Assessment of genetic diversity in Shorea robusta: an economically important tropical tree species Giridara-Kumar Surabhi, Subhankar Mohanty, Rajesh Kumar Meher, Arup Kumar Mukherjee, Lakshmi Narayana R.Vemireddy203_pdf.pdf110-117AMOVA; genetic diversity; ISSR; molecular marker; polymorphism; Shorea robusta.81116The present investigation reports an elucidation of genetic diversity among four-populations of most economically and ecologically important tree species, sal (Shorea robusta Gaertn.) for the first time in India, using ISSR markers. A total of one-hundred individual S. robusta trees were sampled from four different populations, considering twenty-five individuals from each population. In total, twenty-ISSR primers were screened with S. robusta DNA, and out of twenty, sixteen-primer produced reproducible amplicons. Sixteen selected ISSR markers were amplified a total of 118 alleles and the total number of amplicons for individual primers ranged from 5 to 12, with a mean of 7.37 alleles per primer, of which 74 were polymorphic with an average of 4.62 alleles per primer. The ISSR primer (GA)8YG yielded highest number of alleles (12) and primers (CA)8RG and (CT)8G yielded lowest number of alleles (5), with an average alleles size between 200-3500bp. The percentage of polymorphic alleles ranged from 40 [(CA)8RG] to 83.33 [(AC)8C]. A dendrogram based on UPGMA analysis grouped the four populations into two major clusters, having Keonjhar population into first cluster and rest three populations into second cluster. It was notable that the second major cluster was further divided into three-separate sub-clusters, representing population from each three locations. Analysis of molecular variance (AMOVA) revealed that the majority of genetic variation exists within populations, compared to the variation that exists among the populations. The present study is a clear-cut indication that inter- and intra-population genetic variation exists in S. robusta and ISSRs appears to be an efficient marker system in quantifying genetic variability in different populations of S. robusta. Surabhi GK, Mohanty S, Meher RK, Mukherjee AK, Vemireddy LNR. Assessment of genetic diversity in Shorea robusta: an economically important tropical tree species. J App Biol Biotech. 2017; 5 (02): 110-117. DOI: 10.7324/JABB.2017.502181. Ashton PS. Flora malesiana. Series I-Spermatophyta, Flowering plants. vol 9, part 2, Dipterocarpaceae, Martinun Nijhoff Publicatons, The Netherlands; 1982.2. Tewari DN. A monograph on sal (Shorea robusta), International book distributors. Dehra Dun; 1995.3. Champion HG, Seth SK. A revised survey of the forest types of India. The manager of publications. Delhi; 1968.4. Momose K, Yumoto T, Nagamitsu T, Kato M, Nagamasu H, Sakai S, Harrison RD, Itioka T, Hamid AA, Inoue T. Pollination biology in a lowland dipterocarp forest in Sarawak, Malaysia. I. Characteristics of the plant-pollinator community in a lowland dipterocarp forest. Am J Bot. 1998; 85:1477-1501.5. Jackson JK. Manual of Afforestation in Nepal, Forest Research and Survey Centre, Kathmandu, Nepal; 1994.6. Pawar GV, Singh L, Jhariya MK, Sahu KP. Regeneration status in relation to anthropogenic disturbance in tropical deciduous forest of Chhattisgarh, The Ecoscan. 2012; 1:281-286.7. Pattanaik S, Dash A, Mishra RK, Nayak PK, Mohanty RC. Seed germination and seedling survival percentage of Shorea robusta Gaertn. f. in buffer areas of Similipal biosphere reserve, Odisha, India. Journal of Ecosystem Ecography. 2015; 5:1-4. 8. Kandya AK. Seed viability in Sal (Shorea robusta). Journal of Botanical Society. 2006; 41: 16-24.9. 'O'Malley LSS. Provincial geographies of India: Bengal, Bihar, and Orissa, Sikkim; 2011. 10. Negi SS. Indian forestry through Ages, Indus Publishing Company, New Delhi; 1994, p. 42-43.11. Narayanamurti D, Das N. A preliminary note on adhesives, building boards, and moulding powder from tree's bark. Indian For. 1951; 77:706-708.12. Chitale VS, Behera MD. Can the distribution of sal (Shorea robusta Gaertn. f.) shift in the northern direction in India due to changing climate?. Current Science. 2012; 102:1126-1135.13. Lee SL, Ng KKS, Saw LG, Lee CT, Muhammad N, Tani N, Tsumura Y, Koskela J. Linking the gaps between conservation research and conservation management of rare dipterocarps: A case study of Shorea lumutensis. Biol Conserv. 2006; 131:72-92.14. Suoheimo J, Li CH, Luukkanen O. Isozyme variation of natural populations of sal (Shorea robusta) in the Terai region, Nepal. Silv Genet. 1999; 48:199-203.15. Jena RC, Samal KC, Pal A, Das BK, Chand PK. Genetic diversity among some promising Indian local selections and hybrids of cashew nut based on morphometric and molecular markers. International Journal of Fruit Science. 2016; 16:69-93.16. Pandey M, T. Geburek T. Genetic differences between continuousand disjunct populations: some insights from sal (Shorea robusta Roxb.) in Nepal. Conserv Genet. 2010; 11: 977-984.17. Pandey M, Geburek T. Fine-scale genetic structure and gene flow in a semi-isolated population of a tropical tree, Shorea robusta Gaertn. (Dipterocarpaceae). Current Science. 2011; 101: 293-299.18. Karp A, Edwards K (1998) DNA markers: A global overview in: DNA markers: protocols, applications and overviews, Anolles GC, Gresshoff PM, editors. Willy-Liss Inc, New York; 1998, p.1-14.19. Senapati SK, Das GK, Aparajita S, Rout GR. Assessment of genetic variability in the Asoca tree of India, Biodiversity. 2012; 13:16-23.20. Rawat S, Jugran AK, Bhatt ID, Rawal RS, Nandi SK. Genetic diversity analysis in natural populations of Roscoea procera Wall. from West Himalaya, India. Braz J Bot. 2016; 621-630.21. Sunar S, Yildirim N, Sengul M, Agar G. Genetic diversity and relationships detected by ISSR and RAPD analysis among Aethionema species growing in Eastern Anatolia (Turkey), Comptes Rendus Biologies. 2016; 339(3-4):147-151. 22. Selyutina IY, Konichenko ES, Dorogina OV, Sandanov DV. Genetic diversity of the endangered endemic milkvetch Astragalus sericeocanus Gontsch., Fabaceae from the Lake Baikal region. Biochemical Systematics and Ecology. 2016; 68:163-169.23. Gupta PK, Varshney RK. The development and use of microsatellite markers for genetic analysis and plant breeding with emphasis on bread wheat. Euphytica. 2000; 113:163-185.24. Staub JE, Serquen FC, Gupta M. Genetic markers, map construction, and their application in plant breeding. Hort Science. 1996; 31:729-739.25. Halldén C, Hansen M, Nilsson NO, Hjerdin A, Säll T. Competition as a source of errors in RAPD analysis, Theort Appl Genetics. 1996; 93:1185-1192.26. Hansen M, Halldénand C, Säll T. Error rates and polymorphism frequencies for three RAPD protocols. Plant Molecular Biology Reporter. 1998; 16:139-146.27. Gupta M, Chyi YS, Romero-Severson J, Owen JL. Amplification of DNA markers from evolutionarily diverse genomes using single primers of simple-sequence repeats. Theor Appl Genet. 1994; 89:998-1006.28. Zietkiewicz E, Rafalski A, Labuda D. Genomic finger-printing by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics. 1994; 20:176-183.29. Hu Y, Wang L, Xie W, Yang J, Li Y, Zhang H. Genetic diversity of wild populations of Rheum tanguticum endemic to China as revealed by ISSR analysis. Biochem Syst Ecol. 2010; 38: 264-274.30. Reddy MP, Sarla N, Siddiq EA. Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica. 2002; 128:9-17.31. Gherardi M, Mangin B, Goffinet B, Bonnet D, Huguet T. A method to measure genetic distance between allogamous populations of alfalfa (Medicago sativa) using RAPD molecular marker. Theoretical Applied Genetics. 1998; 98:406-412.32. Feyissa T, Nybom H, Bartish IV, Welander M. Analysis of genetic diversity in the endangered tropical tree species Hagenia abyssinica using ISSR markers. Genet Resour Crop Evol. 2007; 54:947-958.33. Ballesta P, Mora F, Contreras-Soto RI, Ruiz E, Perret S. Analysis of the genetic diversity of Eucalyptus cladocalyx (sugar gum) using ISSR markers. Acta Scientiarum. 2015; 37:133-140.34. Liu J, Liao K, Nasir M, Zhao S, Sun Q, Peng X. Analysis of genetic diversity of the apricot germplasm in the southern region of the Tianshan Mountains in Xinjiang, China using the ISSR technique. Eur J Hortic Sci. 2016; 81:37-43.35. Peterson G, Seberg O (2002) Molecular evolution and phylogenetic application of DMC1, Molecular Phylogenetic Evolution. 2002; 22:43-50. 36. Surabhi GK, Pattanayak S. Deciphering the genetic identity and fidelity of banana through inter simple sequence repeats fingerprinting, Horticultural Biotechnology Research. 2015; 1:16-22.37. Arroyo MTK, Squeo F. Relationship between plant breeding systems and pollination, In: Kawano, editors. Biological approaches and evolutionary trends in plants, Academic Press, London, 1990, p. 205-227.38. Hamrick JL, Godt JW. Allozyme diversity in plant species, In: Brown AHD, Clegg MT, Kahler AL, Weir BS, editors. Plant population genetics, breeding, and genetic resources, Sinauer, Sunderland, Mass; 1989, p. 43-63.39. Bhat KV, Babrekar PP, Lakhanpaul S. Study of genetic diversity in Indian and exotic sesame (Sesamumindicum L.) germplasm using random amplified polymorphic DNA (RAPD) markers, Eupytica. 1999; 110:21-33.40. Nybom H. Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants. Mol Ecol. 2004; 13:1143-1155.41. Hamrick JL, Godt JW. Allozyme diversity in plant species, In: Brown AHD, Clegg MT, Kahler AL, Weir BS, editors. Plant population genetics, breeding and genetic resources, Sinauer Associates, Sunderland; 1990, p. 43-63.