Development and mapping of simple sequence repeat markers for pearl millet from data mining of expressed sequence tags

Senthilvel, S and Jayashree, B and Mahalakshmi, V and Sathish Kumar, P and Nakka, S and Nepolean, T and Hash, C T (2008) Development and mapping of simple sequence repeat markers for pearl millet from data mining of expressed sequence tags. BMC Plant Biology, 8 (119). 9pp.

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Abstract

Background: Pearl millet [Pennisetum glaucum (L.) R. Br.] is a staple food and fodder crop of marginal agricultural lands of sub-Saharan Africa and the Indian subcontinent. It is also a summer forage crop in the southern USA, Australia and Latin America, and is the preferred mulch in Brazilian no-till soybean production systems. Use of molecular marker technology for pearl millet genetic improvement has been limited. Progress is hampered by insufficient numbers of PCR-compatible co-dominant markers that can be used readily in applied breeding programmes. Therefore, we sought to develop additional SSR markers for the pearl millet research community. Results: A set of new pearl millet SSR markers were developed using available sequence information from 3520 expressed sequence tags (ESTs). After clustering, unigene sequences (2175 singlets and 317 contigs) were searched for the presence of SSRs. We detected 164 sequences containing SSRs (at least 14 bases in length), with a density of one per 1.75 kb of EST sequence. Di-nucleotide repeats were the most abundant followed by tri-nucleotide repeats. Ninety primer pairs were designed and tested for their ability to detect polymorphism across a panel of 11 pairs of pearl millet mapping population parental lines. Clear amplification products were obtained for 58 primer pairs. Of these, 15 were monomorphic across the panel. A subset of 21 polymorphic EST-SSRs and 6 recently developed genomic SSR markers were mapped using existing mapping populations. Linkage map positions of these EST-SSR were compared by homology search with mapped rice genomic sequences on the basis of pearl millet-rice synteny. Most new EST-SSR markers mapped to distal regions of linkage groups, often to previous gaps in these linkage maps. These new EST-SSRs are now are used by ICRISAT in pearl millet diversity assessment and marker-aided breeding programs. Conclusion: This study has demonstrated the potential of EST-derived SSR primer pairs in pearl millet. As reported for other crops, EST-derived SSRs provide a cost-saving marker development option in pearl millet. Resources developed in this study have added a sizeable number of useful SSRs to the existing repertoire of circa 100 genomic SSRs that were previously available to pearl millet researchers.

Item Type: Article
Divisions: UNSPECIFIED
CRP: UNSPECIFIED
Subjects: Mandate crops > Millets
Depositing User: Users 6 not found.
Date Deposited: 26 Aug 2011 09:53
Last Modified: 26 Aug 2011 10:14
URI: http://oar.icrisat.org/id/eprint/578
Official URL: http://dx.doi.org/10.1186/1471-2229-8-119
Projects: Plant Sciences Research Programme R8183
Funders: Department for International Development, Consultative Group on International Agricultural Research
Acknowledgement: The authors acknowledge Dr MK Reddy from the International Centre for Genetic Engineering and Biotechnology (ICGEB) and Dr AR Reddy from the Central University of Hyderabad for providing the EST sequences, and Mr M Narsi Reddy of ICRISAT for technical assistance throughout this study. This document is an output from a project (Plant Sciences Research Programme R8183) funded by the UK Department for International Development (DFID) and administered by CAZS Natural Resources for the benefit of developing countries. The views expressed are not necessarily those of DFID. Financial support from the Generation Challenge Programme (GCP) of the Consultative Group on International Agricultural Research (CGIAR) is also gratefully acknowledged.
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