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Research Article

volume 41 issue 2 (april 2018) : 175-181,   Doi: 10.18805/LR-3401

Assessment of genetic diversity in black gram [Vigna mungo (L.) Hepper] genotypes based on ISSR

D
Divya Vyas
A
Arunabh Joshi
G
Ganesh Rajamani
D
Devendra Jain
1Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur–313 001, Rajasthan, India.

  • Submitted24-12-2014|

  • Accepted21-03-2017|

  • First Online 20-01-2018|

  • doi 10.18805/LR-3401

Cite article:- Vyas Divya, Joshi Arunabh, Rajamani Ganesh, Jain Devendra (2018). Assessment of genetic diversity in black gram [Vigna mungo (L.) Hepper]genotypes based on ISSR. Legume Research. 41(2): 175-181. doi: 10.18805/LR-3401.

ABSTRACT

A study was carried out among 22 black gram genotypes to study the genetic diversity using 20 ISSR primers. Screening of the primers revealed that only 15 out of the 20 primers produced amplification.  A total of 84 amplified bands were obtained, out of which 72 were polymorphic 85.71 percent polymorphism. The total number of amplified bands varied between 1 (UBC-813 and UBC-878) and 9 (UBC-826) with an average of 5 bands per primer. The overall size of PCR amplified products ranged between 250 bp to 2000 bp. PIC values ranged from 0.00 to 0.51 with an average of 0.285 across all genotypes. Five unique bands were detected in four genotypes, out of which the genotype U-9 gave maximum number of distinct bands. The size of these unique bands ranged from 450 bp to 2000 bp. Based on the UPGMA derived dendrogram and PCA, the 22 genotypes could be divided into four main clusters. While Cluster I included 16 genotypes, the Clusters II, III and IV included two genotypes each. Genotypes IC-16511 and UTTARA, UH-177 and IPU2K-21, STY-2834 and UH-177 were found to be genetically distant from each other with a minimum similarity value of 0.42. The results are encouraging with the suggestion that the ISSR marker could prove to be a versatile tool in further screening of the Vigna germplasm pool for study of genetic divergence and the establishment of phylogenetic relationship amongst accessions.

REFERENCES

  1. Abd El-Hady, E.A.A., Haiba, A.A.A., Abd El-Hamid, N.R., Al-Ansary, A.M.F. and Mohamed, A.Y. (2010). Assessment of Genetic Variations in Some Vigna Species by RAPD and ISSR Analysis. New York Science Journal, 3(11): 120-128.
  2. Ajibade, S.R., Weeden, N.F. and Chite, S.M. (2000). Inter simple sequence repeat analysis of genetic relationships in the genus Vigna. Euphytica, 111(1): 47-55.
  3. Das, S., Das, S.S. and Ghosh, P. (2014). Analysis of genetic diversity in some black gram cultivars using ISSR. European Journal of Experimental Biology, 4(2): 30-34.
  4. Despeiger, A., Goubely, C., Lenoir, A., Cocherel, S., Picard, G., Rayanl, M., Greltel, F. and Delseny. (1995). Identification of the most represented repeat motif in Arabidopsis thaliana microsatellite loci. Theoretical and Applied Genetic, 91: 160-168.
  5. 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.
  6. Jaccard, P. (1908). Nouvelles researches surla distribution florale. Societe Vaudoise des Science Natulelles Bulletin, 44: 233-170.
  7. Kanimozhi, M., Jayamani, P. and Nadarajan, N. (2009). Genetic diversity as assessed by ISSR markers in Black gram [Vigna mungo (L.) Hepper]. Electronic Journal of Plant Breeding, 1: 12-17.
  8. Lukoki, L., Marechal, R. and Otoul, E. (1980). Les ancetres sauvages des haricots cultives: Vigna radiata (L.) Wilczek et V. mungo (L.) Hepper. Bull. Jard. Bot. Nat. Belgique., 50: 385-391.
  9. Nisar, M., Ghafoor, H., Ahmad, K.M., Rashid, A.S., Qureshi, H. and Ali, H. (2008). Evaluation of genetic diversity of pea germplasm through phenotypic traits analysis. Pakistan Journal of Botany, 40(5): 2081-2088.
  10. Pardhe, D.D. and Satpute, R.A. (2011). A Comparative Analysis of Genetic Diversity in Genus Vigna savi Genotypes using ISSR. International Journal of PharmaTech Research, 3(1): 464-470.
  11. Reddy, M.P., Sarla, N. and Siddiq, E.A. (2002). Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica, 128: 9-17.
  12. Rocha, R., Abad, J., Pires, I. and Araujo, E. (2002). Fingerprint and genetic diversity analysis of Eucalypts spp.genotypes using RAPD and SSR markers, Scientia Forestalis., 62: 24-31.
  13. Rohlf, F.J. (2004). NTSYS-PC. Numerical taxonomy and multivariate analysis system. Exeter Software, New York.
  14. Russel, J.R., Fuller, J.D., Macaulay, M., Hatz, B.G. and Jahoor, A. (1997). Direct comparison of levels of genetic variation among barley accessions detected by RFLPs, AFLPs, SSRs and RAPDs. Theoretical and Applied Genetics, 95: 714-722. 
  15. Sambrook, J., Fritschi, E.F. and Maniatis, T. (1989). Molecular cloning: a Laboratory Manual, Cold Spring Harbor Laboratory Press, New York. 
  16. Shafique, S., Khan, M.R., Nisar, M. and Rehman, S.U. (2011). Investigation of genetic diversity in black gram [Vigna mungo (L.) Hepper]. Pakistan Journal of Botany, 43(2): 1223- 1232.
  17. Singh, R., Heusden, A.W.V. and Yadav, R.C. (2011). A comparative genetic diversity analysis in mungbean (Vigna radiata L.) using inter-simple sequence repeat (ISSR) and amplified fragment length polymorphism (AFLP). African Journal of Biotechnology, 12(47): 6574- 6582.
  18. Sneath, P.H.A. and Sokal, R.R. (1973). Numerical Taxonomy. San Francisco: Freeman.
  19. Souframanien, J. and Gopalakrishna, T. (2004). A comparative analysis of genetic diversity in black gram genotypes using RAPD and ISSR markers. Theoretical and Applied Genetics, 109: 1687-1693.
  20. Sultana, T., Ghafoor, A. and Ashraf, M. (2006). Geographic pattern of diversity of cultivated lentil germplasm collected from Pakistanas assessed by protein assays. Acta Biologica Cracoviensia, series Botanica, Poland, 48: 77-84. 
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    Research Article

    volume 41 issue 2 (april 2018) : 175-181,   Doi: 10.18805/LR-3401

    Assessment of genetic diversity in black gram [Vigna mungo (L.) Hepper] genotypes based on ISSR

    D
    Divya Vyas
    A
    Arunabh Joshi
    G
    Ganesh Rajamani
    D
    Devendra Jain
    1Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur–313 001, Rajasthan, India.

    • Submitted24-12-2014|

    • Accepted21-03-2017|

    • First Online 20-01-2018|

    • doi 10.18805/LR-3401

    Cite article:- Vyas Divya, Joshi Arunabh, Rajamani Ganesh, Jain Devendra (2018). Assessment of genetic diversity in black gram [Vigna mungo (L.) Hepper]genotypes based on ISSR. Legume Research. 41(2): 175-181. doi: 10.18805/LR-3401.

    ABSTRACT

    A study was carried out among 22 black gram genotypes to study the genetic diversity using 20 ISSR primers. Screening of the primers revealed that only 15 out of the 20 primers produced amplification.  A total of 84 amplified bands were obtained, out of which 72 were polymorphic 85.71 percent polymorphism. The total number of amplified bands varied between 1 (UBC-813 and UBC-878) and 9 (UBC-826) with an average of 5 bands per primer. The overall size of PCR amplified products ranged between 250 bp to 2000 bp. PIC values ranged from 0.00 to 0.51 with an average of 0.285 across all genotypes. Five unique bands were detected in four genotypes, out of which the genotype U-9 gave maximum number of distinct bands. The size of these unique bands ranged from 450 bp to 2000 bp. Based on the UPGMA derived dendrogram and PCA, the 22 genotypes could be divided into four main clusters. While Cluster I included 16 genotypes, the Clusters II, III and IV included two genotypes each. Genotypes IC-16511 and UTTARA, UH-177 and IPU2K-21, STY-2834 and UH-177 were found to be genetically distant from each other with a minimum similarity value of 0.42. The results are encouraging with the suggestion that the ISSR marker could prove to be a versatile tool in further screening of the Vigna germplasm pool for study of genetic divergence and the establishment of phylogenetic relationship amongst accessions.

    REFERENCES

    1. Abd El-Hady, E.A.A., Haiba, A.A.A., Abd El-Hamid, N.R., Al-Ansary, A.M.F. and Mohamed, A.Y. (2010). Assessment of Genetic Variations in Some Vigna Species by RAPD and ISSR Analysis. New York Science Journal, 3(11): 120-128.
    2. Ajibade, S.R., Weeden, N.F. and Chite, S.M. (2000). Inter simple sequence repeat analysis of genetic relationships in the genus Vigna. Euphytica, 111(1): 47-55.
    3. Das, S., Das, S.S. and Ghosh, P. (2014). Analysis of genetic diversity in some black gram cultivars using ISSR. European Journal of Experimental Biology, 4(2): 30-34.
    4. Despeiger, A., Goubely, C., Lenoir, A., Cocherel, S., Picard, G., Rayanl, M., Greltel, F. and Delseny. (1995). Identification of the most represented repeat motif in Arabidopsis thaliana microsatellite loci. Theoretical and Applied Genetic, 91: 160-168.
    5. 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.
    6. Jaccard, P. (1908). Nouvelles researches surla distribution florale. Societe Vaudoise des Science Natulelles Bulletin, 44: 233-170.
    7. Kanimozhi, M., Jayamani, P. and Nadarajan, N. (2009). Genetic diversity as assessed by ISSR markers in Black gram [Vigna mungo (L.) Hepper]. Electronic Journal of Plant Breeding, 1: 12-17.
    8. Lukoki, L., Marechal, R. and Otoul, E. (1980). Les ancetres sauvages des haricots cultives: Vigna radiata (L.) Wilczek et V. mungo (L.) Hepper. Bull. Jard. Bot. Nat. Belgique., 50: 385-391.
    9. Nisar, M., Ghafoor, H., Ahmad, K.M., Rashid, A.S., Qureshi, H. and Ali, H. (2008). Evaluation of genetic diversity of pea germplasm through phenotypic traits analysis. Pakistan Journal of Botany, 40(5): 2081-2088.
    10. Pardhe, D.D. and Satpute, R.A. (2011). A Comparative Analysis of Genetic Diversity in Genus Vigna savi Genotypes using ISSR. International Journal of PharmaTech Research, 3(1): 464-470.
    11. Reddy, M.P., Sarla, N. and Siddiq, E.A. (2002). Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica, 128: 9-17.
    12. Rocha, R., Abad, J., Pires, I. and Araujo, E. (2002). Fingerprint and genetic diversity analysis of Eucalypts spp.genotypes using RAPD and SSR markers, Scientia Forestalis., 62: 24-31.
    13. Rohlf, F.J. (2004). NTSYS-PC. Numerical taxonomy and multivariate analysis system. Exeter Software, New York.
    14. Russel, J.R., Fuller, J.D., Macaulay, M., Hatz, B.G. and Jahoor, A. (1997). Direct comparison of levels of genetic variation among barley accessions detected by RFLPs, AFLPs, SSRs and RAPDs. Theoretical and Applied Genetics, 95: 714-722. 
    15. Sambrook, J., Fritschi, E.F. and Maniatis, T. (1989). Molecular cloning: a Laboratory Manual, Cold Spring Harbor Laboratory Press, New York. 
    16. Shafique, S., Khan, M.R., Nisar, M. and Rehman, S.U. (2011). Investigation of genetic diversity in black gram [Vigna mungo (L.) Hepper]. Pakistan Journal of Botany, 43(2): 1223- 1232.
    17. Singh, R., Heusden, A.W.V. and Yadav, R.C. (2011). A comparative genetic diversity analysis in mungbean (Vigna radiata L.) using inter-simple sequence repeat (ISSR) and amplified fragment length polymorphism (AFLP). African Journal of Biotechnology, 12(47): 6574- 6582.
    18. Sneath, P.H.A. and Sokal, R.R. (1973). Numerical Taxonomy. San Francisco: Freeman.
    19. Souframanien, J. and Gopalakrishna, T. (2004). A comparative analysis of genetic diversity in black gram genotypes using RAPD and ISSR markers. Theoretical and Applied Genetics, 109: 1687-1693.
    20. Sultana, T., Ghafoor, A. and Ashraf, M. (2006). Geographic pattern of diversity of cultivated lentil germplasm collected from Pakistanas assessed by protein assays. Acta Biologica Cracoviensia, series Botanica, Poland, 48: 77-84. 
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