PROCEEDINGS ISBN: 978-81-920945-4-0 National Seminar “Next Generation Sciences : Vision 2020 and Beyond” (NGSV) March 08, 2014 Organized by Department of Zoology Maharshi Dayanand University Rohtak 124001 Haryana (India)Proceeding of National Seminar (ISBN : 978-81-920945-4-0) “NEXT GENERATION SCIENCE : VISION 2020 AND BEYOND” (NGSV) MARCH 08, 2014 Reviewers Committee Dr. Hari Parkash Sharma, Virginia Dev Dutt Sharma, Raleigh NK Chadda, Mumbai PP Bakre, Jaipur Editorial Committee Dr. Vineeta Shukla Dr. Minakshi Sharma Dr. Sudhir K Kataria* Dr. Sudesh Rani Dr. Ranjana Jaiwal Dr. Vinay Malik a Organizer Department of Zoology, Maharshi Dayanand University, Rohtak-124001 (Haryana) INDIA ww mdurohtak.com Note : The authors as contributors in publication/s are responsible for their research work '& dispute, if any, on account of the same shall be within Rohtak jurisdiction, * Presently at RIE, AimerNATIONAL SEMINAR “Next Generation Sciences : Vision 2020 & (NGSV) MARCH 08, 2014 Patron-in-chief Er. H.S. Chalal Vice-Chancellor, MDU, Rohtak Patron Prof. S.K. Gakhar Dean (Former), Faculty of Life Sciences Convener Dr. Minakshi Sharma Head, Department of Zoology Co-Convener Dr. Vineeta Shukla Professor, Deptt. of Zoology Organizing Secretary Joint Organizing Secretaries Dr. Vinay Malik: Dr. Sudesh Rani Dr. Ranjana Jaiwal Organized by Department of Zoology Maharshi Dayanand University, Rohtak, HaryanaTABLE OF CONTENTS ‘SIN, [ TITLE & AUTHORS [Page No. Preliminary Evalvation of Wastewater Efuents from Food and 1 Beverage Industries Located in DelhL-NCR Abhinav K Srivastava, Mayur Mangik and Rachana Singh 2. | Microsatellites : An Overview with Reference to Fish Stock Identification 8 Aatag Nazir and M- Afzal Khan 3, | Hepatotoxicity of Widely used Neonicotinold insecticide Imidacloprié 9 ‘Ajay Kumar, Monika Tomar, Kavita Sharmaand Sudhie Kumar Kataria “4, | Gut Microbiota, Major Health Conconr : A Review 28 Amita Suneja Dang and Pooja Suneja | Analysis of Nitrate in Ground Water of Rohtak District, 36 Anju Rana, At Singh and J. 8. Laura “| Fluoride Contaminationin the Groundwater of Gohana Town (NCR). a (Sonepat) Haryana, India ‘Anuradha and Bhupinger Singh 7, _ | Antidepressant Activity of Various Extracts from Leaves of Murraya eo koenigi! Spreng Basan Bra, Joginder Singh Duhan, Panks| Rakha, Minakshi Prasa ‘and Anjali Singh ‘solation, Sereening and Identification of Laccases from Gut Bacteria 55 ‘Deeptt Singh, Krishna Kant Sharma, Amit Kumar. Ramnik Singh, ‘and Shen R'Jacob 9. _| indian Perspective of Patenting Biotechnology Inventions. 2 ‘Acohna Tayal 10. | Reproductive Histotoxicty of Neo-Nicotinoide insecticide Imidacloprié cy Ajay Kumar, Monika Tomar, Kavita Sharma and Sudhir Kumar Kataria 71,_ | Role of Nitric Oxide in Reproductive Biology 7 ‘Amita §.Dang and Richa Bhatnagar 42, | Antinypertipidaemic And Antiatherosclerotic Activities of Different 86 Fruit Extracts, Pectin of Lagenaria siceraria and Formulation With Terminalia arjuna Bark In Hyperlipidaemic Rats Prabhakar Kumar Verma, Aru Ohanda, Vipan Kumar Kambo] ané Suit Ranjan 48, | DESIGNER BABIES: Designed by in Vitro Fertilization (IVF) ry Drishti Sehgat 74. | Emerging Nanocomposites for Biosensor Technology : A Review 104 Minakhi Sharma oné Mamta Sharma 75, | Fish and its Positive Impact on Human Health 72 Sudesh Rani 16. | Ameliorating effect of Turmeric (Curcuma longa L.) on pathology "7 of Aeromonas hydrophila infection in Common carp (Cyprinus carpio L) Ravikant, Gahlawat SK and Sinag, R. C 77. | impact Analysis of Profenofos on the Genetic Material of Fish 128 Channa Punetatus (BLOCH) Sunita Dhatiarwal and Vinay Malik18 Migratory Birds of Family Anatidae in Ottu Lake, SIRSA Vivek Goyal, iiay Singh, Poonam Dalal and Vinay Mali 131 10. ‘New Strategy: Removal of Arsenic From Contaminated Soil and Water Gocta Dhania 136 ‘Biological Degradation of Water Hyacinth, Water Lettuce and Cattail by Vermiprocess Experiments Geetanjal Dhawan, Kavita Dhingr@ and Meena Sharma 148 7A Effect of Cd and NaCl salt on the shoot length of Solanum tuberosum ‘Govinda, Sharma Asha, Rohit Jasbir and Singh Sandoep 149 722. Ethnobotanical uses of Some Plantsfrom Jind District of Haryana Jagmahender Singh and Balkar Singh 153 23. Production of Cellulolytic Enzymes under Solidsstate Fermentation ‘AReappraisal Joginder Singh, Meenakshi Suhag, Anil Kumar 160 mA ‘A Comparative Study of Physiochemical Changes of Different Organic Waste During Vermicomposting yet, Seema Rani and Vinoeta Shukla 168 25, “Gonotoxicity Assessment Due to Pesticides by Micronuclous Assay in Humans : In vitro, A Review Kavita Sharma, Ajay Kumar, Monika Tomar and Sudhir Kumar Kataria 7 Importance of Age and Growth Studies in Fisheries Management ‘Salman Khan and M. Afzal Khan 108 27. ‘Antibody Engineering and Production of mAb for Development of New Drugs khushbu Gupta 202 28. Understanding Pollen Allergy-Need of the Hour Manisha Ahlawat and Pushpa Dahiya 210 Importance of Prabioties in Aquaculture : A Review Manju Dew, Sudesh Rani and Poonam 26 Effect of Dietary Proten Supplementation on Nutritive Value of Fish Labeo Rohita \Vieeta Shukla and Meena Kumah Sharma 220 34 ‘OTOLITHS - AN Important Tool for Fish Population Studies Kalsh Miyan and M. Afzal Khan 2a 32, Review: Single cell gel electrophoresis: A reliable biomonitoring tool in in vitro genotoxicity studies Monika Tomar, Ajay Kumar, Kavita Sharma and Sudhir K. Katala 27 33, ‘Advanced Molecular Blology As A Significant Forensic Toot Neelam Arya 7248 Copper induced Toxicity in Labeo Rohita (Hamilton) Neha Dhankhar, Monika Onankhar and Vineeta Shukla 35: InVitro Models of Traumatic B Models of Brain Injury Nai and Govind Singh Injury: AReviewin Vitro 250 ‘Survey of Wetland Birds in Rural Ponds Around Kurukshetra (Haryana) india Parmesh Kumar 265Hematological Change in Thiram Induced Diabetic Male Wistar (Albino) Rats Phool Singh Yadav and R. K.Avasth a PROBIOTICS: Good Bacteria- Are they? Pooja Suneja and Amita S.Dang 279 Plant Products as Fish Meal Replacer in Aquafeods -A Review Poonam, Sudesh Ran and Manju Dov 284 40, leolation and Sore Fung! from Haryana Poonam Sangwan, Sakshi Bidra, Vikram Mor, Viisha and Rajesh Dhankhar ing of Collulase & Amylase Producing 296 4 ‘Screening and Production of Xylanase from Various Ascomycetes Fungi Preeti Chutan, Krishna Kant Sharma, Anil Ohaka, Anu, Deepa and Vinay Malik 2. In Silico PCR-RFLP Based on GS Locus for Diflerentiation of Six Aspergillus Species Rahul Prakash, VK Mishra, S. N. Jha, and Dinesh Kumar 307 Genetic reprogramming of animals: Animal Cloning Anjali Jaiwal and Ranjona Jawal 32 Genetically Modified Crops for Developing countries: [Anjali Janwal, Darshna chaudhary and Ranjana jival 324 Environment and Health Hazards of Plastics Renu Sharma and Savita Agarwal 338 ‘ABC (ATP Binding Casette) Multidrug Transporter CORIP of Candida Albicans Prefer to be within Membrane Rafts Ritu Pastiia Deepika Kunduand Mohammad Naime 340 a “Tranemission Blocking Malarial Vaccines : Parasite and Vector Based Amita 8.Dang and Ritu Deswal 352 ‘Synthesis and Characterization of Metal Oxide Nanoparticle -A Review ‘Minakshi Sharma, Rity Hooda and Mamta Sharma 360 40, Role of Functional Foods, Medicinal Herbs and Nutraceuticals In Promotion of Healthy Lifo Rajesh Kumar, Tanu Aggarwal, Chetna Chugh and Sonia Kapoor 367 30, ‘Constructed Wetlands an alternative Technology for Wastewater ‘Treatment: Review ‘Sandeap Kumar and Sunil Kumar a7 51 Effects of Antioxidants and Plant Growth Regulators on Clonal Propagation of Sugareane (SACCHARUM OFFICINARUM L.) Sarina Panghal and §.§.Son! 300 = “Secondary Screening of Various Fungal nolates for Extracellular Amylase Production ‘Santa Sheoran, Rajesh Ohankhar, Ani Kumar and C.Lavanya ‘396 53 "To Study The Effect of Heavy Metal Toxicity on Earthworm Species Vinesta Shukla, Seema Ran and Jyot 401 Pesticide exposure and its effect on development of children ‘Shetali and Sudhir Kumar Kataria ‘10 ‘Holoptelea integrfolia(Roxb.) Planch: An Important Antidiabetic Plant Showkat Ahmad Ganie and Surender Singh Yadav a438, | Pulmonary Tuberculosis Pathophysiology and Treatment 320 ‘Shakti Goel, Nidhi, Govind Singh '57._| Allelic variation in high molecular weight gluteain subunits in ‘indian wieat landraces and thelr effect on bread making quality Sania Goel, Sapria Grewal and Nagendra Kumar Singh ‘Boernavia diffusa Linn.:An Ethnobotanical and Phytochersical Rewew 88. Yada, Surender S, Gulla, Showkat A. Ganie, TB.S. Raghay, M3. Bhandoca, Neelam ané Rajesh Kumar ‘Wetlands: A Review on its Ecosystem services, Degradation, Conservation and Management S.S. Yaday and Vinay Malik, Surender S. Gulia, Showkat A Ganie, TBS. Raghav and MS. Bhandoria 60, [Physicochemical and Pharmacological Assessment of Calcium Based 459 Ayurvedic Drug of Marine Origin : PRAVAL BHASAM ‘Suman lata and Sarina Panghal Fluoride Problem and its Health Effects: A Review 408 ‘Suni Kumar 162. | In Vitro Multiple Shoots cageneration in Salvadora oleoides Decne ‘Surender Kumar, .S. Laura and Narender Singh ‘Nutrient Managemantin Fenugreek > AReviow Surender Singh and VP. Panghal 64. [Effect of Food adetives inthe Sheltie and Quality ofthe Fresh Vegetables Sushila Dabas, H.K.Dabas and Acha Rani Effect of Eight Host Plant Extracts on Orientation Response and Parasitism of Trichogramma Japonicum Ashmead dai Pratap Singh, Sakshi Gupta, Mayur Marah and Aetna Kumar ‘Antinyperlipidaamis and Antiatherosclerotic Effect of Different Fractions ‘of Lagenaria Siceraria Stem and Laaves Jwice in Hypertipidaernic Rat ‘Prabhakar Kumar Verma, Vipan Kumar Kamboj, Anu Dhanda en Suthin Ranjan “ar 482 487 495Department of Zoology ox Reviewed Proceedings of National Maharshi Dayanand University (C2) Seminar on NGSV (2014) Rohtak = 124001 (Haryana) ey) ISBN = 978-81-920045-4.0 ‘wonwimdurohtak, com Ne In Vitro Multiple Shoots Regeneration in Sa/vadora oleoides Decne Surender Kumar’, J.S. Laura and Narender Singh? "Department of Botany, K.M. Govt. College Narwana *Department of EVS, Maharshi Dayanand University Rohtak Department of Botany, Kurukshetra University Kurukshetra "Corresponding Author : jsimdu@gmail.com Publication data Received 12.4.14 Revised received 186.14 Accepted 23.8.14 Introduction Abstract A highly efficient micropropagation protocol was developed for Salvadora oleoides using shoot tip explants obiained from 12- 15 days old aseptically grown seedlings. Multiple shoots were produced by using shoot tip explants on Murashige and Skoog's medium fortified with different concentrations of cytokinins (Kn and BAP) alone and in combination with auxins (IAA and NAA). ‘Among individual treatments of cytokinins tested, 3.0 mg 1 BAP supplemented in MS medium supported better results. Maximum shoots were induced on MS medium containing 3.0 mg 1-1 BAP in combination with 0.5 mg I-t NAA. Rooting of the plantlets was achieved on MS medium supplemented with 1.0 mg I1 NAA The plantlets showed 80% survival rate during acclimatization and hardening Key words: Shoot tip explants, Salvadora oleoldes, Cytokinins, Auxins Abbreviations: BAP-6-benzylaminopurine, Kn- 6-furfury- laminopurine, IAA- Indole-3-acetic acids, NAA - a-Naphthalene acetic acid, IBA - Indole-3-butyric acid also used in the treatment of piles, tumors, bronchitis, low fever, disease Salvadora oleoides is a multipurpose tree found in the arid tracts in the state of Punjab, Haryana, Gujarat, Rajasthan and south-western parts of Uttar Pradesh in India. It is locally known as Pilu and Jhal. This species has great ethno-medicinal value. Leaves are used in treatment of cough and enlarged spleen and are given to horses as purgative. The fruits are sweets and edible. The fruits contain glucose, fructose, and sucrose and are good source of calcium. The fruits are 476 of spleen and fed to cattles to increase their milk yield (Khan, 1996).The fruit are relished by a variety of insects, birds and rodents. Leaves and fruits are used during child birth and as antidote of snake bites. The seeds are rich in non edible oil and their fat is used in the treatment of rheumatic pains and as a base of ointment (Kirtikar and Basu, 2012). Seed oil is widely used in commercial production of cosmetics, paints, varnish and lubricants. Seed oil of S.oleoides can Next Generation Sciences : Vision 2020 and Beyondbe used as mosquito repeliants and oil of this plant also shows larvicidal activities in the form of hundred per cent mortality against mosquitoes like Culex fatigans and Anopheles stephensi (Tare and Sharma,1991). This species is drought hardy, moderately salt tolerant, sand binder, useful in restoration of fragile arid tracts, stabilization of sand dunes and afforestation of degraded lands Salvadora aleoides provides ecological services such as habitat, food and shelter for varieties of mammals, reptiles, birds and hence maintain an ecological balance (Khan, 1996). This species is decreasing very rapidly due to indiscriminate exploitation and low rate of seed set, poor seed viability and inefficiency to propagate by vegetative means. Therefore, keeping in view the economic and ecological importance of this plant species; efforts were made to develop an efficient method for rapid in vitro propagation using axenic shoot tip explants via optimization of basal media, growth regulators and followed by successful outdoor establishment of regenerated Plants. Materials and Methods All experiments were conducted in plant tissue culture laboratory, Department of Environmental Science. Maharshi Dayanand University, Rohtak Seeds were collected from’ mature Plants. The seeds were washed with detergent in running tap water to remove all traces of fruit pulp, followed by selection of healthy seeds. The seeds were then surface sterilized in 90% ethanol for 3 minutes followed by a treatment of freshly prepared 0.1% HgCl, solution for five minutes. Finally, seeds were washed 4-5 times in sterilized distilled water to remove all traces of mercuric chloride. MS Next Generation Sciences : Vision 2020 and Beyond medium was prepared with 3% sucrose and 0.8% agar-agar. The pH ot medium was adjusted 5.8 prior to adding of agar-agar. The medium was autoclaved for 20 minutes at 15 psi pressure and 121°C temperature. Seeds were inoculated in culture vessels containing half strength MS medium. Culture conditions were Maintained in a culture room at 25+ 2°C temperature under 16/8 hours (light/dark) period with 50 ymol m-2 s- 1 photon flux density provided by cool white fluorescent tubes (Philips, India) and 60% relative humidity. Similar culture conditions and compositions of MS basal medium were used for further investigation to regenerate plantlets using axenic shoot tip explants Shoot tip explants were excised from twelve to fifteen days old aseptically grown seedlings. Explants were inoculated in culture vessels containing MS medium with and without growth regulators. MS basal medium without growth regulator served as control Different cytokinins (Kn and BAP) in various concentrations were used. Individual concentrations of cytokinins which gave best results were used in combinations with auxins (IAA and NAA) to enhance the shoots multiplication and proliferation rate. In vitro regenerated shoots along with explants were transferred to fresh Medium at regular interval for maintaining the healthy growth of shoots. In vitro regenerated shoots after attaining the height of 2- 4 cm were implanted in rooting medium augmented with IBA and NAA. In vitro regenerated complete plantlets were taken out from medium and their roots were washed with the help of fine brush to remove traces of medium by dipping the roots in tap water. Then individual plantlets were transplanted in a pot having sterile soil and vermiculite (1:1). arrInitially the plantlets were kept in laboratory with high humidity and finally these were shifted to field conditions after acclimatization. Data were recorded after every 5 weeks. Each experiment was repeated thrice containing ten replicates for each hormonal treatment. The data were analyzed (mean +S.E.) for each treatment, Results and Discussion The regeneration of multiple shoots is considered as an important step in standardization of micropropagation protocol. But optimum requirement for shoot proliferation stage may differ from species to species. Production of plantiets starting from shoot tip explants taken from aseptically grown seedlings of Salvadora oleoides has been achieved in present study. Shoot formation from the shoot tip was seen in all the treatments of Kn and BAP with variable frequency of regeneration Table 1.- Effects of different concentrations of Kn in vitro raised seedings. Generally, cytokinins are required for in vitro axillary shoot induction and Proliferation but type of cytokinins and its optimal concentration varies with the species. Cytokinins are supposed to play a significant role in DNA synthesis as reported by Park (2008). During Present investigation, shoot tip explants responded morphogenetically to a growth free MS medium with less number of shoots. MS medium augmented with 3.0 mg I? BAP individually was found optimum for shoot induction and proliferation as this concentration gave superior results individually. BAP showed superior results than Kn in present investigation (Table-1). Higher concentration of Kn and BAP than 3.0 mg I" produced less number of shoots. These observations further support the well known inhibitory influence of higher concentration of cytokinins on shoot elongation and regeneration as reported by (Koroch et al. (1997) in Hedeoma multifolium. and BAP on shoot tip explants taken from ‘Media [Concentrations of | Per cont bud break] No. of shoots regens- | Shoat length (on ‘growth regulators rated per explant | (25-6) (mg ¥) GSE) ‘Controt : 40 12023 0820.04 MS¥Kn 08 70 220.43 1,020.08 10 20 2aoaa 1220.16 15. 20 250.16 1.32002 20 100 “420.24 1520.41 25 100 520.63 1,720.56 30 300 620.28 2.080.54 40 70 420.45 1,920.35, MS¥BAP os 20 220.44 1.220.24 10 0 310-18) 1.420.60) 15. 100 320.26 1,520.38 20 100 520.14 1620.32 25 100 20.62 1,820.25 3.0 100 820.36 240.34 40, 20 550.08) 2.00.04 478 Next Generation Sciences : Vision 2020 and BeyondA stimulatory effect of cytokinins (Kn and BAP) with auxins (IAA and NAA) was also evaluated. Auxins are known to promote cell division. So combinations of cytokinins with auxins may proliferate shoots most significantly as observed by Heide (1965), Skoog and Miller(1957). Optimized concentrations of Kn and BAP (3.0 mg I") were used with low concentrations. (0.25-1.0 mg I") of auxins. Combined effect of cytokinins with auxins has been demonstrated by many workers such as Sudha et al.(1998) in Hollostemma annulare, Sreekumar et al.(2000) in Hemidesmus indicus and Arya et al. (2003) in Leptadenia reticulata. Low levels of IAA and NAA (0.5 mg I") in combination with cytokinins had promotional effects ‘on shoot bud induction whereas further higher concentrations (1.0 mg I or more) were not found beneficial Among two cytokinins optimized, combined effects of BAP with IAA and NAA were better than Kn with IAA and NAA. Similarly NAA was evaluated better than IAA when used in combination with cytokinins (Kn and BAP), Since the maximum number of shoots were produced in the presence of MS + BAP (3.0 mg I") and NAA (0.5 mg I-1), this medium was designated as best shoot induction and multiplication medium (Table-2). Similar results were obtained by Ajith et al (1999) in Syzygium travancorium. According to Gasper and coumans (1987), rooting of the developed shoots is usually achieved in auxins containing medium. Root formation is an energy demanding process and thus exogenous supply of carbohydrates is required. However, this being the last stage of in vitro culture, it is important to transform the plant from Table 2 - Combined effects of cytokinins (Kn and BAP) with different concentrations of auxins (IAA and NAA) on shoot tip explants. taken fom in vitto raised seedlings. ‘Media| Concentrations of | Per cent bud break | No. of shoots regene- | Shoot tength (em) ‘growth regulators rated per explant | (#5.E.) (ma F) ese) Msexn | 300025 100 020.64 242087 saa 3005 100 720.72 2a074 3010 100 520.64 2.23062 MSeKn | 30025 100 720.87 222048 NAA 3.005 100 010.58 2.4206 rer) 709) e037 232094 svear | 3.01025, 100 820.37 2.22050 SIAR 3005 700 320.46 2420.56 3010 "100 2072 222032 separ | 3000.25 00 920.35 252018 “NAA. 30-05 700 1020.36 2.0207 30+10 700 20.60 2a2040 Next Generation Sciences : Vision 2020 and Beyond 479heterotrophic to autotrophic mode of nutrition. Thus the supply of exogenous. sugars should be reduced at this time. The rooting response differed according to different concentrations and combinations of auxins used The in vitro-regenerated shoots (2.5- 4 cm), having at least 2-4 leaves and 2-3 nodes were excised from shoot clumps and transferred to rooting medium with and without growth regulators. Presence of NAA (0.51.0, 1.5 and 2.0 mg F') in MS full strength medium facilitated better rhizogenesis as compared to similar concentrations of IBA. In present study maximum rooting (80%) was achieved on full strength MS medium containing 1.0 mg I NAA with fairly good length and number of roots (Table-3). Chand and Table-3 : Effects of different concentrations of auxins (IBA and NAA) on in vitro regenerated shoots for root induction ‘Auxins Concentrations of auxins Percent root formation No. of regenerated roots per shoot (25.E) Control MMS ful strength (mg 1) é MBA 05 7 10. : 18 40 320.40 20 30 340.82 TSeNAA os a0 42054 10 80 620.16 15 60 420.45 20 60 420.45 ‘Singh (2004) reported that NAA was very effective auxins for rhizogenesis in Pterocarpus marsupium . Intermittent callus formation at the junction of root and shoot was reported. To reduce the callogenesis, the sucrose concentration was Fig.-1-3 Multiple shoots regeneration from shoot tip explants. (1) MS+3.0 mg BAP, (2) MS+3.0 mg F*Kn+0.5 mgLINAA, (8) MS + 3.0 mg MBAP +0.5 mg I NAA, (4) Root mg NAA, formation on MS+1.0 (5) Acclimatized plant under field ‘conditions. 480 Next Generation Sciences : Vision 2020 and Beyondreduced to 20gI" from 30gh". Sucrose at reduced concentrations has also been found optimal by Das et al.(1993) for rooting in Acacia auriculiformis. During acclimatization and hardening, shoot elongated and leaves turned green and expanded. In vitro regenerated complete plants were transferred to small thermoco! cups containing sterilized soil and vermiculite (1:1) and maintained under high humidity in culture room by covering them with inverted glass beaker and polythene bags. Equal ratio of Soil and vermiculite has been used by Rao et al. (2006) in Capsicum annum. Further the plantlets were transferred to poly house followed by field conditions with eighty per cent survival rate. In conclusion, BAP individually as well as in combination with NAA can be used for rapid multiplication of true to type plants of Salvadora oleoides using shoot tip explants. The present paper describes a prime and easy to use protocol for large scale production of plants through axenic shoot tip explants of Salvadora oleoides. References Ajith A, Srinivasa RC & Balkrishna P:1999: In vitro propagation of Syzygium travancorium Ganble_an endangered Plant Cell Tissue and 56:59-63, Arya V, Sekhawat NS & Singh RP, 2003: Micropropagation of Leptadenia reticulata - a medicinal plant. In Vitro Cellular and Developmental Biology of Plant 39:180-185, Chand & Singh K, 2004: In vitro shoot regeneration from cotyledonary node explants of a multipurpose leguminous tree, Pterocarpus marsulpium Roxb. In Vitro Cellular and Developmental Biology of Plant 40:167-170. Das PK, Chakravar VI & Maity S, 1993: Plantiots formation in tissue culture from cotyledon of Acacia auriculformis A. Cunn. ex Benth. Indian J Forestry 16: 182-192. Next Generation Sciences : Vision 2020 and Beyond Gaspar T& Coumans M, 1987: Root formation, In: Coll and tissue culture in forestry Voll! Ed: JM Bong and OJ Durzan, Marlinus Nijhof Publishers Dordrecht pp 202-217. Heide OM, 1965: Interaction of temperature, auxins and kinins in the regeneration ability of Begonia leaf cuttings. Physiol Plant 18:891-920, Khan AU,1996: Appraisal of ethno-ecological incentives to promote conservation of Salvadora oleoides Decne. The case for creating a resource area, Biological Conservation 75: 187-190, Kirikar KR. & Basu BD, 2012: indian Medicinal Plants,Vol.2, Periodical Expert Book Agency . Delhi pp 1539. Koroch AR, Hector RJ & Victori SI,1997: ‘Micropropagation and acclimatization of Hedeoma multifolium. Plant Cell Tissue and Organ Culture 48:213-217. Murashige T & Skoog F, 1962: A revised medium for rapid growth and bioassays with tobacco tissue culture, Physiol Plant 15:473-497 Park SY, Kim YW, Moon HK, Murthy HN, Choi YH & Cho HM, 2008: Micropropagation of Salix psudolasiogne tram nodal explants. Plant Cell Tissue and Organ Culture 93:391-346, Rao , Pratibha GS, Parashuram YJ & Kaviraj CP, 2006: ‘High frequency plant regeneration from shoot tip explants chilli (Capsicum annum). Plant Cell Biotechnology and Molecular Biology 7163-166. ‘Skoog F & Miller CO,1957: Chemical regulation of growth and organ formation in plant tissues cultured in vitro, Symp Soc Exp Biol 11:118-131 ‘Sree Kumar S, Seeni S & Pushpagadan P, 2000: | Microproapagation of Hemicesmus indicus for cultivation and production of 2-hydroxy 4 methyle benaldehyde. Plant Cell Tissue and Organ Culture 2:211-218, Sudha GC, Krishnan PN & Pushpangadan P. 1998: In vitro propagatiuon of Holostemma annulare, a rare medicinal plant. In Vitro ‘Cellular and Developmental Biology of Plant 33:57- 63, Tare V & Sharma RN.1991: Larvicidal activity ‘of some tree oils and their common chemical constituents _ against mosquitoes. Pesticides Research Journal 3:169-172. 481