Assessment of genetic diversity in clusterbean using nuclear rDNA and RAPD markers

Authors

  • RAKESH PATHAK Central Arid Zone Research Institute, Jodhpur - 342 003, Rajasthan, India Author
  • S.K. SINGH Central Arid Zone Research Institute, Jodhpur - 342 003, Rajasthan, India Author
  • MANJIT SINGH Directorate of Mushroom Research, Solan - 173 213, H.P., India Author

DOI:

https://doi.org/10.59797/jfl.v24i3.1264

Keywords:

Clusterbean, Cyamopsis tetragonoloba, Genetic diversity, ITS sequencing, RAPD, SNP

Abstract

The extent of genetic variability among 18 genotypes of clusterbean using genetic markers was assessed. Multi-locus genotyping by 9 RAPD primers detected 87.6% intra-specific polymorphism. UPGMA dendrogram clearly delineated the genotypes into four clusters irrespective of their place of origin. Analysis of molecular variance indicated that a greater proportion of total genetic variation exists within population rather than among populations. PCR-based amplification of ITS regions encompassing 5.8S rRNA gene yielded a single band of uniform length of 618 bp in all the samples sequenced. However, multiple sequence alignment of ITS regions of representative genotypes belonging to major RAPD clusters exhibited single nucleotide polymorphism. Thus, these two marker systems validated the existence of genetic variation in studied clusterbean genotypes which can be used in crop improvement.

References

Bisht IS, Mahajan RK and Kawalkar TG. 1998. Diversity in mung bean (Vigna radiata L. Wilczek) germplasm collection and its potential use in crop improvement. Annals of Applied Biology 132: 301-312.

Carvalho A, Guedes-Pinto H and Lima Brito J. 2009. Genetic diversity among old Portugese bread wheat cultivars and Botanical varieties evaluated by ITS rDNA PCR-RFLP markers. Journal of Genetics 88: 363-367.

Casiva PV, Saidman BO, Vilardi JC and Cialdella AM. 2002. First comparative phenetic studies of Argentinean species of Acacia (Fabaceae), using morphometric isozymal and RAPD approaches. American Journal of Botany 89: 843-853.

Cottrell JE, Forrest GI and White MS. 1997. The use of random amplified DNA markers to identify and estimate the relatedness of clones belonging to the genus Populus. Botanical Journal of Scotland 49:89-102.

Doyle JJ and Doyle JL. 1990. Isolation of plant DNA from fresh tissue. Focus 12: 13-15.

Dwivedi NK, Bhandari DC, Bhatnagar N. and Dabas BS.1999. characterization of clusterbean [Cyamopsis tetragonoloba (L) Taub.] germplasm for yield and quality traits. Annals of Arid Zone 38: 151-156.

Esselman EJ, Crawford D, Brauner S, Stussy IF, Anderson GJ and Silva MO. 2000. RAPD marker diversity within and divergence among species of Dendroseris (Asteraceae: Lactuceae). American Journal of Botany 87: 591-596.

Esselman EJ, Jianqiamg L, Crawford DJ, Windus JL and Welfe AD. 1999. Clonal diversity in the rare Calamagrostis porteri spp. Insperata (Poaceae): comparative results for allozymes and random amplified polymorphic DNA and inter-simple sequence repeat markers. Molecular Ecology 8: 443-453.

Kakani RK, Singh SK, Pancholy A, Meena RS, Pathak R and Raturi A. 2011. Assessment of genetic diversity in Trigonella foenum-graecum based on nuclear ribosomal DNA, internal transcribed spacer and RAPD analysis. Plant Molecular Biology Reporter 29: 315–323.

Lavanya GR, Srivastava S and Ranade SA. 2008. Molecular assessment of genetic diversity in mung bean germplasm. Journal of Genetics 87: 65-74.

Nalini E, Bhagwat SG and Jawali N. 2007. Identification and Characterization of some ITS variants from hexaploid wheat (Triticum aestivum L.). Plant Science 173: 262-268.

Pathak R, Singh M and Henry A. 2009. Genetic divergence in clusterbean (Cyamopsis tetragonoloba) for seed yield and gum content under rainfed conditions. Indian Journal of Agriculture Sciences 79: 559-61.

Peakall R and Smouse PE 2006. GENALEX 6: genetic analysis in Excel. Population software for teaching and research. Molecular Ecology Notes 6: 288-295.

Ray Choudhury P, Singh IP, Verma AK, George B and Datta S. 2007. Identification of genotype specific markers and assessment of genetic relatedness among pigeonpea cultivars using RAPD. Journal of Food Legumes 20: 12-15.Rohlf FJ. 1997. NTSYS pc: Numerical Taxonomy and Multivariate analysis system Version 2.02h. Exeter software, New York.

Saini A, Reddy SK and Jawali N. 2008. Intra individual and intra species heterogeneity in nuclear rDNA ITS region of Vigna species from subgenus Ceratotropis. Genetical Research 90: 299-316.

Singh NP, Choudhary AK and Chaudhary SPS. 2003. Genetic divergence in clusterbean. Indian Journal of Agricultural Sciences 73: 356-357. Sneath PHA and Sokal R.R. 1973. Numerical Taxonomy. W.H. Freeman, San Francisco, California.

Sogin ML. 1990. Amplification of ribosomal RNA genes for molecular evolution studies. In: MA Innis, DH Geltand, TT Sninsky and TJ White (Eds), PCR protocols: A guide to methods and applications, Academic press, San Diego, California. Pp: 307-314.

White TJ, Bruns SL and Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: MA Innis, DH Geltand, TT Sninsky and TJ White (Eds), PCR protocols: A guide to methods and applications, Academic press, San Diego, California. Pp: 315-322.

Williams JGK, Kubelik AR, Livak KJ, Rafalski JA and Tingey V. 1990. DNA Polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Research 18: 6531-6535.

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Published

2025-01-04

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How to Cite

Assessment of genetic diversity in clusterbean using nuclear rDNA and RAPD markers. (2025). Journal of Food Legumes, 24(3), 180-183. https://doi.org/10.59797/jfl.v24i3.1264