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Theoretical and Applied Genetics

, Volume 112, Issue 2, pp 347–357 | Cite as

Identification of microsatellite markers from Cicer reticulatum: molecular variation and phylogenetic analysis

  • Niroj Kumar Sethy
  • Shalu Choudhary
  • Bhumika Shokeen
  • Sabhyata BhatiaEmail author
Original Paper

Abstract

Microsatellite sequences were cloned and sequenced from Cicer reticulatum, the wild annual progenitor of chickpea (C. arietinum L.). Based on the flanking sequences of the microsatellite motifs, 11 sequence-tagged microsatellite site (STMS) markers were developed. These markers were used for phylogenetic analysis of 29 accessions representing all the nine annual Cicer species. The 11 primer pairs amplified distinct fragments in all the annual species demonstrating high levels of sequence conservation at these loci. Efficient marker transferability (97%) of the C. reticulatum STMS markers across other species of the genus was observed as compared to microsatellite markers from the cultivated species. Variability in the size and number of alleles was obtained with an average of 5.8 alleles per locus. Sequence analysis at three homologous microsatellite loci revealed that the microsatellite allele variation was mainly due to differences in the copy number of the tandem repeats. However, other factors such as (1) point mutations, (2) insertion/deletion events in the flanking region, (3) expansion of closely spaced microsatellites and (4) repeat conversion in the amplified microsatellite loci were also responsible for allelic variation. An unweighted pairgroup method with arithmetic averages (UPGMA)-based dendrogram was obtained, which clearly distinguished all the accessions (except two C. judaicum accessions) from one another and revealed intra- as well as inter-species variability in the genus. An annual Cicer phylogeny was depicted which established the higher similarity between C. arietinum and C. reticulatum. The placement of C. pinnatifidum in the second crossability group and its closeness to C. bijugum was supported. Two species, C. yamashitae and C. chorassanicum, were grouped distinctly and seemed to be genetically diverse from members of the first crossability group. Our data support the distinct placement of C. cuneatum as well as a revised classification regarding its placement.

Keywords

Microsatellite Sequence Microsatellite Motif Cicer Species Crossability Group STMS Marker 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

The authors would like to thank NCPGR, an autonomous institute of Department of Biotechnology, Government of India, for providing financial support to this work. NKS and BS are recipients of fellowships from the Council for Scientific and Industrial Research (CSIR), Government of India. SC is a recipient of a fellowship from the University Grants Commission (UGC), Government of India. We are grateful to ICRISAT, India, and the Indian Agricultural Research Institute (IARI), India, for providing seed material.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Niroj Kumar Sethy
    • 1
  • Shalu Choudhary
    • 1
  • Bhumika Shokeen
    • 1
  • Sabhyata Bhatia
    • 1
    Email author
  1. 1.National Centre for Plant Genome ResearchJawaharlal Nehru University CampusNew DelhiIndia

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