DEVELOPMENT OF STRATEGIES FOR CONSERVATION OF S.ASOCA: AN IMPORTANT GLOBALLY VULNERABLE TREE SPECIES IN

1SUMANGALA R C., 2GANESH D., 3SACHIN ROSARIO., 4VASUDEVA R., 5UMA SHAANKER R., 6RAVIKANTH G.

1,2Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Palkalai Nagar-625 021 Madurai, Tamil Nadu, India. 1,5,6Conservation Genetics Laboratory, Ashoka Trust for Research in Ecology and the Environment, Royal Enclave, Srirampura, Jakkur Post, 560 064, India. 3,5Department of Crop Physiology and School of Ecology and Conservation, University of Agricultural Sciences, Bangalore 560 065, India. 4Department of Forest Biology & Tree Improvement, College of Forestry, University of Agricultural Sciences, Sirsi, 581 401, India.

Abstract - In recent years, indiscriminate and rampant extraction of the wood to meet the ever-increasing herbal market demand has led to a sharp decline in naturally occurring populations of the species. In many pockets within the Western Ghats, the population size is critically low. The continuous exploitation of several medicinal plant species from the wild and substantial loss of their habitats during the past 15 years have resulted in the population decline of many high value medicinal plant species over the years The present study is examining the genetic diversity and population structure of this species across Western Ghats, one of the global biodiversity hot-spots. A total of 78 alleles with an average of 15.6 alleles per locus were found across the 12 populations. The allelic richness per locus ranged from 2.6 to 5.03. The results obtained from the present investigation is discussed for development of viable conservation strategies for S. asoca in the Western Ghats.

Keywords - Niche Suitability, Microsatellite Markers, Genotyping, Globally Vulnerable

I. INTRODUCTION in the country or is simply unlikely to be S. asoca. It is also found in countries such as Pakistan, Sri Lanka, Saraca asoca (Roxb.) W. J. de Wilde or Saraca Bangladesh, Burma and in Malay where it grows at indica L. is one of the most medicinal plants known an altitude of ~750 m. There is an another closely in India. It belongs to the family Caesalpiniaceae. resembling species of Saraca with a botanical name This popular tree is known by many names in Saraca declinata, known as Red Saraca present in different languages such as Kankeli (Sanskrit), Florida, Burma and Sri Lanka. Ashoka (Assamese, Bengali, Gujrati, Oriya, Hindi), S. asoca is considered to be the most important and Ashok adamara (), Ashok (Kashmiri), vital ingredients in a variety of indigenous medicine Asokam (Malayalam), Ashok (Marathi, Punjabi), systems, including Ayurvedic, Unani, and Siddha in Ashoka (Oriya) , Asogam (Tamil), Ashokapatta India. Its use in health care has been documented in a (Telugu). This plant has been regarded as a universal number of classical Indian Ayurvedic texts such as panacea in old Indian Ayurvedic texts. The tree Charaka Samhita dating to 1000 BC , Susruta dating reaches height of 7-10 m. Ecologist considered this as to 500 BC and Chakradatta dating to eleventh century an evergreen and deciduous tree and produces AD (Singh et al., 2015). Because of its overwhelming profuse branching pattern with paripinnate leaves and medicinal importance, the tree is revered in the Hindu orange to scarlet fragrant flowers arranged in dense tradition. Almost all parts of the plant, including the lateral corymbs. The geographical distribution of bark, flowers, leaf, and seeds, have been attributed to Saraca is mainly in Asia and some parts of North have medicinal value (Singh et al., 2015). America. The tree is found mainly throughout India S. asoca occupying a narrow ecological niche and the especially in West Bengal, Assam, Odisha, Tamil decline in natural population of this species is largely Nadu, , Kerala, Andhra Pradesh, due to unsustainable methods of harvesting. Meghalaya and Maharashtra. It is also widely Primarily, the harvesting of the bark causes distributed across the Western Ghats (both South and irreversible damage to the plant (Ankur, 2016). Central) and Sahyadri regions in Himalayas. Recognizing the importance of this species and the The recent information on the distribution of this imminent threat to its existence, S. asoca was species is sparse, mainly in parts of the Western assigned ‘vulnerable’ status by IUCN (Anonymous, Ghats of Maharashtra, Goa and Karnataka and in 2015) and ‘endangered’ status (Anonymous, 2001). some parts of Tamilnadu, Kerala, Eastern Ghats of The ashoka is a rain-foresttree. Its original Odisha and Meghalaya (Begum et al., 2014). Ved and distribution was in the central areas of the Deccan Goraya (2008) believe that the huge demand is either plateau, as well as the middle section of the Western met from some hitherto unknown source of S. asoca Ghats in the western coastal zone of the Indian

Proceedings of ARSSS International Conference, 09th September, 2018, Bengaluru, India 21 Development of Strategies for Conservation of S.Asoca: An Important Globally Vulnerable Tree Species in Western Ghats India subcontinent. Saraca indica is reported to be Rnase digestion at room temperature for one hr. distributed in Thailand, Vietnam, Indonesia, Java, incubation and electrophoresis was carried out using Sumatra and Malaysia. It is said (Flora of Ceylone) to 0.8% agarose gel in 1 x TAE (Tris - acetic acid - be introduced in India (along with Saraca declinata EDTA) as running buffer pre stained with ethidium (Jacq.) Miq. and Saraca thaipingensis Cantley ex bromide. Purified genomic DNA is quantified using Prain). However, genus Saraca includes shrubs and bio photometer at 280 nm and the values are recorded trees distributed in Central Southeast Asia from India in Nano gram units. Each sample is diluted to 25 to China through Malaysia, Sumatra and eastwards to ng/µl in separate vials for using as template DNA for Celebes. Indian Saraca asoca is separated from PCR amplification. Remaining genomic DNA were Malaysian Saraca indica based on the nature of stored at -800C . bracteoles. Species with clasping bracteoles were PCR amplification was set using fluorescent dye described as Saraca asoca, while Saraca indica labelled primers (Table8) with template DNA in an possessed spreading bracteoles. aliquot of 22.5 µL reaction mixture containing: 10X The continuous exploitation of several medicinal PCR reaction buffer ( 2.5 µl), 5pmole/µl plant species from the wild and substantial loss of forward primer(1.0 µl),5pmole/µl reverse primer(1.0 their habitats during the past 15 years have resulted in µl),1mM dNTP’s (2.5µl),Distilled water(12.0 µl),Taq the population decline of many high value medicinal DNA (0.20 µl),DNA template(1.50 µl).To amplify plant species over the years (Shah and Kapoor, 1978; the template DNA, the following PCR condition was Kala, 2003, 2006; Mishra et al., 2009, 2011). There set. Initial denaturation - 94oC for 3 minutes, are several other potential causes for rarity in followed by 35 cycles of denaturation - 94oC for 1 medicinal plant species, such as habitat specificity, min, annealing of primer for1 min 30 sec, Extension narrow range of distribution, land use disturbances, for 72o for 2 minute and final extension for 72o for 10 introduction of non-natives, habitat alteration, min. climatic changes, heavy livestock grazing, explosion of human population, fragmentation and degradation III. RESULTS of population, population bottleneck, and genetic drift (Weekley and Race, 2001; Kala, 2005). Additionally, Vegetative characteristic features of S. asoca:Saraca natural enemies (i.e., pathogens, cattle and forest asoca is a medium sized, evergreen tree grows to a fires) could substantially limit the abundance of rare height of 7–10 m, distributed up to an altitude of medicinal plant species in any given area (Bevill et 750 m MSL. Leaves are 15–20 cm long, rachis al., 1999). Habitat loss and degradation are believed glabrous, corky at the base; petioles very short; to threaten 91% of the rare plants in the world. Many stipules intra-petiolar, completely united, 4–6 pairs of of the world’s rare plant species are edaphic leaflets which are oblong- lanceolate, obtuse or acute, endemic, whose unique soil needs, habitats, and quite glabrous, base rounded or cuneate. The bark is restricted distribution make them especially rough and uneven, dark brown or grey or almost vulnerable to human activities (Hopper et al., 1990 black with warty surface. Bark channeled, smooth andBriggs and Leigh, 1996). with circular lenticels and transversely ridged, sometimes cracked (Ali, 2008; Rastogi, 2003). II. MATERIALS AND METHODS Reproductive characteristic features:The flower buds were yellowish green and commenced opening from Based on the distribution of current study species in December end and continue until May with peak India, four study sites namely Jambani, Patoli, flowering season between February and March. The Urabail and Palda present in excellent habitat trees remained in bloom for 2–3 months. Flowers are suitability; five medium habitat suitability sites viz., fragrant, apetalous, large, snowy and yellow to Devimane, Banavasi, , Honnavara and orange yellow in colour when just open, but change kakkalli are selected. Three sites of good habitat to red gradually and are borne in dense corymbose suitability area such as Subramanya, Jog and inflorescence. The pods are flat, leathery 10-25 cm were selected for the present study. long 3.5-5 cm wide, reddish brown when immature Young fully expanded leaf tissues of S. asoca were and turning to green when the seeds are mature. Each collected from approximately 277 individual trees of pod contains 4-8 seeds, which are ellipsoid oblong S. asoca distributed in 12 natural occurrence sites and compressed with brown smooth seed coat and (Table 1) and stored at -200C (deep white cotyledens (Ved and Ravikumar, 2000). freezer) and these samples were used as source of Fruiting commenced from March end to July with materials for isolation of genomic DNA throughout peak during May–June. The fruits attained maturity the study. from last week of May to July. Seeds dispersed from Total genomic DNA from fresh leaf samples of S. the pod on the ground upon complete maturity. Test asoca was isolated by following CTAB method with weight of hundred seeds ranged from 633.00 to modification of procedure suggested by Glenn and 969.00 g. Seed size was highly variable, which Schable, 2005.Genomic DNA isolated from various ranged from 2.06 to 11.56 g. Average length and leaf samples were subjected to purification using

Proceedings of ARSSS International Conference, 09th September, 2018, Bengaluru, India 22 Development of Strategies for Conservation of S.Asoca: An Important Globally Vulnerable Tree Species in Western Ghats India breadth of the seeds varied from 2.80 to 5.80 cm and individuals of S.asoca are more diverse within 1.60 to 4.10 cm respectively. population level with high percentage of The measure of genetic variation in population is the variation(91.38 %) than among populations (8.62 %) amount of heterozygosity. The Ho is found ranging with less variation using 1023 permutation. and low from 0.638 -0.879 and He found ranging 0.506 - 0.76 fixation index Fst value found is 0.0862. The percent numerical value respectively with the mean values of molecular variance among niche suitability was 0.789 and 0.669 for 12 population of S. asoca. The 2%, followed by 17% among population and 81% observed heterozygosity was higher than the expected within population (Table4). heterozygosity among 12 populations. Populations viz., Patoli, Urabail, Banavasi, Kuppalli and Jog DISCUSSION showed higher Ho. The occurrence sites, namely Jambani, Urabail, Palda, Banavasi, Kuppalli, S. asoca is one of the popular medicinal plants with a Honnavara and Jog showed higher He (Table 2). Two potential of curing many human disease and populations, namely Kakkalli and Subramanya physiological disorders .This tree is considered to be showed lesser Ho and He among the 12 populations. one of the potential sources of herbal formulations The number of alleles in 12 population of S. asoca owing to its wide range of medicinal properties. S. was ranged from 2.6 to 8.2 and allelic richness was asoca finds a prominent place in many Hindu ranged from 2.6 to 5.03. From the data, the mean literatures, since ancient time and proven its number of allles for all the 12 population of S. asoca importance as one of the traditional herbal medicines was 6.53. The ppulations Jambani, Urabail, over a period of several years. The perennial tree Devimane, Banavasi, Kuppalli, Honnavara and although reported to be distributed in Southern part of Kakkalli showed higher number of alleles. Urabail India(Singh et al. 2015)substantial evidences site showed highest allelic richness. The effective indicated its distribution to other parts of the world. It number of alleles (ne) is also used to measure the is reported that the genus extends its distribution in amount of genetic variation, Urabail and Honnavara South East Asia from India to South West China and showed highest effective number of alleles with 4.20 in Malaysia from Sumatra to Celebes, except in and 4.05 respectively. Less number of alleles was Philippines. The phenotypic character such as shape found in Subramanya (2.30). Of the 12 populations of the leaflets and their venation, colour and size of studied, the mean number of effective alleles among the flowers are too variable to be trusted. It was the population was 3.48., eleven populations showed reported that both S.asoca and S.indica were 100% polymorphism except Subramanya with 80% described for the first time in the year 1968 and 1869 polymorphism. This results indicates that S. asoca is respectively. Though S.asoca was reported to be distributed in different geographical location with synonymous of S.indica the plant samples identified wider genetic variability within population for its for the present study was subjected to taxonomical better adaptation with its more tendency towards observation and compared with the published heterozygosity. description by Zujiderhoudt(1967). The correlation between geographical distance and While comparing the vegetative characteristics of S. leniarizes Fst was positive. However, the inbreeding asoca with S.indica. it was inferred that both the coefficiency and heterozygosity was negatively species exhibits with very close resemblance with correlated. The low level of genetic differentiation respect to the distribution, shape of leaves, leaflets, with GST=0.109(anything>0.15 is high according to leaf size and shape, texture, leaf venation and Nei,1978) and The high level of gene flow reticulation. The reproductive characteristic features value(Nm= 2.0353), FST=0.1350 of S.asoca and S.indica was also compared in the present study. It was found that, a large amount of Population genetic variability across habitat variation exists between the two species based on suitability: The observed heterozygosity of 12 floral, fruit and seed characterestics. However, none population of S. asoca was more than expected of the report explaining the occurrence and heterozygosity across three niche suitability distribution of S. indica in South india. In the absence parameters. Allelic richness was found to be more in of S. indica in southern part of India could be due to the excellent niche and less in good niche suitability. geographical barrier and several other unknown Number of alleles and allelic richness was found reasons. more in medium suitability, the other parameters Intensive disturbances of natural population of S. such as heterozygosity and the gene diversity was asoca through the indiscriminate use and destructive more in excellent niche (Table 3). The private alleles harvesting process of ashoka bark has led to acute were found to be present more in excellent niches. scarcity of the genuine raw drug and this in turn, has led to cost escalation and widespread adulteration or Molecular variance: In the present study, molecular substitution in a variety of herbal formulation (Beena variance for within and among populations for S. and Radhakrishnan 2010; Pradhan et al., 2009). This asoca in South India was assessed. The result of has resulted in drastic reduction in population of AMOVA a significant (p> 0.01) showed that many of the medicinal plants and S. asoca has no

Proceedings of ARSSS International Conference, 09th September, 2018, Bengaluru, India 23 Development of Strategies for Conservation of S.Asoca: An Important Globally Vulnerable Tree Species in Western Ghats India exception. Presently, S. asoca has been included in REFERENCES the ‘vulnerable’ category of the ICUN list (IUCN 2011). The unscientific management practices of S. [1] Ankur, P., Makarand, P., Monali, M., Prerna, A., Narayani, asoca coupled with ever increasing demand for its B., Rajesh, G., Amit, M., Chandrakant, S., and R. Vasudeva. (2016). Distribution and population status of threatened bark and phytochemicals, poor seed viability medicinal tree Saraca asoca (Roxb.) De Wilde from (Pushpagandhan et al., 2004) and over-exploitation of Sahyadri– ecological corridor. Curr. Sci. 111(9): the plant parts such as bark, flowers and seeds has 1500-1506. contributed greatly for the declining population of [2] Anonymous (2007). World Wildlife Fund (content partner); Mark McGinley (Eds). North Western Ghats mountain rain this species. Presently, the high demands for the raw forests. Encyclopaedias of Earth. http:// www.eoearth.org materials for S. asoca is met by widespread [3] Ali, M. (2008).Pharmacognocy. CBS Publishers and adulteration with other plant materials. Distributors. New Delhi. pp 668-669. There is presence of more effective number of alleles [4] Beena, C.and Radhakrishnan, VV. (2010). Haemagglutination as a rapid tool to differentiate Saraca within a population which is indicative of suitable asoca bark from the adulterant Polyalthia longifolia. J adaptability to the environment and evolutionary Progress Agric 1(1):1–3. changes. The microsatellites characterized and used [5] Begum, SN., Ravikumar, K. and Ved, D K. (2014). Asoka- for the present study showed significantly high an important medicinal plant, its market scenario and conservation measures in India. Curr. Sci. 107: 26-28. polymorphism. There is evidence of presence of [6] Bevill, RL., Louda, SM. and Stanforth, LM., (1999). private alleles approximately in half of the population Protection from natural enemies in managing rare plant studied.The microsatellite data as analysed implied species, Cons. Biol.13: 1323-1331. strong geneflow between S.asoca populations (Nm= [7] Briggs, JD. and Leigh, JH. (1996). Rare and Threatened Australian Plants. Revised edition, CSIRO publishing, 2.0353). The low level of genetic differentiation with Melbourne, Australia, p446. GST=0.109 are generally less adoptive to [8] Hopper SD., Van-Leeuwen, S., Brown, AP.and Patrick, SJ. environmental stress. In the present study , Nm value (1990). Western Australia's Endangered Flora and other suggests that genetic drift may not have been a plants under consideration for declaration, Department of Conservation and Land Management. p 139. dominant factor determining the genetic structure of [9] Kala, CP. (2003). Commercial exploitation and conservation S. asoca populations. Increase in geographical status of high value medicinal plants across the borderline of distance between the population restricted the gene India and Nepal in Pithoragarh. Ind. For. 129: 80-84. flow significantly. [10] Kala, CP. (2006). Medicinal plants of the high altitude cold desert in India: Diversity, distribution and traditional uses. The overall observed heterozygosity (0.789) is higher Int. J Biodiv. Sci. Manag.2:1-14. than the expected heterozygosity (0.65). We found [11] Mishra, M. and Kotwal, PC. (2009).Traditional harvesting that genetic differntaition is 13 % among the and processing methods of Dioscorea daemona (Baichandi) populations studied of the total genetic diversity. tubers in the forests of Madhya Pradesh, India. J. Trop. Med. Plants10(1): 113-118 AMOVA results similarly showedthat the proportion [12] Mishra, A., Kumar, A., Rajbhar, N. and Kumar, A. (2013). of genetic diferentiation among populations Phytochemical and accounted for about 8.6% of the total genetic [13] pharmacological importance of Saraca indica. Int. J. Pharm. diversity which also explains the higher genetic Chem. Sci. 2: 1009–1013. [14] Mishra, M. (2011). Conservation of biodiversity in the variation within population. The present results show natural forests of central India: a case of critically endangered that S.asoca maintains relatively high genetic medicinal species Safed musli in Bhopal forest, Madhya diversity. The major reason for globally Pradesh, India. Bios. Disc. 2(3): 299-308. vulnerable/locally endangered may be due to over [15] Nei, M. (1973). Analysis of gene diversity in subdivided populations. Proc. Natl. Acad. Sci. USA. 70: 3321–3323. exploitation from wild population to meet high [16] Pradhan, P., Joseph, L., Gupta, V., Chulet, R., Arya, H., demand of pharmaceutical value and traditional uses Verma, R. and Bajpai, A. (2009). Saraca asoca (Ashoka): A rather than lack of overall genetic diversities. review. J. Chem. Pharm. Res.1: 62-71. [17] Rastogi., VD. Pharmacognosy & Phytochemistry, Career Publication, Nashik. (2003). 269-270. CONCLUSION [18] Satpal Singh, TH., Krishna, A., Kamalraj, S., Kuriakose, GC., Jinu Mathew, V. and Jayabaskaran, C. (2015). The high medicinal value of S. asoca has resulted in Phytomedicinal importance of Saraca asoca (Ashoka):an over-exploitation of the species across its natural exciting past, an emerging present and a promising future. Curr. Sci. 109 (10):1790-1801. range of distribution. Therefore, policies must enact [19] Shah, NC. and Kapoor, LD. (1978). Depletive medicinal ed to forbid unlawful felling and peeling of this tree plants of Kumaon Himalayas. J. Res. Ind. Med. Yoga Homeo. in natural population. Artificial cultivation may be an 13(3): 38-43. effective approach to securing medicinal and bark [20] Ved, DK. and Goraya, GS. (2008). Demand and supply of medicinal plants in India. Foundation for Revitalisation of supply which generates beneficial ecological effect Local Health Traditions. Bangalore, India. and considerable economic benefit. In order to [21] Ravikumar, and Ved,( 2000), In: 100 Red-listed Medicinal maintain local variation , plant materials used in plants of conservation concern in South India. Foundation for reforestation efforts should be obtained from within Revitalization of Local Health Tradition, Bangalore, pp.203- 206. the same conservation unit. Some populations with [22] Weekley, CW. and Race, T. (2001). The breeding system of higher genetic variation, eg Urabail, Jambani, should Ziziphus celata Judd and D.W. Hall (Rhamnaceae), a rare be given high priority in in-situ conservation. endemic plant of the Lake Wales Ridge, Florida, USA: implications for recovery. Biol. Conserv. 100: 207-213.

Proceedings of ARSSS International Conference, 09th September, 2018, Bengaluru, India 24 Development of Strategies for Conservation of S.Asoca: An Important Globally Vulnerable Tree Species in Western Ghats India

a) Habit of S. asocain natural habitat b) Girth at breast height

c) Compound leaf d) Flowers of Saracaasoca e) Red flowers.

f) Pod of Saracaasoca g) Seeds of Saracaasoca Figure1: Field survey photographs of S.asoca in its natural habitat(a),GBH(b) and tree parts(c-g).

Name of the Sample Niche Population Id Latitude Longitude Population Size suitability 1 Jambani(JM) 14.5983 74.5514 28 Excellent

2 Patoli(PT) 15.1878 74.5535 25 Excellent

3 Urabail(UB) 14.8038 74.7418 25 Excellent

4 Palda(PL) 14.5205 74.5604 23 Excellent 5 Devimane(DV) 13.6179 74.3089 25 Medium 6 Banavasi(BN) 14.591 74.9976 24 Medium

7 Kuppalli(KP) 14.2231 74.8112 25 Medium environmental variables used. variables environmental 8 Honnavara(HN) 13.8781 74.8644 13 Medium 9 Kakkalli(KK) 14.7686 74.6583 29 Medium Annual temperature and annual rain fall are the the are fall rain annual and temperature Annual 10 Subramanya(SB) 13.5492 75.789 06 Good

Proceedings of ARSSS International Conference, 09th September, 2018, Bengaluru, India 25 Development of Strategies for Conservation of S.Asoca: An Important Globally Vulnerable Tree Species in Western Ghats India 11 Jog(JG) 14.2701 74.5995 15 Good 12 Kodachadri(KD) 13.8923 74.8216 14 Good Table 1: Study sites, Geographic coordinates, sample size and density of S. asocapopulations in the Central Western Ghats, Karnataka, India.

Number Effective Observed Expected No. Population Allelic Fis per %Poly of number heterozy- Heterozy- Fst Private name richness population morphism alleles of alleles -gosity -gosity Alleles Jambani 8.200 3.777 0.767 0.671 4.5962 -0.146 0.097 4 100

Patoli 6.000 3.254 0.832 0.654 3.9826 -0.28 0.119 0 100

Urabail 8.200 4.209 0.879 0.760 5.037 -0.161 0.065 4 100

Palda 6.000 3.546 0.783 0.685 4.176 -0.146 0.097 0 100

Devimane 7.000 3.069 0.772 0.645 4.1122 -0.202 0.086 4 100

Banavasi 7.000 3.540 0.796 0.694 4.3954 -0.149 0.073 0 100

Kuppalli 7.200 3.858 0.872 0.753 4.6968 -0.161 0.078 2 100

Honnavara 6.800 4.059 0.769 0.687 4.8714 -0.126 0.094 0 100

Kakkalli 7.600 3.213 0.638 0.593 4.0316 -0.077 0.088 3 100

Subramanya 2.600 2.301 0.767 0.506 2.6 -0.597 0.236 0 80

Jog 6.400 3.888 0.813 0.725 4.7222 -0.127 0.102 4 100

Kodachadri 5.400 3.135 0.786 0.656 4.1054 -0.207 0.097 0 100

Mean 6.533 3.487 0.789 0.669 0.1037 98

SE 0.346 0.195 0.027 0.022 0.044 1.67

Table2: Over all genetic variance of 12 populations of S.asoca across South India.

Figure2: Estimates of the proportion of ancestry Q, in each of K=5clusters for 252 individuals of S.asoca Western Ghats India.

Table3: Genetic diversity assessment across niche suitability of populations of S. asoca in South India.

Proceedings of ARSSS International Conference, 09th September, 2018, Bengaluru, India 26 Development of Strategies for Conservation of S.Asoca: An Important Globally Vulnerable Tree Species in Western Ghats India Source Df Sum of Squares Estimated Variance Total variance Among niche suitability 2 41.493 0.073 2% Among Populations 9 142.696 0.615 17% Within populations 240 701.18 2.922 81% Table 4: Molecular variance analysis (AMOVA) for 252 individuals of S. asoca.

Figure3: The correlation of extinction factors gene flow, geographic distance and pair wise Fst of 12 populationsof S.asoca in South India.



Proceedings of ARSSS International Conference, 09th September, 2018, Bengaluru, India 27