Assessment of molecular diversity of greengram [Vigna radiata (L.) Wilzek] through RAPD

DOI: 10.18805/lr.v0iOF.9442    | Article Id: LR-3507 | Page : 1-8
Citation :- Assessment of molecular diversity of greengram [Vigna radiata (L.) Wilzek] through RAPD .Legume Research-An International Journal.2017.(40):1-8

Gunnjeet kaur*, Arunabh Joshi, Devendra Jain, Ganesh Rajamani and Divya Vyas

itsgunjeet@gmail.com
Address :

Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, M.P.U.A.T., Udaipur-313 001, India.

Submitted Date : 20-04-2015
Accepted Date : 13-10-2015

Abstract

A total twenty three genotypes of green gram (Vigna radiata) were subjected to Randomly amplified polymorphic DNA (RAPD) analysis for molecular characterization. A total of 25 randomly selected decamers were screened, out of which only 15 generated 126 amplification products from which 117 bands were found polymorphic, the average polymorphism being 93.48%. The total number of amplified bands varied between 2 (primer OPP-09) to 17 (primer OPA-1) with an average of 9.5 bands per primer. The overall size of PCR amplified products ranged between 200 bp to 2900 bp. The average  Polymorphism Information Content(PIC)  was 0.32 ranging from 0.17 to 0.46. Primer OPA-01 and OPP-06 detected two unique bands ranged between 250 bp to 2500 bp in two genotypes (PUSA-672 and HUM-12). Jaccard’s similarity coefficient values ranged from 0.28-0.90 with an average of 0.59. Based on dendrogram generated through UPGMA method and PCA, most of the genotypes got divided into four main clusters. Genotype EC-398885 lay far apart and thus showed maximum genetic distance. The assessment of genetic diversity is a prerequisite and important step for the improvement of any legume crop. Thus, present results of the present study could be further extrapolated to other green gram accessions in Vigna germplasm.

Keywords

Diversity analysis PCA Polymorphism RAPD Similarity coefficient UPGMA.

References

  1. Afzal, M.A., Haque, M.M. and Shanmugasundaram, S. (2004). Random Amplified Polymorphic DNA (RAPD) Analysis of Selected Green gram [Vigna radiata (L.)Wilczek] Cultivars. Asian Journal of Plant Sciences, 3: 20-24. 
  2. Arumuganathan, K. and Earle, E.D. (1991). Nuclear DNA content of some important plant species. Plant Molecular Biology Report , 9: 208-218.
  3. Bernatzky, R. and Tanksley, S.D.(1989).Restriction fragments as molecular markers for germplasm evalution and utilization, in The use of plant genetic resources. Cambridge University Press, New York: 353-362.
  4. Bhat, K.V., Lakhanpaul, S. and Chadha, S. (2005). Amplified fragment length polymorphism (AFLP) analysis of genetic diversity in Indian green gram [Vigna radiate (L.) Wilczek] cultivars. Indian Journal of Biotechnology, 4:56-64.
  5. Bhuyan, S.I., Hossain, M.S., Islam, M.M., Begum, S.N., Urbi, Z. and Hossain, M.S. (2014).Molecular Assessment of Genetic Diversity and Relationship in Selected Green gram Germplasms. Biotechnology, 13: 126-134.
  6. Datta, S., Gangwar, S., Kumar, S.,Gupta, S.Rai, R, Kaashyap, Singh ,P., Chaturvedi, S.K., Singh ,B.B. and Nadarajan, N.(2012).Genetic Diversity in Selected Indian Green gram [Vigna radiata (L.) Wilczek] Cultivars Using RAPD Markers. American Journal of Plant Sciences, 3:1085-1091.
  7. Doyle, J.J. and Doyle, J.L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf material. Phytochemical Bulletin, 19: 11-15.
  8. Economic Survey. (2012-2013). Ministry of Finance, Department of Economic Affairs, Economics Division Government of India, New Delhi.
  9. Jena, B., Panigrahi, K. K., Baisakh, K., Das, T. R. (2015). Genetic Diversity as Assessed by Agronomical Traits and Molecular Markers in Local Land Races of Green gram (Vigna radiata L.) Under Rice Fallow Condition. Molecular Plant Breeding, 6: 1-7.
  10. Gepts, P. (1993). The use of molecular and biochemical markers in crop evolution studies. Evolution Biology, 27: 51-94.
  11. Gherardi ,M., Mangin, B., Goffinet, B., Bonnet, D. and Huguet, T. (1998). A method to measure genetic distance between allogamous populations of alfalfa (Medicago sativa) using RAPD molecular markers. Theory Application Geneticia, 96: 406-412.
  12. Gwag, J.G., Dixit, A., Park, Y.J., Ma, K.H., Kwon, S.J., Cho, G.T., Lee, G.A., Lee, S.Y.,Kang ,H.K. and Lee, S.H. (2010).Assessment of genetic diversity and population structure in green gram. Genes & Genomics , 32: 299-308.
  13. Jaccard, P. (1908). Nouvelles researches surla distribution florale. Societe Vaudoise des Science Natulelles Bulletin, 44: 233-170.
  14. Lakhanpaul, S., Chadha, S. and Bhat, K.V. (2000). Random amplified polymorphic DNA (RAPD) analysis in Indian mung bean (Vigna radiata (L.) Wilczek) cultivars. Genetica , 109: 227-234.
  15. Lavanya, G.R., Srivastava, J. and Ranade, S.A. (2008). Molecular assessment of genetic diversity in green gram germplasm. Journal of Genetics, 87: 65–74.
  16. Reddy,K.S., Souframanien, J., Reddy, K.S. and Dhanasekar, P. (2008). Genetic diversity in mung bean as revealed by SSR and ISSR markers. Journal of Food Legumes, 21: 15- 21.
  17. Rohlf, F.J. (2004). NTSYS-PC. Numerical taxonomy and multivariate analysis system. Exeter Software, New York.
  18. Saini, M., Singh, S., Hussain, Z. and Yadav, A. (2010). RAPD analysis in green gram (V. radiata (L.) Wilczek): I. Assessment of genetic diversity. Indian Journal of Biotechnology, 9:137-146.
  19. Sambrook, J., Fritschi, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York.
  20. Santala, M., Power, J.B. and Davey, M.R. (1998). Genetic diversity in green gram germplasm revealed by RAPD markers. Plant Breeding, 117: 473-478. 
  21. Singh, A.K., Mishra, A. and Shukla, A. (2010). Genetic evaluation and identification of genetic donors in black gram (Vigna mungo) revealed by agro-morphological traits and seed storage protein analysis. World Applied Sciences Journal, 10: 477-488.
  22. Singh, A., Dikshit, H.K., Jain, N., Singh, D. and Yadav, R.N.(2013).Efficiency of SSR,ISSR and RAPD markers in molecular characterization of green gram and other Vigna species. Indian Journal of Biotechnology, 13: 81-88.
  23. Sneath, P.H.A. and Sokal, R.R. (1973). Numerical Taxonomy. San Francisco: Freeman. 
  24. Smith, J.S.C., Chin, E.C.L., Shu, H., Smith, O.S., Wall, S.J., Senior, M.L., Mitchell, S.E., Kresovich, S. and Zeigle, J. (1997). An evaluation of the utility of SSR loci as molecular markers in maize (Z. mays L.): comparisons with data from RFLPs and pedigree. Theoretical and Applied Genetics, 95: 163-173.
  25. Sony, S.K., Habib, M.A. and Islam, M.N. (2012). Genetic diversity analysis of thirteen green gram (V. radiata L.) cultivars using RAPD markers. Bangladesh Journal of Botany, 41: 169-175.
  26. Srivastava, P., Pandey, A. and Sinha, D.P. (2013). Genetic diversity analysis in different varieties of black gram using RAPD markers. International Journal of Biological Research, 1: 43-48.
  27. Undal, V.S., Thakare, P.V., Chaudhari, U.S., Deshmukh, V.P. and Gawande, P.A. (2011). Estimation of Genetic Diversity among wild Vigna species revealed by RAPD Markers. Annals of Biological Research, 2: 348-354.
  28. Yadav, K. Yadav, S.K., Yadav ,A., Pandey, V.P. and Dwivedi, U.N.(2014). Comparative Analysis of Genetic Diversity among Cultivated Pigeonpea (Cajanus cajan (L) Millsp.) and Its Wild Relatives (C. albicans and C. lineatus) Using Randomly Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) Fingerprinting. American Journal of Plant Sciences, 5: 1665-1678.
  29. Williams, G.K., Kubelik, A.R., Livak, K.L., Rafalshi, J.A. and Tingey, S.V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Research, 18: 6531-6535.

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