flavonoids such as quercetin, kaempherol, catechin,
33
rutin, apigenin and naringin and sugars, tannins and Vellarine. Among the triterpenes, the most important
biologically active compounds are the Asiatic acid, madecassic acid, asiaticoside and madecassoside.The
development of plant tissue culture technology holds great promise for conservation and enhancement of
valuable medicinal plants
Material and Methods:
(A) Materials:
Plants of Centella were procured from Forest Research Institute, Dehradun. The procured plantlets of Centella
were allowed to multiply in damp and shaded area of garden. Plantlets of equal size and age were selected for
transfer to experimental pots for in vivo propagation. For the present study leaves and petiole of Centella
asiatica were used as experimental materials.
C. asiatica L. is a small stonoliferous perennial creeping aromatic herb belonging to the family
Apiaceae (Umbelliferae). The leaves are 2-5 cm wide, hastate or cordate or palmately lobed or reniform,
arranged in an alternate fashion in form of clusters at stem nodes having long stalk and sheathing leaf bases. The
petiole is long and stipules are small in size.
Fig.1 showing C. asiatica L. (Urb.) plant
C. asiatica L. has been used for several hundred years in folk medicines. In Indian Ayurveda literature,
C. asiaticais considered as one of the recognised drugs used for Rasayana purpose. In Chinese medicine, C.
asiaticais used for treatment of vomiting, epistaxis, urinary calculi, scabies and jaundice. In homeopathic
medicine, it is used for treating ascariasis, elephantiasis and in granular cervicites. Clinical tests have formulated
several benefits of C. asiatica extracts in terms of wound healing, burns and in skin diseases in gastrointestinal
disorders and in treatment of leprosy, lupus, scleroderma, eczema, veins diseases. It is also used for treatment of
psoriasis. It gives protection against diseases by enhancing immunity of the body. The extract of the whole plant
is reported to have anticancerous activity and the methanolic extract of aerial parts of C. asiatica inhibit the
growth of human uterine carcinoma, human gastric carcinoma, and murine melanoma cells in vitro . C. asiatica
is also used as nervine tonic, along with antibacterial, antifeedant and antileptic property. It is also efficient in
promoting fast growth of skin and keratinization. It also possesses anti-inflammatory and memory enhancing
property. It also finds application in controlling anxiety and thereby imparting mental calmness. Wound healing,
memory enhancement, neuroprotective, immunomodulatory, hepatoprotective, cardiovascular, antidepressant
and anticancer.
METHODS:
STANDARDIZATION OF CULTURE ESTABLISHMENT MEDIUM:
34
Chemicals: All the chemicals used for preparation of basal medium as well as for biochemical analyses, were
procured from Qualigens Pvt. Ltd., Mumbai, India and vitamins and plant growth regulators from Merck India
limited and Sigma Chemicals
Preparation of culture media: Standard medium [6] was used as basal medium throughout the present tissue
culture studies. Young leaves and petioles of Centella were surface sterilized with 0.1% HgCl2 and 70% alcohol
before inoculation.
Explant: In Centella young leaves and their petioles were used as explant. The explants were inoculated for
establishing callus culture. The callus induced from different explants was maintained on same medium for
minimum of 2-3 passages and was used for subsequent experiments. The undifferentiated stock calli were
routinely subcultured at an interval of every 4 weeks for subsequent experimentation.
Incubation of cultures: The cultures were incubated in culture room and provided with light of 1400 lux
intensity with a photo period of 16/8 h light/ dark cycles at 25 2oC temperature maintained by automated
photoperiod controlled device and air conditioner or room heater.
Standardization of basal medium: For culture initiation MS medium supplemented with growth regulators
was studied for culture establishment. BAP, Kn individually or in combination with NAA and IBA along with
30 g/l sucrose and 8.0 g/l agar agar were used.
Sub culture procedure : The calli were regularly transferred every 3-4 weeks in their exponential phase of
growth on the fresh media. In subsequent passages, the non-morphogenic callus was transferred on auxin or
cytokinin alone or in combination of both, with three salt treatments (50, 75 & 100 mM NaCl) and without salt
added media (as control). Actively dividing cells of calli were treated with Colchicine (Ctrl and 0.4%) in liquid
medium for 24 hours. Then these were analyzed with their respective controls for growth and type of callus after
fourth week as treated calli showed various growths under different hormonal treatments, salts as well as
colchicine treatment.
PHYSICOCHEMICAL ANALYSIS:
Weight analysis (Fresh weight, Dry weight) and Moisture content- Moisture percent was calculated
according to the following formula: -
Growth Index: Growth index of callus was calculated by the following formula:-
Determination of Total Proteins [2], Reducing, Non-reducing and Total sugar [8], Proline
content [1] and Total Phenolic content [9] was carried out by standard methods mentioned in
parentheses.
35
Extraction Procedure for Asiaticoside and its estimation: Each of the dried 1.0 g samples was
dried and soxhlet extracted in Methanol for 24 hours. Each of the Methanol extracts of various test
samples was separately dried in vacuo [4]and taken up in methanol for further analysis of
Asiaticoside estimation. Further quantitative estimation was done by HPLC. (Kind courtesy of
Natural Remedies Private Ltd., Bangalore).
Every estimation was done in three extractions done in triplicates and subjected to statistical analysis in terms of
mean and standard deviation for testing significance of data.
RESULTS AND DISCUSSION:
As explants, leaves were selected for callus initiation because of the superiority of callus from the same, over
callus obtained from petioles. Callus growth index was best in medium supplemented with combinations of
auxins and cytokinins, instead of when they were used alone. BAP (2.0mg/l)+ NAA (0.1mg/l) supplementation
to MS medium using 7 days old leaves as explant produced superior callus in C.asiatica, that was used for
further studies.
Callus Growth index under 50 to 100 mM salt and 0.5 to 2.0 mg/l concentrations of various Auxin (Fig.1-a,b,c),
Cytokinins (Fig.2-a,b) and their combinations (Auxin+ Cytokinin) supplemented media (Fig.3)
Fig-1(a) Effect of NAA and Salt on

1.5 four weeks old callus of C. asiatica
Growth Index
0.5
0
DW 50mM 75mM 100mM
0.5 NAA 1.0 NAA 2.0 NAA
Fig-1 (b) Effect of 2,4-D and Salt on

1 four weeks old callus of C. asiatica
Growth Index
0.5
0
DW 50mM 75mM 100mM
0.5 2,4-D 1.0 2,4-D 2.0 2,4-D
36
Fig-1 (c) Effect of IAA and Salt on

four weeks old callus of C. asiatica
1.5
Growth Index
0.5
0
DW 50mM 75mM 100mM
0.5 IAA 1.0 IAA 2.0 IAA
Fig-2 (a) Effect of BAP and Salt on

1.5
Growth Index
0.5
0
DW 50mM 75mM 100mM
0.5 BAP 1.0 BAP 2.0 BAP
Fig-2 (b) Effect of Kinetin and Salt on

1.5
Growth Index
0.5
0
DW 50mM 75mM 100mM
0.5 Kn 1.0 Kn 2.0 Kn
Fig-3 Effect of Kinetin and Salt on

2 four weeks old callus of C. asiatica
Growth Index
0
DW 50mM 75mM 100mM
0.5 2,4D+ 1.0 2,4D+ 2.0 2,4D+ 2 BAP+
0.5Kn 0.5Kn 0.5Kn 0.1NAA
High protein content in callus of C.asiatica indicates its good nutritional value that is much higher than 60 days
old leaves grown in vivo. Callus is able to maintain good growth index even in salt treatment by accumulating
Osmoprotectants as proline as well as phenolics in it as shown in Table- 1. With Salt stress Proline content
significantly increased in callus in comparison to control (DW). Phenolic content also significantly increased
with salinity stress as well as in colchicines treated callus. This indicates that callus has significant antioxidant
activity which contributes towards its medicinal and nutritional value for human beings, yet not as good as
found in in vivo grown 60 days old leaves. Dietary antioxidant supplementation is a promising mean to
strengthen the antioxidant defense and repair systems [5]. However, antioxidants from natural sources are of
37
great value as most commonly used synthetic antioxidants (e.g. butylatedhydoxyanisole,

butylatedhydoxytoluene and propylgallate) have health hazardous side effects like liver damage and
carcinogenesis. Several reasons may explain the differences in concentration of bioactive compounds; genetic
diversity, environmental conditions, and nutrients present in the soil/culture medium may alter the metabolic
activity of plants [7]. Alternatively, climatic conditions and bioavailability of nutrients, may alter the growth
rate of explants, and thereby plant metabolic activity.
Table:1
S. Proline (mg Phenolics ( mg trans Protein (mg

No. Media for Proline/ g f wt) cinnamic acid eq g/ f casein eq /g fwt)
growth wt)
1. 2BAP+0.1NAA 0.017 1.17 20.98
(DW)
2. 2BAP+0.1NAA
(50Mm) 0.036 1.466 21.46
3. 2BAP+0.1NAA
(75mM) 0.026 1.254 9.19
4. 2BAP+0.1NAA
(100mM) 0.025 0.868 12.96
5. 0.4% 0.007 1.31 10.49
Colchicines
treated callus
6. Soil grown 60
days old leaves 0.004 4.64 16.36
With increasing salt stress callus accumulated osmoprotectants (Table -2) that is apparent in the form
of Total sugar content accumulated in callus. Non reducing Sugars also accumulated in callus in 100mM NaCl
treatment in comparison to control that signifies increased level of triterpenoid saponins that are responsible for
medicinal properties of C. asiatica plant. This fact is confirmed by the Asiaticoside content measured by HPLC,
where 100mM NaCl treated callus accumulated, yet remained lower in comparison to in vivo grown 60 days old
leaves. In comparison to salt treated calli, Colchicine treatment of callus for Asiaticoside content proved to be
beneficial as it showed accumulation of sugars as well as asiaticoside content equitable with soil grown 60 days
old leaves (in vivo). This may be due to the stress experienced by the callus and tolerance response exhibited.
Climatic conditions, bioavailability of nutrients, may alter the growth rate of explants, and thereby plant
metabolic activity [3].
Table-2
S.No. TS RS NRS Asiaticoside %

Media used (mg glucose eq g/ dw SD)
38
1 2BAP+0.1NAA (DW) treated callus 0.19 0.155 0.035 0.0045
2 2BAP+0.1NAA (50Mm) treated callus 0.144 0.122 0.022 -

3 2BAP+0.1NAA (75mM) treated callus 0.154 0.131 0.023 -
2BAP+0.1NAA (100mM) treated callus 0.237 0.186 0.051 0.07
4 0.4% Colchicines treated callus 2.05 1.114 0.91 0.107
5 Soil grown 60 days old leaves (in vivo) 0.837 0.031 0.806 0.118
CONCLUSION:
Thus, it can be stated that research on plants have been enthralled throughout the world to emblematize the
tremendous potential of medicinal plants in recent years. Due to its wide prospects and potential, its demand has
led to a quantum increase which plays a vital role in alleviating human sufferings due to lesser side effects, easy
availability at affordable cost and being non-narcotic. Sometimes, it is the only source of health care available to
the poor. C. asiatica L. (Urb.) is one such medicinally important herb with well known biological activities in
terms of immunomodulatory, memory enhancer, antidepressant, etc., proved by clinical studies. Thus, present in
vitro studies of C. asiatica provide a promising strategy to conserve the green cover of Mandookparni, without
overexploiting it and providing callus with medicinal, antioxidant activity, nutritional value as good as given by
soil grown 60 days old leaves. However, further research on C. asiatica must be explicit in terms of exploring
its immense potential as nutraceutical. In addition to this, studies should be premeditated regarding the
investigation of underutilized green leafy vegetables, substantial to permeate nutritional ailments.
ACKNOWLEDGEMENT:
Authors are thankful to Late Professor C.M. Govil. and Prof A. K. Srivastava for their valuable
advice. One of the authors (Shalini Sharma) extends her gratitude to C.S.I.R. for the award of Senior Research
Fellowship.
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utilizing the principles of dye binding. Annal.Biochem.72 : 248-254(1976).
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Solvent PolarityJournal of Pharmacognosy and Phytochemistry, 1(6): 27(2013).
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39
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40
Role of Nanotechnology in Pollution Control: Review
Shalini Singh
Department of Zoology, M.K.R. Govt. Degree College, Ghaziabad
shalinsin@gmail.com
Human activity and industrialization leads to the environment filled with different kinds of pollutants. Air is
filled with cabonmonooxide (CO), Cholofluorocarbons (CFC), heavy metals (lead, arsenic, chromium,
cadmium, mercury, zinc), hydrocarbons, nitrogen oxide, sulphur oxide and particulates. Water pollution is
caused by numerous factors including sewage, oils spills, leaking of fertilizers, herbicides and pesticides from
land etc. Contaminants are most often measured in parts per million (ppm) or parts per billion (ppb) and their
toxicity defined by a toxic level. The toxic level for arsenic, for instance is 10 ppm in soil whereas for mercury
is 0.002 ppm in water. Therefore, very low concentrations of a specific contaminant can be toxic. There is need
for technologies are capable of monitoring recognizing and ideally treating such small amount of contaminants
in air, water and soil. Environmental nanotechnology is considered to play a key role in the shaping of current
environmental engineering and science. The nanotechnological applications and products can lead to a cleaner
and healthier environment [1]. Maintaining and re-improving the quality of water, air and soil, so that the Earth
will be able to support human and other life sustainably, are one of the great challenges of our time.
Nanotechnology can play a vital role in providing clean air, water and soil in an efficient and cheap way [2].
Nanoscience allows designing and manipulating materials at the atomic and molecular level.
Nanomaterials can be fabricated with specific properties that can recognize a particular pollutant within a
mixture. The small size of nonmaterials together with their high surface to volume ratio can lead to very
sensitive detection. These properties will allow developing highly miniaturize, accurate and sensitive pollution-
monitoring devices (nano-sensors). Nanomaterials can also be engineered to actively interact with a pollutant
and decompose it in less toxic species. Thus, in the future nanotechnology could be used not only for detecting
contaminated sites but also treating them.
POLLUTION DETECTION AND SENSING

Fortification of the human health and protection of the environment requires the rapid, sensitive
detection of pollutants and pathogens with molecular precision. Sensors are needed for in situ detection, as
miniaturized portable devices, and as remote sensors, for the real-time monitoring of large areas in the field. A
sensor is a device built to detect a specific biological or chemical compound, usually producing a digital
electronic signal upon detection. Sensors are now used for the identification of toxic chemical compounds at
ultra low levels (ppm and ppb) in industrial products, chemical substances, water, air and soil samples, or in
biological systems. Nanotechnology can improve current sensing technology in various ways. First, by using
nanomaterials with specific chemical and biological properties, the sensor selectivity can be improved, thus
allowing isolating a specific chemical or biological compound with little interference. Hence, the accuracy of
the sensors is improved. As with other nano-engineered products discussed in this document, the high surface-
to-volume ratio of nanomaterials increases the surface area available for detection, which in turn has a positive
effect on the limit of detection of the sensor, therefore improving the sensitivity of the device. Scaling down
41
using nanomaterials allows packing more detection sites in the same device, thus allowing the detection of
multiple analytes. This scaling-down capability, together with the high specificity of the detection sites
obtainable using nanotechnology, will allow the fabrication of super-small multiplex sensors, this way
lowering the cost of the analysis and reduce the number of devices needed to perform the analysis with an
economic benefit. Advancements in the field of nanoelectronics will also allow the fabrication of nanosensors
capable of continuous, real time monitoring [3].
Various nanostructured materials have been explored for their use in sensors for the detection of
different compounds [4]. An example is silver nanoparticle array membranes that can be used as flow-through
Raman scattering sensors for water quality monitoring [5]. The particular properties of carbon nanotubes
(CNTs) make them very attractive for the fabrication of nanoscale chemical sensors and especially for
electrochemical sensors [6-9]. A majority of sensors described so far use CNTs as a building block. Upon
exposure to gases such as NO2, NH3 or O3, the electrical resistance of CNTs changes dramatically, induced by
charge transfer with the gas molecules or due to physical adsorption [10, 11]. The possibility of a bottom-up
approach makes the fabrication compatible with silicon microfabrication processes. The sensor is made of an
array of electrode pairs fabricated on a silicon chip and separated by few nanometres. When the electrodes are
exposed to a solution of water containing metal ions, these deposit inside the nano-gap in between the
electrodes. Once the deposited metal bridges the gap a jump in conductance between the electrodes is
registered. The size of the gap, being only few nanometres, allows the detection of a very low concentration of
metal ions. This type of sensor is called nanocontact sensor. [12]. The connection of CNTs with enzymes
establishes a fast electron transfer from the active site of the enzyme through the CNT to an electrode, in many
cases enhancing the electrochemical activity of the biomolecules [8]. In order to take advantage of the properties
of CNTs, they need to be properly functionalized and immobilized. CNT sensors have been developed for
glucose, ethanol, sulfide and sequence-specific DNA analysis [8]. Trace analysis of organic compounds, e.g. for
the drug fluphenazine, has also been reported [13]. Nanoimmunomagnetic labeling using magnetic nanoparticles
coated with antibodies specific to a target bacterium have been shown to be useful for the rapid detection of
bacteria in complex matrices [14]. Materials that are more environment-friendly fabricated using
nanotechnology include biodegradable elf-cleaning glasses, such as Activ Glass [15], the glass is composed
of a special coating made of nanocrystals of TiO2 which, once exposed to daylight, reacts in two ways. First, it
breaks down any organic dirt deposits on the glass and second, when exposed to water, it allows rain to 'sheet'
down the glass easily and washes the loosened dirt away.
NANOCATALSIS
A catalyst is a substance that increases a chemical reaction rate without being consumed or chemically
altered. One of the most important properties of a catalyst is its active surface where the reaction takes place.
The active surface increases when the size of the catalysts is decreased . The higher is the catalysts active
surface, the greater is the reaction efficiency. Also, research has shown that the spatial organization of the active
sites in a catalyst is important as well [16]. Both properties (nanoparticle size and molecular
structure/distribution) can be controlled using nanotechnology. In the environmental field, nanocatalysis is being
investigated for desulphurizing fuels, with the aim of developing clean fuels containing very low sulphur
products (produced in the fuel during its refining process and responsible for generating sulphuric acid upon fuel
42
combustion).
Another area where nanotechnology is making a contribution is the development of fertilizers and
wood treatment products that are more stable and leach less into the environment. For instance, researchers at
the Michigan State University have incorporated biocides for wood treatment inside polymeric nanoparticles.
Their small size allows them to efficiently travel inside the very fine, sieve-like structure of wood. At the same
time, the biocide, being safely trapped inside a nanoshell, is protected from leach and random degradative
processes [17].
GREEN MANUFACTURING
Manufacturing processes are always accompanied by the production of diverse waste products, many
of which pose a threat to the environment and thus need to be removed and treated. Green manufacturing
includes the development of new chemical and industrial procedures (for instance water-based rather the
solvent-based processes); reduction in the use of unsafe compounds (such as metals); development of green
chemicals that are more environment-compatible; and efficient use of energy. In terms of its application to the
reduction of manufacturing waste, nanotechnology can contribute in two ways: by directing the manufacturing
to be more controlled and efficient, and by using nanomaterials (such as catalysts) that can raise the
manufacturing efficiency while reducing or eliminating the use of toxic materials. Overall, nanotechnology has
the potential of making industrial processes more efficient in terms of energy usage and material usage, while
minimizing the production of toxic wastes. The application of green nanotechnology [18] to manufacturing
includes bottom-up, atomic-level synthesis for developing improved catalysts; inserting information into
molecules to build new materials (such as DNA) through highly specific synthetic routes; scaling down material
usage during chemical reaction by using nanoscale reactors; and improving manufacturing to require less energy
and less toxic materials.
An example of green nanotechnology is the development of aqueous-based microemulsions to be
used in alternative to volatile organic compounds (VOCs) in the cleaning industry. These toxic and potentially
carcinogenic compounds, such as chloroform, hexane, percholoroethylene, are conventionally used in the
cleaning industry (like the textile industry) as well as in the oil extraction industry. Microemulsions contain
nano-sized aggregates that can be used as receptors for extracting specific molecules at a nanoscale level.
Other example is microemulsions having water-attractive and water-repellent linkers inserted between the
head and tail parts of a surfactant molecule [19]. The result is a surfactant that has a very low interfacial tension
with a wide range of oils. When tested for cleaning textiles from motor oil residues, as well as for extracting
edible oil from oilseeds, the microemulsions were found to be very competitive with conventionally used VOCs,
both in terms of extraction yield and simplicity of the process.
Nanowires of semiconductors such as silicon has established knowledge for the chemical modification
of their surface. Boron-doped silicon nanowires (SiNWs) have been used for the sensitive real-time electrical
detection of proteins, antibodies the metabolic indicator calcium[20, 21] and glucose in water [22]. The small
size and the capability of these semiconductor nanowires to detect in real-time a wide range of analytes could be
used for developing sensors for detecting pathogens, chemical and biological agents in water, air and food.
Nanotechnologys potential and promise have steadily been growing throughout the years. The world is
quickly accepting and adapting to this new addition to the scientific toolbox. Although there are many obstacles
43
to overcome in implementing this technology for pollution control, science is constantly refining, developing,
and making breakthroughs.
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(2005)
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nanotubes/(3-mercaptopropyl) trimethoxysilane bilayer modified gold electrodes, Talanta 64 (2), 380-386
(2004)
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fluids, Environ. Eng. Sci. 24 (1), 58-72 (2007)
15. Activ Glass, Pilkington, www.pilkington.com
16. Gemming S. and Seifert G. Catalysts on the edge, Nature 2, 21-22 (2007)
17. Liu Y., Yan L., Heiden P. and Laks P. Use of nanoparticles for controlled release of biocides in solid wood,
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44
18.For a review on Green nanotechnology covering definition, concepts and applications see K. Schmidt,
Green Nanotechnology (PEN8), Woodrow Wilson Center International Center for Scholars, free to download
from www.nanotechproject.org/reports .
19. Acosta E.J. , Nguyen T. , Witthayapanyanon A. , Harwell J. H., and Sabatini D.A. Linker-based bio-
compatible microemulsions, Environ. Sci.Tecnol. 39 (5), 1275-1282 (2005)
20. Cui Y., Park H. and Lieber C.M. Nanowire nanosensors for highly sensitive and selective detection of
biological and chemical species, Science 293 (5533), 1289-1292 (2001)
21.Patolsky F. and LieberC.M. Nanowire nanosensors, Materials Today 8 (5) 20-28(2005)
22 Shao M., Shan Y, Wong N. and Lee S. Silicon nanowire sensors for bioanalytical applications: glucose and
hydrogen peroxide detection, Adv. Func. Mater. 15 (9), 1478-1482 (2005)
45
Identification of Geminivirus in Cowpea (Vigna unguiculata) Through Amplification of

Selective DNA Fragment Using Degenerate Primers
Shail Pande
Mahatma Gandhi P G College, Gorakhpur
ABSTRACT
Mostly Gemini viruses have been characterized by symptomatology and host range tests,reliance on symptoms
only; may impede diagnosis, as symptoms are affected by several variables such as virus strain, the growth stage
of the plant at the time of infection, the plant cultivar and environment conditions.The particles of gemini
viruses occur in plants in only low to moderate concentrations, which can be inadequate for detection by the
conventional serological tests but are adequate for some of the newer, more sensitive methods like polymerase
chain reaction (PCR).DNA probes can be valuable especially those derived from the DNA-2 of bipartite
genomes they are used to detect virus in homologous sequences.PCR is especially useful for detecting gemini
viruses, because of its sensitivity to the viral template in low titers. In addition, taxonomically informati ve
domains may be amplified by PCR. Annealing temperature, type and concentration of PCR additives and MgCl 2
concentration were first adjusted to improve sensitivity and reproducibility. DNA concentration required for
amplification was assessed by detection of the initial and end points of dilution and the intensity of the amplified
bands.
KEYWORDS- Geminivirus, degenerate primer, PCR, DNA dilutions.
INTRODUCTION
Historically, Gemini viruses have been characterized by symptomatology and host range tests. A
reliance on symptom development may impede diagnosis, as symptoms are affected by several variables such as
virus strain, the growth stage of the plant at the time of infection, the plant cultivar and environment conditions
[1]. In general, the particles of gemini viruses occur in plants in only low to moderate concentrations, which can
be inadequate for detection by the conventional serological tests but are adequate for some of the newer, more
sensitive methods for e.g. Immunosorbent electron microscopy is possible, although it is not straight forward
because the virus particles are disrupted or damaged in some buffers and they show relatively poor contrast in
most electron dense strains. DNA probes can be valuable especially those derived from the DNA-2 of bipartite
genomes they are used to detect virus in homologous sequences [2]. Among several diagnostic tools available,
the polymerase chain reaction (PCR) using degenerate primers has been the most useful for the detection of
Gemini viruses. PCR is especially useful for detecting Gemini viruses, because of its sensitivity to the viral
template in low titers. In addition, taxonomically informative domains may be amplified by PCR and used in
identification and phylogenetic studies [3-4].
MATERIALS & METHODS
Experimental Strategies for PCR -
The isolation of DNA from plant is often a limiting factor; this is because of the special nature of plant
tissue and cells. A tough cell-wall, abundant secondary metabolites, presence of several classes of chemical
compounds with varying properties are the feature unique to plant cells and these have led to the development of
several plant specific DNA isolation procedures. Once the DNA is isolated it can be subjected to PCR analysis.
46
The PCR in general is a technique requiring several rigorous conditions. Thus the experimental strategy for
carrying out studies in case of plants using PCR-based approach is mainly influenced by the PCR-conditions.
As a technique, though PCR is simple, it is also influenced by a number of factors, these include.
(i) Commercial and biological source and quality of the thermostable polymerase.
(ii) Length and base composition of the primers.
(iii) Concentration of the template and primer.
(iv) Concentration of Mg++ions.
(v) Presence or absence of K+ ions.
(vi) Stringency of Primer annealing conditions.
(vii) Thermocycler parameters such as ramping and its duration.
(viii) Cycle parameters such as number and duration of cycles.
(ix) Cleanliness of work, especially the presence or absence of other contaminating DNAs.
Several workers have described the role of above parameters and have also detailed the optimization of
the experimental protocols [5-6].
After the PCR has been carried out, the amplification product can be analysed by gel electrophoresis on
1.4% agarose gel stained with ethidium bromide and visualized by UV lamp. The patterns of bands revealed are
used for comparison amongst healthy and diseased plants. This technology is proving to be invaluable in
pathogen analysis, especially in case of viruses. This technology is being used for detection of pathogens present
in low concentration.
Reaction setup for PCR amplification
DNA isolated was checked on 0.8% agarose get by electrophoresis for quality and yield.
Isolated DNA was given RNAase treatment to remove RNA by treating it with RNAase (100mg/ml
stock - Solution of which 2l was used in 5l of DNA) at 370C for 1 hour.
Yield was again checked on 0.8% agarose by electrophoresis.
Optimization of the PCR amplification was done in order to maximize sensitively and reproducibility
of detection of virus templates in DNA from symptomatic leaves using degenerate primers.
Annealing temperature, type and concentration of PCR additives and concentration of PCR additives
and MgCl2 concentration were first adjusted to improve sensitivity and reproducibility sensitivity was
assessed by detection of the initial and end points of dilution and the intensity of the amplified bands.
Table-1: Detection of initial and end points of dilution and respective amplified band intensity
Replicates Dilutions
30ng 20ng 10ng 5ng
1 +++ ++ + -
2 ++++ ++ + -
3 +++ +++ + -
4 +++ ++ + -
++++ - Very intense amplified band
47
+++ - Intense amplified band

++ - Visible amplified band
+ - Barely visible band
When DNA was used at concentration above 30ng/l, shearing in bands were seen.
From this table it is evident that 30ng/l of DNA is most suitable for the PCR so we used 30ng/l concentration
of DNA for PCR.
DNA was diluted to get 30ng/l of concentration and following PCR reaction was set.
dNTPs - 1.0l
Taq DNA Polymerase - 0.5l
10X Buffer - 2.5l
Primer A (Forward) - 1.0l (~ 25ng/l)
Primer B (Reverse) - 1.0l (~ 25ng/l)
DNA from diseased and healthy leaf - 1.0 l
The rest of the volume (18l) was made up by water up to 25l.
This reaction mixture was put in DNA engine (PCR machine) for the following cycles.
940C - 5 Minute
0
94 C - 1 Minute
0 One Cycle 35 times
52 C - 1 Minute
720C - 1 Minute
0
72 C - 5 Minute
40C - for ever
When reaction cycle is completed then amplified product was checked in 1.2% agarose gel by taking 10l of
PCR product with 2l of dye, bromophenol blue.
This PCR product is run along the marker -DNA ECQRI and Hind-III double digest to determine
the size or base pairs of bands.
For control, DNA from the healthy leaf is also given the same reaction mixture for PCR
amplification and run on the same gel.
The PCR amplification product were resolved in 1.2% agarose gel stained with ethidium bromide
and screened on UV transilluminator Gel documentation system for analysis was used to take
photograph of gel for analysis.
Degenerate primer pairs [7] is used for DNA amplification. The two degenerate oligonucleotide primers
had the following sequences.
Primer PA - 5' TAA TAT TAC CGG AGG AGG CCC CC3'
PB - 5' TGG ACC TAA CAA GGG CCT TCA CA 3'
These degenerate primers permit detection of sub-group-III Gemini viruses by annealing to sequences that
flank the conserved core region of the coat protein gene, yielding a diagnostic about 565bp virus fragment [8].
This means primer will bind to the DNA of virus only and amplification of that particular DNA will be done by
PCR.
48
Plate-I of gel shows marker in first and last lane. Second and third lane represents control plant. Fourth,
fifth and sixth lane contain amplification product of diseased leaf. As it is evident from the photograph of gel
that the lane containing amplification product of diseased plant are showing a specific band of about 500 bp, no
such band is seen in control lane making it clear that the virus under study belongs to family Gemini viridae and
subgroup-III genus begomovirus species cowpea golden mosaic virus.
Figure: 1 First and last lane- Molecular weight markers, second and third lane- control plants, fourth fifth and
sixth row- diseased plant with 500bp band.
DISCUSSION :
For identification of gemini viruses, serology has never been a preferred tool for the simple reason that
the particles are difficult to purify, making it hard to produce good antisera. On the other hand begomo virus is
the close serological relationship due to conservation in coat protein gene [9], as a result polyclonal antibody
raised against one begomovirus could detect all the begomoviruses in double antibody sandwich ELISH or
ISEM.
Because of the difficulty in purifying begomoviruses virion particles, nucleic acid based approaches
like PCR are being widely preferred for the diagnosis. Advent of PCR technique revolutionized research on
begomoviruses. Full length genome or specific regions in the viral genome are amplified with degenerate
primers or specific primers.
Diagnostic PCR should fulfill several qualitative characteristics, of which the most important are
specificity, sensitivity, efficiency and reproducibility. Designed degenerate primers flank the 5' terminal region
of the coat protein gene to specifically detect begomoviruses and when tested with degenerate primers it showed
the specific band in the diseased plant DNA corresponding to the coat protein gene of begomovirus confirming
the presence of begomovirus in diseased plants.
REFERENCE
1. 1.Goodman, R.M. Geminivirus. In Hand book of Plant virus Infections and comparative Diagnosis, ed.
E.Kurstak Amsterdem : Elsevier / North-Holland Biomed. 879-910,(1981).
2. Harrison, B.D. Advances in Geminivirus research, Ann. Rev. Phytopathol., 23, 55-82. (1985).
49
3. Fauquet, G.M., Stanley, J. Geminivirus classification and nomenclature-progress and problems, Ann
Appl. Biol., 142, 165-189 (2003).
4. Malathi, V.G., Usharani, J.S., Sivalingam, P.N., Rouhibaksh, A., Padma Latha, K.V. and Periasamy,
M. Diversity and complexity of Begomoviruses, Annu. Rev. Plant Pathal., 3, 225-270 (2004).
5. Samec, P. DNA Polymorphism and RAPD technology, Priloha Casopisu Genetika a Slechteni 29 , 291-
230. (1993).
6. Williams, J.G.K., Manafey M.K., Rafalkl J.A. and Tingey S.V. Genetic analysis using random
amplified polymorphic DNA markers, Methods in Enzymology, 218, 704-740 (1993).
7. Deng D., McGrath P.F., Robinson D.J., Harrison B.D. Detction and differentiation of whitefly
transmitted geminiviruses in plant and vector insect by the polymerase chain reaction with degenerate
primers, Ann. Appl. Biol., 125 , 327 - 336. (1994).
8. Wyatt S D & Brown J K. Detection of subgroupIII Geminivirus isolates in leaf extracts by degenerate
primers and polymerase chain reaction, Phytopathology ,86(12) ,1288-1293(1996).
9. Harrison, B.D. and Robinson, D.J. Natural genomic and antigenic variation in whitefly-transmitted
geminiviruses, Ann. Rev. Phytopathal., 37, 369-398 (1999).
50
Science and Technology in Rural India
Richa Atreya
Department. of Botany, M.S. College, Saharanpur (India)
e-mail address - richaatreya@rediffmail.com
ABSTRACT
The importance for science and technology for rural India was appreciated by eminent leaders and scientists in
1930s giving rise to the centre for science for villages and advanced institutions for education. However post
independence steps were biased towards urban areas and science & technology turned their attention for rural
areas about 40 years later in 1970s. The most well known step was from Indian Institute of Sciences by its
program for the application of science and technology to rural areas also called by its acronym ASTRA.
Present scenario poses promising future of S&T in rural areas with many governmental and nongovernmental
organizations participating in process of its development. Ministry of human resource development has started
Rashtriya Avishkar Abhiyan that works in conduit with Sarva Shiksha abhiyan, Madhyamik Shiksha Abhiyan
and also promotes study of S&T in higher education among rural children. Likelihood for science education can
be fostered among rural Indians by making it affordable, technologically assisted, developing rural
infrastructure, and creating awareness through rigorous campaigns and fairs. Our present National Education
Policy shall be revised for promotion of science and innovations in India.
KEYWORDS - S&T, infrastructure, ASTRA, biomethanation, biogasification, STEM.
INTRODUCTION
Many pioneer leaders like Bal Gangadhar Tilak, Raja Ram Mohan Roy, Swami Dyanand Saraswati along with
eminent Indian scientists like J.C. Bose, P.C. Roy and C.V. Raman realized the need of acquaintances with
science and technology (S&T) for common Indians including rural people. They envisioned the importance of
science aptitude among people for nation building. But post independence era had an upthrust for industrial
development and ideas of mainstream scientific and technological development were marginalized. Research
and development got a mention in Indian budgets, though R&D budget in1957-58 was mere 18.81 crores. Indian
economy is an agro economy with rural areas as its main centers and therefore it is indispensible to promote
science education in rural areas and harnessing S&T for them. Problems of lack of easy access, lack of interest,
common curricula, gender differentiation, and most important lack of infrastructure in rural areas is holding
back promotion of S&T. New & better government policies shall be formulated for rural areas. Yet public
private partnership is paving way for better rural India and now we see rural areas that are tech-savvy and have
more acceptances for science and innovations.
POST INDEPENDENCE S &T IN RURAL AREAS
The post independence establishment was preeminently dominated by scientists and engineers who returned
from Europe and North America and they were prolifically influenced by their studies and sojourns abroad.
They were more open to industrialization and technology and at that time demand of Indian industry and
government were the determining force for development. There can be meticulous mention of post
51
independence growth of S&T in India as the R&D budget increased; a number of science and technology
institutions like DRDO, IARI, ISRO etc. established; an enormous increase in technically trained personnel;
international collaboration for S&T was also achieved.
Rural areas faced biased attitude- By and large all scientific development shifted to urban needs and rural areas
were unnoticed. Conclusions for such polar development of S&T can be drawn from anti rural bias in-
R&D expenditure
Establishment of institutions
Distribution of technical personnel
Development of infrastructure
Government policies and future plans
Even five years plans of India addressed primary education, health facilities, sanitation, agriculture and
irrigation in these areas and to add setbacks to these initiatives many of five year plans collapsed due to natural
and political upheaval in the country. So there was no major breakthrough in creating scientific awareness and
harnessing S&T for rural people. Later in 1970s when Indian institute of sciences presented a program called
ASTRA for use of S&T for rural people, there arose hopes of creating scientific aptitude among rural Indians
[1]. ASTRA an institutional experiment was against above mentioned backdrops that there arose a need to
reorient Indian science and technology towards need for rural India. In 1970 Indian Institute of Sciences
Bangalore made a presentation and translated it into a program called Application of Science and Technology
to Rural Areas, also called by its acronym ASTRA. The ASTRA institutional experiment was based on a
model of technology & rural society interactions. The rural studies of energy, building, water, health etc. carried
out by ASTRA provided an important step for technology development. ASTRA is now known as Center for
Sustainable Energy (CST). Energy efficient wood burning devices, biomethanation, biogasification, green
building, bioenergy and climate change studies are few examples of fruitful interventions of ASTRA [1].
CAPART (Council for Advancement of Peoples Action and Rural Technology), an autonomous body
registered in 1986 under aegis of Ministry of Rural Development is envisioned to play dynamic role to
strengthen the voluntary movement in the country and facilitate the promotion of innovative rural technology[2].
Several other steps to promote science education among rural people were adopted but yet rural infrastructure
was poor and incapable to support S&T for rural Indians.
PRESENT SCENARIO FOR S&T IN RURAL AREAS
STEM refers to the academic disciplines of science, technology, engineering and mathematics. The term is
typically used when addressing education policy and curriculum choices in schools to improve competitiveness
in science and technology development. STEM crisis has implications for work force development, national
security concerns, immigration policy and S&T development in a country. Some educationist, technocrats,
academicians, scientists and economists believe there exist a STEM crisis in India while others do not agree to
it. But if we see analytical and statistical details we find STEM crisis does occur in rural India. Annual reports
and surveys may be seen as an effort towards strengthening the S&T statistical network within our country. A
survey by NCAER (National Council of Applied Economic Research) reveals engineering was the favorite
52
subject chosen by maximum number of students (22%) as the one in which they will like to complete higher
education, medicine was next (18%) and pure sciences was marginally low [3]. FIG. 1
An another NCAER survey found that a fourth of those in rural areas said they would like to complete their
higher education in arts as compared to 15% in urban areas. Since curiosity is basic nature of human, rural
attention and interest towards S&T can be easily nurtured. It is believed that development of rural infrastructure
and more awareness for S&T will help in socio-economic as well as technological development of rural areas.
Moreover the agro economy of India works with agriculture and the rural people, who constitute more than 70%
of our total population, still plays a front role. Today farmers use S&T for agriculture; some of these
techniques/technologies included the use of manure/fertilizers, the use of water harvesting or green manuring
[3]. FIG. 2. Facilities like 24 hours satellite channel and phone helplines along with internet services are
working for farmers and these prove highly beneficial to them. With such instances of S&T use by rural people
the spread of mainstream science education may also be imagined.
Other silent reforms to enhance associations between rural society and S&T are underway. Recently Planning
Commission of India was discarded and a new institution called NITI Aayog was established. NITI Aayog shall
provide a critical directional and strategic input into developmental processes. It will work by providing better
inter ministry and center-state coordination. It will develop mechanisms to formulate credible plans to the
village levels and aggregate these progressively at higher levels. These plans include the rural areas.
In 2015 Ministry of Human Resource Development set up Rashtriya Avishkar abhiyan (RAA) to nurture a
spirit of enquiry and creativity, love for science and mathematics and effective use of technology among
children. RAA will span across MHRDs schematic interventions of Sarva Shiksha Abhiyan, Rashtryia
Madhyamik Shiksha Abhiyan in Department of School Education and Literacy and programs of Department of
Higher Education to encourage science, mathematics and technology. Institutes like ICAR, CSIR, DBT, CPRI
etc. have established their local institutes in many towns of India. Many private medical and engineering
institutes are established in rural areas due to low cost of settlement in these areas. NGOs like Pratham are
working on direct instruction model for creating aptitude for science and math education among rural children.
Aser-A nongovernmental survey reveals improvement in science and math learning through education camps
[4]. FIG. 3. Agastya International Foundation works for developing science aptitude among rural children. They
foster mobile labs, conduct science fairs, establish science centers in rural areas and run programs targeting drop
out students and communities. Azim premji Foundation helping 2 million students across 16000 students from
14 states, Vidya gyan foundation and Samudaya are few other to mention.
CONCLUSIONS
Today the socio-economic conditions of India present an environment where rural India may S&T developed.
Efforts to create their access to resources and a better infrastructure with proper facilities of electricity and water
etc. shall be structured . Since technology prompts interest among rural student, E-learning may be a correct way
for it. Results presented in surveys and reports will essentially be an important input for entire scientific
community and policy makers to set achievable goals and work out action plans towards S&T development in
rural areas. Since 1986 social, economic and scientific atmosphere of India has changed drastically, so our
53
education policy needs to be reviewed. A National Education Policy shall be formulated that work on soul
motto of science and innovations and consider every child as an integral part of our nation building. This may
fulfill Indian aspirations of being a developed economy someday. Promotion and development of S&T in India
with special emphasis on rural areas may help on the path of national development.
REFERENCES
1. Reddy AKN. Choices of Alternative Technologies. Economic and political weekly. 3(25), 1109-1114.
(1973).
2. CAPART (Council for advancement of peoples action and rural technology). Avail from:
http://www.capart.nic.in. 2010. [cited 1 oct 2015].
3. Rajesh Shukla. Indian Science Report - Science education, HR & Public attitude towards S&T. NCAER,
New Delhi. ISBN - 81-88830-07-0. (2005).
4. Pratham Education Foundation. Annual Status of Education Report (Rural) 2010. Pratham resource centre,
New Delhi. (2011).
FIGURES
Figures mentioned above are represented here in ascending order.
6-8th
9th Least informed
10th Moderately
11-12th Most informed
SCIENCE ENGINN MEDICINE ARTS COMMERCE OTHERS agriculture household communication health
FIG. 1- Preferred subject for higher education by level of students FIG. 2- Distribution of public by awareness of technology related to
Source: NCAERs National Survey-2004 selected sectors. Source: NCAERs National Survey- 2004
54
FIG. 3- Special learning camps in U.P. - Math progress-The percentage of children that can recognize 2-3 digit
numbers has increased by 69% in 10 day model. Source: A Non Governmental Survey by Pratham Education
Foundation.
55
Effects of Distellery Effluent Irrigation on Mustard (Brassica campestris)
RENU CHOUDHARY, NARESH KUMAR* AND HARENDRA MALIK *
Department of Biology, G.G.I.C. Gagalheri, Saharanpur(U.P.), India.
*Department of Botany, C.C.R. (P.G.) College, Muzaffarnagar (U.P.), India.
Email- harendra_ghoni@rediffmail.com
Abstract
A large network of distilleries has been established in India which have been recognized as one of the
most polluting agro-based industries, generating huge quantities of distillery effluent. The present study was
conducted to observe the impact of different concentrations i.e. 0.5,1.0,2.0,5,10,25,35,45 and 55% of different
distillery effluents i.e. raw spent wash (RSW), biomethanated spent wash (BSW) and lagoon sludge (LS)
collected from Shamli distillery and chemical works, Shamli, District-Shamli) on Mustard (Brassica campestris)
cv. PAC-401. Distillery effluents (RSW, BSW and LS) considerably affected all the growth parameters in
treated plants. Growth parameters improves up to 5% concentrations of RSW and up to 10% concentrations of
BSW and LS, while above these concentrations growth is significantly reduced. The improvement or reduction
in different biochemical content was observed according to the concentrations of effluents. The chlorophyll a
and b, carotenoids and oil contents of plants were increased initially upto 5% concentrations of RSW and upto
10% concentrations of BSW and LS effluents. Higher concentrations (>5% of RSW and >10% of BSW and LS)
of distillery effluents were detrimental to all the treated crop plants.
Keywords : Distillery effluent, Raw spent wash, Biomethanated spent wash, Lagoon sludge, Germination,
Growth, Mustard.
Introduction
During the last two decades, India has emerged as the largest sugar producer in the world. The sugar
factories after extraction of crystalline sugar from cane juice, sell the molasses to distilleries, where it is
converted into alcohol by yeast fermentation. The raw spent wash coming out as a result of fermentation process
from the distilleries is almost 100 times more concentrated as compared to sewage with regards to its pollutional
characteristics. Uttar Pradesh is the highest sugar producing state in India and therefore, here about 126 sugar
factories and 61 distilleries are established in public as well as private sector mainly in western Uttar Pradesh.
Most of these distilleries are highly polluting units. The effluents of almost all of these units are discharged
either directly into water bodies or nearby rivers through drains. Some distilleries discharge their effluents into
the adjoining crop areas and farmers use this effluent directly for irrigation. Therefore, in the present study, an
attempt has been made to assess the effects of different concentrations (0.5,1.0,2.0,5,10,25,35,45 and 55%) of
distillery effluents i.e. raw spent wash (RSW), biomethanated spent wash (BSW) and lagoon sludge (LS)
irrigation on three important oil yielding crop plants grown in this locality viz. Mustard (Brassica campestris)
cv. PAC-401.
Materials and Methods
56
The main object of the present investigation is to study the effects of different distillery effluents
(RSW, BSW and LS) irrigation at different concentrations on various aspects of Mustard (Brassica campestris)
cv. PAC-401. All three pure effluents of distillery were diluted with tap water to get 0.5%, 1.0%, 2.0%, 5%,
10%, 25%, 35%, 45%, and 55% concentrations. The tap water was used as control. The soil of research plots or
pots was irrigated with different concentrations of effluent (RSW, BSW and LS) on alternate days or according
to the requirement of the crop. For seed germination studies, healthy seeds were selected for uniformity (criteria
being the size and colour of seeds) and were surface sterilized with 0.1% mercuric chloride for two minutes and
thoroughly washed with distilled water. Plants grown in research plots were taken out and rinsed their roots
repeatedly with unionized water to eliminate undesirable nutrients from the root surface. Excess of moisture,
thus created, was wiped out with the help of clean blotting paper and absorbent towels. Ordinary scale was used
to measure the length of root and shoot in centimeters. For phytomass determination (g dw/plant), different plant
parts were separated and oven dried at 800C for 24 hours or until a constant weight was achieved. Dry weight of
different plant parts were added to get the phytomass of each plant. Chlorophyll, carotenoid and oil content
calculated using several deeds [1, 3 & 4].
Result and Discussion
A slight increase in germination percentage was recorded upto 5% concentration of RSW and 10%
concentration of BSW and LS. 100% seed germination occurred at above concentrations of RSW, BSW and LS
in Brassica campestris cv. PAC-401. Reduction in seed germination percentage was 42.42, 37.37 and 34.34
percent under 55% concentration of RSW, BSW and LS respectively. The speed of germination index increased
upto 10.35 percent in 5% RSW concentration and 15.26 and 12.80 percent in 10% BSW and LS concentration
treatments respectively, The reduction percentage was 20.00, 16.84 and 15.61 percent under 55% of RSW, BSW
and LS concentration [7,8].
Table 1 : Effect of different concentrations of distillery effluents on seed germination percentage and
speed of germination index of 8 days old seedlings of Brassica campestris cv. PAC-401.
Treatments
RSW concentration (%)
Parameters
Contro
0.5 1.0 2.0 5.0 10 25 35 45 55
l
Seed
germination 99 99 100 100 100 95 84 74 66 57
percentage
Speed
germination 570 576 591 608 629 541 520 503 478 456
index
BSW concentration (%)
Seed
99 100 100 100 100 100 86 79 70 62
germination
57
percentage
Speed
germination 570 581 597 615 636 657 542 520 503 474
index
LS concentration (%)
Seed
germination 99 99 99 100 100 100 91 83 72 65
percentage
Speed
germination 570 573 588 605 626 643 556 529 511 481
index
Growth parameters improves up to 5% concentrations of RSW and up to 10% concentrations of BSW

and LS, while above these concentrations growth is significantly reduced. Results of exposure to different
concentrations of distillery effluents on various growth parameters of both cultivars are presented in tables 2, 3
and 4. The root length was increased upto 22.00% at 5% of RSW and 25.40 and 23.78% at 10% concentrations
of BSW and LS treatments. The reduction percentage were 32.03, 26.05 and 24.43 percent in RSW, BSW and
LS respectively at 55% concentration in 25 days old plants. In 50 days old plants the root length increased upto
16.52% at 5% of RSW and 19.80 and 17.90 percent at 10% of BSW and LS treatments respectively, while
reduction percentage were 28.58, 24.12 and 22.07 percent at 55% concentrations of RSW, BSW and LS
treatments. In 75 days old plants similar pattern was observed. Shoot length also show the similar pattern of
increase and decrease in 25, 50 and 75 days old plants. The biomass production (phytomass accumulation) of
both cultivars were found to be showing similar changes as found in root and shoot length in 25, 50 and 75 days
old plants. Decrease in total dry weight or phytomass accumulation eventually lead to decrease in net primary
productivity (NPP) of treated plants [5,6].
Table 2 : Effect of different concentrations of distillery effluents on different growth parameters of 25

days old plants of Brassica campestris cv. PAC-401.
Treatments
Parameters RSW concentration (%)
Control 0.5 1.0 2.0 5.0 10 25 35 45 55
6.180. 6.360. 6.780. 7.110. 7.540. 5.790. 5.340. 5.010. 4.580. 4.200.
Root length (cm)
84 81 77 72* 79** 82 74* 67* 65** 69**
9.880. 10.27 10.76 11.29 11.85 9.190. 8.180. 7.760. 7.380. 7.010.
Shoot length (cm)
62 0.83 0.88 0.89* 0.82** 60 56* 59** 73** 76**
Biomass 2.660. 2.770. 2.900. 3.090. 3.260. 2.510. 2.360. 2.230. 2.120. 1.970.
production(g) 46 52 67 59* 61** 45 42* 38* 36** 31**
NPP (g/plant/day) 0.106 0.110 0.116 0.123 0.130 0.100 0.094 0.089 0.084 0.078
58

Root length (cm) 6.180. 6.420. 6.880. 7.220. 7.640. 7.750. 5.670. 5.240. 4.820. 4.570.
84 76 85 73* 88** 81** 75 71* 67** 65**
Shoot length (cm) 9.880. 10.50 10.98 11.54 12.04 12.17 8.620. 8.180. 7.760. 7.340.
62 0.68 0.60 0.67* 0.70** 0.63** 59 55* 63** 54**
Biomass 2.660. 2.790. 3.030. 3.190. 3.350. 3.440. 2.430. 2.300. 2.180. 2.050.
production(g) 46 48 44* 49** 52** 43** 39 41* 37** 33**
NPP (g/plant/day) 0.106 0.111 0.121 0.127 0.134 0.137 0.097 0.092 0.87 0.082
Root length (cm) 6.180. 6.300. 6.680. 7.010. 7.430. 7.650. 5.800. 5.370. 4.940. 4.670.
84 81 87 76* 89** 82** 78 83* 72** 76**
Shoot length (cm) 9.880. 10.23 10.72 11.22 11.69 12.12 8.890. 8.300. 2.390. 2.270. 2.100.
62 0.65 0.60 0.58* 0.64** 0.69** 71 66* 41* 37* 35**
Biomass 2.660. 2.730. 2.850. 3.010. 3.190. 3.300. 2.510. 0.095 0.090 0.084

production(g) 46 43 49 52* 56** 58** 44
NPP (g/plant/day) 0.106 0.109 0.114 0.120 0.127 0.132 0.100
Values are in mean standard deviation.

Significance of difference from control; P* < 0.05; P** < 0.01 and non-significant
59

days old plants of Brassica campestris cv. PAC-401
Treatments
Control 0.5 1.0 2.0 5.0 10 25 35 45 55
13.681 14.051 14.611 15.291 15.941 13.261 12.281 11.391 10.561 9.771.
Root length (cm)
.44 .21 .28 .42* .48** .36 .31* .22** .26** 09**
42.301 43.171 44.881 46.741 48.111 41.531 37.651 35.831 34.191 31.521
Shoot length (cm)
.78 .65 .82 .89* .91* .56 .50* .43** .33** .15**
Biomass 8.560. 8.710. 9.080. 9.460. 9.920. 8.370. 7.980. 7.510. 6.900. 6.320.
production(g) 66 61 65 67* 71** 58 53* 62** 59** 52**
NPP (g/plant/day) 0.171 0.174 0.181 0.189 0.198 0.167 0.159 0.150 0.138 0.126
Root length (cm) 13.681 14.161 14.831 15.581 16.201 16.391 13.111 12.431 11.651 10.381
.44 .34 .38 .30* .26** .15** .11 .24* .32** .19**
Shoot length (cm) 42.301 43.461 45.501 47.61. 48.911 49.381 41.691 38.521 36.151 33.811
.78 .81 .86 92* .94** .26** .32 .39* .40** .33**
Biomass 8.560. 8.780. 9.220. 9.680. 10.130 10.350 8.240. 7.830. 7.390. 6.750.
production(g) 66 56 63 65* .69** .61** 58 54* 51** 55**
NPP (g/plant/day) 0.171 0.175 0.184 0.193 0.202 0.207 0.164 0.156 0.147 0.135
Root length (cm) 13.681 13.961 14.461 15.071 15.731 16.131 13.191 12.541 11.811 10.661
.44 .31 .42 .39* .46** .36** .39 .28* .24** .29**
Shoot length (cm) 42.301 43.091 44.721 46.511 47.741 48.611 41.831 39.261 36.691 34.531
.78 .74 .81 .85* .69* .86** .52 .46* .62** .65**
Biomass 8.560. 8.670. 8960.7 9.310. 9.780. 10.190 8.310. 7.970. 7.540. 6.980.
production(g) 66 70 4 78* 72** .81** 68 62* 58** 56**
NPP (g/plant/day) 0.171 0.173 0.179 0.186 0.195 0.203 0.166 0.159 0.150 0.139

60

days old plants of Brassica campestris cv. PAC-401.
Treatments
Control 0.5 1.0 2.0 5.0 10 25 35 45 55
16.081 16.291 16.851 17.431 18.261 15.521 13.241 12.471 11.611 10.521.
Root length (cm)
.36 .39 .42 .44* .45* .33 .28* .31** .24** 27**
123.42 125.80 128.63 132.02 136.48 118.65 104.73 98.813 93.693 88.113.
Shoot length (cm)
3.48 3.56 3.81 3.73* 3.79* 3.56 3.42* .36** .29** 48**
Biomass 44.631 45.381 46.781 48.621 51.271 42.381 38.141 36.791 35.411 33.641.
production(g) .16 .24 .34 .48* .53** .21 .26* .18** .11** 16**
NPP (g/plant/day) 0.595 0.605 0.623 0.648 0.683 0.565 0.508 0.490 0.472 0.448
Root length (cm) 16.081 16.381 17.031 17.691 18.521 18.961 13.891 12.961 12.071 11.321.
.36 .41 .54 .39* .48** .51** .33* .27** .29** 26**
Shoot length (cm) 123.42 126.49 129.52 133.22 137.29 140.633 111.64 102.883 96.753 90.823.
3.48 3.51 3.59 3.77* 3.68* .76** 3.29* .15** .31** 28**
Biomass 44.631 45.641 47.261 49.361 51.941 52.881 40.191 37.531 36.041 34.771.
production(g) .16 .19 .27 .35* .39** .42** .18* .14** .08** 12**
NPP (g/plant/day) 0.595 0.608 0.630 0.658 0.692 0.705 0.535 0.500 0.480 0.463
Root length (cm) 16.081 16.201 16.671 17.151 17.891 18.451 14.201 13.281 12.461 11.831.
.36 .34 .48 .54 .31* .35** .46* .39** .33** 28**
Shoot length (cm) 123.42 125.61 128.19 131.17 135.81 137.523 112.36 103.433 97.383 91.883.
3.48 3.56 3.68 3.85* 3.76* .82** 3.37* .61** .48** 39**
Biomass 44.631 45.271 46.561 48.091 50.681 52.141 40.781 38.211 36.691 35.491.
production(g) .16 .27 .31 .37* .44* .50** .29* .14** .22** 21**
NPP (g/plant/day) 0.595 0.603 0.620 0.641 0.675 0.695 0.543 0.509 0.489 0.473

Considerable increase and decrease in chlorophyll a and chlorophyll b content of leaves were observed
in different concentrations of RSW, BSW and LS treatments. For example, chlorophyll a and b content
increased upto 21.48 and 22.47% at 5% concentration of RSW respectively, while at 10% concentration of BSW
and LS these values were 25.65, 27.75 and 21.61, 23.85% respectively in 25 days old plants. Carotenoids, the
accessory pigments increased by 19.69, 24.57 and 21.21% at 5% concentration of RSW and 10% concentration
of BSW and LS respectively against control of 25 days old plants. Whereas, reduction of carotenoid content
61
occurred as 30.97, 24.91 and 23.40% at 55% concentration of RSW, BSW and LS respectively. Similar trends
of results also obtained from 50 days and 75 days old plants. Oil content of seeds were increased upto 5%
concentration of RSW, i.e. by 6.38% and upto 10% concentration of BSW and LS, i.e. by 8.27 and 7.85%
respectively. Above these concentrations of RSW, BSW and LS, oil content decreased regularly and decrease
was maximum at 55% concentration of RSW, BSW and LS, i.e. by 38.71, 20.67 and 20.38% respectively [ 2,9].
25 Days
1
Chlorophyll-a content
0.8
(mg/g f.wt.)
0.6
RSW
0.4
BSW
0.2 LS
Treatments (%)
25 Days
1
50 Days
contentcontent
1.2
0.8
(mg/g f.wt.) f.wt.)
Chlorophyll-a
0.61
(mg/g
RSW
0.8
0.4
Chlorophyll-a
BSW
0.2
0.6 LS
RSW
0
0.4 BSW
0.2 LS
Treatments (%)
0
Treatments (%)
62
75 Days
1
Chlorophyll-a content
0.8
(mg/g f.wt.) 0.6
0.4 RSW
BSW
0.2
LS
0
Treatments (%)
Figure 1 : Effect of different concentrations of distillery effluents on chlorophyll-a content (mg/g f.wt.) of
25, 50 and 75 days old plants of Brassica campestris cv. PAC-401.
25 Days
0.6
Chlorophyll-b content
0.5
(mg/g f.wt.)
0.4
0.3 RSW
0.2 BSW
0.1
LS
0
Treatments (%)
50 Days
0.8
0.7
Chlorophyll-b content
0.6
(mg/g f.wt.)
0.5
0.4 RSW
0.3
BSW
0.2
0.1 LS
0
Treatments (%)
63
75 Days
0.7
Chlorophyll-b content 0.6
(mg/g f.wt.) 0.5
0.4
0.3 RSW
0.2 BSW
0.1 LS
0
Treatments (%)
Figure 2 : Effect of different concentrations of distillery effluents on chlorophyll-b content (mg/g f.wt.) of
25, 50 and 75 days old plants of Brassica campestris cv. PAC-401.
25 Days
2.5
Carotenoid content
2
(mg/g f.wt.)
1.5
1
LS
0.5
BSW
0
RSW
Treatments (%)
50 Days
2.5
Carotenoid content
2
(mg/g f.wt.)
1.5
1 LS
0.5 BSW
0 RSW
Treatments (%)
64
75 Days
2
Carotenoid content 1.5

(mg/g f.wt.)
1
0.5 LS
BSW
0
RSW
Treatments (%)
Figure 3 : Effect of different concentrations of distillery effluents on carotenoid content (mg/g

f.wt.) of 25, 50 and 75 days old plants of Brassica campestris cv. PAC-401.
cv. PAC-401
50
45
40
35
Oil content (%)
30
25 RSW
20 BSW
15 LS
10
5
0
Treatments (%)
Figure 4 : Effect of different concentrations of distillery effluents on oil percentage of Brassica campestris
cv. PAC-401.
References
1. Arnon, D.I. Copper enzymes in isolated chloroplasts, polyphenol oxidase in Beta vulgaris. Plant
Physiol. 24 : 1-15 (1949).
65
2. Chandra, R., Kumar, K. and Singh, J. Impact of anaerobically treated and untreated (raw) distillery
effluent irrigation on soil microflora, growth, total chlorophyll and protein contents of Phaseolus
aureus L. J. Environ. Biol. 25 (4) : 381-385 (2004).
3. Maclachlan, S. and Zalik, S. Plastid structure, chlorophyll concentration and free amino acid
composition of chlorophyll mutant of barley. Can. J. Bot. 43 : 1053-1062 (1963).
4. Pain, S. K. and Nayek, B. Studies on the physiology of growth and development of Sesamum indicum
cv. 13-9 with special reference to yield of seed and oil : The effect of indolyl-3-propeonic acid used as
foliar spray. J. Indian Bot. Soc. 60 : 202-207 (1981).
5. Pandey, S.N., Nautiyal, B.D. and Sharma. C.P. Pollution level in distillery effluent and its phytotoxic
effect on seed germination and early growth of maize and rice. J. Environ. Biol. 29 (2) : 267-270
(2008).
6. Rath ,P., Pradhan, G. and Mishra ,M.K. Effect of Sugar factory distillery spent wash (DSW) on the
growth pattern of sugarcane (Saccharum officinarum) crop. J. Phytology. 2 (5) : 33-39 (2010).
7. Sharma, V., Sharma, R. and Sharma, K.D. Distillery effluent effect on seed germination, early seedling
growth and pigment content of Sugarbeet (Beta vulgaris Linn. Var. Mezzanau-Poly). J. Environ. Biol.
23 (1) : 77-80 (2002).
8. Sunitha, N. S. and Seenappa, C. Indigenous State-of-The-Art Technology Development for Distillery
Effluents: An organic biochemical reagent for productive soils by means of Aerobic Sponge Method
Vermitechnology (ASMV). J. Chem. Bio. Phy. Sci. 3 (1) : 559-566 (2013).
9. Zengin, F. K. and Kirbag, S. Effects of copper on chlorophyll, proline, protein and abscisic acid level
of sunflower (Helianthus annuus L.) seedlings. J. Environ. Biol. 28 (3) : 561-566 (2007).
66
Role of Transgenic Plants in Agriculture

Renu Rani
Department of Botany, Govt. Degree College, Behat, Saharanpur
Email : renu31panwar@gmail.com
ABSTRACT
Environmental Stresses, population explosion and food shortage have caused serious problems to mankind on
the globe. The world population is increasing alarmingly and is projected to reach 9 billion by 2050. To fulfill
the food demand on every individual from limited natural resources is difficult. This factor has resulted in food
deficiency thereby causing malnutrition, which is a serious health problem these days. Food production will
need to increase at the same rate or more in order to satisfy the needs of such an enormous number of people in
some older centuries. So there is a need to use the genetic techniques to improve crops over the recent decads.
Transgenic breeding uses molecular cloning techniques to identify cloned or synthesized genes of interest and
directly transforms the recipient genome. This process manipulates plant genomes through insertion of gene(s)
from another species. Transgenic plant have been found to have many advantages like, development of high
yielding varieties of crop plants and disease resistant, and are plants with improved tolerance to biotic and
abiotic stress.
Key words: Abiotic stress, Genetically modified Organism, Genetic Engineering and Transgenic plants,.
INTRODUCTION
Increasing world population and food demands require world agricultural production be increased by 50% by
2030 (The Royal Society, 2009). In the meantime, climate change and shrinking environmental resources are
limiting agricultural production over the world [1]. These challenges bring an urgent need to enhance crop
productivity. To breed crops with increased yield and resistance to environment stresses, a pivotal consideration
is how to effectively utilize genetic diversity. Genetic crossing, selection of natural or artificial mutations, and
transgenics, are the main techniques for plant breeding. Traditional plant breeding uses crossing, mutagenesis
and somatic hybridization for genome modification to improve crop traits. It introduces new beneficial alleles
from crossable species. Due to crossing barriers and linkage drag, however, traditional plant breeding is time-
consuming and requires several generations of breeding and selection. Transgenic breeding uses molecular
cloning techniques to identify cloned or synthesized genes of interest and directly transforms the recipient
genome. This process manipulates plant genomes through insertion of gene(s) from another species. An
organism that is generated this way is considered to be a genetically modified organisms (GMOs). Most
genetically modified plants are generated either by particle gun method or by Agrobacterium tumefaciens
mediated transformation method. Since 1990s, the major emphasis of agriculture biotechnology can be found on
traits for improvement in crops related to insect and herbicide resistance, nutritional quality, virus resistance,
shelf life and bio-fuel production [2].Transgenic plant have been found to have many advantages like,
development of high yielding varieties of crop plants and disease resistant, and are plants with improved
tolerance to biotic and abiotic stress [3-6]. Recently, biotechnology has revolutionized crop improvement by
producing GM crops with enhanced availability and utilization of important traits [7]. In agriculture, yield is a
major output and improvement in yield of plants is a major thrust area by counteracting biotic and abiotic
environmental cues. Thus, crop cultivars with enhanced yield and stability are required.
67
ADVANTAGE OF TRANSGENIC PLANTS

Environmental factors are essential components which affect crop yield to a great extent. The introduction of
resistance to abiotic and biotic stress into crop plants has become a topic of major economic interest for
agriculture. Infectious diseases are the most dangerous problems in the present world and each year one third of
all deaths are caused by the infectious agents. Genetically engineered plants offer significant benefits by
improving yield, transportation costs, enhancing the nutritional content, biotic and abiotic stress and biofuel
production. in this review we analyze the role of transgenic plants in combating environmental stress.
INSECT RESISTANCE
Plants are equipped with the natural plant defense system against insects, fungi, bacteria which is provided by
the proteinase inhibitors [8-10]. PIs are ubiquitous in plants especially in Leguminoseae, Gramineae,
Compositeae. They are natural defense related proteins often present in seeds and induced in certain plant tissue
by herbivory and wounding. The activity of PIs is due to their capacity to form stable complexes with target
proteases, blocking, altering or preventing access to enzyme active sites.
Bacillus thuringiensis (Bt) insect resistant crops are one of the most astounding achievements in plant transgenic
technology. Bt is a potent insecticide which comprises crystal protein endotoxin produced by some strains of
soil bacterium B. thuringiensis (a soil bacterium). The Bt crystal (cry) insecticidal protein (-endotoxin) genes
are toxic to lepidopterans [11], dipterans [12], coleopterans [13]. Bt cry protein is non-toxic to humans and
animals, but toxic to insects [14]. When cry protein are ingested by insects, they are dissolved in the alkaline
juice present in midgut lumen. The gut proteases process them hydrolytically to release the core toxic fragment.
The toxic fragment of cry protein have three domains, domain I function in pore and ion channel formations,
domain II is involved in receptor recognition, while domain III bind to receptor. The toxic fragments are
believed to bind to specific high affinity receptors present in the brush border of mid gut epithelial cell. As a
result, the brush border membrane develops pores, permitting influx into the epithelial cell of ion and water,
which cause their swelling and eventual lysis.
HERBICIDE RESISTANCE
The early herbicides were found to be very destructive for most plants and they created undesirable
environmental impacts. New chemicals such as glyphosate have been widely recommended for use because
glyphosate is environmental-friendly as soil microorganisms are able to degrade it rapidly. Plants expressing
transformed herbicide tolerance accounted for 71% of all transgenic crops grown worldwide in 1998 and 1999
[15]. Herbicide tolerant soybean, corn, cotton and canola represent the major transgenic products [16].
Herbicide resistant Amaranthus palmeri developed by expressing glyphosate-insensitive herbicide target site
gene, 5 enol pyruvylshikmate-3 phosphate synthase (EPSP) that is involved in the shikimate cycle where it
catalyzes the reversible addition of enolpyruvyl moiety of phosphor enolpyruvate to shikimate 3 phosphate[17].
Generally two approaches have been used to create herbicide tolerant crops either modify the degree of
sensitivity of the target enzyme so that the plant sensitivity to the herbicide is inhibited, or to engineer the
herbicide-detoxifying pathway into the plant [18]. First approach includes transgenic plants tolerant to the
herbicide acifluorfen, which inhibits chlorophyll biosynthesis, have been produced through over-expression of
the target enzyme involved in chlorophyll biosynthesis [19]. In comparison, resistance to glufosinate and
bromoxynil is based on the second approach. By introducing genes that enhance metabolism of these herbicides
the active compound is converted to products that are non-toxic to the crop [20].
68
VIRUS RESISTANCE
TMV resistant tobacco and tomato plant are produced by introducing viral coat proteins. Other viral resistant
transgenic plants are Potato virus resistant potato, RSV resistant rice, YMV resistant black gram, YMV resistant
green gram etc.
ABIOTIC STRESS TOLERANCE
Abiotic stresses such as salt, drought, flooding, extreme temperature and oxidative stresses often diminish plant
growth and final yield. Agricultural productivity could be increased dramatically if crops were redesigned to
better cope with environmental stresses. Over-production of a superoxide dismutase (SOD) gene resulted in
increased chilling tolerance in plants. This could be due to the reason that different stress environment (high
light intensity, pathogens and cold) produce reactive oxygen species (ROS) which can damage to plants.
Antioxidant enzymes such as superoxide dismutase, catalase and peroxidase have the capacity to neutralize the
effect of ROS [21, 3, 4]. While, observing the constitutive expression of Osmyb4 rice gene in A. thaliana under
salinity, drought, temperature (low and high), and oxidative stress, shows that this gene helps in stress tolerance
by regulating vital metabolites as well as ROS scavengers [22]. The over-expression of strawberry GalUR gene
in transgenic potato resulted in enhanced tolerance to methyl viologen (MV), mannitol and salinity by
increasing chlorophyll pigments and 1.62-fold high accumulation of AsA in transgenic plants as compared to
that in wild type (non-transformed) plants[23]. The levels of AsA in the transgenic potato were significantly
associated with enhanced GalUR activity.
CONCLUSION AND FUTURE PROSPECTIVE
The advent of genetic engineering (GE) and other tools has enabled plant biologists to fight against the
prevailing adversaries. GM plants have been generated for their enhanced tolerance to herbicides and pests. Bt
cotton is one of the best example of insect resistance transgenic plant. In India, transgenic Bt cotton was
approved for commercial cultivation in 2002 and area under Bt cotton increased at the rate of almost 100%
every year. In 2007, 131 different Bt cotton hybrids were in cultivation. The productivity of cotton during this
period (2002-2007) increased from 300kg/ha to around 500kg/ha and as a result, India has now become a net
exporter of cotton from being a net importer till 2003-04. In future, the transgenic crops will be used not only
for improved agronomic traits, but also for traits involving food processing, pharmaceuticals (including edible
vaccines) and specialty chemicals. Transgenic rubber tree has also been produced and will be used for a variety
of purposes. Thus the future of transgenic crop is bright. Undoubtedly, there is a consistent increase in the use of
genetically modified organisms for food or other essential commodities. The promoters of GM foods claim that
they are environment-friendly, have no risk to human health, profitable for farmers as well as well regulated,
many people are still of the firm view that GM foods can be injurious to human and animal health, because they
have not been properly tested. Also it is not certain what types of long-term effects GM foods can cause. Critics
argue that transferring new genes into a food can alter the chemical composition of that food, which may trigger
the human body to respond differently to that food, thereby developing allergies or causing long-term toxicity.
Thus, every country needs to frame well defined rules and regulations for the utilization of GM organisms,
although many developed and some developing countries have already formulated specific regulations.
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69
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70
21. Vannini C., Locatelli F., Bracale M., Magnani E., Marsoni M. and Osnato M. Overexpression of rice
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71
Depression in Serum Zinc Concentration and Elevation in Serum Potassium

Concentration in Chronic Renal Failure Patients
Punam Yadav
Department of Chemistry, M. S. College, Saharanpur U.P.
Corresponding address: Dr. Punam Yadav, New Madhonagar Saharanpur-247001 (U.P.)
Email- yadavpunam200670@gmail.com
ABSTRACT
Abnormal serum zinc and serum potassium levels have been associated with increased mortality in numerous
observational studies. Hypozincaemia is defined as a decrease in the serum zinc concentration to a level below
48-96 g/ 100 ml. Hyperkalaemia occurs when serum potassium concentration is increased in chronic renal
failure patients. Hyperkalaemia is dangerous because cardiac arrest can occur when plasma potassium exceeds 7
mmol/L. Both Hypozincaemia and Hyperkalaemia are common conditions, especially in hospitalized patients
and in patients with various comorbid conditions such as chronic renal failure disease. The present paper
includes the study of serum potassium levels of 200 patients (according to age group and sex) with chronic renal
failure (CRF) before and after the process of treatment and it has been compared with 50 normal healthy
individuals comprising the control group.
Key Words: Serum zinc, Serum Potassium, Hypozincaemia, Hyperkalaemia, Chronic Renal Failure
INTRODUCTION
Zinc is an important cation in the body. In recent times, zinc has been recognized as a constituent of prime
importance in a variety of metallo-enzyme systems and biochemical pathways essential for protein synthesis and
metabolism of carbohydrates, fats and proteins. Zinc is therefore, crucial for growth and development.
Potassium is the major components of the cations of the extracellular fluid and exists in the body in association
with the anion is chloride, bicarbonate, phosphate and lactate. The important functions of potassium are to
regulate acid-base equilibrium and maintenance of the osmotic pressure of the body fluid thus protecting the
body against excessive fluid loss. It also functions in the preservation of normal irritability of muscles and the
permeability of the cells. Conclusive evidence of the essentiality of zinc to the normal growth and development
of animals was not reported until 1934 [1]. The first concrete demonstration of a specific biologic function was
published in 1939 [2]. More recently DNA department, RNA polymerase has been shown to be zinc dependent
enzymes [3].The activity of ribonuclease has been demonstrated to increase in zinc deficient tissue, suggesting
that RNA catabolism is regulated by zinc. The chronic renal failure (CRF) is one of the most severe diseases
worldwide[4]. The renal failure occurs when the kidneys cannot properly remove wastes that causes buildup of
waste and fluid in the body [5].It was first reported that uremics had a factor in plasma that could reduce Na +-K+
ATPase activity of normal erythrocyte using a cross incubation method [6]. The potassium balance is usually
maintained in the early stage of chronic renal failure through the increased potassium excretion per functioning
nephron and the colon by aldosteron induced increase in Na-K ATPase activity as long as urine output remains
adequate [7]. The evidence is also available to suggest a contribution of Potassium recycling to the overall
handling of potassium along the loop of henle [8]. The thick ascending limb of the loop of henle is an important
site of sodium, potassium, bicarbonate and ammonium transport[9,10]. In patients of chronic renal insufficiency,
fractional potassium excretion is greatly increased [11]. The deficiency in the pumps energy substrate the ATP
72
itself may perhaps be ruled out since the pump is saturated with ATP under physiological conditions [12]. The
serum potassium values in the renal failure patients were not significantly different from normal values (4.5
0.2 meq/l vs 4.2 0.1 meq/l) [13].The relationship between changes in potassium balance and electrolyte and
fluid transport, particularly with respect to potassium along the loop of henle remains to be fully elucidated. He
also observed that no significant changes in plasma sodium concentration were observed. Moderately elevated
plasma potassium levels were observed in another study of chronic hyperkalemia [14]. Chronic kidney disease
is known to affect by the disturbance in the concentration of serum urea, serum creatinine, serum electrolytes
and serum uric acid [15-17].
EXPERIMENTAL
Materials and methods:
The present study was carried out on 200 adult patients of chronic renal failure attended in the S.V.B.P. hospital
attached to L.L.R.M. Medical College, Meerut and also 50 normal healthy individuals with age, sex matched
who had no history of renal failure to serve as controls. All the known cases of chronic renal failure were
included in this study on the basis of clinical and biochemical criteria. After confirmation of diagnosis on the
above parameters, blood samples were drawn from these patients for the estimation of serum zinc and serum
potassium levels.
Observations:
TABLE I
SHOWING DISTRIBUTION OF C.R.F. CASES ACCORDING TO AGE GROUP AND SEX
Age Groups Number of cases Total

(Years) Males Females
10-30 5 2 7(3.5%)
31-50 50 25 75(37.5%)
51-70 63 40 103(51.5%)
71-above 10 5 15(7.5%)
Total 128(64.0%) 72(36.0%) 200(100.0%)
Out of 200 individuals, 128 (64%) controls were males individuals and rest 72 (36%) were females. All the 200
individuals were between the age group of 10 above 70 years. The maximum number of cases, 103 (51.5%),
were observed in the age group of 51-70 years followed by 75 (37.5%) cases in the age group of 31-50 years,
15(7.5%) cases in the age group of above 70 years and 7 (3.5%) cases in the age group of 10-30 years. It is
observed that the incidence of chronic renal failure reaches its maximum strength during middle age and later
part of life.
TABLE II
SHOWING DISTRIBUTION OF CONTROL CASES ACCORDING TO AGE GROUP AND SEX
73
Age Groups Number of cases Total

(Years) Males Females
10-30 4 1 5(10.0%)
31-50 12 6 18(36.0%)
51-70 18 9 27(54.0%)
Total 34 16 50(100.0%)
Out of 50 control cases, 34 (68.0%) cases were males and 16 (32.0%) were females. 54.0% were found in the
age group 51-70 years, 36.0% were 31-50 years age group and 10.0% were 10-30 years age group.
TABLE III
DISTRIBUTION OF C.R.F. CASES ACCORDING TO DURATION OF ILLNESS
Duration of illness No. of cases Percentage %

3 months-6 months 42 21.0%
6 months-1 year 114 57.0%
More than 1 year 44 22.0%
Total 200 100.0%
The majority of chronic renal failure cases were among more than 6 months- 1 year duration (114 cases, 57.0%)
and then more than I year children (44 cases, 22.0%).
TABLE IV
SERUM ZINC AND SERUM POTASSIUM LEVEL IN NORMAL HEALTHY CONTROLS
ZINC POTASSIUM
Age in years Age in years
MALE 10-30 31-50 51-70 Total 10-30 31-50 51-70 Total
No. 4 12 18 34 4 12 18 34
Range 76-115 80-114 83-115 76-115 4.5-6.0 2.5-5.9 4.2-6.0 2.5-6.0

MeanS.D. 103.75 98.38 97.61 98.59 5.25 4.22 5.14 4.82
16.08 28.86 9.26 10.41 0.56 0.97 0.52 0.86
FEMALE 10-30 31-50 51-70 Total 10-30 31-50 51-70 Total
No. 1 6 9 16 1 6 9 16
Range 73-80 85-110 80-109 73-110 3.5-4.5 3.0-4.0 3.8-4.9 3.0-5.0

MeanS.D. 73.00 97.58 97.33 95.88 4.10 3.86 4.40 4.18
9.32 8.46 9.44 10.84 0.34 0.98 0.37 0.58
TOTAL 10-30 31-50 51-70 Total 10-30 31-50 51-70 Total
74
No. 5 18 27 50 5 18 27 50
Range 73-115 80-114 80-115 73-115 3.5-6.0 2.5-5.9 3.8-6.0 2.5-6.0

MeanS.D. 97.60 98.11 97.52 97.72 5.02 4.10 4.89 4.61
18.92 9.49 9.29 10.79 0.67 0.92 0.59 0.84
The level of serum zinc in healthy subjects was 73-115 g/100ml (mean 97.7210.79 g/100ml). In males, the
range was 76-115 g/100ml (mean 98.5910.41 g/100ml) and in females, the range was 73-110 g/100ml
(mean 95.8810.84 g/100m)l. The highest serum zinc level was observed in the age group of 31-50 years,
ranged as 80-114 g/100ml (mean 98.119.49 g/100ml) while highest serum potassium level was observed in
the age group of 10-30 years, ranged as 3.5-6.0 mmol/L (mean 5.020.67mmol/L). The lowest serum zinc level
was observed in the age group of 51-70 years, ranged as 80-115 g/100ml (mean 97.529.29 g/100ml) while
lowest serum potassium level was observed in the age group of 31-50 years, ranged as 2.5-5.9 mmol/L (mean
4.100.92mmol/L). No significant difference was seen among the serum zinc and potassium levels of different
age groups and sexes. Our observations are very close to the observations of many workers (Kavukcu et. al.
1993, normal serum potassium level is 4.10.2 mmol/L), (Unwin et. al. 1994, normal serum potassium level is
4.090.06 mmol/L), (Price 1978, normal serum potassium level is 3.4-5.4 mmol/L) and (Harper et. al. 1979,
normal serum potassium level is 2.5-5.0 mmol/L)[14,18-20].
TABLE V
SERUM ZINC AND SERUM POTASSIUM LEVELS BEFORE AND AFTER TREATMENT IN
TOTAL CASES OF CHRONIC RENAL FAILURE
Serum Zinc and Potassium
ZINC POTASSIUM
Interval No. of Range (g/100 Mean S.D. Range (mmol/L) Mean S.D.
Cases ml)
Control 50 73-115 97.7210.79 2.5-6.0 4.610.84

Before treatment 200 48-96 81.2012.84*** 3.2-8.0 6.461.32***
15 days after treatment 186 50-100 85.6013.77*** 3.0-7.5 5.921.15***
30 days after treatment 169 60-108 91.5610.95*** 2.8-6.9 5.270.99**
60 days after treatment 145 66-110 95.368.74 2.6-6.4 4.850.95
90 days after treatment 122 70-114 97.0410.34 2.4-5.9 4.540.89
P- Significance, control vs treatment
*p < 0.05, **p < 0.01, ***p < 0.001.
75
The range of serum zinc level was significantly low before and after thirty days of treatment while range of
serum potassium level was significantly high before and after fifteen days of treatment in the patients of chronic
renal failure as compared to controls. The range of serum zinc before, after fifteen and thirty days of treatment
were 48-96 g/100 ml (mean 81.2012.84 g/100 ml), 50-100 g/100 ml (mean 85.6013.77 g/100 ml) and
60-108 g/100 ml (mean 91.5610.95 g/100 ml) respectively. Serum zinc level after sixty and ninety days of
the treatment ranged as 66-110 g/100 ml (mean 95.368.74 g/100 ml) and 70-114 g/100 ml (mean
97.0410.34 g/100 ml) respectively. The range of serum potassium level before treatment was 3.2-8.0 mmol/L
(mean 6.461.32 mmol/L). After fifteen, thirty, sixty and ninety days of treatment serum potassium ranged as
3.0-7.5 mmol/L (mean 5.921.15 mmol/L), 2.8-6.9 mmol/L (mean 5.270.99 mmol/L), 2.6-6.4 mmol/L (mean
4.850.95 mmol/L) and 2.4-5.9 mmol/L (mean 4.540.89 mmol/L) respectively.
After sixty and ninety days no significant difference was observed in serum zinc and serum potassium as
compared to controls.
DISCUSSION
Hypozincaemia in uremia is rather a result of a shift of zinc from plasma into tissue due to zinc while
Hyperkalemia is generally occur in the patients of severe chronic renal failure due to potassium imbalance. The
mean whole blood zinc concentration of male and female hemodialysis patients was significantly below control
values while mean whole blood potassium concentration of male and female patients was significantly high than
control values during chronic renal failure. The treatment of both these conditions is tricky, as over-treatment
can lead to potentially dangerous complications and under-treatment is associated with significant mortality and
morbidity. It is therefore essential to monitor the serum sodium and serum potassium concentration every 2 4
hours to prevent treatment related complications. The present study is conducted on a total of 250 individuals,
out of which 50 are normal healthy individuals comprising the control group and rest 200 is of chronic renal
failure. Results of biochemical parameter like serum uric acid from this study are discussed below-
Out of 200 individuals, 128 (64%) controls were males individuals and rest 72 (36%) were females. All the 200
individuals were between the age group of 10 above 70 years. The maximum number of cases, 103 (51.5%),
were observed in the age group of 51-70 years followed by 75 (37.5%) cases in the age group of 31-50 years,
15(7.5%) cases in the age group of above 70 years and 7 (3.5%) cases in the age group of 10-30 years (Table I).
Out of 50 healthy controls, 34 (68%) controls were males individuals and rest 16 (32%) were females (Table
II). It is observed that the incidence of chronic renal failure reaches its maximum strength during middle age and
later part of life.
Biochemical Studies
The levels of serum zinc and serum potassium were studied in controls and in all cases of chronic renal failure.
In normal healthy subjects serum zinc ranged from 73-115 g/100ml (mean 97.7210.79 g/100ml).In males, it
ranged from 76-115 g/100ml (mean 98.5910.41 g/100ml) and in females, 73-110 g/100ml (mean
95.8810.84 g/100ml) (Table IV). The range of serum potassium in healthy subjects was 2.5-6.0 mmol/L
(mean 4.610.84mmol/L). In males, it was 2.5-6.0 mmol/L (mean 4.820.86mmol/L).and in females, it was 3.0-
5.0 mmol/L (mean 4.180.58mmol/L) (Table IV). No significant difference was observed in the serum zinc
level of different age groups and sex. In cases of chronic renal failure serum zinc was found to be depressed in
72% cases while serum potassium was found to be elevated in 84% cases. Before treatment serum zinc level
76
was 81.2012.84 g/100 ml which was significantly low (p < 0.001) as compared to that of controls
(97.7210.79 g/100 ml) and serum potassium level was 6.461.32 mmol/L which was significantly high (p <
0.001) as compared to that of controls (4.610.84 mmol/L). Generally Hypozincaemia occurs due to lower
serum zinc level while hyperkalaemia occurs due to high serum potassium level in the patients of chronic renal
failure. In Hypozincaemia the decreased intake of zinc due to low protein diet is regarded as a relevant factor in
the development of zinc deficiency in chronic renal failure. The plasma zinc concentration is dependent on the
balance between anabolic and catabolic processes.
SERUM ZINC LEVELS IN TOTAL CASES OF CRF
120
100
SERUM ZINC LEVELS
80
60 CRF CASES
40
20
0
Control Before After 15 After 30 After 60 After 90
Treatment Days Days Days Days
TIME INTERVAL (DAYS)
Hyperkalaemia is caused by various metal disorders, shift of potassium from tissues etc. Hyperkalaemia may
also develop rapidly if the potassium load is increased or excretory capacity is limited. Hyperkalaemia is
dangerous because cardiac arrest can occur when plasma potassium exceeds 7 mmol/L. So, the corrections of
sodium and potassium electrolytes are very important for the improvement of the condition of patients.
77
SERUM POTASSIUM LEVELS IN TOTAL CASES OF

CRF
7
SERUM POTASSIUM LEVELS
4
CRF CASES
3
0
Control Before After 15 After 30 After 60 After 90
Treatment Days Days Days Days
TIME INTERVAL (DAYS)
SUMMARY AND CONCLUSIONS
The decreased levels of serum zinc caused hypozincaemia and increased levels of serum potassium levels
caused Hyperkalaemia in chronic renal failure patients. In the study group, the levels of serum zinc were found
decreased ranging between 48-96 g/ 100 ml and mean 81.2012.84 g/ 100 ml however the levels of serum
potassium were found highly increased, ranging between 3.2-8.0 mmol/L and mean 6.461.32 mmol/L.
Significant difference (p < 0.01) was observed among the chronic renal failure patients and controls. The serum
zinc and serum potassium levels are closely related to the severity of the disease.
The following conclusions are derived from this study:
1- There was insignificant difference in the levels of all the above mentioned parameters as regards to age
or sex of the healthy controls included in this study.
2- Maximum probabilities of chronic renal failure were found in the age group of 51-70 years (51.5%).
3- Minimum probabilities of chronic renal failure were found in the age group of 10-30 years (3.5%).
4- The levels of serum zinc and serum potassium were found to be significantly depressed and elevated
respectively in cases of chronic renal failure as compared to that of controls.
5- The levels of serum zinc and serum potassium remained low and high respectively after thirty days of
treatment and then returned to normal.
6- The fall in the levels of serum zinc and serum potassium is related to the extent of the disease.
7- The levels shifted to normal range as the condition of patients improved clinically.
REFERENCES
1. Tood W.R., Elvehjem C.A. and Hart E.B. Amer. J. Physiol. (107) 146 (1934)
2. Keilen D. and Mann T. Nature. 144, 442.(1939)
78
3. Kichgesser H., Roth H.P. and Weigand E. Biochemical changes in zinc deficiency in trace
elements in human health and disease. Vol. I. Prasad A.S. ed. Acad. Press, New York. P-189
(1976)
4. Stenvinkel P. and Alvestrand A. Inflammation in end-stage renal disease: Sources,
Consequences, and Therapy. Semin Dial, 15(5) 329-337 (2002)
5. Lise M., Cassio L. and Richar M.G. "Kidney Failure". The Journal of the American Medical
Association, 301(6) 686 (2009)
6. Cole C.H., Balfe J.W. and Welt L.G. Induction of a Quabain Sensitive ATPase defect by uremic
plasma. Trans Assoc. Am. Phys. (81) 213-220 (1968)
7. Bastl C., Hayslett J.P. and Binder H.J. Increased large intestinal secretion of potassium in renal
insufficiency. Kidney Int., (9) 12 (1977)
8. Jamison R.L., Work J. and Schafer J.A. New pathways for potassium transport in the kidney. Am.
J. physiol. (242) 297-312 (1982)
9. Good D.W., Kneppar M.A. and Burg M.B. Ammonia and bicarbonate transport by thick
ascending limb of rat kidney. Am. J. Physiol. ( 247), 35-44 (1984)
10. Greger R., Schlatter and Lang F. Evidence for electroneutral sodium chloride co-transport in the
cortical thick ascending limb of Henles lop of rabbit kidney. Pfluger Arch., (396) 308-314 (1983)
11. Hene R.J., Koomans H.A., Boer P., Roos J.C. and Mees E.J.D. Relation between plasma
Aldosterone concentration and Renal handling of Sodium and Potassium in particular in patients
with CRF. Nephron. (37) 94-99 (1984)
12. Mujais S.K., Sabatini S. and Kurtzman N.A. Pathophysiology of the uremic syndrome in the
kidney, edited by Brenner BM, Rector FC, (3rd ed.) phiadelphia, Saunders WB. P- 1950 (1986)
13. Ray S., Piraino B., Chong T.K., Shanawy M.E. and Puschett J.B. Acid excretion and serum
electrolyte in particular in patients with Advanced CRF. Miner elect. Metab. (16)355-361 (1990)
14. Unwin R., Capasso G. and Giebisch G. Potassium and sodium transport along the loop of Henie,
effect of altered dietry potassium intake. Kid Int. (46)1092-1099 (1994)
15. Yadav Punam, Malik Dinkar, Kumar Sandeep, Malik Vijai. A role of serum uric acid in Chronic
Renal Failure Patients and its effects. Int. J. Sci. Res. & Edu. 2(3), 434-442 (2014)
16. Yadav Punam, Malik Dinkar, Kumar Sandeep, Malik Vijai. Effect of elevated creatinine level in
blood serum of Chronic Renal Failure Patients. Biological Forum, 6(1) 48-52 (2014)
17. Yadav Punam, Malik Dinkar, Kumar Sandeep, Malik Vijai. Study of serum urea in the patients of
chronic renal failure. Medical Science, (4) 76-80 (2014)
18. Kavukcu S., Saatci U. and Ozean S. Effects of recombinant human erythropoietin on sodium
balance in non dialysed children with CRF. Int. Urol. Nephrol. 144,442 (1993)
19. Prices. Text book of the practice of Med. Scott, R.B.E.L.B.S. 12 th ed. Oxford, 1948-1950, 388
(1978)
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Maruzen Asian ed. Lange Medical Publications. ( 215) 579 (1979)
79
Enhancement of Fe and Zn in Cowpea grain by applying soil and foliar application
Namita Yadav* & Y.K. Sharma

Plant Physiology lab, Botany Department, University of Lucknow, Lucknow
ABSTRACT
This pot experiment was carried out in green house condition to investigate Fe, Zn and Fe+Zn soil and foliar
application effects on their concentration and protein in cowpea (vigna unguiculata) grain. The experiment was
designed with three replicate. In soil application concentration of Fe and Zn (10ppm) were applied with N: P:
K and farm yard manure in presowing soil for proper status of soil. In foliar application 0.5% of Fe and Zn
were sprayed at four different stages of plant growth. The results showed that seed protein and tissue
concentration of elements were more significant in individual and combined application of Fe and Zn than
control. The study of results explains that soil and foliar application with micronutrients may have an important
role in increasing cowpea yield.
Keyword: cowpea, soil application, foliar application, tissue concentration and protein
*Corresponding Author
namita0788@gmail.com
Introduction:
Cowpea is an important crop of legume family. Legumes are rich source of protein and energy for human food.
Cowpea seeds are a nutrition component in the human diet as well as a nutritious livestock feed. The protein in
cowpea seeds is rich in lysine and tryptophan amino acids compared to cereal grains [1]. Due to nutritional
value and economic importance, it is necessary to focus on improving crop yield and production by applying
fertilization strategies. Using soil and foliar application of micronutrients is one of most important strategies
involved in improving plant growth, yield and quality of cowpea crop. Iron and zinc are essential micronutrient,
need in small amount for plants. They can maintain crop physiology balance and play vital role in plant
metabolism such as photosynthesis respiration, nitrogen fixation, chlorophyll development and function and
reproductive physiology [2]. El-Fouly [3] reported that the availability of micronutrients such as Fe, Mn and Zn
is much affected by pH and CaCO3 content as well as soil texture. The availability and uptake of micronutrients
by plants decrease with increasing soil pH [4]. The mobility of microelements in soil and their translocation in
plants as well as interaction among themselves play an important role in plants nutrition [2]. The deficiency of
microelements in higher plants impaired the important metabolic functions and resulted in poor growth and
yield of crops [5]. They also play vital role in enzyme metabolism being the cofactor and constituent of some
enzymes and protein [2]. To sustain high crop yield the application of nutrients are required. In soil application
nutrients are applied to plants with soil mixing while in foliar application nutrient are applied to plants in soluble
form by spray. The aim of this study is to investigate the effect of micronutrient foliar and soil application on
cowpea yield and quality of grain in soil.
Materials and methods:
80
Cowpea (Vigna unguiculata) var. rituraj plants were grown in pots containing 10 Kg soil in a glass house at
ambient temperature (15.C30.C). The crop was raised in soil prepared with recommended dose of nitrogen (N),
phosphorous (P) and potassium (K) 120: 60: 60 and farmyard manure. Experiment included a soil application
for proper status of soil and four foliar treatments of iron and zinc at different stages of plant growth as follows:
Soil application
1) NPK + farmyard manure ( control)

2) FeSO4 (10ppm) with NPK + farmyard manure
3) ZnSO4 (10ppm) with NPK + Farmyard manure
4) ZnSO4 (10ppm) + FeSO4 (10ppm) with NPK + farmyard manure
Foliar application
ZnSO4 (0.5%) and FeSO4 (0.5%) applied on 35D, 45D, 55D and 65D.
The experiment was designed with three replicate. Initially two plants were maintained in each pot. Apart from
the visual effect of Fe and Zn, the concentrations of iron, zinc and protein were observed at 35D, 45D, 55D and
65D in cowpea seeds. Tissue concentration was determined in oven dried seed material after wet digestion [6]
and estimated in AAS and seed protein was estimated by Lowry et al [7] method.
Results and Discussions:
Vegetative growth of crop was promoted with the foliar application of iron and zinc. Plant height, pod length,
number of pods per plant, number of seed per pod, weight of 100 seeds were more significantly increase as
compare to control.
Cowpea
Results showed that individual and combined treatment of iron and zinc significantly increased nutrient
concentration of cowpea seeds as compared to control treatment (Figure 1, 2). The results indicate that highest
concentration of iron was accumulated at vegetative phase (35D) in individual treatment but in combined
treatment iron concentration suppressed by zinc application. Zinc concentration was more accumulated at
reproductive phase (65D) in individual application and in combined application zinc concentration was more as
compared to control treatment. Seed protein result showed significant differences among treatments as well as
difference for interaction of iron and zinc on the protein percentage of seed. The highest protein percentage was
obtained when cowpea treated with iron and zinc individual treatment and in combined treatment of iron and
zinc the protein concentration was more as compared to control treatment (Figure 3). Zeidan [8] observed that
Zn application significantly increased the grain Zn concentration, while simultaneously reduced the grain P
concentration.
81
Graph Values are mean of 3 replications-
120
100
ppm Fe in dry matter

80
60 Fe(contol)
40 Fe
20 Fe(Zn+Fe)
0
35D 45D 55D 65D
Days of foliar application
Figure 1: Effect of foliar concentration of iron in Cowpea seed at different days
40
ppm Zn in dry matter
35 Zn(control)
30
25
20
Zn
15
10
5
0 Zn(Zn+Fe)
35D 45D 55D 65D
Figure 2: Effect of foliar concentration of zinc in cowpea seed at different days
20
% protein in dry matter
19
18
control
17
Zn
16
Fe
15
Zn+Fe
14
35D 45D 55D 65D
Figure 3: Effect of foliar concentration of Fe and Zn on protein percentage in cowpea seed
Conclusion:
Zinc and iron deficiency in human are important malnutrition problem worldwide. Human Zinc deficiency is
major cause of disease and deaths in developing countries. Zinc deficiency is associated with poor growth and
82
development and impaired immune system. By applying soil and foliar application can improve the quality and
value of grain. This is good implication for human health because these elements are required in human.
Acknowledgements:
Authors are very grateful to Dr. Joba Chatterjee, Department of Botany, University of Lucknow, Lucknow for
her valuable help and guidance. Corresponding author is grateful to CSIR, New Delhi for financial help (ref no
21/12/2014(ii) EU-V).
References:
1. Esia G. S. A and Ali T. B. Impact Spraying of Some Microelement on Growth, Yield, Nitrogenase Activity
and Anatomical Features of Cowpea Plants, World Journal of Agricultural Sciences, 10(2), 57-67 (2014).
2..Marschner, H., Mineral Nutrition of Higher Plants. 2nd ed. Academic Press, Harcourt Brace Company,
Publisher Lodon, pp. 313- 396 (1995).
3. Fouly, M.M., Micronutrients in arid and semiarid areas: Level in soils and plants and the need for fertilizers
with reference to Egypt. Proc. 17th Colloquium of the International Potash Institute Bern, Switzerland, pp: 163-
173 (1983).
4.. Mortvedt, J .J., F.R. Cox, L.M. Shuman and R.M. Welch, Micronutrients in Agriculture,Published by Soil
Soc. Amer. Inc. Madison, Wisconsin, USA, pp: 760 (1991).
5. Srivastava, P.C. and Gupta, U. C. Trace Elements in Crop Production. Science Pub. Inc. Lebanon, NH03766
USA, pp: 366 (1996).
6.. Piper, C. S. Soil and Plant analysis monograph from waite Agric. Res.Inst.TheUniv. Adelaide .Adelaide
(1942).
7. Lowry, O. H., Rosenbrough, N. J., farr,A.L. and Randell,R.J. Protein measurement with Folin-Phenol. J. Biol.
Chem. 193, 265-275 (1951).
8. Zeidan, M.S.,. Response of wheat plants (Triticum aestivum L) to different methods of Zinc fertilization in
reclaimed soils of Egypt. Plant Nutrition - Food Security and Sustainability ofAgro-ecosystems (Eds W.J. Horst,
et al.), Kluwer,Dordrecht, The Netherlands, pp: 1048-1049 (2001).
83
Adoption of Improved Technologies in Black Gram Crop

LOKENDRA KUMAR SINGH
Department of Agricultural Extension
Janta Vedic Collage, Baraut, Baghpat.
ABSTRACT
India is largest producer of pulses in world with 25 per cent share in global production.
Chickpea, Pigeon Pea.MungBean, UradBean, lentil and Field Pea are important pulses crop
contributing 39.00 per cent,21.00 per cent, 11.00 per cent, 7.00 per cent and 5.00 per cent to the
total production of pulses in the country. The total production was estimated 14.56 million tones
and an area of 23.63 million hectares with average producti vity 625kg/ha. (www agro pedia. Nic
in.2015) Over 75% of the total of Black gram is generally known as dal milling or, dehulling. Milling means
removal of the outer husk and splitting the grain into two equal halves. Dalmilling is one of the major food
processing industries in the country, next only to rice milling. Black gram used as daal and as ingredient in
snacks like idly, dosa, vada and papad etc. Adoption of Improved practice is the more essential part of
increasing the production, because the population is increase day to day and holding size is decrease, so dal
price is highly explosion The first week of October 2015 noted of cost of Black Gram dal 150- 180 Rupees per
kg in the retails just before one year the cost October 2014 was 75-85 Rupees per kg(sources individual market
survey 2015 ). We need the more production of pulses, we solve this problems by the applied more technologies
in their field .
Key Word : Black Gram. Improved Practice, Agriculture.
Introduction
No doubt India lives in the villages and about 50 per cent of the 6.41 lac villages of the country are
situated in different terrain characterized by poor socio-economic condition. Even a casual glimpse at the
sub continent of India is sufficient to convin ce that ours is a land of villages. Good majorities of her
people i.e. nearly 68.84 per cent lives in villages and are occupied in the agriculture. According to
the latest census figures, there are only 7936 towns in India; whereas the numbers of villages are
6.41 Lac.
India is largest producer of pulses in world with 25 per cent share in global production.
Chickpea, PigeonPea.MungBean,UradBean, lentil and FieldPea are important pulses crop
contributing 39.00 per cent,21.00 per cent, 11.00 per cent, 7.00 pe r cent and 5.00 per cent to the
total production of pulses in the country.The total production was estimated 14.56 million tones and
an area of 23.63 million hectares with average productivity 625kg/ha. (www agro pedia. Nic
in.2015) Over 75% of the total of Black gram is generally known as dal milling or, dehulling. Milling means
removal of the outer husk and splitting the grain into two equal halves. Dalmilling is one of the major food
84
processing industries in the country, next only to rice milling. Black gram used as daal and as ingredient in
snacks like idly, dosa, vada and papad etc. Rhyzobium bacteria are present on the root nodules of Black gram.
The Black-Gram crop fixes atmospheric nitrogen in symbiotic association with Rhyzobium bacteria and
maintains the soil fertility Black gram is a member of the Asiatic Vigna crop group. It is an annual pulse grown
mostly as a fallow crop in rotation with rice. Similar to the other pulses, black gram, being a legume, enriches
soil nitrogen content and has relatively short (90-120 days) duration of maturity.
Materials and Methods:
Research methodology is of paramount importance in any scientific study as the validity and reliability
of the facts depend upon the system of investigation. It provides the details of the various aspects concerning the
research methodology. The scientific steps required to carry out the research are being discussed below in depth.
A. Selection of the state:
In 1947, when India gained independence, the state of United Provinces was renamed as Uttar Pradesh.
Uttar Pradesh is the biggest of 31 States in India. Uttar Pradesh is now divided into seventy five districts under
eighteen divisions .
B. Selection of the district :
There are 75 districts in the state of U.P. The Research was conducted in the District of Baghpat (U.P.).
The District of Baghpat was purposively selected.
C. Selection of the block :
Baghpat District has the privilege of having six blocks namely Baghpat, Baraut, Binauli, Chaprauli,
Khekra and Pilana. Out of six blocks, Baraut blocks were selected purposively keeping in view of the nature of
study.
D. Selection of the village :
There were 54 villages in the C.D. block Baraut, but the the five villages were selected by randomly
the selected villages are 1 Malakpur,Sinoli,Hilwari, Baoli and Lohari.
E. Selection of the respondents :
From each Villages 30 farmers were finally selected for the study. Who had adopted improved farm
practices of Black-Gram.The total 150 respondents to be interviewed under the study.
RESULTS AND DISCUSSION
The finding of the research of investigation is presented in the following table.
Adoption of Improved Agricultural Practice by the Black Gram growers.
S.No. Practice Numbers Per cent
85
(%)
1 Knowledge about Improved variety. 122 81.33
2 Sowing of Improved Variety
Type-9, Pusa-1&Pant-19. 102 68.00
3 Field Preparation
(I) By Scientific methods.- Summer Plowing, Leveling, 2- 68 45.33
Plowing of harrows &2-4 Plowing of Cultivators.
(II) By traditions methods. 82 54.64
4 Sowing time
(I) Early Time-(February-march) 86 57.33
(II) Timely-(May-June) 64 42.66
5 Soil treatment
(I) Weed control-At the time of field preparation and before the 35 23.33
germination of seed. Spray 1.00kg Basalineadd 1000liter
water.
(II) Nutrient test- NO2,P2O5 ,Ca,Mg,etc
(III) Insect treatment 5% Aldrin dust @-25 kg to add in the soil 22 14.66
at the time of field preparation,to control the eggs and pupa of
insects. 55 36.66
6 Seed rate
(I) Correct- 10-12 kg/ha. 65 43.33
(II) In Correct 12-16kg/ha. 85 56.66
7 Seed treatment Practice
(I) Sowing treated seed(packets seed) 110 73.33
(II) Sowing Un treated seed 40 26.66
8 Methods of sowing
(I) Sowing in the line (furrow method) 58 38.66
(II) Traditions method By broad casting 92 61.33
9 Irrigation
(I) Spring season 2-3 Irrigation apply 85 58.66
(II) Rainy season 1-2 Irrigation apply 68 45.33
10 Fertilizers
(I) 20-30 Kg Nitrogen 90 60.00
(II) 40-50 Kg Phosphorus 55 36.66
(III) 30-40 Kg potash 34 22.66
11 Crop Protection
(A)Weed Control
(I) Physical methods (by hands) 135 90.00
86
(II) Chemical methods(by herbicides) 15 10.00

(B) Insect Control
(I) To control Bihar hairy Cutter Pillar and Aphid to spray 57 38.00
Indosulphan 35 EC@0.15% and 800 liter water two spray to interval
of 8-10days.
12 Harvesting of Crop
(I) Maturity of pod -75-85% 64 42.66
(II) Maturity of pod 85-95% 86 57.33
13 Threshing of Crop
(I) By threshing machine 65 43.33
(II) By Bullocks and mens 85 56.66
Above table reveal that 81.00 per cent respondents have well awareness of about improved varieties of
black, but only 68.00 per cent farmers finally apply of improved varieties on their field. In case of field
preparation 54.64 per cent farmers are use of tradition methods and 45.33 per cent respondents have apply
scientific methods in their field preparation of Black Gram. While the 36.66 per cent, 23.33 and 14.66 per cent
respondents have apply soil treatment practice under the Insect treatment, weed control, and Nutrient test in
their field before the sowing. In case of seed rate and seed treatment majority i e 56.66 per cent , 73.33 per cent
respondents have adopted correct seed rate and sowing treated seed available in the market and other seed
stores. Only 38.66 per centrespondents have applied scientific methods of sowingof black Gram crop, majority
of respondents do not apply the scientific recommendation of sowing methods. In case of irrigation practice
58.66 per cent and 45.33 per cent respondents provide irrigation facilities their crop 2-3 irrigation in spring
season and 1-2 irrigation in rainy season in lack of rain. While 60.00 per cent,36.66 per cent 22.66 per cent
respondents have applied 20-30kg Nitrogen,40-50 kg Phosphorus and 30-40 Kg Potash in their field. The
very low amount of respondents is 22.66 per cent to have applied Potashic fertilizer in their crops of Black
Gram. 90.00 per cent respondents have applied physical methods ( by hands) to control weed their won
standing crops and 38.00 per cent respondents have applied chemical methods recommended by the scientist to
control hairy cutter pillar or aphid .While in case of harvesting and threshing of crop the majorities of i.e.57.33
per cent respondents harvest our crops when the 85-95 per cent pod mature, and 42.66 per cent respondents have
harvest won crop when the pod mature is 75-85 per cent, majorities of respondents 56.66 per cent have thresh
their crop by the mens power and Bullock and 43.33 per cent respondents are applied threshing machine in
duration threshing of our crops in case of Black Gram.
CONCLUSION
Overall majorities i.e. 81.00 per cent, respondents have knowledge about Improved varieties,68.00 per
cent respondents have applied Improved varieties wne their field,57.00 per cent and 57.00 per cent,respondents
adopted Early sowing and incorrect seed rate of Black Gram, 73.00per cent and 61.00 per cen,t respondents
87
have sowing the treated seed, and were used traditional methods at the time of sowing, 60.00 per cent, and 90.00
per cent, grower apply the recommended dose of Nitrogen fertilizer and farmers control of weeds in our crop by
hands helps by the Khurpy,(weeding tools) 57.00 per cent respondents harvesting of their crop when the pod
mature 85-95 per cent and threshing of their crop by men hands ,Bullock and tractors.
REFERENCES
1. Jeswani, L.M. and Baldev, B., (1988). Advances in Pulse Production Technology, Indian Council of
Agricultural Research Publication
2. Pandey ,P.H.(1988). Principles and Practices of Post Harvest Technology,
3. Acharya, S.S.andAgarwal,N.L.(1999). Agricultural Marketing in India,
4. Chakraverty, A.(1988). Post Harvest Technology of Cereals, Pulses and Oil seeds,
5. Annual Report, 2003-2004, National Cooperative Development Corporation, New Delhi.
6. Annual Report, 2004-2005,Central Warehousing Corporation, New Delhi.
7. Agmark Grading Statistics, 2003-2004 and 2004-2005, Directorate of Marketing and, FaridabadInspection.
8. Agarwal, P.K., Agricultural Marketing, ( 2002).. Establishing Regional and Global Marketing Network for
Small holders Agricultural Produce / Products with reference to Sanitary and Phytosanitary (SPS)
Requirement, PP.15-23
88
Study on SO2 Induced Effects and their Amelioration in Arhar (Cajanus cajan)
HARENDRA MALIK, NARESH KUMAR AND RENU CHOUDHARY*
Department of Botany, C.C.R. (P.G.) College, Muzaffarnagar (U.P.), India.
* Department of Biology, G.G.I.C. Gagalheri, Saharanpur(U.P.), India.
Email- harendra_ghoni@rediffmail.com
Abstract
Sulphur dioxide is a well known wide spread air pollutant and most harmful to plants. The main aim of
present investigation is to study the extent and magnitude of damage induced by sulphur dioxide and
amelioration of this damage with the help of some chemical agents like calcium hydroxide and sodium benzoate
on Cajanus cajan cv. Manak. The present study was conducted to observe the impact of exposure of four
different concentrations of sulphur dioxide i.e. 653, 1306, 2612 and 3918 g m-3 alone and after calcium
hydroxide treatment and sodium benzoate treatment on Arhar (Cajanus cajan) cv. Manak. Exposure to SO
2
inhibited the seed germination, foliar injuries, root, shoot and whole plant growth, chlorophyll a, chlorophyll b
and carotenoids contents of leaves. However, the accumulation of anthocyanin and total phenolics contents of
leaves were increased significantly in higher SO concentrations. After treatment of two ameliorating agents i.e.
2
calcium hydroxide and sodium benzoate, better growth and development was observed in SO treated plants.
2
Keywords : Pollutant, Sulphur dioxide, Amelioration, Germination, Growth, Calcium hydroxide, Sodium
benzoate, Arhar.
Introduction
Major air pollutants are oxides of sulphur (SOx), oxides of nitrogen (NOx), carbon monoxide (CO),
hydrogen sulphide (H2S), hydrocarbons, ozone (O3 ), fluorides, lead, mercury and particulates etc. Excessive
quantity of these all air pollutants can impair normal healthy life of all organisms. Amongst these air pollutants
oxides of sulphur including sulphur monoxide (SO), sulphur dioxide (SO2), sulphur trioxide (SO3), sulphur tetra
oxide (SO4), sulphur sesquioxide (S 2O3) and sulphur heptaoxide (S 2O7). SO2 is one of most toxic air pollutant
and major sources of SO2 emission are automobile exhaust, burning of fossil fuels in thermal power plants,
smelting industries, other process such as manufacture of sulphuric acid and fertilizers and refining of crude
petroleum. The hazardous effects of SO2 create a big threat to the survival and sustenance of the living systems.
As the total control of SO2 air pollution is technically and economically not feasible, various crop management
practices, such as nutrient supplementation and spraying of chemical protectants have been used to reduce air
pollution injury in plats.
Germination studies were carried out on selected plants. Seeds were selected uniformly (criteria being
the size and colour of seeds). For long-term treatments the exposure to SO2 was initiated when the seedlings
were 11 days and stop at 60 days old plants. The seedlings of each cultivars of the plants were grouped into
thirteen research plots so that four of them treated with. 653, 1306, 2612 and 3918 g m-3 concentrations of
89
SO2, remaining four treated with these four SO2 concentration and then sprayed with calcium hydroxide
(denoted by TC ) and last four sprayed with sodium benzoate after SO2 treatments (TS), while the last one was set
aside as control. The plants were given the treatments of SO2 for 2 h. on alternate days. All the normal
agronomic practices, other than fertilizer and pesticide application, were followed. Chlorophyll, carotenoid,
anthocyanin and phenolics content calculated using several deeds [1, 6, 7 & 9].
Result and Discussion
Exposure to SO2 inhibited the seed germination of the Cajanus cajan cv. Manak. Reduction in seed
germination percentage was as 0.96, 5.73, 11.20 and 16.02 percent under 653, 1306, 2612 and 3918 ug m-3
concentration of SO2, respectively. Exposure of seeds to varying concentrations of SO2 with treatment of
calcium hydroxide (Tc) and sodium benzoate (Ts) exhibited lesser reduction as compared to SO2 alone. These
results are in agreement with the findings of [3,11] in different plants.
Table 1. Seed germination percentage, mean germination percentage and seedling survival percentage of
Cajanus cajan cv. Manak exposed to different concentrations of SO 2 alone and treated with calcium
hydroxide and sodium benzoate (Tc and Ts).
Parameters SO2 treatment (g m-3)

Tc/Ts Control 653 1306 2612 3918
Seed - 94.24 93.33 88.84 83.68 79.14
germination Tc - 93.85 91.23 88.42 86.04
percentage Ts - 93.92 91.86 89.28 86.57
Mean - 20.86 20.24 19.63 18.83 17.88
germination Tc - 20.46 20.13 19.56 19.01
percentage Ts - 20.49 20.18 19.63 19.07
Seedling - 87.82 86.78 85.58 84.24 82.12
survival Tc - 87.08 86.73 86.32 85.89
percentage Ts - 87.15 86.81 86.54 86.11
Plants exposed to varying concentrations of SO2 and with calcium hydroxide and sodium benzoate
showed detectable foliar injuries. Such type of observations were also made by [2,13]. In 30 days old plants,
chlorotic patches at the margin of the lamina and at the tips of the lower leaves were exhibited. But when the
SO2 treatment duration was prolonged these patches become dark brown bifacial necrotic lesions. The injury
was mostly confined in mature leaves and new leaves develop normally or less affected. The extent of injury
increased with increasing concentration of SO2.
90
Table 5 : Growth responses of 30 days old plants of Cajanus cajan cv.

Manak exposed to different concentrations of SO2 alone and
treated with calcium hydroxide and sodium benzoate (Tc and Ts).
Cajanus cajan cv. Manak
Parameters Tc SO2 treatment (g m-3)
Control
Ts 653 1306 2612 3918
- 89.66+4.72 85.88+4.19* 79.32+2.78** 70.19+3.34** 64.31+3.73**
Plant height
Tc - 87.37+4.42 82.00+3.76* 77.89+4.17** 75.86+3.42**
(cm)
Ts - 87.87+5.11 82.60+4.63* 78.64+3.25** 76.56+3.35**
- 13.28+1.24 12.62+1.47 11.76+1.19* 10.48+1.51** 9.42+1.40**
Length of root
Tc - 12.78+1.31 12.08+1.28* 11.58+1.22* 11.09+1.33**
(cm)
Ts - 12.84+1.54 12.17+1.37* 11.76+1.13* 11.18+1.52**
- 76.38+3.48 73.26+2.72* 67.56+1.59** 59.71+1.83** 54.89+2.33**
Length of
Tc - 74.59+3.11 69.92+2.48* 66.31+2.95** 64.77+2.09**
shoot (cm)
Ts - 75.03+3.57 70.43+3.26* 66.88+2.12** 65.38+1.83**
- 3.94+0.62 3.79+0.71 3.52+0.56* 3.08+0.80** 2.68+0.59**
Fresh weight
Tc - 3.83+0.58 3.63+0.42 3.49+0.74* 3.28+0.34**
of root (g)
Ts - 3.85+0.62 3.67+0.33 3.53+0.67* 3.33+0.28**
- 24.57+1.22 23.44+1.51 21.84+1.27* 19.26+1.38** 17.13+1.12**
Fresh weight
Tc - 23.76+1.36 22.68+1.45* 21.18+1.60* 20.11+1.58**
of shoot (g)
Ts - 23.84+1.43 22.81+1.66* 21.37+1.83* 20.35+1.72**
- 0.436+0.072 0.428+0.063 0.396+0.048* 0.347+0.039** 0.303+0.045**
Dry weight of
Tc - 0.431+0.041 0.403+0.052* 0.371+0.044* 0.354+0.038*
root (g)
Ts - 0.432+0.069 0.408+0.029* 0.379+0.061* 0.359+0.074*
- 3.88+0.58 3.75+0.36 3.48+0.61* 3.07+0.49** 2.73+0.35**
Dry weight of
Tc - 3.80+0.52 3.68+0.31 3.42+0.52* 3.11+0.48**
shoot (g)
Ts - 3.81+0.47 3.71+0.41 3.48+0.73* 3.19+0.41**
- 56.08+1.29 54.22+1.15 51.92+1.24** 46.73+1.36** 42.41+1.46**
No. of leaves
Tc - 55.47+0.95 53.61+1.31* 50.64+1.27** 49.09+1.31**
per plant
Ts - 55.80+0.89 53.93+1.21* 50.97+0.98** 49.78+1.24**
- 7.10+1.48 6.96+1.53 6.77+1.34 6.40+1.46** 6.19+1.72**
No. of primary
branches per Tc - 7.01+1.31 6.82+1.18 6.68+1.12* 6.58+1.28*
plant
Ts - 7.02+1.26 6.84+1.22 6.71+1.08* 6.63+1.19*
- 24.52+1.89 23.92+2.16 21.83+2.25* 20.63+1.68** 19.80+1.51**
No. of nodules
Tc - 24.08+1.69 22.65+1.58* 22.09+1.46** 21.67+1.36*
per plant
Ts - 24.13+1.82 22.81+1.76* 22.15+1.70** 21.82+1.22*
Dry matter - 4.31+1.30 4.17+0.99 3.87+1.09* 3.41+0.88** 3.03+0.80**
production
Tc - 4.23+1.21 4.08+0.83 3.79+0.96* 3.46+0.86**
(phytomass
accumulation) Ts - 4.23+0.88 4.11+0.70 3.85+1.34* 3.54+1.15**
- 0.143 0.139 0.129 0.113 0.101
Net primary
productivity Tc - 0.141 0.136 0.126 0.115
(g/plant/ day)
Ts - 0.141 0.137 0.128 0.118
Values are in mean + standard deviation

It was observed that root, shoot and whole plant growth reduced in SO2 exposures. At SO2
concentration of 653, 1306, 2612, and 3918 g m-3, the reduction percentage in plant height were 4.21, 11.53,
21.71, and 28.27 in 30 days old plants. In Tc plants, the reduction were 2.55, 8.54, 13.12 and 15.39 percent
similarly in Ts plants, these reductions were 1.99, 7.87, 12.29 and 14.61 percent, respectively. Similar pattern of
reduction was observed in plant height of 60 days old plants. The reduction values for root length were 4.96,
11.44, 21.08 and 29.06 percent, respectively in 30 days old plants. In comparison to control, 60 days old plants
exhibited maximum reduction in root length at 3918 g m-3 concentration of SO2.
91
Table 7 : Growth responses of 60 days old plants of Cajanus cajan cv.

Manak exposed to different concentrations of SO2 alone and
treated with calcium hydroxide and sodium benzoate (Tc and Ts).
Cajanus cajan cv. Manak
Parameters Tc SO2 treatment (g m-3)
Control
Ts 653 1306 2612 3918
- 175.21+6.01 168.90+6.93* 153.99+6.50** 138.40+6.18** 123.32+4.98**
Plant height
Tc - 171.81+7.25 160.27+5.92* 150.30+5.68** 147.34+5.26**
(cm)
Ts - 172.25+6.45 160.74+5.57* 151.22+5.83** 149.67+5.56**
- 22.86+1.33 21.46+1.81* 19.68+1.52** 17.76+1.55** 15.95+1.37**
Length of root
Tc - 22.13+1.78 20.54+1.47* 19.72+1.14** 19.00+1.49**
(cm)
Ts - 22.19+1.69 20.62+1.63* 19.81+1.68** 19.15+1.65**
- 152.35+4.68 147.44+5.12* 134.31+4.98** 120.64+4.63** 107.37+3.61**
Length of
Tc - 149.68+5.47 139.73+4.45* 130.58+4.54** 128.38+3.77**
shoot (cm)
Ts - 150.06+4.76 140.12+3.94* 131.41+4.15** 130.52+3.91**
- 4.13+0.88 3.96+0.73 3.68+0.56* 3.10+0.60** 2.81+0.83**
Fresh weight
Tc - 4.04+0.58 3.81+0.76 3.56+0.47* 3.43+0.74**
of root (g)
Ts - 4.06+0.81 3.83+0.67 3.59+0.51* 3.44+0.69**
- 65.42+2.36 62.34+2.41* 56.81+2.48** 50.73+2.58** 46.09+2.33**
Fresh weight
Tc - 63.85+2.53 59.44+2.54* 56.65+2.64** 54.63+2.29**
of shoot (g)
Ts - 63.91+2.23 59.68+2.61* 56.82+2.50** 54.76+2.46**
- 0.762+0.037 0.749+0.056 0.676+0.073** 0.589+0.064** 0.519+0.037**
Dry weight of
Tc - 0.753+0.048 0.698+0.075* 0.668+0.052** 0.623+0.033**
root (g)
Ts - 0.754+0.063 0.703+0.067* 0.673+0.049** 0.626+0.041**
- 12.65+0.73 11.89+0.69* 10.74+0.86** 9.31+0.65** 8.66+0.47**
Dry weight of
Tc - 12.09+0.57 11.29+0.61* 10.54+0.58** 9.80+0.76**
shoot (g)
Ts - 12.14+0.82 11.35+0.90* 10.66+0.72** 9.89+0.63**
- 193.11+5.61 186.34+5.19* 161.58+4.87** 129.36+3.54** 103.65+4.66**
No. of leaves
Tc - 188.45+4.33 171.92+5.26* 149.88+3.71** 133.51+3.51**
per plant
Ts - 189.72+3.93 172.69+4.36* 151.43+5.12** 139.72+3.28**
- 12.41+2.27 12.08+2.16 11.51+1.96* 10.89+2.11** 10.35+2.46**
No. of primary
branches per Tc - 12.21+1.53 11.91+1.32 11.55+1.14* 11.29+1.81*
plant
Ts - 12.23+1.37 11.93+1.28 11.58+1.21* 11.38+1.64*
- 34.52+2.11 32.21+1.79* 29.49+2.31** 26.88+1.77** 25.02+1.56**
No. of nodules
Tc - 33.09+1.93 31.28+1.81* 29.53+1.69** 28.35+1.65**
per plant
Ts - 33.19+2.25 31.44+2.07* 29.68+1.62** 28.79+1.72**
Dry matter - 13.41+1.10 12.63+1.25 11.41+1.56* 9.89+1.20** 9.17+0.84**
production
Tc 12.84+1.05 11.98+1.36* 11.20+1.10* 10.42+1.09**
(phytomass
accumulation) Ts 12.89+1.45 12.05+1.57* 11.33+1.21* 10.51+1.04**
- 0.223 0.210 0.190 0.164 0.152
Net primary
productivity Tc - 0.214 0.199 0.186 0.173
(g/plant/ day)
Ts - 0.214 0.200 0.188 0.175
Values are in mean + standard deviation

The reductions in shoot length in 30 days old plants were 4.08, 11.54, 21.82 and 28.13 percent,
respectively. In Tc plants reduction was observed upto 15.20 percent at 3918 g m-3 SO2 concentration.
Corresponding values in Ts plants were 14.40 percent. Reductions in fresh weight of shoot were 30.28, 18.15
and 17.17 percent against 3918 g m-3 concentration of SO2 alone, Tc and Ts plants, respectively in 30 days old
plants. 60 days old plants also showed similar pattern of reduction in fresh weight of shoot. Reductions in dry
weight fractions of both root and shoot were recorded in all treated plants. Similar findings were also made by
[12,10,4,11] in Vigna radiata.
92
Chlorophyll a, chlorophyll b and total chlorophyll contents of leaves reduced considerably in all the
treatments. The reduction percentage in 30 days old plants in total chlorophyll content were 16.74, 9.88 and 9.10
in 3918 g m-3 of SO2 treated, Tc and Ts plants, respectively. Almost similar trend of reduction was observed
in 60 days old plants. Carotenoids, the accessory pigments also exhibited reduction in their content in all
treatments. Such studies relate to the findings of [8,5]. SO2 induced the accumulation of anthocyanin. Both, the
age of plant and concentration of fumigant (SO2) were found to have a direct relation with the amount of
anthocyanin accumulated in the cultivars. The increase in anthocyanin content was found to be maximum at
3918 g m-3 of SO2 treated plants. In comparison to control, total phenolic contents of leaves were increased
significantly in higher SO2 concentrations. In 60 days old plant, percent increase in total phenolic contents, were
64.33, 29.72 and 31.46 percent in 3918 g m-3 of SO2 treated, Tc and Ts plants, respectively.
cv. T-21
1.2
Chlorophyll content
1.0
(mg/g f.wt.)
0.8
0.6
0.4
0.2
653 g m -3
1306 g m -3
2612 g m -3
3918 g m -3
Control
Control
Control
Control
Tc
Ts
Tc
Ts
Tc
Ts
Tc
Ts
cv. Manak
1.2
Chlorophyll content
1.0
(mg/g f.wt.)
0.8
0.6
0.4
0.2
653 g m -3
1306 g m -3
2612 g m -3
3918 g m -3
Tc
Ts
Tc
Ts
Tc
Ts
Tc
Ts
Control
Control
Control
Control
Chl. a
Chl. b
Figure 2 : Effect of different concentrations of SO 2alone, after calcium

hydroxide treatment (Tc) and sodium benzoate treatment (Ts)
on Chl. a, Chl. b and total chlorophyll content of 30 days old
plants of Cajanus cajan cv. T-21 and Manak.
93
cv. T-21
1.4
1.2
Chlorophyll content
1.0
(mg/g f.wt.)
0.8
0.6
0.4
0.2
653 g m -3
1306 g m -3
2612 g m -3
3918 g m -3
Tc
Ts
Tc
Ts
Tc
Ts
Tc
Ts
Control
Control
Control
Control
cv. Manak
1.4
1.2
Chlorophyll content
1.0
(mg/g f.wt.)
0.8
0.6
0.4
0.2
653 g m -3
1306 g m -3
2612 g m -3
3918 g m -3
Control
Control
Control
Control
Tc
Ts
Tc
Ts
Tc
Ts
Tc
Ts
Chl. a
Chl. b
Figure 3 : Effect of different concentrations of SO 2alone, after calcium

hydroxide treatment (Tc) and sodium benzoate treatment (Ts)
on Chl. a, Chl. b and total chlorophyll content of 60 days old
plants of Cajanus cajan cv. T-21 and Manak.
Figure 4 :Effect of different concentrations of SO 2 alone, after calcium hydroxide treatment (Tc) and
sodium benzoate treatment (Ts) on carotenoid content (mg/g f.wt.) of 30 and 60 days old plants of
Cajanus cajan cv. Manak.
30 Days Carotenoid content (mg/g f.wt.)
0.5
0.45 Series1
0.4
0.35
0.3 Series2
0.25
0.2
Series3
0.15
0.1
0.05 Series4
0
1 2 3 4
60 Days Carotenoid content (mg/g f.wt.)
94
0.7
0.6 Series1
0.5
Series2
0.4
0.3
Series3
0.2
0.1
Series4
0
1 2 3 4
Figure 5 : Effect of different concentrations of SO2 alone, after calcium hydroxide treatment (Tc) and
sodium benzoate treatment (Ts) on anthocyanin content (mg/g f.wt.) of 30 and 60 days old plants of
Cajanus cajan cv. Manak.
30 Days Anthocyanin content (mg/g f.wt.)
0.4
0.35
Series1
0.3
0.25
Series2
0.2
0.15 Series3
0.1
Series4
0.05
0
1 2 3 4
95
60 Days Anthocyanin content (mg/g f.wt.)
0.7
0.6
Series1
0.5
0.4 Series2
0.3
Series3
0.2
0.1 Series4
0
1 2 3 4
Figure 6 : Effect of different concentrations of SO2 alone, after calcium hydroxide treatment (Tc) and
sodium benzoate treatment (Ts) on phenolics content (mg/g f.wt.) of 30 and 60 days old plants of Cajanus
cajan cv. Manak.
30 Days Phenolics content (mg/g f.wt.)
3.5
Series1
3
2.5
Series2
2
1.5 Series3
Series4
0.5
0
1 2 3 4
60 Days Phenolics content (mg/g f.wt.)
96
3.5 Series1
3
2.5 Series2
2
Series3
1.5
1
Series4
0.5
0
1 2 3 4
Conclusion
The present study was conducted to observe the impact of exposure of four different concentrations of
sulphur dioxide i.e. 653, 1306, 2612 and 3918 g m-3 alone and after calcium hydroxide treatment and sodium
benzoate treatment on Arhar (Cajanus cajan) cv. Manak. The effect of SO2 alone and with treatments of
chemicals (calcium hydroxide and sodium benzoate) were studied in respect of seed germination, visible foliar
injury, different growth parameters and some biochemical changes like chlorophyll content, carotenoid content,
anthocyanin content and phenolics content. Thus, all the treated plants under study were sensitive to SO2
pollution, though their responses vary with SO2 concentration, age of plant and stage of development. SO2
causes reduction in all growth and biochemical components (except anthocyanin and phenolics content). After
treatment of two ameliorating agents i.e. calcium hydroxide and sodium benzoate, better growth was observed
in all Tc and Ts plants under study.
References
1. Arnon, D.I. Copper enzymes in isolated chloroplasts, polyphenol oxidase in Beta vulgaris. Plant
Physiol. 24 : 1-15 (1949).
2. Ayer, S.K. and Bedi, S.J. Effect of artificial fumigation of sulphur dioxide on growth and yield of Zea
mays L. var. American Sweet Corn. Pollut. Res. 9 : 33 -37 (1990).
3. Goswami, R. Toxicity of air pollution to plants. A. Ph.D. Thesis, C.C.S. University, Meerut, India
(2002).
4. Jeyakumar, M., Jayabalan, N. and Arockiasamy, D.I. Effect of sulphur dioxide on maize (Zea mays L.)
var. (CO-1) seedlings at lethal dose 50. Physiol. Mol. Biol. Plants. 9 (1) : 147-151(2003).
5. Joshi, P.C. and Swami, A. Air pollution induced changes in the photosynthetic pigments of selected
plant species. J. Environ. Biol. 30(2) : 295-298 (2009).
97
6. Maclachlan, S. and Zalik, S. Plastid structure, chlorophyll concentration and free amino acid
composition of chlorophyll mutant of barley. Can. J. Bot. 43 : 1053-1062 (1963).
7. Mancinelli, A.L., Yang, C.P.H., Lindquist, T., Anderson, O.R. and Robion, I. Photocontrol of
Anthocyanin III. The action of streptomycin on synthesis of chlorophyll and anthocyanin. Plant
Physiol. 155 : 251-257 (1975).
8. Ranieri, A., Pieruccetti, F., Panicucci, A., Castagna, A., Lorenzini, G. and Soldatini, G.F. SO 2-induced
decrease in photosynthetic activity in two barley cultivars. Evidence against specific damage of the
protein-pigment complex level. Plant Physiol. Biochem. 37 : 919-929 (1999).
9. Sadasivam, S. and Manickam, A. Phenolics : In biochemical methods for agricultural sciences. Wiley
Eastern Limited, New Delhi, India. pp. 187-189 (1992).
10. Saxena, D.K., Saxena, A., Gupta, P. and Kamakshi. Effect of short-term fumigation of SO2 on seedling
growth of Cicer arietinum L. J. Indian Bot. Soc. 80 : 6366 (2001).

11. Tyagi, A. Sulphur dioxide damage to plants. A Ph.D. Thesis, C.C.S. University, Meerut, India (2006).
12. Verma, M. and Agrawal, M. Response of wheat plants to sulphur dioxide and herbicide interaction at
different fertility regiones. J. Indian Bot. Soc. 80 : 67-72 (2001).
13. Wang, C., Da Xing, D., Zeng, L., Ding, D. and Chen. Effect of artificial acid rain and SO 2 on
characteristics of delayed light emission. Luminescence. 20 : 51-56 (2005).
98
Synthesis, Characterization and Biological Activities of Some Metal Complexes with 2-

Amino -4- (p- Ethoxy Phenyl) Thiazoline Ligand
Dinkar Malik
Department of Chemistry, M. S. College, Saharanpur U.P.
Corresponding address: Dr. Dinkar Malik, 1800/1, Mission Compound, Saharanpur-247001 (U.P.)
Email- dinkar_malik@rediffmail.com
ABSTRACT
The ligand complexes of Ni(II), Co(II) and Cu(II) with 2 - amino -4- (p- ethoxy phenyl) thiazoline have been
synthesized and characterized with the help of their elemental analysis, IR, electronic and magnetic
susceptibility studies. From the analytical and spectral data the stoichiometry of these complexes have been
found to be of the type ML2X2 (where M = Cu (II), Co (II) and Ni (II)}. It is found that Ni(II), Cu(II) and Co(II)
complexes exhibit octahedral and square planar geometry. The fungicidal activities of ligands and metal
complexes were screened by growth method against various fungi i.e. Drechslere setramera, Fusarium
oxyporum, Macrophomera phaseoli at different concentrations. It is found that the activity decreases with
decrease of concentration and the metal complexes are less toxic than the parent ligand.
Key Words: Thiazoline Complexes, Fungicidal Activity, Heterocyclic Compounds, Toxicity, Fungicidal
activity,
INTRODUCTION
Complexes of transition metals ions containing ligands with N, S and N, S, O donors are known to exhibit
interesting stereo chemical, electrochemical and electronic properties. Semicarbazones and thio semicarbazones
are amongst the most widely studied nitrogen and oxygen/sulphur donor ligands. Besides, thio semicarbazones,
in the last two decades, have emerged as an important class of sulphur ligands particularly for transition metal
ions . The real impetus towards developing their co-ordination chemistry i.e. their physiochemical properties
and significant biological activities. Thiazolines and their derivatives have created an interest due to their wide
range of activity. Thiazole and their derivatives possess anti-malarial, anti-theminitic, anti-fungal, anti-bacterial,
anti-sparodic and anti-tubercular activities. Such compounds can also be used as local anaesthetic, anti-radiation
drugs,anti-viral and anti-protozoan agents and also in the rubber industry as vulcanization accelerators.
Transition metal complex formed by organic ligands are essential in plant nutrition, they have been studied
which induced several amines containing sulphur and mercaptoacetates. The survey of literature revealed that
metal complex play an important role in biological activity of drugs. The mechanisms of the activity of such
drugs have been explained on the basis of complex formation. Attempts have been made to study their structure
with the help of elemental analysis, megnatic mesurments, spectral studies and conductance measurements. The
synthesis, spectral characterization and biological activity of Schiffs base derived metal complexes were
studied by many workers[1-3]. Similar experiments on fungicidal and antimicrobial activites of Cu (II), Co (II)
and Ni (II) Complexes with O, N, and S donor, their EPR and electronic spectral studies were also conducted by
many workers [4-8] Schiffs base derived complexes of derivatives of DHA, their spectra and synthesis under
microwave irradiation were also studied by many workers [9.10]. The present paper deals with the preparation
99
and characterization of Cu(II),Co(II) and Ni(II) complexes with 2-amino-4-(p-ethoxy phenyl) thiazoline. Metal
complexes play an important role in biological activity. In many cases metal complexes are more potent than
free ligands. These newly synthesized complexes were also screened for their antifungal activity against fungi
viz. Drechslera-tetramera, Fusarium-oxysporum and Macrophomera-phoseoli at different concentrations.[11].
EXPERIMENTAL
MATERIALS AND METHODS:
All the chemicals and reagents used were of analytical grade; otherwise they were purified before use. Organic
solvent used was absolute alcohol. IR spectra of the ligand and complexes are recorded in nujolmull. The
fungicidal activity of ligands as well as complexes was determined by using the Growth method. The electronic
spectra were recorded in MgO at room temperature on VSU-22 spectrophotometer. The measurements were
carried out Guru Nanak Dev University, Amristar. Metal and sulphur contents of these complexes were
estimated using the standard procedures reported in literature [12,13]. The estimation of carbon, hydrogen,
sulphur and nitrogen were carried out at BHU, Varanasi and CDRI, Lucknow and results are given in Table 1.
Magnetic measurements were carried out at IIT Roorkee at room temperature using Co [Hg (CNS) 4] as a
calibrant.
TABLE 1
ELEMENTAL ANALYSIS DATA
Complexes %Calc./ Obs.

C H S N O M
C23H20ON2S 74.19 5.37 8.60 7.52 4.30 -----

74.17 5.31 8.56 7.51 4.29
[Cu(C23H20ON2S)2Cl2] 62.83 4.55 7.28 6.37 3.64 7.22
62.73 4.52 7.26 6.33 3.61 7.21
[Ni(C23H20ON2S)2Cl2] 63.17 4.56 7.35 6.42 3.65 6.77
63.14 4.52 7.29 6.39 3.61 6.73
[Co(C23H20ON2S)2Cl2] 63.15 4.57 7.32 6.40 3.66 6.75
63.13 4.52 7.26 6.39 3.64 6.71
[Cu(C23H20ON2S)2(CH3COO)2] 64.82 4.97 6.91 6.05 10.37 6.87
64.76 4.94 6.90 6.02 10.34 6.78
-
[Ni(C23H20ON2S)2(CH3COO )2] 65.16 4.98 6.97 6.11 10.45 6.43
65.14 4.94 6.95 6.05 10.39 6.41
[Co(C23H20ON2S)2(CH3COO-)2] 65.14 4.99 6.94 6.09 10.42 6.40
65.12 4.92 6.91 6.06 10.36 6.38
The ligand 2-amino-4-(p-ethoxy phenyl) thiazoline was prepared using the procedure reported in the
literature[14].
100
TABLE 2
CHARACTERISTIC IR BANDS OF LIGANDS AND COMPLEXES
Complexes IR Bands (cm-1)

N-H C-S C-H C=C C=N M-S
C23H20ON2S 3468- 834-682 3090- 1620- 1639- --
3286 3082 1590 1620
[Cu(C23H20ON2S)2Cl2] 3381- 855-690 3108- 1623- 1633- 315-309
3262 3098 1592 1618
[Ni(C23H20ON2S)2Cl2] 3391- 859-677 3106- 1621- 1630- 326-318
3264 3099 1597 1619
[Co(C23H20ON2S)2Cl2] 3399- 860-683 3105- 1618- 1627- 335-322
3283 3097 1599 1611
[Cu(C23H20ON2S)2(CH3COO)2] 3406- 866-690 3107- 1602- 1628- 319-315
3287 3091 1582 1609
[Ni(C23H20ON2S)2(CH3COO-)2] 3396- 853-684 3103- 1618- 1632- 330-321
3267 3094 1599 1607
-
[Co(C23H20ON2S)2(CH3COO )2] 3391- 867-685 3104- 1604- 1625- 338-325
3276 3096 1597 1613
A shift in the C-S and N-H band frequencies is observed in all the complexes. This shows that the lone pair of
electron presents on the sulphur atom of thiazoline ring and nitrogen atom of free amino group is taking part in
co-ordination (Table 2).
TABLE 3
(a) ELECTRONIC REFLECTANCE SPECTRAL DATA AND THEIR ASSIGNMENTS OF
NI(II) COMPLEX
Complexes 1 2 3 Dq B 2/ 1 3(Calc.)
[Ni(C23H20ON2S)2Cl2] 8523 14518 24448 1285.6 600 1.70 28966
[Ni(C23H20ON2S)2(CH3COO)2] 8515 14511 24417 1283.5 662.4 1.70 28997
1 = 3A2g (F) 3T2g (F), 2 = 3A2g (F) 3T1g (F) and 3 = 3A2g (F) 3T1g (P).
(b) ELECTRONIC REFLECTANCE SPECTRAL DATA AND THEIR ASSIGNMENTS OF

CO(II) COMPLEX
101
[Co(C23H20ON2S)2Cl2] 8810 14245 20080 1198.1 667 1.61 27226

[Co(C23H20ON2S)2(CH3COO)2] 8798 14242 20081 1195.4 666 1.61 27199
1 = 4T1g (F) 4T2g(F), 2 = 4T1g (F) 4 A2g (F) and 3 = 4T1g (F) 4T1g (P).
(c) ELECTRONIC REFLECTANCE SPECTRAL DATA AND THEIR ASSIGNMENTS OF

CU(II) COMPLEX
[Cu(C23H20ON2S)2Cl2] 15320 19118 -- -- -- -- --
[Cu(C23H20ON2S)2(CH3COO)2] 15325 19124 -- -- -- -- --
1 =2B1g 2A1g and 2 = 2B1g 2Eg
CZ-record UV-Viz. spectrometer provided with an automatic recorder was used to record the electronic spectra
of the complexes in ethanol at room temperature (Table 3).
PREPARATION OF METAL COMPLEXES
In general all these complexes were synthesized by refluxing the respective metal salts with ligand 2-amino-4-
(p-ethoxy phenyl) thiazoline in 1:2 molar ratio in ethanolic medium on water bath for one hour. The solution
was concentrated to half of its volume then it was kept for some time. The crystals of complexes separated out
which were filtered, washed with alcohol and dried in vacuum. Similarily some complexes of thiazoline were
also synthesized by many workers [15-21].
Adducts of all the complexes were prepared by refluxing the respective metal salts with ligands in 1:2 molar
ratio in ethanolic medium. The crystals of complexes separated out which were filtered, washed with alcohol
and dried in vacuum.
IR Studies: The (C=N) band frequencies in the free ligand are completely unaffected on complexation. The
unchanged position of the band indicates that the ring nitrogen does not take any part in the coordination. The
band observed at 834 cm-1 in the free ligand assigned to asymmetric (C-S) is shifted to lower frequency after
complexation. But the symmetric (C-S) frequency completely disappears or intensity of this band is reduced
after complexation. These facts confirm that the ring sulphur is taking part in complex formation. The (N-H)
asymmetric and symmetric stretching frequencies appearing in the region 3468 and 3286 cm-1 respectively, also
decreases in the complex. This shows that the lone pair of electron available on nitrogen atom took part in
coordination. From the above observation it is clear that the nitrogen of the NH2 group and ring sulphur take
part in coordination.
Electronic Reflectance Spectral Studies:
102
In the electronic spectra of Ni (II) complexes three bands at 8515-8523, 14511-14518 and 24417-24448 cm-1
were observed which may be assigned for 3 A2g (F) 3T2g (F) (1), 3 A2g (F) 3 T1g (F) (2) and 3A2g (F) 3T1 g
(P) (3) which are characteristic of octahedral Ni(II) ion. The magnetic moment values are found in the range
2.90-3.20 B.M. This is in support of high spin octahedral complex. The value is however is raised only to a
small extent suggesting that the splitting is weak and that the environment is quite close to an octahedral one
[22].
The observed value of magnetic moment is found in the range 2.97-3.55 B.M. which is expected for octahedral
Co(II) complex. Three bands were observed at 8798-8810, 14242-14245 and 20080-20081 cm-1 which may be
assigned to 4T1g (F) 4 T2g (F) (1), 4T1g (F) 4 A2g (F) (2) and 4 T1g (F) 4 T1g (P) (3) respectively for
octahedral complexes.
Two bands were observed in the electronic spectra of Cu (II) complexes in the region 15320-15325 and 19118-
19124 cm-1 which may be assigned to 2B1g 2A1g and 2B1g 2Eg respectively in a planar field. The magnetic
moment value foe the Cu (II) complexes lie in the range 1.54-1.58 B.M. which support square planar geometry.
The fungicidal activities of the ligand as well as of metal complexes were screened against different fungi at
different concentrations 100, 50 and 20 ppm in Czapeks dox agar medium. It has been observed that the
fugitoxicity of the metal complexes are lesser than the free ligand. This might be due to the fact that the group
which is responsible for toxicity is not free in complexes due to co-ordination however it is free in ligand. The
ligand as well as the metal complexes is most toxic at higher concentration i.e. the fungicidal activity decreases
with the decrease of concentration.
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4. Chandra S. and Sangeetika J., EPR and electronic spectral studies on copper(II) complexes of
some N-O donor ligands, J. Indian Chem. Soc., (81) 203(2004)
5. Shriodkar S.G., Mane P.S. and Chondhekar T.K. Synthesis and fungitoxic studies of Mn(II),
Co(II), Ni(II) and Cu(II) with some heterocyclic Schiff base ligands, Indian J. Chem, (40A) 1114-
1117(2001)
6. Belaid S., Landreau A., Benali-Baitich O., Khan M.A. and Bouet G., Synthesis, characterisation
and antifungal activity of a series of cobalt(II) and nickel(II) complexes with ligands derived from
reduced N, N'-ophenylenebis (salicylideneimine), Trans. Met. Chem., (33) 511(2008)
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7. Mapari A.K. and Mangaonkar K.V. Synthesis, Characterization and Antimicrobial Activity of
Mixed Ligand Complexes of N-(2-ethoxy-1-naphthylidene)-2,6-diisopropylaniline and N-(2-
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Journal of ChemTech Research., 3(2) 636-641(2011)
8. Ravanasiddappa M., Sureshg T., Syed K., Radhavendray S. C., Basavaraja C. and Angadi S. D.,
Transition Metal Complexes of1, 4(2'-Ethoxyphenyl-1-yl) di-iminoazine: Synthesis,
Characterization and Antimicrobial Studies, E-J. Chem., 5(2) 395-403(2008)
9. Manch W. and Conard Fernelius W., The Structure and Spectra of Nickel(II) and Copper(II)
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10. Naik B. and Desai K., Novel approach for the rapid and efficient synthesis of heterocyclic Schiff
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11. Bharti S.K., Nath G., Tilak R. and Singh SK. Synthesis, anti-bacterial and anti-fungal activities of
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12. Vogal A.I. Quantitative Organic Analysis., (1958)
13. Vogal A.I. A Text Book of Quantitative Inorganic Analysis. 3rd ed. (English Language Book
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14. Dodson R.M. and King L.C. The reaction of ketones with halogens and thiourea J. Am. Chem.
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15. Adibpour N., Khalaj A. and Rajabalian S. Synthesis and antibacterial activity of
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105
Nutritional sink formation in galls of Alstonia scholaris Linn. (Apocynaceae) by the

insect Pauropsylla tuberculata (Homoptera: Psyllidae).
Deepak Kumar, Vijai Malik and S.C. Dhiman*
Department of Botany, Department of zoology*,
M.S. College, Saharanpur (U.P.)
Email- dk_arya117@rediffmail.com
ABSTRACT
Galls are the localized outgrowth of various plant organs in which host tissue stimulated by host
parasite interaction. Galls act as nutritional sinks provide essential nutrients for insect growth and development.
Gall former have the ability to alter the developmental process of plant tissue to cause the formation of tumour
like growth that surround the insect to protect it from the environment and supply it with a source of food. Galls
caused by insect Pauropsylla tuberculata on Alstonia scholaris tree were studied to examine the different
changes resulting from the biotic stress caused by insect feeding. It is supposed that Ist nymphal instar initiates
gall formation during feeding by injecting its proteins and lytic enzymes rich saliva. This leads to hypertrophy
and hyperplasia in the localized area of feeding site causing mobilization of nutrients such as free amino acids,
reducing sugars, total soluble sugars, total phenols and protein to the gall from the un-galled region of plant.
Key words - Pauropsylla tuberculata, Alstonia scholaris, gall, nutritional sink.
INTRODUCTION
Alstonia scholaris R.Br. (Apocynaceae) commonly known as Saptaparni is an evergreen, tropical tree
with white funnel-shaped creamish flower and milky sap. It grows upto 40m tall with a spread of 10m. It is well
known remedy for the treatment of various types of disorders in the Ayurveda, homoeopathic and folklore
system of medicine in India [1, 2]. Various parts of the A. scholaris tree have been widely used in treatment of
several aliments. It barks is used for medicinal purpose, ranging from malaria and epilepsy to skin conditions
and asthma [3]. In Ayurveda it is used as a bitter and as an astringent herb for treating skin disorders, malarial
fever, urticarial, chronic dysentery, diarrhoea and in snake bite. The milky juice from the different part of tree is
used for the treatment of ulcers [4].
A. Scholaris is also a beautiful foliage tree with large canopy and because of this; it has become a popular
ornamental tree in landscapes and gardens. Now a days it is preferred as roadside plant in cities as it is highly
affected to pollutants.
However, this plant of great economic value suffers with the gall infested by Pauropsylla tuberculata
(Homoptera: Psyllidae). Galls have been reported on the various parts of the tree and this causes serious damage
to A. scholaris tree. Gall insects have developed highly specialized and nutritional relationship with their host
plant because these insects spend a major part of their life within galls and interact with galls through
modifications that range from simple tissue removal or damage to vascular tissue to complex manipulation of
synthesis and transport of host-plant nutrients [5, 6, 7, 8]. Insects can use the galls as sinks for nutrient to
employ in the growth and reproduction of larvae [9, 10]. Growth of gall tissue is associated with the changes in
the levels of their cellular contents such as carbohydrates, proteins, nucleic acids phenols and IAA enzymes
106
[11]. This work was undertaken to find out the formation of nutritional sink by identifying the different
chemicals level in the galled and ungalled leaf tissue.
MATERIALS AND METHODS
A. scholaris tree having galls of different stages were collected at Saharanpur district and adjacent areas.
Comparisons of different phytochemicals among various stages of gall development namely young, mature and
old gall were carried out. Total free amino acids were extracted and determined by following the method of
Sugano et al. [12] with slight modification. Total protein was extracted by the Acetone-TCA precipitation
method as described by Parida et al. [13] and estimated by following the method of Lowery et al. [14] using
defatted bovine serum albumin (BSA) as standard. The amount of total phenolic in normal and galled tissue
were determined with folin ciocalteu reagent according to the method of Singalton and Rossi [15] with slight
modification using tannic acid as a standard. Estimation of reducing sugar was done by phenol sulphuric acid
reagent method [16, 17]. The determination of different enzyme activity viz., peroxidase, IAA-oxidase,
Invertase, -amylase were estimated using the methods suggested by Birecka et al., [18], Mahadeven and
Sridhar [19], Harris and Jaffcoat [20] and Bernfeld [21] respectively.
Galls are specialized plant structures formed when a galling organism alters the development
of normal plant tissue [22]. The development of gall is a function of cell inhibition differentiation, growth or
suppression of host plant tissues, regardless of their induction agents. However, the initiation of galling and the
subsequent manipulation mechanism of gall forming process, such as phenolic compounds, protein, plant
hormone analogues and genetic manipulation are still unclear [23, 24, 25, 26].
The amount of total amino acid was recorded to be more in gall tissue as compared to normal leaf
tissue (Table- 1). Since, insect derive their nutrition from gall tissue, the gall becomes a sink for different
nutrients and energy that will be vital for the insects growth. Miles and Lloyd [27] and Miles [28] suggested
that increase in quantity of amino acids in gall tissues may be due to break down of proteins into utilizable
unites by the enzyme protease secreted by the salivary glands of the insects. Koyama et al. [29] observed high
concentration of amino acids in aphid gall and this support the nutrition hypothesis for gall formation.
Total protein show high values at the start of gall growth and decline progressively with ageing.
Secondly quantity of total protein was low when compared with the values obtained from normal leaf (Table-1).
These observations were corroborates with the observations of Arora & Patni [30], El-Akkad [31], Scareli-
Santos & Varanda [32], and Kumar [33]. The formation of gall requires mechanical and chemical stimulus.
According to Rosenthal and Jenzen [34] an interaction between the offensive stimuli involving growth
substances released by insects and defensive response by appears to be the hallmark of gall production. The
action of stimulus leads to the formation of new tissues, which cover the nymph. Kinsella [35] suggested that
proteins are the primarily building block sources for new tissue, in plants, animal and human beings. Thus
stimulus of gall forming insects redirects the growth and differentiation of cells which act as a sink of nutritive
substances [36]. On the other hand defensive importance of diverse plant proteins was also suggested by
107
Reinbothe et al., [37]. Defensive proteins are to be believed that they block the action of proteolytic enzymes
from herbivores. These proteins known as proteinase inhibitors [38]. So, it can be concluded that when plants
are attacked by insects they generate signals and one of these signals is the initiator of expression of certain
polypeptides that may be useful in providing the basis for new crop protection strategies.
Total phenol was high in galled tissues as compared to healthy leaf (Table- 1). The increased quantity
of total phenol might be attributed to defence mechanism. This resistance to disease caused by pathogen was
attributed to the presence of high amount of phenol [39, 40, 41, 42]. Insects can stimulate plants to produce galls
and secrete high levels of phenolic compounds [43, 44]. Kaur et al., [45] reported potent antioxidant activity of
ethanolic extract in Quercus infectoria galls contained high levels of polyphenols. Hence, the increased quantity
of phenols in the galled tissues may be contributing to the resistance against pathogen (P. tuberculata).
According to Kraus and Spiteller [46] different classes of phenolics substances act as plant defensive agents
against microorganisms and herbivores. The resistance to disease caused by the pathogens is due to the presence
of high amount of phenol [47]. The increase in phenolics in relation to resistance was reported in Brassica [48].
Subsequent increase in phenol contents were general responses associated with resistance mechanism in plants
as reported by Ghosal et al., [49]. Gupta [50] also found higher phenolic (total phenol and orthohydroxy
phenols) as compared to their normal counterparts, in gall tissues of Dalbergia sissoo, Salvodora oleoides and S.
persica. Thus, increased quantity of total phenolics in the galled tissue of A. scholaris is basically for providing
resistance against insect infestation.
The quantity of soluble sugar was considerably high in gall tissue as compared to normal tissue (Table-
1). Sugar has large numbers of stereo-isomers, because they contain several asymmetric carbon atoms [51].
Increase in sugar content might be due to accumulation of these substances. This accumulation may involve the
translocation of soluble sugars from the neighboring healthy tissues to physiological sinks. Findings of Shaw
and Samborski [52] also support this view. Sugars are very important biochemical nutrients for the plants. If
abundantly existing, these are detected naturally by various micro-organisms like bacteria, fungi, insects etc. So
once infected, inside the host tissue these organisms utilize the excess sugar present in the plant for their growth
and substance [53, 54]. High sugar contents in young and mature galls may be due to increased metabolic
activity under feeding stress of nymph, which in turn may be responsible for addition of sugar.
In Alstonia scholaris, IAA- oxidase activity was higher in galled tissue as compared to normal leaves
tissue (Table- 2). IAA (Indole Acetic Acid) the main auxin in higher plants has profound effects on plant growth
and development. Both plants and some plant pathogens can produce IAA to modulate plant growth [55]. A
higher level of phenol affects adversely the IAA- oxidase activity in plant tissue resulting in a higher level of
IAA [50, 56], thus leading to hyperauxinity and gall formation. Kumar [57] also observed higher amount of free
auxin concentration and IAA- oxidase in gall tissue. High level of IAA will support cell expansion
(Hypertrophy) and Cell division (Hyperplasia).
Gall tissues showed increased peroxidase activity as compared to normal leaves tissue (Table- 2).
Increase in peroxidase activity is due to phenol concentration, which plays an important role in oxidating
enzymes [30, 50, 57]. Peroxidases are responsible for oxidation of phenolics [58]. Increase activity of these
108
oxidative enzymes indicates a state of high catabolism induced during pathogenesis. Devnathan et al., [59]
observed high peroxidase activity in bunchy top banana virus infected cultivars of banana. Meena et al., [42]
suggested that increased peroxidase activity was associated with resistance reaction which would be due to
increased phenol concentration, where phenols were cofactor of peroxidase and hence influenced.
Increase activity of alpha-amylase was found in gall tissues (Table- 2). Due to larval feeding stress,
metabolic activity increased galled tissues which in turn stimulate the synthesis of sugar. Carbohydrates may
also accumulate by depletion of starch due to the activated alpha-amylase activity and other enzymes [60, 61,
62]. Shekhawat [61] and Purohit [63] also reported increased activity of alpha-amylase along with increased
sugar contents.
Invertase activity was also higher in Alstonia gall tissues ((Table- 1)). As in crown galls formed by
Agrobacterium [64] and galls of aphid Hormaphis hamameludis [65]. Our results implicate elevated invertase
activity as a means by which insect galls become sinks. Galls are known to act as a sinks for plant assimilates
[66] and high level of soluble invertase are associated with the establishment of sink characteristics [67, 68, 69].
Within the tissue, vacuolar acid invertase may hydrolyze sucrose to provide the hexoses required for elevated
metabolic activity [70, 71] and invertases in nutritive tissue are known to hydrolyse sucrose to glucose and
fructose [66]. While gypsy moth wounding may also elicit invertase activity [72], the increase brought about by
gallers was much greater than that caused by 1 week of gypsy moth feeding.
CONCLUSION
Plant and insects interact at various levels and insect-induced galls are the one of the most exciting and
deepest relationship between them. Galling insects are unique in controlling the within plant movement of
photo-assimilates and tissue morphogenesis. The nutritional status of host plant tissues favours the induction and
the establishment of the insect gall. Our analyses led us to the conclusion that the gall inducing insect
(Pauropsylla tuberculata) use host plant resources (Alstonia scholaris) to its own benefit
ACKNOWLEDGEMENTS
The authors are grateful to the Principal M. S. (P.G) College Saharanpur for providing the laboratory
research facilities, and encouragement.
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TABLE -1 Quantitative estimation of different metabolites
PARTS IAA- - PEROXIDASE INVERTASE

OXIDASE AMYLASE A/gm.dw/min Sucrose/
starch hydrolysed
hydrolysed mg/min. dwt
mg/min.dwt
NL 2.440.52 2.30.2 0.80.1 3.60.4
YG 2.900.30 3.10.1 1.80.4 4.40.2
MG 3.60.42 3.50.3 1.40.3 4.20.1
OG 2.500.02 2.80.3 1.30.3 3.90.1
113
TABLE - 2 Quantitative estimation of different enzymes

*NL=Normal leaf, YG=Young gall, MG=Mature gall, OG=Old gall
PARTS TOTAL TOTAL TOTAL TOTAL TOTAL

SOLUBLE REDUCING PHENOL PROTEIN FREE
SUGARS SUGAR (mg/gm dw) (mg/gm dw) AMINO
(mg/gm dw) (mg/gm dw) ACIDS
(mg/gm dw)
NL 3.3 0.08 1.3 0.1 0.62 0.02 1.8 0.22 3.0 0.71
YG 4.2 0.01 2.8 0.2 1.8 0.46 3.6 0.35 4.6 0.52
MG 3.7 0.40 3.5 0.2 1.02 0.03 2.8 0.34 5.3 0.30
OG 2.5 0.22 2.30.2 0.82 0.02 2.5 0.33 4.2 0.21
114
Perspective of Stem Cell Research in India

Bindu Sharma
Associate Professor,
DN PG College, Meerut (UP)
Email: bindu2502@rediffmail.com
ABSTRACT
Stem cell therapy has shown immense potential by treating diseases that were traditionally considered
degenerative and incurable such as diabetes, Parkinson's, Alzheimer's disease. In India, stem cell programmes
have been initiated with the aim of promoting both basic and translational research in view of its potential
applications. Techniques have been developed for the in vitro culture of stem cells. As a result, scientists can
now carry out experiments aimed at determining the mechanisms underlying the conversion of a single,
undifferentiated cell, the fertilized egg, into the different cells comprising the organs and tissues of the human
body. National agencies are pro-active in supporting and promoting this area. Over 30 institutions, hospitals and
industry are involved in SCR in the country. However, there are many challenges in current stem cell research
such as non-availability of human resources of adequate expertise; very few indigenous hESC lines generated;
inter disciplinary structure is yet to be created; derivation of ES cells from early human embryos, and EG and
fetal stem cells from aborted, fetal tissues raise ethical, legal, religious, and policy questions. Further, the
potential use of stem cells for generating human tissues and, perhaps, organs, is a subject of ongoing public
debate.
Key words: Stem cell, India, embryonic stem cell research.
INTRODUCTION
Stem cells are one of the human bodys master cells with the ability to grow into any one of the bodys more
than 200 cell types Stem cells are undifferentiated biological cells that can differentiate into specialized cells
and can divide to produce more stem cells. In mammals, there are two broad types of stem cells: embryonic
stem cells, which are isolated from the inner cell mass of blastocysts, and adult stem cells, which are found in
various tissues. In adult organisms, stem cells and progenitor cells act as a repair system for the body,
replenishing adult tissues Stem cells are precursor, unspecialized, undifferentiated cells capable of self-
proliferation, migration and differentiation. In the simplest form, the stem cell is an immature cell that has the
capability to differentiate into any possible mature cell. Much like the bone marrow, cord blood is one of the
richest sources of stem cells. Cord blood stem cell research is being conducted for potential future use in the
treatment of certain auto-immune disorders, neurological disorders, muscular/cartilage diseases, stroke, etc.
Stem Cell Research in India
Indians have also started to develop stem cell lines, including at least three human embryonic stem cell (hESC)
lines to date (UK Stem Cell Bank and National Institutes of Health (NIH) Human Embryonic Stem Cell
Registry).Two hESC lines derived by the Jawaharlal Nehru Centre for Advanced Scientific Research
(JNCASR), in Bengaluru, have been accepted for deposition and distribution by the UK Stem Cell Bank. The
cell lines, derived from low-quality embryos discarded post-IVF procedures, will be part of the International
115
Stem Cell Initiative 2 (ISCI2) project to identify the common genetic changes that occur in hESC lines on
prolonged culture. This will be the first time that India is represented on an ISCI project. Indias Department of
Biotechnology (DBT), has listed more than 30 research institutes, hospitals, and firms involved in stem cell
research in India [1-5].
Research Institutes
Public research institutes are the largest group active in stem cell research with in India. Some institutes focus
primarily on basic research. JNCASR studies ESC differentiation into cardiovascular cells. Other institutes
balance basic research with applied activities such as animal modeling, clinical trials, or pilot treatments. NCCS
has conducted animal and preclinical analyses of bone marrow stem cell injections for pancreatic regeneration.
Research efforts from this institute succeeded in rescuing mice with experimentally induced diabetes after a 30
day follow up [5-7], and scientists at NCCS hope to extend this work to an autologous clinical trial in human
diabetic patients. The institute is working to establish a team of clinicians, scientists, and patients to act as a
platform for the trial, a process they estimate will take 34 years. Their general business model is to conduct the
basic research and find a private partner for further development. This is a model they have successfully applied
in the past, as illustrated in their transfer of collagen sheet wound dressing technology to a local company,
Euchre Pharmaceutical Pvt. Ltd. (Chennai), for production.
Hospitals and Clinics
Unlike Indias biotechnology sector as a whole [4, 7], few Indian companies are involved in stem cell research.
Instead, Indian hospitals and clinics are key players. Indias large research-intensive hospitals, such as AIIMS,
conduct basic and applied research and have clinical trial and pilot treatment capabilities. This combination of
resources creates a bridge between research and therapy and makes hospitals pivotal for Indian stem cell
innovation. AIIMS works on a wide spectrum of clinical applications in cardiology, ophthalmology, neurology,
and hematology and also carries out basic research on, for example, stem cells and biopolymers aimed at
treatments for orthopedic, ocular, and skin diseases. Christian Medical College and Hospital (CMC) in Vellore
has funded a center for stem cell research in collaboration with DBT to promote translational research with stem
cells. Sankara Nethralaya, an eye hospital based in Chennai, has similarly built a research building to house the
Kamalnayan Bajaj Institute of Research in Vision and Ophthalmology. Stempeutics, in Bengaluru, is the
research arm of Manipal Education and Medical Group (MEMG). CryoStem Cell, in partnership with Sri
Bhagwan Mahaveer Jain Hospital (in Bengaluru), conducted a pilot stem cell treatment for Bergers disease, a
severe form of peripheral vascular disease, which is relatively rare in the Western hemisphere but common in
India If successful, CryoStemCells pilot therapy has the potential to address a very real health need in India.
Reliance Life Sciences is also involved in treatment development and is investing in an animal facility to
conduct toxicology and preclinical efficacy studies for cell based therapies.
Universities
Few Indian universities appear active in stem cell research at this time. The main exception is the University of
Delhi, which, together with the Indian Institute of Nuclear Medicine and Allied Sciences, is examining basic
mechanisms of stem cell function. However, many of the hospitals, firms, and research institutes active in stem
116
cell R&D are also involved in education. Both CMC and AIIMS are teaching hospitals that provide postgraduate
degrees. Many of Indias research institutes are collocated with a university and informally train students within
their research labs.
Government Support
The Indian government has been a key factor in encouraging stem cell R&D activity and growth. Stem cell
engineering is seen as an important area for the government and is identified as a strategic biotechnology area in
the DBTs 2007 Biotechnology Strategy (available online). Four government departments are key supporters of
stem cell R&D: DBT, the Indian Council for Medical Research (ICMR), the Department of Science and
Technology, and the Council of Scientific and Industrial Research. Stem cell R&D promotion is driven largely
by the DBT Stem Cell Taskforce and by ICMR through its affiliated institutes and regulatory capacity. DBT
provides direct funding to targeted initiatives in this field and supports both infrastructure building and
operational activities such as clinical trials. It has begun to support large-scale strategic programs, such as a
phase III multicenter trial using bone marrow cells to treat myocardial infarctions, and sponsors stem cell
research centers like the Centre for Stem Cell Research (CMC, in Vellore) and a soon to-be-established stem
cell research center in Bengaluru, the Institute for Stem Cells and Regenerative Medicines. As has been shown
elsewhere for the case of health biotechnology [10-14], government support is often key to capability building
within emerging economies. Building Stem Cell R&D on Indias Strengths Indias capacity to participate in a
cutting-edge field such as stem cell research is, in part, built on skills and infrastructure previously developed by
the nations pharmaceutical and biotechnology sectors [17-19]. One such skill is the proven ability of Indian
firms to develop process innovations in order to lower prices [20]. Innovations that lower process costs have
already occurred in the stem cell field. Researchers do not necessarily copy blindly the techniques used in
developed countries but create their own cost-efficient cell and tissue culturing and storage techniques that use
fewer disposable devices. Indian companies have also begun to produce materials, such as growth factors, that
are needed in stem cell research at significantly lower rates. In addition, Indias pharmaceutical and
biotechnology sectors have helped India develop an expertise in conducting clinical trials. Bolstering this
expertise, Indias large, diverse, and treatment-nave population provides a valuable resource for clinical trials,
especially for rare diseases where the Indian population could provide sufficient patients for trial groups. As a
result, India has great potential to act as a clinical trial destination of choice for stem cell therapies. This could
help India develop and test stem cell therapies for a variety of diseases. This strength is likely to encourage more
international ties and joint ventures, a trend already exemplified in the collaborations of United States/Indian
firm StemCyte India Therapeutics and the Japanese/Indian venture NCMR.
Overcoming Indias Stem Cell R&D Challenges
Promoting Linkages
As a translational research field, stem cell development requires a high degree of linkage between basic and
clinical expertise. Indias most successful research institutes and hospitals, such as AIIMS and LVPEI, are those
that integrate the efforts of basic scientists with clinicians. Unfortunately, this integration has historically been
weak at most research sites elsewhere in India. Increasing coherence and connectivity between these different
117
sectors is the cornerstone of Indias new biotechnology strategy (DBT Strategy, 2007), and specific institutions,
as well as the government, have initiated steps to promote interactions between basic and clinical stem cell
researchers. The construction of a joint research center between the academic Centre for Cellular and Molecular
Biology and the hospital-based Nizam Institute of medical Sciences, both in Hyderabad, is an example of an
Indian institutional initiative aiming to increase clinician/ scientist interaction. The DBT is promoting improved
integration with its stem cell cluster initiative, which encourages publicly and privately funded stem cell
research groups to share ideas, facilities, and research. Four clusters are already established: around the CMC in
Vellore, the NCCS in Pune, in Bengaluru, and in Hyderabad. The project is expected to expand and promote an
additional cluster in Delhi.
Strengthening Training
For India to increase its stem cell research capacity, it will need to strengthen the quality of its current scientific
education. The DBT is helping Indian scientists gain expertise abroad by offering overseas stem cell fellowships
and travel bursaries for conferences. Research institutes are increasing their involvement in training through
postgraduate supervision and by coordinating workshops, such as the stem cell training workshops currently run
by JNCASR in partnership with the NCBS, both in Bengaluru. Universities are also becoming more involved in
stem cell training programs, such as Manipal Universitys efforts to establish an Institute of Regenerative
Medicine with a related graduate program in the summer of 2007. NCMR, in collaboration with Acharya
Nagarjuna University (in Chennai), launched a stem cell PhD program in April 2008. The program will be
focused on bringing clinicians together with scientists. Indians can also strengthen their capacity for stem cell
research by attracting Indian experts who are currently active in this field in developed countries. Members of
the Indian scientific diaspora (expatriates working in industrially developed countries) have begun to return to
India in greater numbers, encouraged by economic prosperity and active recruitment initiatives from firms and
research institutes. More proactive strategies could strengthen this flow of returning scientists, with targeted
efforts designed to attract expatriate experts in stem cell research.
Increasing Public Awareness
While Indians who have heard of stem cell research and therapies are generally supportive, this field is still
relatively unknown to the general public, except to a small subpopulation of educated urbanites. There is some
risk that this lack of broad understanding may lead to uninformed mistrust of the field, particularly if scandals or
slow results help to destroy public support and lead Indians to mobilize against new stem cell therapies. This
mobilization against new technologies has already occurred in India with the introduction of genetically
modified cotton [21]. While this risk is not unique to India, it is particularly challenging to educate the extensive
Indian population about stem cells, due to the logistical difficulty of communicating the details of scientific
advances to a vast population with typically low levels of general education. Another risk that is nonspecific to
India, but that might be unusually challenging to overcome in this country, is that expectations for stem cell-
based therapies may be overly inflated to the ultimate detriment of the field. There is considerable potential for
the exploitation of patients who see stem cell therapy as a magic bullet to solve their health needs. This
exploitation has already been described in anecdotes of Indian clinics offering stem cell therapies without a
strong scientific basis or proper safety and efficacy tests.
118
Developing the Regulatory Framework
Until recently, India lacked comprehensive stem cell research and therapy guidelines, an omission that
compounded the potential for patient exploitation. To address this barrier, ICMR led the development of stem
cell research and therapy guidelines (see ICMR National Guidelines) with the participation of DBT and others.
Newly finalized, the guidelines are relatively permissive, allowing hESC and adult stem cell research and cell
line development under close monitoring. Experimental treatments, embryo creation for research, and chimera
studies are permissible subject to approval. The guidelines create two levels of stem cell research review and
monitoring: a National Apex Committee for Stem Cell Research and Therapy as well as institutional
committees. Depending on the research topic, projects will be approved nationally or institutionally; research
will be categorized into permissive, restricted, and prohibitive areas for research, and all projects will have to
register nationally. Indias guidelines are relatively permissive when compared to other countries. Regarding
hESC research, one study surveyed 50 countries and found that hESC research was allowed under strict
conditions in 23 countries and banned in five, while the rest had no explicit policy [14-18]. The new Indian
guidelines are not legally binding; however, many Indians remain optimistic that their existence will encourage
researchers to begin working in the area of stem cells while simultaneously stopping unethical R&D. Its unclear
how successful the guidelines implementation will be. India has a poor track record at monitoring IVF clinics
and enforcing its guidelines for biomedical research using human subjects [12-15].
Ethical issues
The extraction of HESCs from inner cell mass for research purpose leads to the destruction of the embryo. The
major source of human embryonic stem cell tissues are the spare or supernumerary embryos created during in
vitro fertilization as a part of infertility treatment. The other source is creating embryos with somatic cell nuclear
transfer techniques (SCNT). The legislation of most countries including India allows use of spare or
supernumerary embryos either fresh or frozen created during in-vitro fertilization. Some countries with more
liberal view have allowed creation of human embryos with SCNT as a source of embryonic tissues.
Conclusion
As Indias capacity in stem cell research continues to develop, it can draw upon the nations numerous strengths
to actively expand its involvement in this field. Based on its track record in information technologies and
biotechnology, it is likely that India will be successful in building its capacity for implementing stem cell
therapies. All ethical principles applying to research must also be ensured in stem cell research: Principles of
essentiality, of voluntariness, informed consent and community agreement, of non-exploitation, of privacy and
confidentiality, of precaution and risk minimization, of professional competence, of accountability and
transparency, of maximization of public interest and distributive justice, of public domain and the principle of
totality of responsibility and compliance.
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1. Becker A. J., McCulloch E.A. and Till J. E. Cytological demonstration of the clonal nature of spleen colonies
derived from transplanted mouse marrow cells. Nature. 197:452-4 (1963)
119
2. Siminovitch L, McCulloch E. A. and Till J. E. The distribution of colony-forming cells among spleen
colonies. J Cell Physiol. 62:327-36 (1963)
3. Velu N. Stem cell transplantation. API medical update. 14:366-77(2004)
4. Friedenstein A. J., Gorskaja J. F. and Kulagina N. N. Fibroblast precursors in normal and irradiated mouse
hematopoietic organs. Exp Hemato. l 4(5):267-74 (1971)
5. Murrell W., Feron F. and Wetzig A. Multipotent stem cells from adult olfactory mucosa. Dev Dyn.
233(2):496-515 (2005)
6. Caveleri F. and Scholar H. R. Nanog: a new recruit to embryonic stem cell orchestra. Cell. 113:551-2 (2003)
7. Wang X., Yang Y. J. and Jia Y. J. The best site of transplantation of neural stem cells into brain in treatment
of hypoxic-ishemic damage: experiment with newborn rats. Zhonghua Yi Xue Za Zhi. 27;87(12):847-50 (2007)
8. Molofsky A. V., Pardal R. and Iwashita T. Bmi-1 dependence distinguishes neural stem cell self-renewal from
progenitor proliferation. Nature. 425 (6961):962-7 (2003)
9. Park I. K., Qian D. and Kiel M. Bmi-1 is required for maintenance of adult self-renewing haematopoietic stem
cells. Nature. 423(6967):302-5(2003)
10. Beachy P. A., Karhadkar S. S. and Berman D. M. Tissue repair and stem cell renewal in
carcinogenesis. Nature. 432(7015):324-31 (2004)
11. Rosenthal N. Prometheuss vulture and the stem-cell promise. N Engl J Med. 349(3):267-74 (2003)
12. Korbling M. and Estrove Z. Adult stem cells for tissue repair-a new therapeutic concept? N Engl J
Med. 349(6):570-82 (2003)
13. Marshall G. P., Laywell E. D. and Zheng T. In vitro-derived "neural stem cells" function as neural
progenitors without the capacity for self-renewal. Stem Cells. 24 (3):731-8 (2006)
14. Lavker R. M. and Sun T. T. Epidermal Stem cells: properties, markers, and location. Proc Natl Acad
Sci USA. 97(25):13473-5 (2000)
15. Liu S., Dontu G. and Wicha M. S. Mammary stem cells, self-renewal pathways, and carcinogenesis.
Breast Cancer Res. 7(3):86-95 (2005)
16. Shackleton M., Vaillant F. and Simpson K. J.. Generation of a functional mammary gland from a single
stem cell. Nature. 439:84-8 (2006)
17. Tuch B. E. Stem cells--a clinical update. Aust Fam Physician. 35(9):719-21 (2006)
18. Rideout W. M., Hochedlinger K. and Kyba M. Correction of a genetic defect by nuclear transplantation
and combined cell and gene therapy. Cell. 109(1):17-27 (2000)
19. Evans M. J. and Kaufman M. H. Establishment in culture of pluripotential cells from mouse embryos.
Nature. 292(5819):154-6 (1981)
20. Reynolds B. A. and Weiss S. Generation of neurons and astrocytes from isolated cells of the adult
mammalian central nervous system. Science. 255(5052):1707-10 (1992)
21. Vawda R., Woodbury J. and Covey M. Stem cell therapies for perinatal brain injuries. Semin Fetal
Neonatal Med. 12(4):259-72 (2007)
120
Study of Medicinal Angiosperms of Rampur District (UP), India With Special

Reference to Their Sustainable Use
BEENA KUMARI
Botany Department, Hindu College (PG),Moradabad.244001
Email: beenakumari.botany@gmail.com
ABSTRACT
Sustainable development may be defined as development that is economically sound, socially relevant and
environment friendly. There are more than two thousand five hundred plant species in India having documented
medicinal value. These medicinal plants and their raw materials are used in the prevention, treatment and cure of
health disorders since ancient time. The knowledge of plants and medicines has undergone various stages till
today. Many of todays drugs have been derived from plant sources. Rampur district is located at Longitude 78 0
54 to 690 28E and Latitude 280 25 to 29010 N and spans an area of 2,367 km. It was incorporated into the
state of U.P. in 1949. Due to rapid increase in industrial area and clearing of urban forest area for residential use
green plants facing the danger of extinction. The maximum population of the district Rampur resides in rural
areas. They use folk methods to cure different diseases through wild plants. But due to urbanization of the area,
the traditional knowledge of wild plants is now disappearing day by day. In the present study 58 angiosperm
plant species belonging to 32 families have been enumerated. According to one estimate more than 25% of
medicinal plants used by pharmaceutical companies are collected from wild and much of this is illegal. 70% of
this involves destructing harvesting. Therefore, in order to maintain a sustainable supply of the raw material
from the forest, their cover exploitation should be stopped and strict law should be enforced by the government
to converse these valuable medicinal plants. In this way we can save our plant wealth.
Keywords: Angiosperms, Biodiversity, Medicine, Sustainable use, Rampur.
Introduction
Medicinal plants are considered very important in primary health care system. A large number of plants are
known for their medicinal properties. There is a large demand for medicinal herbs due to increase in the use of
herbal formulations. Herbal medicines are used by about 75-80% of the worlds population for primary health
care because of better cultural acceptability, better compatibility with human body and lesser side effects
[1,2]Wild harvesting- medicinal plants at risk. The negative impact of commercial collection of medicinal
plants cause tremendous short and long term damage to plant resources, vegetation communities and ecosystem
[3,4]. Earlier, medicinal plants were obtained from the forests. At that time in the forests they were in abundance
and the consumption was in milligrams or grams. But now, the situation has reverseddue to deforestation,
uprooting of the plants for fulfilling the requirements and the craze for herbalglobalization[5,2].So the medicinal
plants have become endangered. Therefore, the rates have also increased and are unable to fulfil the requirement
of the genuine material in the world.
Study Area
Rampur district is located at Longitude 780 54 to 690 28E and Latitude 280 25 to 29010 N. Spread in area of
2,367 km2, Rampur is 192 meter above sea level in north and 166.4 meter in south. During summers the
temperature is usually from 43 C to 30 C and during winters it is from 25 C to 5 C. The average rainfall
varies between 800 to 900 mm.The relative humidity is up to 90% in monsoon season and in drier part of the
121
year it decreases to less than 20%. As per the 2011 Census of India Rampur had a population of 325,248
[compared to 281,549 in 2001] showing 16% growth in 2001-11.People living in the rural areas of Rampur
district of Rohilkhand region of U.P. are greatly dependent on medicinal plants for variety of uses. It proved to
be a good income source and cheap source of curing diseases at local level. Currently medicinal plants are under
severe threat of extinction due to rapid deforestation, over and improper collection, over grazing etc. All the
plant material comes from thewild and little effort has been made for their cultivation[6-8]. The present study is
an effort to analyse the current status of medicinal angiosperms in the context of its value for the local people of
Rampur district.
Methodology
The field studies were carried out in 2011 - 2012. To collect information on medicinal angiospermsand their
uses for different ailments, local traders, Vaidyas and old women wereidentified and interviewed. Plant
specimens were collected from study area and shown to them for authentic information. Available floras [9,10]
were consulted for identification ofplants. The survey area map is enclosed for reference [fig.1].
Results &Discussion
In the present study 58 angiosperm plant species belonging to 32 families have been enumerated [Table 1.].
Herbs [29 genera & 36 species] were collected from 21 families, shrub [9 genera & 10 species] from 6 families,
trees [5 genera & 5 species] from 5 families and climbers [7 genera & 7 species] from 6 families. Asteraceae
family is dominant with 6 species [fig.2& 3]. Total number of plant parts used in different ailments are shown in
fig. 4.
Medicinal plants collectors are untrained, and almost half of the material collected by untrained manpower is
wasted. Therefore, there is a need to find ways to harvest medicinal plants sustainably from the wild. This
includes training local collectors in proper collection techniques, training people to grow medicinal plants, and
removing some of the middlemen from the trading chain [11,12]. More than 25% of the medicinal plants used
by Indian Industry are collected from wild and much of this is illegal. 70% of the collection involves destructive
harvesting practices like overexploitation, fragmentation of natural habitats and introduction of exotic species
[13,4]. Medicinal plants can save lives, livelihoods and cultures until they themselves are saved. Therefore, in
order to maintain a sustainable supply of the raw materials from the forests their overexploitation needs to be
stopped and strict laws should be enforced by the Government for the conservation of forests [12,14]. The
conservation program can be strengthened by using the people who have knowledge and also respect for the
Mother Nature. Women can be great contributors in the conservation program as they manage most of the plant
resources that are used by humans [5,15,16]. Thus, with the involvement of the collectors, producers and traders
including ultimate users, the motive of sustainable use of the resources can be achieved [fig. 5].
Acknowledgement
Author is thankful to UGC for providing financial assistance (F.No.8-2 (302)/2011 (MRP/NRCB) and local
people for providing necessary information regarding the uses of plant species for various ailments.
References
1. Singh, Akanksha, Singh, B. N., Singh, S.P., Chaudhary, Anita and Sharma, B.K. Sustainable use of
medicinal plant biodiversity for poverty alleviation. National Conference on Biodiversity,
Development and Poverty Alleviation, U. P. State Biodiversity Board, Lucknow, 24- 29, (2010)
2. Uniyal, R.C., Uniyal, M.R. and Jain, P. Cultivation of Medicinal Plants in India: A Reference Book.
122
TRAFFIC India, Delhi, India, (2000)

3. Astaw, D., Abebe, D. and Urga, K. Traditional medicine in Ethiopia: Perspective and development
efforts, J Ethiop MedPrac, 1, 114-117, (1999)
4. Tewari, D. N. Report of the task force on conservation and sustainable use of medicinal plants. Govt.
of India, Planning Commission, New Delhi, 175, (2008)
5. Pandey,S.S., Srivastava, B. K., Pandey, V. S., Diwedi, S. and Shriya. Medicinal plant resources of
Uttar Pradesh : An urgent need for conservation,Plant Archives,10 (2),861-864,(2010)
6. Schippmann, U., Leaman, D.J. and Cunningham, A.B. Impact of cultivation and gathering of
medicinal plants on biodiversity: global trends and issues. In: Biodiversity andthe Ecosystem
Approach in Agriculture,Forestry and Fisheries. Ninth Regular session of the commission on Genetic
Resources for Food and Agriculture. FAO, Rome, Italy, 1-21, (2002)
7. Sharma, J., Painuli, R.M. and Gaur, R.D. Plants used by the rural communities of district
Shahjahanpur, Uttar Pradesh, Indian Journal of Traditional Knowledge, 9(4), 798-803, (2010)
8. Siddiqui, M.B. & Husain, W. Medicinal plants of wide use in India with special reference to Sitapur
district (Uttar Pradesh), Fitoterapia, 3, 65-67, (1994)
9. Duthie, J.F. Flora of Upper Gangetic Plain and of the Adjacent Siwalik and Sub Himalayan Tracts, 3
vols, (Botanical Survey of India, Calcutta), (1903-1929)
10. Maheswari, J. K. Flora of Delhi, CSIR Publication, India, (1963)
11. Chang, L., Hua, Y. and Chen, Shi Lin. Framework for Sustainable Use of Medicinal Plants in China.
Plant Diversity and Resources, 33 (1), 65-68, (2011)
12. U., Manjkhola, S. and Joshi, M. Current status and future strategy for development of medicinal plants
sector in Uttaranchal, India, Curr. Sci., 83, 956-964, (2002)
13. Mullikem, T. Sustainable use of medicinal plants -A multi sectoral challenge and opportunity,
TRAFFIC International, Cambridge, U. K.(www.traffic.org), (2012)
14. Hamilton, A. C. Medicinal plants conservation and livelihoods, Biodiversity Conservation, 13, 1477-
1517. (2004)
15. Khanna, K.K. Unreported ethnomedicinal uses of plants from the tribal and rural folklore of Gonda
district, Uttar Pradesh, Ethnobotany, 14, 52-56, (2002)
16. Taylor, J.L.S., Rabe, T., McGaw, L.J., Jager, A.K. and Staden, J. Van. Towards the scientific validation
of traditional medicinal plants, PlantGrowth Regul., 34, 2337, (2001)
123
Fig.1. Map of Rampur district
Lamiaceae
Chenopodiaceae
Amaranthaceae
Solanaceae
Malvaceae
Euphorbiaceae
Asteraceae
0 1 2 3 4 5 6 7
Fig.2 Dominant families with species of the study area
40
30
20 Family
10
0 Genera
species
124
Fig.3. Total medicinal plants family, genera and species in different taxa.
Latex Fruit Tuber Bark

3 6 1 3
Seed Leaf
5 19
Root
16
Flower whole
3 plant
19
Fig.4. Number of plant parts utilized for medicinal purpose.
Export
middle man Traders Vaidyas
collectors
Drug
shop manufacturer
consumers
keepers
Fig. 5. Chain of people involved in sustainable use of medicinal resources
Table-1. Medicinal plant diversity of Rampur district of Rohilkhand region of U.P.
S.no Family & Botanical name Local Name Part used Uses
.
Acanthaceae
1. Andrographis paniculata Kalmegh Leaf Diabetes & snake bite
(Burm. f.) Wallich ex Nees
2. Peristrophe paniculata Burm Chirchit Whole Wounds, gout & rheumatism
plant
Aizoaceae
3. Trianthema portulacastrum L. Santhi Root Constipation & asthma
(Santhi)
Amaranthaceae
4. Achyranthes aspera L. Latjeera Leaf Cuts & Wounds
5. Amaranthus spinosus L. Katelichaula Root Eczema
i
6. Amaranthus viridis L. Chaulai Whole Vermifuge
plant
125
Asclepiadaceae
7. Calotropis procera (Willd) Akua Flowers Constipation
Drey.
8. C. gigantia (L.) R. Br. Madar Root bark Dysentery
Asteraceae
9. Acanthospermum hispidum Gondhichedi Whole Skin disease& fever
DC. plant
10. Ageratum conyzoides L. Gundrya Flower, Cuts &sores,kidney
seed stone,diarrhoea,leprosy & uterine
disorders
11. EcliptaprostrataL. Bhangra Whole Constipation,Livercomplaints,asthma&H
plant air growth
12. SphaeranthusindicusL. Mundi Whole Skin diseases & piles
plant
13. TridaxprocumbensL. Kanphuli Leaf Cuts & Wounds.
14. Xanthium strumarium L. Gokhuriya Whole Malaria, piles, ulcer & rheumatism
plant
Bombacaceae
15. Adansonia digitata L. Balamkheera Fruit Kidney stones
Caesalpiniaceae
16. Bauhinia racemose Lamk. Kachnar Bark& Dysentery & diarrhoea
Fruit
17. Cassia toraL. Chakunda Leaf& Eczema, cuts & jaundice
Root
Commelinaceae
18. Commelina diffusa Burm.f. Kansura Root Snake bite
Chenopodiaceae
19. Chenopodium album L. Bathua Leaf Constipation& Urinary problems
&Seed
20. Chenopodium ambrosioides Bathua Whole Dysentery, pneumonia and piles
L. plant
21. Chenopodium murale L. Bathu Leaf Asthma
Convolvulaceae
22. Evolvulusalsinoides L. Shankhpuspi Whole Fever, cough and cold, stomach ache,
plant ulcer, dysentery, asthma
bronchitis & brain tonic
Cucurbitaceae
23. Momordica dioica Roxb. ex Jangli- Root Fever, piles, asthma, bronchitis, head
Willd. Karela ache,
126
dysentery& Diphtheria.
24. Coccinia grandis (L.) J. Kanduri Leaf Skin disease

Voigt.
Dioscoreaceae
25. Dioscorea bulbiferaL. Ratalu Tuber Dysentery, abdominal pain, jaundice &
boils, piles
Crassulaceae
26. Bryophyllum pinnatum Ajuba Leaf Swelling & Kidney stone
(Lamk) Oken
Euphorbiaceae
27. Euphorbia hirta L. Duthi Latex Wounds & lip crakes
28. Euphorbia tirucalli L. Kharsani Latex Eczema, wounds, toothache, earache,
scabies, rheumatism & warts.
29. Phyllanthus fraternus Web. Bhuiamla Whole Allergy, diarrhoea, dysentery, dropsy
plant &jaundice.
30. Ricinus communis L. Arandi Leaf, root, Skin diseases, sores, gum trouble,
seed cholera, boils, constipation, dysentery,
joint pain,
muscular pain, head ache & burns
Fabaceae
31. Abrus precatorius L. Ratti Root scorpion sting & snakebite
Fumariaceae
32. Fumaria indica Pugsley Pitpapra Whole blood purifier
plant
Lamiaceae
33. Leucas aspera (Willd.) Link. Gubba Leaf head ache & fever.
34. Ocimum basilicum L. Ramatulsi Leaf & Cold & cough, fever,
seed stone complaints, dropsy & cholera
35. Ocimum sanctum L. Shymatulsi Whole Antiseptic, cold & cough, fever, urinary
plant troubles, vomiting, bronchitis, chicken-
pox, cholera, constipation, headache,
diarrhoea, dropsy, ear complaints,
malaria, colitis gastric complaints &
antidote to poison leprosy
Malvaceae
36. Abutilon indicumL Kanghi Leaf, root Dental problems
37. Abelmoschus manihot Janglibhindi Root Pneumonia
(L.) Med.
127
38. Malvastrum Suchi Leaf Wounds & sores

coromandelianum (L) Garcke
39. Urena lobata L. Chatkura Leaf, Diarrhoea, Dysentery,
Flower, expectorant, body pain &rheumatism
Root &
Bark
Menispermaceae
40. Cissampelos pareiera L. Parha Leaf Antidote to snake & scorpion bite
Mimosaceae
41. Albizia lebbeck (L.) Benth Safed siris Bark Piles
Moraceae
42. Ficus palmate Forsk. Fagu Latex, Sores, constipation &
fruit Stomachache
Nyctaginaceae
43. Boerhaavia diffusa L. Pundra Root & Jaundice & constipation
whole
plant
Oleaceae
44. Jasminum multiflorum Chameli Leaves Pimples & eczema
(Burm. f.) Aners
Oxalidaceae
45. Oxalis corniculata L. Khatibuti Whole Diarrhoea, dysentery
plant epilepsy, piles, fever & jaundice
Papaveraceae
46. Argemone maxicana L. Pili kateli Root Snake bite
Passifloraceae
47. Passiflorafoetida L. Gharibel Seed, Tonic, headache,
leaves& cold & cough, wounds, Inflammation,
fruit itching& asthma
Polygonaceae
48. Polygonum plebejum R.Br. Jayanti Root Baldness
49. Polygonum barbatumL. Whole Snake bite
plant
Portulacaceae
50. Portulaca oleracea L. Kulfa Whole Constipation
plant
51. Portulaca grandiflora L. Luaniya Leaf Eczema
Scrophulariaceae
128
52. Veronica anagallis-aquatica Sadevi Whole Healing burns

L. plant
Solanaceae
53. Datura metel L. Dathura Leaf Ear-ache
54. Physalis minima L. Damphu Fruit Dropsy
55. Solanum nigrumL. Makoi Whole Cough & cold
plant
56. S. surratense Burm.f Barkatali Root Gum trouble & tooth decay.
Verbenaceae
57. Phyla nodiflora (L.) A. Rich Jal booti Whole Menstrual complaints.
plant
Zygophyllaceae
58. Tribulus terrestris L. ChotoGokhru Fruit & Urinary diseases
Root
129
Water Quality Control of Industrial Effluents Using Crosslinked Chitosan Hydrogel Beads.
Anuja Agarwal*& Vaishali**
*Associate Professor, Department of Chemistry, J.V. Jain College, Saharanpur, India.
E mail: anujaagarwaljvjc@gmail.com Mobile no.: +919411484839
**Research Scholar, Department of Chemistry, J.V. Jain College, Saharanpur.
ABSTRACT
Polymers from natural resources have been studied in the recent past as an important material for
biotechnological application owing to their unique characteristics such as biological compatibility with natural
environment, non-toxicity and biodegradability. Chitosan, deacetylated product of chitin obtained from skin of
crustacean fish is(1 4) 2- amino 2-deoxy D glucan and is one of the well known biodegradable polymers
metabolized by human enzymes. It can be prepared as hydrogel micro and nano sized beads, having a positive
charge at wide pH range. These beads are known to adsorb a number of metals and some of other pollutants.
Chitosan beads crosslinked with glutaraldehyde have been prepared by us to study their removal efficiency for
dyes and beads have been characterized by SEM, FTIR, XRD and DSC analytical methods. Swelling behavior
of beads has also been studied at pH 2.0, 7.4 and 10.0 which proves that swelling is more in acidic medium than
in basic. Percent Color removal efficiency for dye effluents has been determined and found to be dependent of
dye initial concentration, pH and temperature. The results concluded that chitosan beads can be used to remove
color in effluents containing dyes and is able to improve water quality of effluents from industries.
Key words: Dye, Effluent, FTIR, SEM, XRD, CRE
Introduction
Polymers from natural resources have been studied in the recent past as an important material for
biotechnological application owing to their unique characteristics such as biological compatibility with natural
environment, non-toxicity and biodegradability1,2 and also possess gel forming ability at low pH3. Deacetylated
product of chitin obtained from crustacean fish is(1 4) 2- amino 2-deoxy D glucan and is known as the
chitosan and is one of the well known biodegradable polymers metabolized by human enzymes 4. Chitosan can
be prepared as hydrogel beads, having a positive charge at wide pH range5. Three dimensional hydrophilic
polymer network of hydrogel beads are capable of retaining large amount of water for an extended time period.
Hydrogels are thermodynamically compatible with water and exhibit swelling in aqueous media. Cross linked
hydrogel polymer network can be obtained by cross linking chitosan using a cross linker like glutaraldehyde.
Their properties depend mainly on the cross linked density(the ratio of moles of cross linking agent to the moles
of polymer repeating units). Formation of hydrogel network requires a critical number of cross links per chain
and it forms porous structure whose pore size depends upon swelling of beads which in turn depend on external
environment like pH, temperature etc. Chitosan is a highly basic polysaccharide. Beads obtained from chitosan
after crosslinking with glutaraldehyde are solid, spherical, micron or nano sized constituting a matrix type of
structure. The beads obtained from hydrophobic polymers have found to be higher uptake as compared to the
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beads prepared from more hydrophilic surfaces6. So nano/micro beads surface charges increased hydrophobicity
of polymeric matrix have been found effective in a positive sense to utilize them as an adsorbent for removing
pollutants from water effluents. Our study is an attempt to develop chitosan beads cross linked with
glutaraldehyde for adsorbingdyes as a model adsorbent to investigate itsmodeling color removal efficiencyand
adsorbing properties. We planned a study to obtain beads of chitosan crosslinked with glutaraldehyde which
may be fruitful for further studies. We made an effort to study characteristics, swelling behavior and adsorbing
capacity of the cross linked chitosan beads for dyes.
Experimental
Chitosan, a natural polymer of animal origin was purchased by India Sea Food, Kerala, and was used as
received. Its percentage of deacetylation after drying was 89%.Glutaraldehyde was procured from Loba Chemie
Pvt.Ltd ,India and used as a crosslinking agent between chitosanchain units of polymer. All other chemicals like
acetic acid, methanol, NaOH, HCl, KCl, KH2PO4 etc. were used of analytical grade. Double distilled water was
used throughout the studies.
Preparation of chitosan beads
Chitosan (1.0 g) was dissolved in 40 ml of 2% acetic acid under stirring condition for 3h at room temperature.
The homogeneous mixture was extruded in the form of droplets using a syringe into NaOH-methanol solution
(1:20 (w/w)) under stirring condition at 400 rpm. The resultant beads were then placed in a water jacket
containing glutaraldehyde maintained at 50oC for about 10 minutes. Finally the beads were washed with hot and
cold water successively and then vacuum dried 7.
Swelling studies
Swelling behavior of chitosan beads was studied in different pH (2.0, 7.4 and 10.0) solutions. A known weight
(2.0 g) of the prepared beads was placed separately in the conical flask containing media for required period of
time. After predecided time period the swollen beads were collected and their net weight were determined by
first blotting the beads with filter paper to remove adsorbed water on the surface. The percentage of swelling for
each sample at time t was calculated using the following formula-
Percentage of swelling = {(WtWo)/Wo} x 100
Where, Wt = weight of the beads at time t after emersion in the solution.
Wo= weight of the dried beads.
Color removal efficiency (CRE)
50 ml dye solution of known concentration with 1.0 g of beads was taken in conical flask at desired temperature
and pH. It was shaken for 10 min and then kept for 24 h and lastly the remaining concentration of dye was
estimated spectrophotometrically at max of used dye. The CRE (%) for CS beads was calculated by given
formula (9)-
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CRE (%) = (1 Ac/Ai) X 100
Where, Ai and Ac are the absorbance of the dye solution before and after the adsorption process.
Fourier transformed infra red (FTIR) spectroscopy
Dried samples were ground into powder and crushed with KBr for homogenization. A Nicolet economy sample
press was used to obtain optically clear pellets. Pellets were analyzed using transmission FTIR using a Thermo
Nicolet Avatar 370 FT-IR spectrometer system. Dry air was used as the chamber purge stream for all samples.
The FTIR spectra were obtained at room temperature over a spectral frequency range of 400-4000 cm-1. IR
bands are expressed in terms of frequency (cm-1). The background was obtained against a pure KBr pellet and
the data was analyzed by Omnic software. Functional groups present in the raw material and products were
determined.
Scanning electron microscopy (SEM)
The shape and surface morphology of the beads were examined using FESEM QUANTA 200 FEG model
(FEI, The Netherlands make) with operating voltage ranging from 200 V to 30 kV. FESEM micrographs were
taken after coating the surfaces of bead samples with a thin layer of gold by using BAL-TEC-SCD-005 Sputter
Coater (BAL-TEC AG, Balzers, Liechtenstein Company, Germany) under argon atmosphere to make the
sample conducting. The surface appearance, shape and size of scanning electron micrograms were used to
perform textural characterization of full and cross sectioned IPN beads. Magnifications were applied to each
sample in order to estimate the morphology and interior of the bead.
Thermal analysis
Thermal gravimetric analysis (TGA), Differential thermal gravimetric (DTG) and Differential thermal analysis
(DTA) were carried out simultaneously by using a (PYRIS Diamond). TG/DTA thermal analyzer model DSC-7,
supplied by Perkin Elmer and the data was processed and analyzed by PYRIS muse measure and standard
analysis software (V. 3.3U; #. 2002 Seiko instruments inc). The sample was kept in Alumina pan, the reference
material was Alumina powder and study was carried out at heating rate 10C/min under 200 ml/min flow rate of
air or nitrogen atmosphere. Indium and gallium were used as standards for temperature calibration. The
measurements were run from room temperature to 600oC. The thermal stability of produced beads was assessed.
X-ray diffraction (XRD)
X-ray diffraction studies were performed by using Bruker AXS D8 Advance using CuK Nickel filter and
Copper as target at wavelength of 1.54 with goniometer and speed was kept at 2/min. Wide angle X-Ray
scattering patterns of the samples were obtained using DIFFRAC plus XRD commander software and analysis
was done by DIFFRAC plus (version 8.0) software. The range of scanning angle for the sample was kept in the
range 2 of 10 60o.
Results and Discussion
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The CS beads are almost round in shape when freshly prepared but drying results them in uneven shape and also
decreased their volume. The original shape of beads was restored after swelling. The color of uncrosslinked
beads was creamy which was changed into yellow after crosslinking with glutaraldehyde. The yellow colour of
crosslinked beads was turned into brown after air drying or oven drying.
(a) (b)
Figure 1Photographs of freshly prepared (a) and dried (b) chitosan beads
Fourier transform infra red spectroscopy (FTIR)
FTIR curve for chitosan exhibited a broad peak at 3450 cm-1 which was assigned to NH stretching vibration
which might be due to deacetylation of chitosan. The peak at around 3500 cm-1 due to hydrogen bonded O-H
vibrational frequencies and O-H bond stretch of gluco pyranose units. Peaks at 1639 cm-1 and 1319 cm-1 were
observed due to >C=O stretching of amide bond. The peak at 1613 cm-1 was assigned to strong N-H bending
vibrations of secondary amide8. Bands at 2919 cm-1 and 2810 cm-1 represent the aliphatic C-H stretching
vibrations. The observed sharp peak at 1384 cm-1 is due to CH3 symmetrical deformation mode9-10.Two peaks
around 894 cm-1and 1171 cm-1 appeared in spectra corresponding to saccharide structure11. A broad band
appearing near 1083 cm-1 indicated the >CO-CH3 stretching vibration of chitosan.
CHITOSAN
Sun Oct 20 22:16:42 2002 (GMT-05:00)
BD
CS
3500 3000 2500 2000 1500 1000 500

Wavenumbers (cm-1)
133
Figure 2-FTIR of pure chitosan powder (CS) and chitosan Beads (BD)
The structural changes in the beads were investigated using infra-red spectroscopy. It was concluded that during
crosslinking, formation of the C=N group of the imine involved a reaction between the amino group of chitosan
chain with aldehyde group of glutaraldehyde.
Thermal analysis
TGA curve for chitosan indicates that it decomposed in two steps. Chitosan has shown approximately 10%
weight loss below 100oC. This step involves a smaller weight loss, may be due to initial loss of water molecules,
after this no weight loss occurs upto 249oC. A sudden weight loss is observed after 249 oC and the total weight
loss at 400oC is about 60 %. This destruction of chitosan moiety occurs in the second step. TGA curves of beads
clearly shows that pure chitosan crosslinked beads lost weight after 2000C and total weight loss at 4000C is
approximately 46%. This indicates that crosslinking of chitosan with glutaraldehyde increases its thermal
stability.
290Cel 4 1 9 Ce l
40.0 2.00 15.0 0 .6 1 0 m g / m i n
1.425mg/m in
2 4 4 Ce l
63Cel
180.00 0 .3 9 5 m g / m i n
0.500
1 59 C el
0.169mg/ min 10.0 0 .1 5 0m g / m in 200.00
20.0 0.00
296Cel 160.00 0.000
12.9 uV
5.0 2 4 0 Ce l
2 .6 u V
268 mJ/mg 140.00
0.0 -2.00 97.5 mJ/mg
-152 mJ/mg 0.0 -0.500
150.00
D T G m g /m i n
D T G m g /m i n
-57.4 mJ/mg
uV
uV
65Cel 120.00 120 mJ/mg 68.6 mJ/mg

%
%
-7.4 uV 1 41 C el
-20.0 17Cel -4.00 -5.0 - 2. 4 u V -1.000
D TA
D TA
TG
TG
99. 95 %
249Cel 100.00 2 3 Ce l
1 0 0 .0 0 %
87.14 % -10.0 2 0 0C e l 5 0 4 Ce l
4 1 2 Ce l
- 7 .8 u V 100.00 -1.500
-40.0 -6.00 8 9 .6 8 % - 8 .5 u V
100Cel
89. 06 %
200Cel 80.00 1 0 0C e l 2 6 4C e l
88.04 % 275Cel 9 9 .5 3 % 7 2 .0 9 %
82.05 % 300Cel -15.0
57.05 % -2.000
589Cel
32.47 %
60.00 -8.00
3 9 8 Ce l
5 3 .7 0 %
-60.0 350Cel
44.09 % 400Cel 450Cel -20.0 5 9 7 Ce l
40.21 % 50.00
37.68 % 500Cel 550Cel
2 2 .6 2 %
325Cel 35.62 % 33.81 % 40.00 5 0 0 Ce l
2 8 .8 1 %
-2.500
47.88 %
-10.00 -25.0
-80.0
50 100 150 200 250 300 350 400 450 500 550 50 100 150 200 250 300 350 400 450 500 550
Temp Cel Temp Cel
Figure 3 Thermal analysis of chitosan. Figure 4 Thermal analysis of chitosanbeads
DTG thermograms of pure chitosan indites one peak at 290 oC corresponding to higher rate of
weight loss about 1.4 mg/ min. In DTG thermograms in case of crosslinked beads showed lesser
rate of weight loss at 2440C. DTA thermograms for chitosan powder showed endothermic peak at
65oC due to loss of free water and one exothermic peak at 296oC due to chemical transformation.In
case of crosslinked beads only one exothermic peak is observed and no endothermic peak is
observed by chitosan powder.
X-Ray diffraction
The diffraction pattern of pure chitosan has the characteristic peaks at 2 of 12 to 16, 20 and 29.X-
ray diffractograms of beads clearly show the similarly peaks as chitosan powder
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BD
CS
Figure 5- X-ray diffractograms of chitosan (CS)& chitosan beads (BD)
Scanning electron microscopy
The surface appearance, shape and size of scanning electron micrograms were used to perform textural
characterization of full and cross sectioned IPN beads. Magnifications were applied to each sample in order to
estimate the morphology and interior of the bead.
(a) (b) (c) (d)
Figure 6 SEM micrographs of full chitosan bead (a), magnified bead (b),
cross sectioned bead (c) and magnified cross sectioned bead (d)
SEM micrographs of full dried beads with their magnified (2000 X) surface morphology are shown in Figure. It
was concluded from these Figures that the beads were nearly spherical or some what oval in shape. The
approximate size of beads is 164 m. They had rough, rubbery fibrous and folded surfaces with wrinkles.
The micrographs showing internal structure of beads with their magnification are obtained by half cut beads
which are presented in Figure. Interior of the beads appeared to have micropores and micro tube like interior
spaces which confirms the highly porous structures of polymeric beads, although they appeared solid externally.
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Swelling studies
The percentage swelling of pure chitosan beads cross linked with glutaraldehyde in solution of pH 2.0, 7.4 and
10.0 is shown in Figure 7. It was observed that swelling rate increases with increasing pH. When the cross
linked beads were placed in the solution, the solution penetrates into the bead and the bead subsequently tries to
swell. Generally, the swelling process of the beads in pH<6 involves the protonation of amino/imine groups in
the beads and mechanical relaxation of the coiled polymeric chains. Initially during the process of protonation,
amino/imine groups of the bead surface were protonized which led to dissociation of the hydrogen bonding
between amino/imine group and other groups. Afterward, protons and counter ions diffused into the bead to
protonate the amino/imine groups inside the beads and dissociating the hydrogen bonds 12-13 .
pH 2.0 pH 7.4 pH 10.0

50
% Swelling
40
30
20
10
0
0 2 4 6 8
Time (h)
Figure 7- Graphs presenting effect of pH on swelling behavior of CS beads.
Color removal efficiency (CRE)
It was clear from table 1 that chitosan beads removed color of studied dyes in aqueous effluents to a reliable
extant and also% CRE of dyes increases with increasing temperature.
Table-1. CRE of dyes on CS beads
Dye max Initial conc. % CRE (pH 5.0) % CRE (pH 7.0)
-4
(nm) x 10 M
30oC 40oC 30oC 40oC

Congo red 497 3.59 13.65 25.88 12.59 22.94
7.18 43.87 78.64 39.66 70.87
Methyl orange 365 1.795 23.3 36.13 18.78 30.23
Methyl red 410 3.59 34.09 52.04 29.98 40.17
Fluorescein 494 3.59 27.8 42.32 24.79 38.22
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It is further notable that % CRE decreases with increasing pH of the dye solution and this behavior is also
indicated by swelling studies. The % CRE increased with increasing initial concentration in case of congo red
dye. Color removal of dyes occurred due to the adsorption of dye molecules by chitosan beads hence adsorption
of dye on chitosan beads increased with increase of initial concentration of dye and temperature while decreased
with increase of pH.
Conclusion
The CS beads are quite suitable for removal of dyes in waste water effluents due to their good adsorbing
capacity towards dye. Further studies are required to study adsorption and adsorption kinetics.
REFERENCES
[1] George.M.; and Abraham T. E.; Journal of Controlled Release, 2006, 114,1-14.
[2] Gan.Q.; WangT.; Cochrane C.; McCarron P.; Colloids and Surfaces B: Biointerfaces, 2005, 44( 2-3),
65-73.
[3] Tseng H.; Furhata K.; and Sakamoto M.; Carbohy- drateResearch, 1995, 27 (2),149-161.
[4] Kumar Ravi. M. N. V.; Reactive and Functional Polymers, 2000, 46(1),1-27.
[5] Chiou, M. S.; Ho, P. Y.; Li. H. Y.; Dyes and Pigments, 2004, 60, 69-84.
[6] Jung T.; Kamm W.; Breitenbach A.; Kaiserling E.; Xiao J. X.; Kissel T.; European Journal of Phar-
maceutics and Biopharmaceutics, 2000, 50(1)147-160.
[7] Rani M.; Agarwal A.; Negi Y.S.; Bioresources, 2010, 5(4), 2765-2807.
[8] Dhanikula, A. B.; Panchagnula R.; AAPS J, 2004, 6(3) 1-2 .
[9] Peng, T.; Yao, K.D.; Chen.; Goosan, M.F.; J. Poly. Sci. Part A Polymerchemistry, 1994, 32, 591-596
[10] Sannan, T.; Kurita, K.; Ogura, K.; Iwakura Y.; Polymer, 1978,19, 458- 459.
[11] Yoshioka, T.; Hirano, R.; Shioya, T.; Kako, M.; 1990, 35, 66-72.
[12] Gupta, K.C.; Kumar Ravi, M.N.V.; Polymer International , 2000a, 49, 141-146.
[13] Gupta, K.C.; Kumar Ravi, M.N.V.; J. Applied Polymer Science, 2001a, 80, 639-649.
137
Clinical Application and Potential Role of Stem Cells
ABHISHEK GUPTA & BINDU SHARMA

Department of Zoology
DN PG College, Meerut (UP), India
Email Id: abhishekgupta2030@gmail.com
ABSTRACT
Stem cells are basic cells of all multicellular organisms having the potency to differentiate into wide range of
adult cells. Self renewal and totipotency are characteristic of stem cells. Though totipotency is shown by very
early embryonic stem cells, the adult stem cells possess multipotency and differential plasticity which can be
exploited for future generation of therapeutic options. The knowledge of regulators of stem cells has opened the
therapeutic usage of stem cells in the form of neuron regeneration, treatment of bone defect, drug testing, gene
therapy and cell based therapy in the form of muscle damage, spinal cord injury, cancer therapy etc. Cell based
therapies might become commercial in coming years.
Keyboards: cells, potency, potential, differentiation, therapy, tissue
INTRODUCTION
Stem cells are primal cells common to all multicellular organisms that retain the ability to renew themselves
through cell division and can be differentiated into a wide range of specialized cell types. Modern therapeutics is
having a lot of hope from stem cell research in the field of organ transplantation and replacement of lost tissue.
By virtue of self renewal and potency, stem cells can form various types of tissue cells [1-3]. The regulators of
stem cell growth at genomic and proteomic level are identified and we might be able to control stem cell in
vitro. In developed countries, stem cell transplant has become a therapeutic option but in developing countries, it
is still under trial phase. There can be two sources of stem cells Autologous and Allogenic. Autologous
embryonic stem cells generated through therapeutic cloning and highly plastic adult stem cells from the
umbilical cord blood or bone marrow are promising candidates. Allogenic stem cells can be derived from
marrow, peripheral blood, cord blood, family donors or HLA typed or untyped unrelated donors. This article
focuses on types of stem cells and stem cell regulation with enlightening comments on clinical application and
future aspects [4-7].
WHAT IS STEM CELL?
Stem cells are primal cells which are considered to be progenitor of more than 200 cell types present in adult
body. All stem cells are unspecialized (undifferentiated) cells that are characteristically of the same family type
(lineage). They retain the ability to divide throughout life and give rise to cells that can become highly
specialized and take the place of cells that die or are lost. The rigorous definition of a stem cell requires that it
possesses two properties: Self renewal and Unlimited potency. Self renewal means the ability to go through
numerous cycles of cell division while maintaining the undifferentiated state. Unlimited potency means the
capacity to differentiate into any mature cell type. In a strict sense, this makes stem cells either totipotent or
pleuripotent. Multipotent and unipotent are also described to define stem cell potency. Two broad categories of
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stem cells exist: embryonic stem cells derived from blastocyst and adult stem cells which are found in adult
tissue. In a developing embryo, stem cells are able to differentiate into all the specialized embryonic tissue. In
adults, stem cells act as a repair system for the body replacing specialized damaged cells [8-12].
Potency specifies the differential potential of the stem cells. Totipotent stem cells are produced from the fusion
of an egg and a sperm cell. Cells produced by the first few divisions of the fertilized egg are also totipotent.
These cells can differentiate into embryonic and extraembryonic cell types. Only the morula cells are totipotent
able to become all tissues including a placenta. Pleuripotent stem cells are the descendents of totipotent cells and
can differentiate into cells derived from 3 germ layers. Pleuripotent stem cells originate as inner cell mass within
a blastocyst (Blastula). Blastocyst is a thin walled hollow sphere made up of an outer layer of cells, a fluid filled
cavity and an inner cell mass containing pleuripotent stem cells. The blastocyst develops after cleavage and
prior to implantation, in approximately 5 days. These stem cells become any type of tissue in the body excluding
a placenta. Multipotent stem cells can produce only cells of a closely related family of cells e.g. hematopoetic
stem cells differentiate into red blood cells, white blood cells, platelets etc. Unipotent stem cells can produce
only one cell type but have the property of self renewal which distinguishes them from nonstem cells.
TYPES OF STEM CELLS
Stem cells are broadly classified into two categories: Embryonic stem cells (ESC) and Adult stem cells (ASC).
Embryonic Stem Cells
These cells are also known as early stem cells. Embryonic stem cells are derived from embryos at a
developmental stage before the time of implantation would normally occur in the uterus. This developmental
stage is the blastocyst stage 32 cell stage, from which these pleuripotent cells can be isolated. Embryonic stem
cells can give rise to cells from all three embryonic germ layers i.e. ectoderm, mesoderm and endoderm, even
after being grown in culture for a long time. In other words they can develop into each of more than 220 cell
types of the adult body when given the sufficient and necessary stimulation for a specific cell type [13, 14]. ES
cells can be maintained in culture as undifferentiated cell lines or induced to differentiate into many different
lineages. Pleuripotency distinguishes ES cells from multipotent cells found in adults, which can only form a
limited number of different cell types.
Adult Stem Cells
Adult stem cells are undifferentiated cells found throughout the body that divide to replenish dying cells and
regenerate damaged tissue. They are also known as somatic stem cells which can be found in children as well as
adults. The rigorous definition of stem cell require that it possesses two properties: Self renewal- the ability to
go through numerous cycles of cell division while maintaining the undifferentiated state and Multipotency- the
ability to generate progeny of several distinct cell type e.g. both glial cells and neurons, opposed to unipotency
restriction to a single cell type. To ensure self renewal, stem cell undergoes two types of cell division:
symmetric division give rise to two identical daughter cells both endured with stem cell properties and
asymmetric division which produces only one stem cell and a progenitor cell properties and asymmetric division
which produces only one stem cell and a progenitor cell with limited self renewal potential. Progenitor can go
139
through several round of cell division before terminally differentiating into a mature cell. It is believed that
molecular distinction between symmetric and asymmetric division lies in differential segregation of cell
membrane proteins (such as receptors) between the daughter cells.
Clinical application and potential role of embryonic and adult stem cells
There are many ways in which human stem cells can be used in basic research and in clinical research. These
are:
Genetic therapy: Embryonic stem cells benefit the gene therapy by the following ways: First human
embryonic stem cells could be genetically manipulated to introduce the therapeutic gene. This gene
may either be active or awaiting later activation, once the modified embryonic stem cells has
differentiated into the desired cell type. Recently published reports establish the feasibility of such an
approach. Skin cells from an immunodeficient mouse were used to generate cellular therapy that
partially restored function in the mouse. This can also be used in treating human patient with immune
deficiency [15].
Drug Testing: Because embryonic stem cells can proliferate without limit and can contribute to any
cell type, human embryonic stem cells offer an unprecedented access to tissue from the human body.
They will support basic research on the differentiation and function of human tissues and provide
materials for testing that may improve the safety and efficacy of human drugs for example, new drugs
are not generally tested on human heart cells because no human heart cell lines exist. Instead
researchers rely on animal models. Because of important species specific differences between animal
and human heart, however, drugs that are toxic to the human heart have occasionally entered clinical
trials, sometimes resulting in death. Human ES cells derived heart cells may be extremely valuable in
identifying such drugs before they are used in clinical trials, there by accelerating the drug discovery
process and leading to safer and more effective treatments [16].
Cell based therapies: It is perhaps the most important potential application of human stem cells. They
generate cells and tissues that could be used for cells based therapies. Stem cells, directed to
differentiate into specific cell types, offer the possibility of a renewable source of replacement cells and
tissues to treat various disease.
Brain Damage: In the case of brain injury although reparative process appears to initiate, substantial
recovery is rarely observed in adults suggesting a lack of robustness. Recently from research conducted
in rats subjected to stroke suggested that administration of drugs to increase the stem cell division rate
and direct the survival and differentiation of newly formed cells could be successful.
Cancer: Researcher at Harvard Medical School caused intracranial tumor in rodents. Then they
injected human neural stem cells. Within days the cells had migrated into the cancerous and produced
cytosine deaminase, an enzyme that convents a non-toxic pro-drug into a chemotherapeutic agent. As a
result, the injected substance was able to reduce tumor mass by 80 percent.
Spinal cord injury: Recently extensive study work is carried out in treating spinal cord injury. Scientist
have treated the patient of spinal cord injury by isolating adult stem cells from umbilical cord blood
and then injected them into damaged part of the spinal cord.
140
Muscle damage: Adult stem cells are also apparently able to repair muscle damaged after heart attacks.
Heart attacks are due to coronary artery being blocked, staring tissue of oxygen and nutrients. Days
after the attack is over, the cells try to remodel themselves in order to become able to pump harder.
However, because of the decreased blood flow this attempt is futile and results in even more muscle
cells dying. Researchers found that injecting bone marrow stem cells, a form of adult stem cells, into
mice which had heart attacks induced resulted in an improvement of 33% in the functioning of heart.
The damaged tissue had regrown by 68% [17].
Heart damage: Several clinical trials targeting heart disease have shown that adult stem cell therapy is
safe. However none of these trials have proven efficacy. Recently the use of patients own bone marrow
derived stem cells and peripheral blood derived stem cells is becoming popular.
FUTURE PERSPECTIVES OF STEM CELL RESEARCH
Low blood supply: Now the method to produce large numbers of Red blood cells has been developed.
In this method precursor Red blood cells, called hematopoietic stem cells are grown together with
stromal cells, creating an environment that mimic the conditions of bone marrow, the natural site of red
blood cell growth. Erythropoietin, a growth factor, is added coaxing the stem cells to complete terminal
differentiation to red blood cells. Further research into this technique will have potential benefits to
gene therapy and blood transfusion.
Baldness: Hair follicles also contain stem cells, and some researchers predict research on these follicle.
Stem cell may lead to successes in treating baldness through "hair multi-placation" and known as "hair
cloning" as early 2011. This treatment is expected to work through taking stem cells from existing
follicles, multiplying them in cultures, and implanting the new follicle cells which have shrunk during
the ageing process, which in turn respond to these signals by regenerating and once again making
healthy hair [18-20].
Missing teeth: The work on tooth generation has reached to a stage that it will be available to the
general population in that decade. In theory, stem cells taken from the patient could be coaxed in the
lab into turning into a tooth bud which, when implanted in the gums, will give rise to a new tooth,
which would be expected to take two months to grow. It will fuse with jaw bones and release chemicals
that encourage nerve and blood vessels to connect with it.
Deafness: Those have been success in regrowing cochlear hair cells with the use of stem cells.
Blindness and vision improvement: Since 2003 research have successfully transplanted retinal stem
cells into damaged eye to restore vision. Using embryonic stem cells, scientists become able to grow
the sheet of top potent stem cells in the laboratory. When these sheets are transplanted over the
damaged retina, the stem cells stimulate neural repair, eventually restoring vision [21].
CONCLUSION
Stem cells pose a bright future for the therapeutic world by promising treatment options for the diseases which
are considered as noncurable now a days. However, because of significant peri and post-transplant morbidity
and mortality further research and trials are required to refine and optimize conditioning regimens and
modalities of supportive care. By virtue of funding of stem cell research, we hope to see new horizon of
141
therapeutics in the form of organ development and replacement of lost tissue such as hairs, tooth, retina and
cochlear cells.
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143
Wild Life and Biodiversity in the Plays of Shakespeare
Rashmi Rana
Department of English
M. S. College, Saharanpur
What is Wildlife ? This is how we refer to the animals of our jungles and to the beautiful birds that brighten our
lives. I wonder sometimes what these animals and birds think of man and how they would describe him. They
had the capacity to do so. I rather doubt if their description would be very complementary to man. Inspite of our
culture and civilization in many ways man continues to be not only wild but more dangerous than any of the so
called wild animals........................................................................................................................J. L. Nehru
Nick Bottom is a character in Shakespeare's A Midsummer Night's Dream who provides comic relief
throughout the play. He is famously known for getting his head transformed into that of a donkey by the
elusive Puck. Bottom and Puck are the only two characters who converse with and progress the three central
stories in the whole play. Puck is first introduced in the fairies' story and creates the drama of the lovers' story by
messing up who loves whom, and places the donkey's head on Bottom's in his story. Similarly, Bottom is
performing in a play in his story intending it to be presented in the lovers' story, as well as interacting with
Titania in the fairies' story.
Nick Bottom is a member of a theatrical troupe of Athens known as the Mechanicals, who perform a play within
the play. They are foolish and clumsy men, all of whom are craftsmen in Athens: Bottom, the weaver; Snout, the
tinker; Snug, the joiner; Starveling, the tailor; Flute, the bellows-mender; and Peter Quince, the carpenter. The
Mechanicalssometimes called the Hempen Homes punsled by Peter Quince, are rehearsing a play, Pyramus
and Thisbe (written by Peter Quince) in hopes of performing for Duke Theseus on his wedding day and perhaps
even being awarded "six pence a day" for life, really a small reward for these six men. Bottom is given the lead
role of Pyramus in the play, and something of a power struggle ensues between Bottom, a charismatic natural
leader, and Quince, the somewhat nervous playwright attempting to direct his own play.
While they are in the woods rehearsing, the fairy Puck, a mischievous sprite and minion of Oberon, king of the
fairies, happens upon their rehearsal. He decides to have some fun with them, carrying out part of Oberon's
orders in the process, and when Bottom (as Pyramus) exits the stage, he transforms his head into a donkey's.
When Bottom returns, unaware of his own transformation, his fellow actors run away from him with Quince
screaming, "We are haunted!" Bottom believes they are playing a prank on him, proclaiming, "This is to make
an ass of me, to fright me if they could." So he stays in the forest by himself and sings loudly to show them he is
not afraid. The Fairy Queen Titania is awakened by Bottom's song. She has been enchanted by a love potion,
which will cause her to fall in love with the first living thing that she sees when she wakes (no matter who, or
what it is), made from the juice of a rare flower, once hit by Cupid's arrow, that her husband, Oberon, King of
the Fairies, spread on her eyes in an act of jealous rage. During his enchantment over her, he utters "Wake when
some vile thing is near." The first thing she sees when she wakes is the transformed Bottom, and she
immediately falls in love with him. She even commands her fairy minions to serve and wait upon him. Later,
Oberon finally releases Titania from her enchantment. After being confronted with the reality that her romantic
144
interlude with the transformed Bottom was not just a dream, she is disgusted with the very image of him and
also seems very suspicious of how "these things came to pass." After Oberon instructs Puck to return Bottom's
head to his human state, which Puck reluctantly does, the fairies leave him sleeping in the woods, nearby the
four Athenian lovers, Demetrius, Helena, Hermia, and Lysander.
He wakes up after the lovers leave. His first thought is that he has fallen asleep in the woods during rehearsal
and has missed his cue. He quickly realizes he has had "a most rare vision". He is amazed by the events of this
dream, and soon begins to wonder if it was in fact a dream at all. He quickly decides that he will "get Peter
Quince to write a ballad of this dream", and that "it shall be called 'Bottom's Dream,' because it hath no
Bottom". Upon being reunited with his friends, he is not even able to utter what has happened and says "For if I
tell you, I am no true Athenian".
Theseus ends up choosing Pyramus and This be as the performance for his amusement, now also the wedding
day of the young Athenian lovers. The play is poorly written and poorly acted, though obviously performed with
a great deal of passion. Bottom performs the famous Pyramus death scene in the play within the play, ironically
one of the most comedic moments in the play.
In performance, Bottom, like Horatio in Hamlet is the only major part that can't be doubled, i.e. that can't be
played by an actor who also plays another character, since he is present in scenes involving nearly every
character.
Bottom's discussion of his dream is considered by Ann Thompson to have emulated two passages from
Chaucer's The Book of the Duchess[1]
Critics have commented on the profound religious implications of Bottom's speech on his awakening without the
ass's head in act 4 of A Midsummer Night's Dream:
"[...]The eye of
man hath not heard, the ear of man hath not seen,
man's hand is not able to taste, his tongue to conceive,
nor his heart to report, what my dream was. I
will get Peter Quince to write a ballad of this
dream: it shall be called 'Bottom's Dream', because
it hath no bottom; and I will sing it in the latter end
of a play, before the Duke. Peradventure, to make it the
more gracious, I shall sing it at her death."(4.1.209216)
This speech seems to be a comically jumbled evocation of a passage from the New Testament's 1
Corinthians 2.910:
"The things which

eye hathe not sene, nether eare hath heard,
nether came into man's heart, are,which
God hathe prepared for them that love him.
But God hathe reveiled them unto us by
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his Spirit: for the Spirits earcheth all

things , yea , the deepe things of God."
Steven Doloff also suggests that Bottom's humorous and foolish performance at the end of "A Midsummer
Night's Dream" mimics a passage from the previous chapter of Corinthians:
"For seing the worlde by wisdom knewe

Not God in the wisdom of God, it pleased
God by the foolishness of preaching
to save them that believe:
Seing also that the Jewes require a signe,
And the Grecians seke after wisdome.
But we preache Christ crucified :unto
the Jewes, even a stumbling blocke, & unto
the Grecians, foolishnes:
But unto them which are called, bothe
Of the Jewes & Grecias we preache Christ,
the power of GOD, and the wisdom of God.
For the foolishnes of God is wiser the men [. . .]." (1 Corinthians 1.2125)
This passage's description of the skeptical reception Christ was given by his Greek audience appears to be
alluded to in Bottom's performance. Just as Christ's preaching is regarded as "foolishnes," Bottom's audience
perceives his acting (as well as the entirety of the play he is a part of) as completely without value, except for
the humor they can find in the actors' hopelessly flawed rendering of their subject matter. Doloff writes that this
allusion is especially likely because, in both texts, the skeptical audience of the "foolish" material is composed
of Greeks, as the spectators of Bottom et al. are Theseus, the duke of Athens, and his court [2].
The origin of Bottom's farewell to Quince in Act i, scene i has become the topic of some disagreement among
Shakespeare scholars. Parting with Quince, Bottom instructs his fellow actor to be at the next rehearsal, saying:
"Hold or cut bowstrings." The debate is centered on whether this phrase arose from military or civilian life.
George Capell is the first to have offered an explanation of the origin of this phrase. He states that it is a
proverbial saying and "was born in the days of archery" When an archery contest was planned, 'assurance of
meeting was given in the words of that phrase'. If an archer did not keep the promised meeting, then the other
archers might cut his bowstring, that is, 'demolish him for an archer'. From this 'particular usage, the phrase had
an easy transition among the vulgar to that general application which Bottom makes of it [3].
However, W.L. Godshalk refutes this theory, stating that no subsequent scholars have been able to confirm
Capell's ideas. Godshalk also states that it is unlikely that this was a common civilian phrase, as there are no
other examples of this exact form of the phrase in the work of any author besides Shakespeare.
Godshalk further cites the work of George Steevens, who was able to find two vaguely parallel examples in
seventeenth-century drama [4]. In George Chapman's The Ball, Scutilla asks Lady Lucina, 'have you devices /
146
To jeer the rest?' Lucina answers, 'All the regiment of 'em, or I'll break my bow-strings' (II.ii.127-9) [5].
Godshalk argues that the context implied by 'regiment' is important, as it implies that the breaking (or cutting) of
bowstrings should be seen in terms of military rather than civilian archery. Steeven's other example is from
Anthony Brewer's The Covntrie Girle: A Comedie: 'Fidler, strike. / I strike you else; -- and cut your begging
bowstrings [6]. Godshalk writes that "the first 'strike' means 'to play upon' the fiddle; the second 'strike' may
again suggest a military context for the cutting of bowstrings, though any reference to military archery is comic
since the 'bow' in this case is the fiddler's bow."
Godshalk argues that, just as these examples indicate a military context, this must also be done with Bottom's
"hold or cut bow-strings." He further cites Jean Froissart's account of the Battle of Crecy, which supports the
military origin of Bottom's line: "When the Genoese felt the arrows piercing through their heads, arms, and
breasts, many of them cast down their crossbows, and cut their strings, and resumed discomfited [7]. " Archers
would cut their bowstrings, thus destroying their weapons, in the midst of a retreat so that the enemy could not
use their own instruments against them. It is the equivalent of striking artillery, rendering the equipment useless.
With this understanding, Bottom's phrase can be interpreted as a military expression for "hold your position, or
give up and retreat." In the context of the play, Bottom is being comically pretentious, saying: "Be present at the
rehearsal, or quit the troupe [8].
Some of the more successful interpretations of Nick Bottom are those of Samuel Phelps, Herbert Beerbohm
Tree, Ralph Richardson, Stewart Wright, Andrew Blake, as well as Paul Rogers. Actors who have played the
role on film include Paul Rogers, James Cagney and Kevin Kline. In the BBC Television Shakespeare version
he is played by Brian Glover.
Bottom has been the subject of several paintings. German composer has Werner Henze has used Bottom twice
as an inspiration: in the second sonata which comprises his Royal Winter Music and in his Eighth Symphony.
References
1. Hale, David G., Bottom's Dream and Chaucer, (http://links.jstor.org/

sici?sici=00373222%28198522%2936%3A2%3C219%3ABDAC%3E2%3E2.0CO%3B2-M)
Shakespeare Quarterly, Vol. 36, No. 2 (Summer, 1985), pp
219220, doi:10.2307/2871197(https:dx.doiorg/10.2307%2f2871197)
2. Doloff,Steven, [1] (http://web.ebscohost.com/ehost/pdf?vid=2&hid=117&sid=4881bde4-
d7e342708be5cf6b8b03396 7%40 sessio nmgr 7) Bottom's Greek Audience: 1 Corinthians 1.2125
and Shakespeare's A Midsummer Night's Dream, doi: 11.3 007 / 2 871197
3. George Capell, Notes and Various Readings to Shakespeare (1780; New York, 1973), ii.102
4. Isaac Reed (ed.), The Plays of William Shakespeare ... Notes, by Samuel Johnson and George
Steevens (London, 1803), iv.342
5. Thomas Marc Parrott (ed.), The Plays of George Chapman (New York, 1961), ii.557
6. Cf. Marlowe's Jew of Malta, ed. N. W. Bawcutt (Manchester, 1978)
7. Jean Froissart, The Ancient Chronicles of Sir John Froissart, trans. John Bourchier, Lord Berners
(London, 1814) i.288.
147
8. Godshalk, W.L. "Bottom's 'Hold or cut bow-strings' (A Midsummer Night's Dream I.ii.106)." Notes
and Queries 42.n3 (Sept 1995): 315(2). Academic One File. Gale. Yale University. 30 Nov. 2007
148
Conservation of Withania somnifera and Commiphora wightii medicinal plants
Anshu Dhaka & Vinit Kumar
Department of Botany, D. N. College, Meerut
Department of Environmental Sciences,
Bundelkhand University, Jhansi
Abstract
The roots of the ashwagandha contains a number of alkaloid range from 0.13 -.013 % such as somniferine,
choline, pseudotopanol,3-tigioyloxytropana and tropanol etc. Ashwagandha is used as general tonic, anti-
inflammatory, Anti-stress, adaptogenic, immune modulator, endocrine, anti-tumor, treating ulcers, bacterial
infection and in conjunctivitis. Commiphora wightii commonly known as guggal, belongs to the family
Burseraceae having neem like leaver. The fluid of bark in air contact changes to a reddish brown known as
myrrh used in incense and perfumes. Guggal contains resin (25-35%) and essential oil (2.5 - 6.5%). Gum is
carminative, stomachic, appetite and improving digestion, disinfectant, expectorant, stimulant also used in
cough and fever. Gum resin has proved to have hypocholesterolemic action. Both the plant species are an
important medicinal plant and have economic value became endangered due to over utilization, habitat
destruction, over exploitation, intensive cultivation, environmental degradation, desertification and destructive
harvesting. These species could be conserved through In-situ conservation by protecting plants in their native
growing areas and Ex- situ conservation by cryopreservation , overcoming the seed dormancy following the
appropriate seed treatment methods and to develop protocol for the tissue culture to protect these species from
loss.
Key words: Ex- situ conservation, Germination, cryopreservation, guggal, ashwagandha,
INTRODUCTION: The world health organization (WHO) has listed over 21,000 plants that have recorded
medical uses around the world. At present about 90 species of higher plants have yielding a total of 119 pure
chemical substances that are used in medicine throughout the world. The pharmaceutical industry is directed
dependent on biodiversity. There are the some important endangered medicinal plants of India such as
Glycyrrhiza glabra , Bacopa monnieri, Withania somnifera , Rauwolfia serpentina, Commiphora wightti, Stevia
rebaudiana and Ginkgo biloba . In these Withania somnifera commonly known as Ashwagsndha belongs to the
family Solanaceae. Withania somnifera holds as ayurvedic tradition similar to Ginseng in Chinese therapies and
hence referred to as the Indian Ginsing in ayurvedic world. Roots of only cultivated variety of Withania
somnifera are used in medicine which contain the alkaloid somniferine and is the source of the drug of
economic importance. The roots of the ashwagandha also contains a number of alkaloid range from 0.13 -.013
% such choline, pseudotopanol, 3-tigioyloxytropana and tropanol etc. Ashwagandha is used as general tonic,
anti-inflammatory, Anti-stress, adaptogenic , immune modulator, central nervous system, endocrine, anti-tumor,
anticancer , treating ulcers , bacterial infection and in conjunctivitis .Bark decoction in used for asthma and its
extract are used in the preparation of ashwagasndha powder , tablets syrup and herbal tea. Ashwagandha also
149
has rejuvenating properties. It helps maintain proper nourishment of the tissue muscles and bones and
supporting the proper function of the adrenals and reproductive system.
Commiphora wightii commonly known as guggal, belongs to the family Burseraceae, a thorny shrub with neem
like leaver. The plant distributed in arid area of India, Pakistan and Bangladesh. The gum resin is collected by
wounding the bark when secretion exudes out in the form a yellowish white fluid. The fluid of bark in air
contact changes to a reddish brown known as Myrrh used in incense and perfumes. Guggal contains resin (25-
35%) and essential oil (2.5 - 6.5%). Gum is bitter in taste, pungent, aromatic, carminative, stomachic,
stimulating, appetite and improving digestion. It is also used as alternative, antiseptic, disinfectant, expectorant,
stimulant and also used in cough, fever, hair falling, sores, ulcers and uterine affection .Gum resin has proved to
have hypocholesterolemic action. (lipid metabolism) and antherosclerosis useful in heart trouble, arthritis,
sclerotosis and coronary artery trouble.
Causes of loss of these species :- Both the plant species are an important medicinal plant and have economic
value became endangered due to over utilization for the medical purpose by the local people , habitat destruction
human causes the majority of threats to species , sites and habitat, over exploitation, and faulty tapping
techniques, intensive cultivation, environmental degradation, human population, desertification, uncontrolled
and unscientific grazing and destructive harvesting also responsible for the loss of these species. Because of the
over use of the guggal in its habitat Rajasthan and Gujrat. The world conservation union (IUCN) has enlisted it
in its Red Data List of endangered species. Endangered species are those which have already become extinct
and the plant species which are on the urge of extinction. According to IUCN the endangered species are
classified as Extinct species which cannot be found in area where they recently been inhabited . Rare species
species with small population restricted geographically with localized habitats. They are not in immediate
action. Vulnerable species are under threat of or actually declining in number. Critically endangered when
a species is facing an extremely high risk of extinction in wild in the immediate future.
In India, Indias National Medicinal Plants Board launched a project in Kutch to cultivate 500 800 hectares of
guggal. While a grass root conservation started to educate guggal growers and harvesters in safe, sustainable
harvesting methods. Both the plant became endangered /threatened because of its slow growing nature , poor
seed setting , poor seed germination rate (due to dormancy) overgrazing by domestic animal , lack of cultivation
and excessive &unscientific tapping for its gum resin . These plant are incorporated in Data Deficient Category
of IUCN Red Data list .On the basis of survey southern part , Central Eastern part of Rajasthan were identified
as Biodiversity Hotspots for Commiphora wightii. Southern part of Rajasthan is facing serious threat of
extinction of this plant.
Conservation :-
Conservation means to preserve the species from harm, decay to protect from loss or consumption.
There are two categories of conservation:-In-situ conservation & Ex-situ conservation.
150
In-situ conservation (means on site conservation): conservation of species in its natural habitat in places where
the species normally occurs. The natural surrounding is protected and maintained so that all the constituent
species known and unknown to us are conserved and benefitted. This conservation method has two aims-
To maintain economic production.
To replant the ecosystem with local sources of propagule.
Ex- Situ conservation means off-site conservation maintenance of species or (sample of living organism away
from their natural habitat /ecosystem in the form of whole plant, vegetative propagules , seed , pollen, tissues or
cell cultures conservation of colonel materials ,tissue culture , by genetic modification (disease resistant plants
pest resistant plants.
National Bureau of Plant Genetic Resource (NBPGR ), New Delhi was established in 1976 with the
responsibility to plan ,undertake and coordinate activities and service related to plant genetic resources including
collection , exchange, quarantine , evaluation , documentation conservation and utilization .It is a nodal
organization for developing , operating and coordinating the Indian plant genetic resource system The system
comprises base collections of germplasm sites located throughout the country. These sites are responsible for
evaluation, multiplication and medium term storage of the germplasm.
Cryopreservation is the another storage technique to freezing of recalcitrant seeds, organ, tissue, cell, pollen
bank etc. at liquid nitrogen at 196 o C. The cryogene bank at NBPGR conserve more than 5000 accession
belong to 473 species of threatened /endangered and wild species.
Methods of preservation of seed storage condition :Orthodox seed included small seeded grain crops and
vegetables dried to very low moisture level and at low temperature can be stored easily for several year means
long term storage at -12 o C and 30 % RH. Medium term storage condition at 0-10 o C and 25-30 % RH. Short
term storage condition refers to temperature of 20 o C or less and no control of related humidity for 50 100
year.
There are some appropriate methods for overcoming seed dormancy such as mechanical scarification, hot water
treatment for small seed and various chemical treatment.
The tetrazolium test was conducted to find out the seed viability of these plants and the seed showed 95-97 %
viability. Seed soaked in nitrate of sodium and potassium reduced the number of day taken for germination
enhanced the germination percentage and vigour index. The germination percentage was significantly lower
when seeds were sown as such followed by seed soaked in water without any soaking treatment seed took more
number of days for germination followed by seeds soaked in water. The seed dormancy was to overcome by
using various methods such as seed scarification by mechanical, hot water and chemical treatment. The hot
water soaking treatment for 10 minutes duration was significantly effective and enhanced germination
percentage .The soaking of seeds in GA3 300ppm, 0.5% KNO3, and 0.5 % thiourea solution for 24 hr. was found
highly effective in Withania for improving germination and vigour index the soaking of seeds in of different
medicinal plants in KNO3 0.5% solution for 24 hr. found to enhance germination vigour index and speed of
151
germination. The Ashwandha seeds soaked in 1 percent sodium nitrate recorded the maximum plant height and
largest roots [3] nitrates improved seed germination performance in Commiphora
Discussion: The regeneration of this plant takes place vegetatively either by stem cuttings of air layering [2-
3], but through seeds is extremely poor in nature [2]. A large number of chemical substances such as various
nitrate solutions have been used for breaking seed dormancy and enhancing their permeability and act as a
stimulator and inhibitor. Various concentrations (5, 10, 15 and 20 mg L-1) of nitrate solution such as NH4 NO3,
CO (NO3)2, Ca (NO3) NO and KNO3 were used for enhancing seed germination in Commiphora [1].
Conclusion: There is need to explore the possibilities for developing protocol for induction and establishment
of plant regeneration from shoot multiplication from in-vitro explants. Through in-vitro micro-propagation it is
possible to produce a large number of plantlets within a very short period.
References:
1. Heera lal , pawan kumar kasera1*. Nitrates improved seed germination performance in commiphora
wightii (guggal), a data deficient medicinal plant from the indian arid zone j. Plant develop. 21, 6373,
(2014).
2. KASERA P. K. & PRAKASH J. Ecology and cultivation practices of guggal (Commiphora wightii):
An endangered medicinal plant of the Thar Desert in India. In: D. K. MAJUMDAR & al. (eds.). Recent
Progress in Medicinal Plants: Plant Bioactives in Traditional Medicine. Studium Press LLC, USA,
Vol. 9. Pp 403-423. (2005).
3. Kattimani and Redy. A note on influence of growth of presowing seed treatments on growth and root
yield of Ashwagandha, Karnataka J. Agri. Sci., 14 (3), 846-848, (2001).
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The zero energy sewage treatment plant (ZESTP). A conventional or alternative method of
sewage treatment to enhance the livelihood of marginal formers in peri urban region of
Gorakhpur (U.P.)
Shyam Singh
Department of botany M.G.P.G. College Gorakhpur
drshyamsingh62@gmail.com
Abstract
The process of urbanization in India since beginning of last century reveals a steady increase in size of its
population of sewage also enhance, which create several problem like health and hygiene, pollution water
logging etc. Gorakhpur is one of the fastest growing city in mid gangetic plains of eastern Uttar Pradesh in
present investigation the sewage water is used after the treatment through ZESTP. This technology clean up the
contaminated site. It is a plant based technology, natural plants contains ability to degrade & remove toxic
chemicals and pollutants from sewage water ; three macrophytes such as Eichhornia crassipes (mart.) solms,
Pistia stratiotes L. and Hydrilla Verticillata Casp., three treatment given in ZESTP, physical, biological and
chemical, at the interval of 2 days, 15 days, 3 days this treatment water used by mariginal farmer in periurban
areas of Gorakhpur to enhance their livelihood .
Introduction:
Sewage is the waterborne waste derived from domestic uses and animal or food processing plants. It
includes human excreta, paper, cloth soap, detergents etc. When quality and composition of water is altered by
anthropogenic interference, such that it no more be safe for direct or indirect consumption then the water is said
to be polluted. Contamination of aqueous environment and poses several health problems which are still in need
of effective and affordable technological solutions.
The source of water to Gorakhpur city is mainly ground water. There are 75 power bore well, 8 mini
power well, 3694 hand pumps and 450 public stand posts. About 82 MLD (Million liters per day) of water
produced from ground water and not a single unit of water is produced from surface water sources. Average
daily water supply is 77.60 LPCD (Liter per capita per day). Approximately 70 % domestic water supplied is
released as wastewater. The total waste water generated in Gorakhpur city is 65.84 MLD.
Gorakhpur is the main terminus for Kushinagar, Kapilvastu and Nepal. Gorakhpur is Located the Terai
belt of Eastern Uttar Pradesh in India. It lies between Lat. 2613N and 2729N and Long. 8305E and 8356E.
Gorakhpur has the most rain during June to September. The city is situated in the plain of Saryu of mid Gangetic
valley and at the confluence of Rapti and Rohini River.
Material and method :
Sewage samples were subjected for analysis as prescribed by APHA (1998) for all the selected
parameters. Colour & odour were analysed by direct manual methods at the study site itself however, for
153
analysis of the other parameter the water sample were bought into laboratory. On the basis of nature of
pollutants, resistance and sensitivity of plant, three aquatic macrophytes such as Eichhornia crassipes (Mart.)
Solms, Pistia stratiotes L. and Hydrilla verticillata Casp. were selected for the present investigation. The
experiment conducted for biological treatment was analysed for a period of 20 days at the interval of 5 days,
during the treatment process. For mixed culture experiments, five aquaria of 25 liter capacity were maintained.
Initial reading of all the variables was also recorded for comparison of the results. In each aquarium 25 liter of
waste water was poured and roots of all selected macrophytes were washed thoroughly in tap water before the
plants were placed in aquarium. Best result were obtained in the case of 15 days treatment, therefore, only the
reading of 15 days is expressed.
In the light of above experiment ZESTP model (Figure 4) was developed which have 2270 liter
purifying capacity in peri-urban part of Gorakhpur city (Jangal bahadur Ali block Chargawa). Zero Energy
Sewage Treatment Plant (ZESTP) is a three chambered Sewage Treatment Plant made up of Brick, Cement,
plastic pipe, barrel pump and Iron net which functions without use of energy input. It treats sewage water with
the help of aquatic plant. ZESTP works in following three steps (Figure 5.1)
1. Step First: First of all collects sewage directly in the first tank and stay for two days.
2. Step Second: In this step the stagnant sewage water (except precipitate) from 1st chamber transfer with the
help of barrel pump (Figure 5.1) in second tank of ZESTP, then implant aquatic macrophyte (Two plant of each
Eichhornia, Pistia and Hydrilla respectively/ square feet) for fifteen days.
3. Step Third: After fifteen days the second tank water transfer in third tank with the help of barrel pump
followed by chlorination (5gm/100 liter) and left water for three days before irrigation or other application of
treated sewage.
Eichhornia crassipes (Mart.) Solms, commonly known as water hyacinth (Figure 5.2) belong to
family Pontederiaceae. It is a perennial, free floating; consisting of horizontal and spongy stem having large
aerial leaves, cluster of brown or pinkish adventitious root with root pockets, growing by the means of offset.
Eichhornia crassipes is monocotyledonous and cosmopolitan in distribution.
Pistia stratiotes L. commonly known as water lettuce (Figure 5.3) belong to monocotyledonous of family
Araceae. Plants are small, free floating, perennial aquatic herbs of gregarious nature with sessile and spatiualate
leaves. They are in close spirals and form cup shaped rosette in the region of nodes. Roots bear root pocket.
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Figure 5.1: Eichhornia crassipes (Mart.) Solms Figure 5.2: Pistia stratiotes L
Figure 5.4: Barrel Pump

Figure 5.3: Hydrilla verticillata Casp
Hydrilla verticillata Casp. is a submerged plant belonging to family Hydrocharitaceae of monocot. It is free
floating and herbaceous in nature. Hydrilla verticillata (Figure 5.4) release high amount of oxygen directly into
the water.
Figure-5.5: Zere Energy Sewage Treatment

Plant (ZE)
155
Sewage treatment
2 Day
15 Day 3 Day
Sedimentation
Eichhornia Chlorination
Pistia
Hydrilla
PHYSICAL BIOLOGICAL CHEMICAL

TREATMENT TREATMENT TREATMENT
Figure 5.6: Flow diagram of ZESTP

Treatment Capacity:
One cubic foot (1x1x1) area have = 28.317 Liters water
= 28 Liters and 317 Milliliters.
Total area covered by ZESTP - = 15 x 4 feet
= 60 square feet
Total requirement of aquatic macrophyte -
=60 x 2 Plant each
= 120 plant (Eichhornia, Pistia and Hydrilla respectively)
Size of one tank-
= 4 Length x 5 Breadth x 4 Depth =80 cubic foot
=80 x 28.371
=2269.68 liter water
Therefore, After complete operation of ZESTP , purify nearly 2270 liter sewage water at every 15 days.
Result and discussion:
Colour, Odour and Temperature
The colours of sewage water was light black (Figure 5.7). This is due to decomposition of organic
pollutant, resultant formation of humic acid which dissolve and give dirty colour. Similar observation was also
reported Singh et al (2004). Ahmad et al (2006) observed that the temperature fluctuate during environmental
change. Study site had rotten water like smell due to decomposition of organic matter. The colour of sewage
water changed to light yellowish (Figure 5.8) from dark blackish by biological treatment. After treatment odour
as well as temperature reduced favorably (Table 5.1)
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Figure 5.7: Sewage sample before Treatment Figure 5.8: Sewage sample after Treatment
pH
The pH is important parameter used to indicate the water quality and treatment efficiency. The pH
always fluctuates due to microbial degradation as well as different aquatic chemical reactions. It was found that
the selected vegetation of water hyacinth helped in effectively reducing the pH of wastewater. The pH of treated
effluent was found near to neutral, though it was decreased in small percentage than inlet. Such reduction in pH
with the help of water hyacinth was reported (Wolverton and McDonald, 1980). pH which is 7.86 before
treatment reduced to 7.12 by using macrophyte Eichhornia, Pistia, Hydrilla in mixed culture experiment
respectively. The mixed culture treatment reduced 9.41% pH. It gives the intensity of acidic or basic character
and not the total acidity or alkalinity.
Turbidity and Conductivity
During June temperature is higher due to which the rate of brownian motion is higher at the same time
TSS of the water sample was also higher indicating the contribution of suspended solid in turbidity. Particle of
colloidal dimension suspended and do not settle easily give a dirty or turbid appearance. Conductivity denotes
the capacity of a substance of solution to conduct the electric current. Inorganic substance show better
conductance while organic compound are poor conductance as they do
Table 5.1: Result of Biological Treatment (initial and after fifteen days),
* Increase Percentage ,* *Standard by BIS, UPPCB, ISI, WHO, ICMR
Before After % Reduction Standard
S. treatment Treatment **
No. Parameter (Mix Culture)

Mg/l Mg/l Mg/l
1 Colour Blackish Light yellowish Colourless
2 Odour Sewer like Light smell Odourless
0 0
3 Temperature 38.5 C 28.8 C 25.19 40 0C
4 pH 7.86 7.12 9.41 6.5-8.6
5 Turbidity 76 12 84.21 5 NTU
6 Conductivity 512 278 45.70 300 mho
7 Salinity 234 134 42.73 --
8 Total solid (TS) 1270 817 35.66 500
9 Total Dissolve solid 815 678 16.80 500
10 (TDS) Suspended
Total Solid 455 139 69.45 <100
11 (TSS)
Acidity 145 37.8 73.93 --
12 Alkalinity NIL NIL NIL 200
13 Free CO2 145 44.5 69.31 300
14 Total Hardness 660 248 62.42 500
157
15 Chloride 130.4 38.2 70.70 250

16 DO* 2.4 8.8 72.72* 4-6
17 BOD 185 50 72.97 <100
18 COD 210 118 43.80 <150
19 Cadmium 0.457 0.187 59.08 0.01
20 Iron 2.00 0.985 50.75 0.3
21 Copper 0.905 0.211 76.68 0.05
22 Lead NIL NIL NIL 0.05
23 Chromium NIL NIL NIL 0.05
not dissociate easily. The initial reading of Tutbidity and Conductivity was recorded to be 76 mg/l and 512 mg/l
respectively, which show tremendous reduction 84.21% and 45.70% respectively by the use of aquatic
macrophytes recorded after treatment (Table 5.1).
TS, TDS and TSS
Total solid (TS) can be determined as the residue left after evaporation of the unfiltered weight. Total
Dissolve Solid (TDS) can be determined as the residue left after evaporation of the filtrate sample. A suspended
solid not only release pollutants from the sediments but also increase the turbidity and reduces the light
penetration. Reduction in suspended solids is helpful in decreasing the turbidity as well as BOD load. The
Eichhornia crassipes (Mart.) Solms through their stems ensure the presence of significant amount of air (O2) in
the zone of their root systems, enabling development of aerobic bacterial colonies in the root zone. The water
hyacinth provides appropriate environment for microbial attachment and growth, preventing flow and retaining
suspended solids. Karthikeyan and Singh (2004) reported that when TS and TDS were quite high may cause soil
sickness due to poor aeration and higher BOD. These also affected the availability of trace elements. Mixed
culture of aquatic plant reduces the concentration of TS (35.66%), TDS (16.80%) and TSS (69.45%) in sewage
water, as given in table 5.11 and figure 5.9.
1500
1000
500
0
Conductiv
TDS
TSS
Turbidity
TS
Salinity
158
Before Treatment After Treatment

Figure 5.9: Graphical representation of TS, TDS and TSS
Free CO2 and Chloride

Free Carbon di oxide in water accumulated due to microbial activity and respiration of organism, this
imparts acidity to the water by the formation of carbonic acid. Free CO2 and chloride which is 145 mg/l and
130.4 mg/l respectively before treatment reduced to 44.5 mg/l (69.31%) and 38.2 mg/l (70.70%) after treatment
by mixed aquaculture, as also shown in table 5.1 and figure 5.2.
Acidity, Alkalinity and Hardness
Acidity is the capacity of water to neutralize a strong base to a fixed pH. It is an aggregate property of
water due to several specific substances but chiefly it is due to the CO 2 dissociating from carbonic acid.
Alkalinity of water sample is its quantitative capacity to neutralize a strong acid to a designated pH. Table 1
shows reducing potential of mixed culture of plants, in respect of acidity (73.93%) and hardness (62.42%).
Acidity which is 145 mg/l before treatment reduced to 44.5 mg/l after treatment in the mixed culture of
Eichhornia, Pistia and Hydrilla. Hardness is generally imparted by Calcium and magnesium ions present in
water.
DO (Dissolve Oxygen)
It is amount of oxygen gas that is dissolved into water at any source. DO is usually a critical function
and at time it may cause anoxia (oxygen deficient) and death of aquatic organism. Dissolved Oxygen (DO) is
important characteristic used to check the quality of water. The low DO levels indicate higher pollution load in
the wastewater which affect the treatment efficiency. In the constructed wetlands, all the inlet samples collected
were devoid of DO. Macrophytes play a very important role in increasing DO levels through the process of
photosynthesis. Such increased DO levels favors the aerobic microflora in the wastewater involved in
degradation activity (Bastviken, 2006). The increased DO helps to reduce BOD load of wastewater. It was also
indicated by reduction of BOD and COD in the wastewater and also increases the mass weight of the vegetation.
In wastewater treatment systems, oxygen plays an important role owing to its direct involvement in the
treatment of a number of pollutants (Hiley, 1995). The experiment carried out in surface flow constructed
wetland system for the treatment of wastewater in an institutional complex showed increased levels of DO
because of the use of Eichhornia crassipes (Yadav et.al, 2011). A tremendous increase in dissolve oxygen by
mixed culture was recorded. Initially, DO was 2.4 mg/l, which increases to 8.8 mg/l through aquaculture of
mixed culture respectively (Figure 5.3). The dissolve oxygen level increased (72.72 %) due to photosynthetic
activity (Table 5.1).
BOD and COD
BOD is measures of oxygen that would be needed by microorganism to decompose the organic and
inorganic pollutant Higher BOD indicates the more microorganisms in water. Biochemical Oxygen Demand
(BOD) was used to check the organic load in the water body. Biological oxygen demand can be used to indicate
the amount of organic matter in the water. High BOD levels reflects that more oxygen will be consumed as
159
organic matter decays leading to low dissolved oxygen levels. Similar reduction in BOD has been reported by
using bio filter for treatment of wastewater (Nadirah et. al, 2008). The reduction in BOD occurred due to
aerobic, anoxic and anaerobic microbial processes in the constructed wetland system.
1000
500
0

Figure 5.10: Graphical representation of BOD and COD.
Water hyacinths remove pollutants directly by assimilating them into their tissue and providing a suitable
environment for microorganisms to transform pollutants and reduce their concentrations (Wolverton et al.,
1980). COD is measure of oxygen consumed during the oxidation of oxidizable organic matter by strong
oxidizing agent. The initial reading of COD and BOD was recorded to be 210 and 185 mg/l respectively (Figure
5.2), which show tremendous reduction by the use of aquatic macrophytes recorded after treatment. Choudhary
(2007) showed 87.17% reduction in COD with Eichhornia crassipes (Mart.) Solms. Such reduction is helpful in
increasing the DO level in the wastewater. Roy et.al. (2010) also reported combination treatment of aquatic
macrophytes and an alga were found to be effective for the reduction of COD. The combination treatment
include Nostoc, E. crassipes and P. stratiotes reduced 69 % COD. It was observed that there was 72.97%
Reduction in BOD and 43.80% in COD.
Heavy metal
The concentration of toxic metals such as Cd (0.457ppm), Fe (2.00ppm) and Cu (0.905ppm) were
significantly higher in sewage water while Pb and Cr are not reported. These heavy metals were reduced
concentration (Cd- 59.08%, Fe- 50.75% and Cu- 76.68%) after biological treatment (Table 5.1 and Figure 5.3).
Enhance level of heavy metal in the aquatic environment cause several health problems. Mohanty et al. (2006)
studied aquatic submerged plant E. crassipes as an effective biosorbent for the removal of chromium. Around
73 to 89 % of removal was observed for initial concentration of 10 mg/l when the dose was changed from 0.05
to 0.2 g/100 ml. The results indicated that the biomass of E. crassipes is suitable for development of efficient
biosorbent for the removal of chromium from waste water. Mishra et al. (2008) observed that combination of E.
crassipes and L. minor was most efficient for the removal of heavy metals while E. crassipes was efficient in
monoculture. This method can be applied on the large scale treatment of waste water where volumes generated
are very high and concentrations of pollution are low. Mishra and Tripathi (2009) found E. crassipes as a good
160
accumulator of Cr and Zn. This macrophyte has accumulated Zn and Cr up to 3.542 and 2.412 mg/g metal
concentration of 10 mg/l after 11 days of exposure. This plant has successfully removed up to 84% of Cr and
94% of Zn (Table 5.2). Miretzky et al. (2006) studied the mechanism of simultaneous metal removal (Cd, Ni,
Cu, Zn, and Pb) by three macrophytes biomass (Spirodela intermedia, Lemna minor and Pistia stratiotes).
10
5
0
Chrom
Cadmi
DO
Lead
Cuppor
pH
Iron
Figure 5.11: Graphical representation of Heavy metals

Table 5.2: Removal efficiency of aquatic macrophyte
Aquatic Removal
Heavy Metal Reference
Macrophyte Capacity
Fe 95%
Cu 96%
Mishra and Tripathi
Pistia stratoites Cd 82%
2008
Cr 81%
Zn 92%
Cd 83.24%
Ni 32.77%
Pistia stratiotes Cu 94.48% Miretzky et al. 2006
Zn 26.08%
Pb 54.77%
Eichhornia
Cr (VI) 73-89% Mohanty et al. 2006
crassipes
Fe 90.1%
Cu 95%
Mishra and Tripathi
Eichhornia cressipes Cd 85%
2009
Cr 89%
Zn 95%
161
The species Lemna minor biomass presented the highest mean removal (84.52%) and Pistia stratiotes recorded
the lowest for all metals tested. Pb and Cd were more efficiently removed by all three of them. The absorption
of diluted heavy metal ions, in particular, Pb and Cd by dead S. intermedia, L.minor and P. stratiotes appears to
be an efficient and low cast alternative to be considered in industrial effluents treatment.
Conclusion:
The colour of waste water change light yellowish from dark blackish by biological treatment. After
treatment odour becomes light smell and temperature reduced. pH which is 7.86 before treatment but reduced
7.12 (9.41%) by using macrophyte. Mixed culture of aquatic plant reduces the concentration of TS (35.66),
TDS(16.80%) and TSS(69.45%) in waste water. Free CO2 and chloride which is 145 mg/l and 130.4 mg/l
respectively before treatment reduced after treatment 44.5 mg/l (69.31%) and 38.2 mg/l(70.70%) by mixed
aquaculture. In initial sample DO was 2.4mg/l which increases to 8.8mg/I (72.42%) through mixed culture.
COD and BOD were 210 mg/l and 185 mg/l before treatments which show tremendous reduction (COD 43.80%
and BOD 72.97%) by the use of aquatic macrophyte. Aquatic macrophytes, having rapid growth rate absorb
large quantity of nutrients would provide practical and economical method for tertiary treatment. As an outcome
of the complete remedial experiments, mix culture of the selected aquatic macrophytes for fifteen days is
suggested to be the best possible solution. The concentration of toxic metals such as Cd (0.457mg/l), Fe (2.00
mg/l) and Cu (0.905 mg/l) were significantly higher in sewage water while Pb and Cr are not reported. These
heavy metals were reduced concentration (Cd- 59.08%, Fe- 50.75% and Cu- 76.68%) after biological treatment.
Acknowledgement:
We are thankful to Dr. S.A.Wajih former principal M.G.P.G. College Gorakhpur and GEAG
Gorakhpur for financial support and helpful suggestions.
References:
1. APHA. 1998. Standard mathods for the examination of water and waste water, 20 th ed. American
Public Health Association, Washington DC-USA.
2. BIS (1991) Drinking water speciation. Bureau of Indian Standars. IS, 10500 (Revised 2003).
3. City sanitation plan, Gorakhpur (2011-2012) Center for energy, environment, urban governance
and infrastructure development, Administrative staff college of India (ASCI), Della vista, Raj
Bhavan road Hyderabad-500062.
4. Das GK, and Datta S, (2006): Managing waters of wetlands in and around Kolkata. Indian Science
Cruiser, 20 (3):22,2006.
5. http://en.wikipedia.org/wiki/gorakhpur.
6. Singh P and Thakur IS (2004) Removal of colour and detoxification of pulp and paper mill
effluent by microorganisms in two step bioreactor. J. Scient. Ind. Res. 63(11) : 944-948.
7. Upadhaya AR (2004) Aquatic plant for wastewater treatment, Daya Publication house, Delhi-
110035.
8. Wajih S, Singh B K, Tripathi S, Bartarya S, Srivastava A, Singh A K and Goyal S (2009)
Vulnerability analysis of Gorakhpur, under the asian cities climate change resilience network
(ACCCRN) process and sponsored by Rockfeller foundation. G.E.A.G. Gorakhpur.
162
Health Issues in Old Women: Reasons and Suggestion for Improvement

Meenakshi
Department of Home Science Mewar University
Gangrar,Chittorgarh (Rajasthan)
Email Id : meenakshis581@gmail.com
An older woman refers to women age 50 and older. Ageing women refers to the same chronological group but
emphasizes that ageing is a process that occurs at very different rates among various individuals and groups.
Privileged women may remain free of the health concerns that often accompany ageing until well into their 70s
and 80s. Others who endure a lifetime of poverty, malnutrition and heavy labour may be chronologically young
but functionally old at age 40. Decision-makers need to consider the contextual differences in how the process
of ageing is experienced in their specific environment, when designing gender-responsive policies and
programmes for ageing women.
Ageing is also both a biological and social construct. Physiological changes such as a reduction in bone density
and visual acuity are a normal part of the ageing process. At the same time, socioeconomic factors such as living
arrangements, income and access to health care greatly affect how individuals and populations experience
ageing. Ageing may also constitute a continuum of independence, dependence and interdependence that ranges
from older women who are essentially independent and coping well with daily life, to those who require some
assistance in their day-to-day lives, to those who are dependent on others for support and care. These groups are
heterogeneous, reflecting diverse values, health status, educational levels and socioeconomic status.
Reasons that increase womens vulnerability to poor health in older age:

Gender discrimination against girls' child leading to inequitable access to food and care by female and
male infants and children.
Restrictions on education at all levels.
Childbirth without adequate health care and support.
Low incomes and inequitable access to decent work due to gender-discrimination in the labour force.
Care giving responsibilities associated with mothering, grand mothering and looking after ones spouse
and older parents that prevent or restrict working for an income and access to an employee-based
pension.
Domestic violence, which may begin in childhood, continues in marriage and is a common form of
elder abuse.
Widowhood, which commonly leads to a loss of income and may lead to social isolation.
Cultural traditions and attitudes that limit access to health care in older age, for example older women
are much less likely than older men to receive cataract surgery in many countries.
To improve this situation we need to practices that:

support reproductive health and safe motherhood programmes;
support girls access to education with a special effort to enable their transition from primary to
secondary and to postsecondary schooling;
163
enable equitable entry to the labour market and to meaningful, protected work;
provide incentives for 'family friendly' policies in the workplace which support pregnancy,
breastfeeding, and caring for children and older family members;
support caregivers of family members who are ill or frail, and ease the financial burden and
employment opportunity costs of this essential role;
support changes in work practice that enable older women to remain in both the formal and informal
labour markets;
support voluntary and gradual retirement as well as incentives to save for retirement and long-term care
needs;
ensure that equal rights to the inheritance of property and resources upon the death of a parent or
spouse are upheld;
ensure the right to health and equal access to health care;
ensure that all older women have an income that satisfies the basic necessities of life, as well as equal
access to required health, social, and legal services;
provide additional support to widows as required, to older women who live alone, to those who are
poor or disabled, and to those who require long-term care in or outside of the family residence; and
support compassionate end-of-life care and help with arrangements for a peaceful death and
appropriate burial required.
Culturally appropriate and gender-responsive guidelines for healthy eating and physical activity, and which are
specific to older people should be developed and taught in the community. Ultimately each country must decide
on the best mix of policies and practices in taxation, income security, health care and social services that are
needed to maintain the economic wellbeing and health of older women. The most urgent need is to ensure that
all older women have access to the basic necessities of life, including food, clean water, shelter, primary health
care and social support. To assist older women who live in rural areas and in urban slums will help to improve
the health status of old women. Governments, employers and civil society must ensure that widows and
divorced women are not left destitute and excluded by enacting and enforcing laws that prohibit gender
discrimination in inheritance practices, access to property, pensions and resources, and cultural practices that
harm women whose husbands die or divorce them.
In very old age (80-plus) women far outnumber men in the same age category. It is prudent therefore to
encourage women to prepare financially for old age (and in many cases to live alone) and to promote a mix of
public and private sources of income in old age. Financial aid and social support should be provided for families
who care for older women and men who are unable to live independently.
Reference:
The World Health Report 2002. Reducing risks, promoting healthy life. Geneva, World Health
Organization, 2002
.Women, Ageing and Health: A Framework for Action, United Nations Population Fund (UNFPA)
Population and Development Branch, WHO
164
Morphological Studies of Certain Plant Parasitic Nematodes Associated with Vegetable

Crops of District Bulnadshahr
Om Datta
Department of Zoology, M. S. College, Saharanpur
ABSTRACT
The present investigation was based on the occurrence of various plant parasitic nematodes infesting some
vegetables such as brinjal (Solanum melongena), onion (Allium cepa), and okra (Abelmoschus esculentus) of
district Bulandshahr. The nematodes were isolated from the collected soil samples and extracted by Cobb (1918)
decanting and sieving method. The findings of the survey revealed that the vegetable crops were infested with
various ectoparasitic nematodes identified as, Hoplolaimus proporicus, Hoplolaimus indicus, Pratylenchus
pratensis and Xiphinema americanum. The Hoplolaimus proporicus was first time reported from India
associated with brinjal plants (Solanum melongena). H. indicus is widely distributed species in India found
associated with okra plants (Abelmoschus esculentus). Pratylenchus pratensis isolated from onion field soil. It is
a migratory parasite, attack only on young roots and then migrate into the soil. The dominant parasite Xiphinema
americanum is cosmopolitan in distribution and infests varieties of vegetable crops. The further investigation is
required regarding Hoplolaimus proporicus because of its first time occurrence in India.
Key words: Plant parasitic nematode, H. proporicus, H. indicus, Pratylenchus pratensis, Xiphinema
americanum
INTRODUCTION
The nematodes are round, thread like microscopic worms. Their body is elongated, non-segmented, cylindrical
and tapering at both the ends especially towards the tail. The females may swell to become spherical. The size of
plant nematode ranges from 0.2mm to 10mm & commonly 0.5 to 1.5 mm range. Nematodes are tube within a
tube. The outside tube is the body wall, which consists of muscle layers that are used as a protective covering.
The inside tube is the digestive system. Based on their feeding habit nematodes are free living, predators, fungal
feeder, algal feeder, bacterial feeder, animal parasitic and plant parasitic. All the plant parasitic nematodes are
equipped with a stylet, which helps to receive the nutrition from the plant cell. Plant-parasitic nematodes are
devastating pathogens that infect most cultivated plant species and cause considerable loss of food and fiber
crops in terms of both yield and quality. The total annual yield losses caused by plant parasitic nematodes are
estimated to be >US$125 billion worldwide (Chitwood, 2003).
The importance of taxonomy is to recognize the nematode groups that are species specific but
underrepresented in existing taxonomic collections i.e. Dorylaimida, Tylenchida and Aphelenchida. The study
of morphological features or characters in range of closely related species has importance in identification. It
includes the study of each character within that group. The concept of phenetic species has been employed in
nematode taxonomy for diagnosing a taxon. The present study also deals with the morphological characters of
plant parasitic nematodes infecting some mainly growing vegetables (onion, brinjal and okra) of two locations
(Beehra and Wairafirozpur) of district Bulandshahr.
165
MATERIALS AND METHODS
Total 5o soil samples were collected from the rhizosperic zone of vegetable crops. Plant parasitic nematodes
were extracted from the soil samples by decanting and sieving technique (Cobb, 1918) and preserved for
morphological studies. The nematodes were killed with hot fixative (FAG) to prevent twisting and contraction
of tissues, and remained in the fixative for 48 hours (De Grisse, 1965). After fixing the nematodes, were
dehydrated for 20 to 24 days in dehydration solution and permanent mounting was done in lactophenol and
sealed with wax.
Drawing of permanent mounts of collected nematodes was done with the help of camera lucida under
10x10X and 10x40X magnification. Measurements of different body parameters of nematodes were done with
the help of calibrated stage micrometer. The measured body parameters are as total body length, body width,
stylet length, and esophageal length, position of vulva in females from the anterior and posterior end, tail length,
spicule length and length of the gubernaculums. Morphological parameters were calculated on following keys.
n = No. of species, l =Total body length, a = Body length/greatest body width, b = Body length / distance from
anterior end to oesophagus, b = Body length /distance from anterior end to posterior end of the oesophagus
gland. c = Body length / tail length, c= Tail length/body width, l = Distance from anterior end to anus, V =
Vulval distance from anterior region x 100/ body length, and V = Distance from anterior end to vulva.
Genus: Hoplolaimus (Daday, 1905)
Generic Diagnosis: Annules extending completely around the body and phasmids located erratically. Lip
region divided into six longitudinal striae. Amphids apertures minute slit like and located at edge of lateral lips.
Lip region offset from body wide anteriorly flattened, stylet massive and tulip shaped. Esophageal gland overlap
intestine dorsally and laterally. Spicule massive, somewhat cylindroids, bursa enveloping tail.
Type species: Hoplolaimus proporicus:
Excretory pore located nearly opposite anterior end of median Oesophageal bulb; hence the specific name
proporicus. Hemizonids posterior to latitude of excretory pore.
Specific Characters: Male reproductive system with thickened and transverse gubernaculums.
Observed Specimen: Male 3, Female- Not Found (Fig. 1)
Morphological Characters of Male:
Measurements: l = 1.085 mm, a = 36.16, b = 14.00, b = 14.09, c = 43.4, c = 0.83, Spicule = 15m, Stylet =
33m.
Discussion: Morphological characters of the studied specimen were first described by J.B. Goodey. The present
specimen compared with the Indian species H. abelmoschi; H cephalus (Mulk and Jairajpuri, 1976); H. indicus
(Sher, 1963); H. tylenchiformis (Daday, 1905); this species mainly found in South Africa and later on reported
166
from Nigeria. It infects lateral roots of oil palm trees by making lesions. It is the first record of H. proporicus
reported from India isolated from the rhizospheric soil of brinjal plants. The presence of this parasite indicates
the host - interaction with brinjal plants. Further study is required to know about the occurrence of this species
associated with vegetable crops in India.
Type species: Hoplolaimus indicus (Sher, 1963)
Observed Specimen: Male 3 (Fig. 2)
Morphological Characters of Male:
Measurements: l = 1.189 mm, a = 30.48, b = 16.06, b = 16.06, c = 39.63, c = 0.76, Stylet = 134m, Spicule
= 22m.
Cuticle coarsely annulated and lip region hemispherical marked by 3-4 annules with longitudinal striation. Basal
nobe of stylet is tulip shaped. Excretory pore present anterior to Esophageal intestinal junction. Spermatheca
filled with sperms and spicule arcuate, cephalated.
Specific Characters of Specimen: Tail pointed and gubernaculum present.
Morphological Characters of Female: (Fig. 3)
Measurements: l = 1.213 mm, a = 27.56, b = 13.04, b = 13.04, c = 0.26, c = 102.72, v = 58.53, Stylet length
= 41m
The body length of the female is within the range (0.99 1.24 mm) that is 1.213-mm. and body width 44m.
within the range of (33 - 48m) diameter. Lip region widely flattened having 2 3 teeth like projections. Stylet
muscular and overlapping the intestine dorsally. Ovaries paired out stretched and equally developed. Vulva
situated 54 60% from the anterior end of the body. Tail short and rounded having closely marked annules.
Specific Characters of Specimen: Ovaries paired, out stretched and equally developed.
Discussion: The studied specimen was compared with H. tylenchifromis (Daday 1905), H. sheri (Suryawanshi,
1971); H. sheshadri (Mulk and Jairajpuri, 1976); H. chambus (Jairajpuri and Baqri, 1973); H. indicus (Sher,
1963); H. citri (Handoo and Golden, 1992). The measurement of the body and other characters are very close to
the H. indicus (Sher, 1963). The minor variation in the measurement may be due to the soil variation and food
availability. H. indicus is most widely distributed species in India associated with various crops. The
investigations required further more experiments to isolate other strains of the species on physiochemical,
biochemical and molecular levels.
Genus Pratylenchus (Filipjev, 1934)
Generic Diagnosis: Generic character of the genus Pratylenchus described by Thorne (1949). Lip region
include the number of annules. Head relatively broad and tail bluntly rounded. Esophageal bulb spheroid, more
than half as wide as neck. Basal bulb extending back over intestine. Esophagus and intestine joined by muscular
wall. Excretory pore prominent intestine contain numerous dark granules.
167
Type species: Pratylenchus pratensis

Steiner (1927) reviewed the works of Cobb (1917) and Rensch (1924) and described that Aphelenchus neglectus
and Tylenchus penetrans were synonyms of Pratylenchus pratensis. First of all Steiner showed the photographs
of P. pratensis within the roots of lily. It was the first record of Pratylenchus inhabiting and depositing eggs in
root tissues so it was established that P. pratensis is a root lesion nematode.
Specific Characters of Specimen: Anterior ovary outstretched and posterior branched, rudimentary.
Observed Specimen: Female 3, Male - Not Found. (Fig. 4)
Morphological Characters of Female:
Measurements: l = 5.50 mm, a = 275, b = 9.16, b = 9.16, c = 3.05, c = 9, v = 69.09, I = 50, v = 760, Stylet
length = 21m
In the present female specimen vulva is depressed and slit like. Anterior ovary outstretched, filled with oocytes,
posterior uterus branch rudimentary. Muscular rectum ending in a transverse slit like anus.
Discussion: Pratylenchus pratensis was first described by DeMan. He included a full length drawing of a
female and illustrated certain diagnostic characters to identify the group of this species. The studied specimen
was compared with Pratylenchoides crenicauda, P. coffeae (Filipjev and Stokhoven, 1941); P. zea (Graham,
1951); P. thorne (Thorne, et al., 1953); P. penetrans (Filipjev and Stokhoven, 1941).
All the measurements of specimen are very close to P. pratensis but male specimen was not observed therefore
further study is required to add the detail description of the species.
Genus Xiphinema (Cobb, 1913 a)
Generic Diagnosis: Spear with long extension and basal flanges. At the base of spear guiding ring present.
Esophagus slender, coiled, only straight in extruded form. Intestinal cell packed with refractive granules. Vulva
transverse, ovary double and reflexed.
Type Species Xiphinema americanum (Cobb, 1913 a)
Presence of spear and guiding ring in pharynx well developed. Flanges of spears well developed.
Specific Characters of Specimen: Female reproductive system amphidelphic. Tail short and conoid.
Observed Specimen: Male- Not Found, Female 3 (Fig. 5)
Morphological Characters of Female:

Measurements: l = 1.229 mm, a = 27.93, b = 7.31, b = 7.31, c = 8.19, c = 3.40,v = 58.17, Stylet = 134m
Spicule = 22m
Discussion: The present specimen was compared with Xiphinema index (Thorne and Allen, 1950); X. citri,
(Siddiqi, 1959); X. diversicaudatum (Thorne, 1939); X. americanum (Cobb, 1913a). Morphological parameters
168
of specimen closely related to X. americanum. Siddiqi (1959) reported the presence of X. americanum and X.
brevicaudatum in India and other 3 new species of Xiphinema. It is a cosmopolitan species commonly found in
India, Europe, Israel and Ceylon. To study the interaction with vegetable plants further study is required because
Xiphinema is a deep soil inhabiting nematode and it has observed that small population found associated with
beans, tomatoes and potatoes.
REFERENCES
Chitwood D. J. 2003. Research on plant-parasitic nematode biology conducted by the United States Department
of Agriculture-Agricultural Research Service. Pest Management Science 59, 748753.
Cobb, N. A.1913a. New nematode genera found inhabiting fresh water and non brakish. Jour.Wash.Acad.Sci. 3
(16): 432-444.
Cobb, N. A.1917. A new parasitic nema found infesting cotton and potatoes. J. Agr. Res., 11 (1) : 27-33.
Cobb, N. A.1918. Estimating the nema population of soil. U.S.D.A. Bur. Plant Ind. Agr. Tech.Cir., 1:1-48.
Daday, E.V., 1905. Untersuchungen Uber die Susswasser, Microfauna. Paraguaya Zoologica Stuttgrat (1844)
374pp.
De Grisse. A., 1965. A labour saving method for fixing transferring eelworms to anhydrous glycerin. Rijsk
Facculteit derland bourweton. Leer stoal voor Dierounde, coupure link. 235 gent.
Filipjev,I.N., 1934. The classification of the three living nematodes and their relation to the parasitic nematodes.
Smithsonian Misc Coll. (3216) 89 (6): 1-63
Filipjev,I.N. and Schuurmans-Stekhoven, J.H., 1941. A manual of Agricultural Helminthology. E. J. Brill,

Leiden, 878pp.
Graham, T.W., 1951. Nematode rot rot of tobacco and other plants. S.C. Agricultural Experimental Station
Bulletin, 390.
Handoo , Z. A. and Golden, A. M. 1992. A. key and diagnostic compendium to species of the genus
Hoplolaimus daday, 1905 ( Nematoda: Hoplolamidae) J. Nematol., 24:45 53.
Jairajpuri, M.S. and Baqri, Q.H., 1973. Nematodes of high altitudes in India. Four new species of Tylenchida.
Nematologica, 19:19-30.
Mulk , M. M. and Jaiirajpuri, M. S. 1976. Nematodes of leguminous crops in India III. Three new species of
Hoplolaimus Daday 1905 ( Hoplolaimidae) Indian J. Nematol., 5:1 8.
Rensch, M., 1924. Eine neue Methods zur Bekampfung der Rubennematode. Mitte. deut. Landwirts. Ges.,
38:412-414.
169
Suryawanshi, M. V., 1971. Studies on Tylenchida (Nematoda) from Marathwada, India, with
description of four new species. Nematologica, 17:399-406.
Sher, S. A. 1963. Revision of Hoplolaimus Daday, 1905 and Aorolaimus n.gen. Nematologica, 9: 267 295.
Siddiqi, M. R. 1959. Basiria graminophila n.g., n. sp. (Nematoda: Tylenchidae) found associated with grass
roots in Aligarh, India. Nematologica, 4:217-222.
Steiner, G., 1927. Tylenchus pratensis and various other nemas attacking plants. J. Agr. Res., 35 (11): 961-981.
Thorne, G. 1939. A monograph of the nematodes of the super family Dorylaimoida. Capita. Zool. 8 (5):1-190.
Thorne, G. 1949. On the classification of the Tylenchida new order (Nematoda: Phasmidia) Proc. Helminthes.
Soc. Wash., 16: 37 73.
Thorne, G. and Allen, M. W. 1950. Pratylenchus hamatus n. sp. and Xiphinema index n. sp. Two nematodes
associated with fig roots, with a note on Pratylenchus anceps Cobb. Proc. Helminth. Soc. Wash., 17
(1): 27-35.
170
Fig. 1 (A) Hoplolaimus proporicus anterior region (B) Hoplolaimus proporicus posterior region; Fig. 2 (A)
Hoplolaimus indicus anterior region (B) Hoplolaimus indicus posterior region, Fig. 3 (A) Hoplolaimus
indicus anterior region (B) Hoplolaimus indicus vulval region (C) Hoplolaimus indicus posterior region;
Fig. 4 (A) Pratylenchus pratensis anterior region (B) Pratylenchus pratensis posterior region; Fig. 5 (A)
Xiphinema americanum anterior region (B) Xiphinema americanum Vulval region (C) Xiphinema
americanum posterior region
171
Genetics of Hg++-Tolerance in Barley
Ritu Aggarwal
Department of Botany, M. S. College, Saharanpur
ABSTRACT
Determining the features of the mechanism and genetics of tolerance is an important aspect of research related
to heavy metals. The present work deals with the amount of genetic variability and the mode of inheritance for
tolerance to Hg++ in barley. Tolerant, partially tolerant and non-tolerant accessions of barley were selected after
screening about 150 accessions, against 10-4M mercuric chloride treatments given to seedlings for seven days.
The amount of tolerance present in an accession was estimated using values for response coefficient (RC) for
radicle length of seven days old seedlings. Crosses were made between tolerant and partially/non-tolerant
accessions to produce 25 hybrids. These hybrids and their segregants were analyzed for understanding the
genetics of Hg++ -tolerance. The analyses revealed that Hg++-tolerance in barley is recessive and is under
polygenic control.
Key Words: Genetics, Hg++, tolerance, Barley

Introduction
A global misery of heavy metal pollution has been coerced on biological world. This led to fascination for
exploring, in the last few decades, relationship between hazardous heavy metal ions and biological processes.
One important aspect of this research is to determine features of the mechanism and genetics of heavy metal
tolerance. This field of research, dealing with heavy metal-tolerance in higher plants, has multi-faceted scopes
like biochemical understanding of metal ion binding with bio-molecules, role of metal ions in gene regulation,
scrutiny of population and species differences in metal ion tolerance, development of metal-tolerant cultivars for
being grown in heavy metal polluted areas, production of metal hyper-accumulating plants for phytoremediation
of soils and waters, etc. It is indisputable that data are meager in this area of research and therefore metal-
tolerance becomes a consequential and advantageous subject for research for salvation of our planet from heavy
metal pollution. The present work aimed towards analyzing the amount of genetic variability and the mode of
inheritance for tolerance to heavy metal mercury.

Mercuric chloride (MC; HgCl2) was used as source for Hg++. Molar concentration, 10-4M was selected for
treating the seedlings of barley (Hordeum vulgare) on the basis of previous experiments conducted in our
laboratory by estimating the toxic and tolerance limits for Hg using several crops as test systems. Treatment was
given to seeds in sterilized petriplates lined with cotton pads, sandwiched between filter papers. Control sets
were raised in Hoaglands solution whereas the treated sets were raised in HgCl 2 solutions prepared in
Hoaglands solution. Both types of sets were raised in Caltons Seed Germinator in total darkness at 22oC up
to seven days. These seedlings were used for radicle length.
172
The amount of tolerance present in an accession was estimated using values for response coefficient
(RC) for radicle length of seven days old seedlings. The negative values for RCs indicated inhibition while
positive values indicated stimulation. Response coefficient (RC) was calculated, using the following formula.
VT VC
RC = -------------
VC
(VT = value of treated set; VC = value of control set).
On the basis of mean RC values, accessions were categorized into five category AE (A = > 0.20; B =
0.20 to 0.39; C = 0.40 to 0.59; D = - 0.6 to 0.79; E = < 0.8). Category A comprised of tolerant (T),
category E of non-tolerant (NT) and categories B-D of partially tolerant (PT) accessions. Ten accessions of
barley (Table 1) comprising five tolerant (T), three non-tolerant (NT) and two partially tolerant (PT), were
selected (after screening more than 150 accessions for the presence of Hg-tolerance) for producing twenty five
hybrids for investigating genetics of Hg-tolerance (Table 2). RCs in T, PT and NT accessions were >-0.20, <-
0.20 to 0.80 and <-0.80, respectively.
For quantifying the variability in the range of tolerance (as phenotype), the treated seedlings were
classified into seventeen classes by splitting the range (-0.85 to -0.01) of RCs using a class interval of -0.05 and
class values were accordingly calculated. The total variation present between the distinct samples of parents,
hybrids (F1s) and their segregants (F2s and F3s) was analyzed using ANOVA.
Table 1. List of tolerant, partially tolerant and non-tolerant accessions of barley.
Tolerant Partially tolerant Non-tolerant

T-1 HBL-231 PT-1 NT-1 K-200
K-204
T-2 K-140 PT-2 K-229 NT-2 K-201
T-3 Lucknow-3 NT-3 K-228
T-4 Q. 27-1
T-5 Ratna
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Table 2. List of crosses made for studying genetics of Hg++ -tolerance.
S. Code S. Code
No. No.
Parent 1 Parent 2 Hybrid Parent 1 Parent 2 Hybrid
1. T-1 NT-1 H-1 14. T-3 NT-3 H-14
2. T-1 NT-2 H-2 15. T-3 PT-2 H-15
3. T-1 PT-1 H-3 16. T-4 NT-1 H-16
4. T-1 NT-3 H-4 17. T-4 NT-2 H-17
5. T-1 PT-2 H-5 18. T-4 PT-1 H-18
6. T-2 NT-1 H-6 19. T-4 NT-3 H-19
7. T-2 NT-2 H-7 20. T-4 PT-2 H-20
8. T-2 PT-1 H-8 21. T-5 NT-1 H-21
9. T-2 NT-3 H-9 22. T-5 NT-2 H-22
10. T-2 PT-2 H-10 23. T-5 PT-1 H-23
11. T-3 NT-1 H-11 24. T-5 NT-3 H-24
12. T-3 NT-2 H-12 25. T-5 PT-2 H-25
13. T-3 PT-1 H-13
Results
The overall frequency (%) distributions of the seedlings, belonging to seventeen different classes in parents,
tolerant x partially tolerant, tolerant x non-tolerant and partially tolerant x non-tolerant hybrids and their
174
segregants are graphically represented in Figure 1. Classes falling towards the extreme left side of the abscissae
depict non-tolerance while those falling towards the extreme right represent tolerance. The classes occupying
the middle zone characterise partial tolerance. Data related to ANOVA and F test are given in Table 3.
Table 3. ANOVA for genetic variability in barley parents, F 1, F2 and F3 for MC tolerance.
Source of Variation SS Df MS
F
115.86 9 12.87 762.07**
Parents
Error 15.05 891 0.02
65.55 4 16.39 496.05**

Tolerant parents
Error 13.08 396 0.03
12.35 4 3.09 5995.18**

Non-tolerant parents
Error 0.20 396 0.00
37.96 1 37.96 1731.82**

T vs. NT
Error 2.17 99 0.02
F 1+F 2+F 3 190.23 74 2.57 103.17**
Error 182.55 7326 0.02
F1 140.25 24 5.84 489.53**
Error 28.36 2376 0.01
F2 10.73 24 0.45 9.98**
Error 106.38 2376 0.04
F3 19.27 24 0.80 59.03**
Error 32.31 2376 0.01
F 1 vs. F 2 vs. F 3 19.98 2 9.99 496.05**
Error 15.49 297 0.05
P vs. F 1 vs. F 2 vs. F 3 126.40 3 42.13 628.65**
Error (interaction) 19.91 297 0.07
Total (Parents+ F1+ F2+ F3) 309.53 84 3.68 151.69**
175
Error 202.01 8316 0.02
The frequencies of the seedlings of the non-tolerant parents were highest in classes located towards the
extreme left side of the abscissae. In case of tolerant parents, the seedling frequencies of the classes present on
extreme right side were highest. In partially tolerant parents, the classes with higher seedling frequencies
occupied intermediate positions on the abscissae. Classes occupying extreme left sides of the abscissae had the
highest frequencies of the seedlings in all F 1s. This pattern indicated that non-tolerance for Hg++ was dominant.
The frequency distributions of the seedlings in various classes of F 2s and F3s exhibited continuous variation
occupying the middle left zone of the abscissae. This pattern of frequency distribution suggested polygenic
inheritance for Hg++-tolerance.
DISCUSSION
Metal tolerant plant species were initially reported from soils with elevated levels of one or more metal
ions. These plant species belonged to wild species. However, Prat [1] was probably the first to discover that a
population of Melandrium silvestre from a copper mine grew better on artificially contaminated soil than did
plants derived from uncontaminated soil and suggested that mine populations have evolved tolerant races.
Various aspects of metal tolerance in higher plants were reviewed time to time by several workers [2-23]. The
current screening of the accessions of barley for Hg-tolerance revealed the presence of tolerance conferring
genes in many of them. The present set of data, related to the distribution pattern of RCs for radicle lengths in
parents and F1 hybrids against a concentration of 10-4M HgCl2, clearly indicated that Hg-tolerance was recessive
over non-tolerance. In contrast to present observations certain workers have demonstrated dominance for the
tolerance for some other heavy metals. For instance, Allen and Sheppard [24] demonstrated copper tolerance to
be completely dominant in Mimulus guttatus but, Macnair [13] disagreed with Allen and Sheppard and
demonstrated that Cu-tolerance in Mimulus guttatus is not completely dominant. Paull et al. [25] observed Cu-
tolerance to be dominant in wheat. Meharg and Macnair [26,27] and Macnair et al. [28] reported dominance for
arsenic-tolerance in grasses. Roy and Prasad [29] reported that Cd ++ tolerance in a wheat variety, HUW 234,
was imparted by single dominant gene.
The number of genes conferring tolerance is often assumed to be high in view of the lack of discontinuous
variation in tolerance levels in the natural populations and lack of clear-cut segregation patterns in progeny of
hybrids. During the present study, the same type of distribution pattern for RCs (a modification of Tolerance
Index, TI, used by other workers for measuring tolerance) was reported in F 2 and F3 segregating populations,
disclosing the presence of polymeric inheritance for Hg++-tolerance. Using TI, continuous distribution and
consequently polygenic inheritance has been demonstrated for Zn tolerance in Silene vulgaris [30],
Anthoxanthum odoratum [31], Agrostis capillaris [32], Pb tolerance in Festuca ovina [33] and Al tolerance in
Zea mays [34]. These observations led early reviewers of metal tolerance [3,35] to argue that the tolerance was
an adaptation generally produced by polygenic inheritance. Moreover, several studies also indicated the
presence of relatively simple genetic system for tolerance. There are a number of cases where major genes have
been decidedly evidenced for the tolerance [36-46,25,28].
176
ACKNOWLEDGEMENTS
The author is grateful to DST for providing financial support and to NBPGR, New Delhi and Directorate of
Wheat Research, Karnal for providing seeds of barley accessions.
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179
Stem Cell Therapy
Pragati
Department of Botany, M.S. College Saharanpur
Stem cell therapy is the use of stem cells to treat or prevent a disease or condition. One of the most popular
clinical studies being researched these days is stem cell transplantation. Until recently, moral issues of states and
countries haven't allowed research to expound deeply into the unknowns. Within the last ten years though,
scientists have made leaps and bounds in finding out concrete facts that this stem cell research has supplied.
Tommy G. Thompson, Secretary of Health Services states, "I believe it will open up a world of opportunity for
scientists, not only at the NIH, but elsewhere, because it demonstrates a cooperative atmosphere among
academia, the private sector, and government that will allow us to move ahead" . New ways of conducting stem
cell research have made the healing and repairing treatment for many diverse applications. For over 30 years,
bone-marrow has been used to treat cancer patients with conditions such as leukaemia and lymphoma; this is the
only form of stem cell therapy that is widely practiced. During chemotherapy, most growing cells are killed by
the cytotoxic agents. These agents, however, cannot discriminate between the leukaemia or neoplastic cells, and
the hematopoietic ste cellswithin the bone marrow. It is this side effect of conventional chemotherapy strategies
that the stem cell transplant attempts to reverse; a donor's healthy bone marrow reintroduces functional stem
cells to replace the cells lost in the host's body during treatment. The transplanted cells also generate an immune
response that helps to kill off the cancer cells; this process can go too far, however, leading to graft vs host
disease the most serious side effect of this treatment. Alexander Smikodub, a doctor of Medical Science at the
National Medical University states, "cells that we use are not considered by the immune system of the recipient
as foreign, therefore, they can survive, multiply, and develop full function in the body of a new host"
Smikodub). These cells can then survive and multiply, capable of lasting for months and years in the body of the
recipient. In the areas where tissue or organs have been damaged or lost, they substitute the lack of functional
activities. These cells can also produce new generations of cells that are needed by the patient. When
strategically placed, they can support, restore and replace the functions of their specialization in the body. These
methods have never received such success with amazing reparative and rejuvenating actions and effects.
Scientists working with this field for the last decade have explained the steps observed from their view in detail.
After the transplantation occurs, there appear positive changes in emotional and psychological sphere of the
patient. In this period, the disappearance of depression, improvement in mental creativity and an inner feeling of
strength are a few of the results immediately noticeable. In all of these changes occurring, there is a substantial
reinforcement of the will and spirit of the person. With some diseases, these changes are very drastic and very
important due to the grim and psychological background of the case. Mental capacity also increases quite soon,
creating better concentration and ability to maintain attention, thinking, speech and memory. Some of the
neurotic manifestations of the patient tend to disappear beginning with irritability, easy fatigue and apathy. At
this time sleep improves and insomnia disappears with sleep becoming refreshing and recuperative.
While their sleep is improving, patients also find that their physical activity likewise increases. Desiring to
move, they persistently increase their physical load of bodily functions and workout. In this process, overweight
people tend to become more in shape and skinny people increase weight. Overall, their appearance gains
freshness and rejuvenation of presence. Some of the biggest changes taking place at this time are movements
180
that the patient could previously not accomplish at any level. The coordination of movement being restored, the
nerve system is directly affected for the better. These changes that are occurring can happen quickly of over an
extended period of time depending on the patients' situation.
On the interior of the patient, recovery commences at a steady pace beginning with weakened or damaged inner
organs or tissues. Active and rapid recovery of the composition of peripheral blood as well as the cell
composition of bone marrow- if it was damaged. Chemotherapy and radiotherapy patients see more increased
recovery than any others do. In the process, more efficient blood circulation occurs, granting nutrition and
function of organs, tissues, skin, etc. Activity of kidneys and liver rise drastically.
What scientists have found is that cell therapy does not create a butterfly from a caterpillar- it undertakes and
fulfills realistic tasks: restoring lost functions, reducing deterioration, and compromising aging. In all the steps
of cell therapy described by the scientists at the National Medical University, no negative side effects were ever
observed in the decade of work performed. This is an incredible realization and important fact in the scientific
world.
The interesting part of stem cell transplantation is that the treatment is mainly not different from any other
pharmaceutical preparations. The idea in the future is to have procedures such as the one's described above to be
common and a normal part of disease prevention. Alexander Smikodub, clearly stated his belief just a few
weeks ago for future stem cell therapy. "Cell therapy combines the advantages of a new branch of
Transplantology with the modern achievements of Therapy and the simplicity of a normal pharmaceutical
preparations administration". These changes that are occurring can happen quickly or over an extended period of
time depending on the patients' situation. The overall time for a patient's treatment usually takes from two to
five days. After the treatment though, the patients' body and stem cells are what determines the outcome. "With
some diseases, engraftment probably takes place for a lifetime period. We have results of several ongoing
observations where engraftment duration exceeds 5,7, and 9 years".
In their studies conducted at the National Medical University, these scientists (along with many others) did not
observe any of their effects with the aide of bioengineering. They used only natural tissues. They also didn't see
any negative effects as Alexander Smikodub stated. "We observe no negative side effects despite limited list of
contraindications". In the future, these scientists and many others see stem cell transplantation as a routine
medicinal preparation as any other use of regular preparations. Their feeling is that they have come a great
distance from where they began but they still have a long path ahead. Soon, they predict doctors will be able to
administer the dosage in pill form with no need of grafting. With technology increasing every day, and the
exposure that this medicinal "miracle" is receiving, they feel it won't be long until many more amazing
discoveries are found in this unique and fascinating field of science.
The use of stem cells for the treatment of liver disease in both humans and animals has been the focus of
considerable interest. The liver has some natural regenerative properties, but is often insufficient to deal with the
extent of some liver diseases. Hepatocytes have been formed from some sources of MSC, but they have not
been applied clinically currently. There is a large effort to create stem cells differentiated along the pancreatic
line as a possible cure for diabetes, but no line has been well established.
181
References :
(1) Bone Marrow transplantation and peripheral blood stem cell transplantation, in national cancer institute fact
sheet wed site, Bethesda , MD ,and human services, 2010
(2) "A Stem-Cell-Based Drug Gets Approval in Canada". 17 May 2012
(3) "Prochymal - First Stem Cell Drug Approved". 22 May 2012.
(4) internet stem cell web
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Transgenic Plants in Agriculture

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Proceeding of the UGC Sponsored National Seminar

11 & 12 October, 2015

Dr. Vijai Malik

All rights reserved. No part of this publication may be reproduced, stored, in a

S. Title Name of the Authors Page

2 Shoving Towards a New Green Revolution Vijai Malik 9-13

8 Identification of Geminivirus in Cowpea (Vigna unguiculata) Shail Pande 46-50

11 Role of Transgenic Plants in Agriculture Renu Rani 67-71

12 Depression in Serum Zinc Concentration and Elevation in Punam Yadav 72-79

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION iii

16 Synthesis, Characterization and Biological Activities of Some Dinkar Malik 99-105

28 Stem Cell Therapy Pragati 180-182

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION iv

Role of Biotechnology and Nanotechnology to revolutionize Agriculture and Food

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION v

Green Revolution for Qualitative Traits in Plants: Biofortification of Nutrients

Rajni Shukla, Rashmi Upadhyay and Yogesh Kumar Sharma

Department of Botany, University of Lucknow, Lucknow (226 007)

* Black Revolution - Petroleum Production : 1970

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 1

* Green Revolution - Food grains : Father of Green Revolution M.S. Swaminathan.

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 2

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 3

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 4

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 5

Improving the crop yield and quality through Biofortification:

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 6

GW-366T1SEED GW-366T2SEED GW-366T3SEED

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 7

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 8

Shoving Towards a New Green Revolution

Department of Botany, M. S. College, Saharanpur

Aim of Second Green Revolution:

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 9

Calls for a Second Green Revolution

Role of Nanotechnology to Revolutionize Agriculture

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 10

Time for a Second Green Revolution

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 11

The Second Green Revolution through Biotechnology:

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 12

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 13

Biopesticide: An Ecofriendly Approach For Second Green Revolution in Agriculture

Keywords: Biopesticide, Ecofriendly, Biofungicide, Bacillus thuringensis.

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 14

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 15

Biopesticide Technology in India

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 16

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 17

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 18

Rhododendron Diversity and their Conservation in Sikkim Himalayas

Vishal Kaushik, Department of Botany, N. R. E. C. College, Khurja.

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 19

Results and Discussion:

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 20

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 21

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 22

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 23

[32] R. Almeida, S. Gonalves and A. Romano, In vitro Micropropagation of Endangered Rhododendron

S. NAME OF THE TAXA ALTITUDE STAT

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 24

30 R. niveum Hook. f. 3080-3700 ED2

'THE ROLE OF BIOLOGY IN BRINGING SECOND GREEN REVOLUTION 25

Changes in Micromorphology of Plant Sida veronicaefolia in Response to Air Pollution

382.4362 662.1813 570.1133 555.9490 545.3257

31.6548 15.3783 18.6826 21.5339** 24.1515*

293.909 180.594 191.218 223.087* 254.957

34.008 20.6632 22.6100 26.346* 30.631