Diversity analysis and genotyping in Pisum with sequence tagged microsatellite site (STMS) primers

Authors: Ford R.1; Le Roux K.1; Itman C.1; Brouwer J.B.2; Taylor P.W.J.1

Source: Euphytica, Volume 124, Number 3, 2002 , pp. 397-405(9)

Publisher: Springer

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Abstract:

Pisum sativum specific sequence tagged microsatellite site primers were used to amplify genomic profiles from 15 accessions of P. sativum L. that represented the genetic base of the Australian field pea-breeding program and five accessions of the wild related species P. fulvum. The STMS primers were used to assess genetic relationships among the Pisum accessions in two ways. Firstly, to produce RAPD-like multiple banding marker profiles using an adapted RAMS method, for intra- and interspecific diversity analysis. From the 14 flanking primer pairs assessed, 133 markers were obtained. Conservation and reproducibility of markers among individuals within accessions was demonstrated. The largest distance observed among P. sativum accessions was 22% and among P. fulvum accessions was 40%, similar to that revealed with other PCR-based methods. The maximum distance between P. sativum and P. fulvum accessions was 46%. Phylogenetic clustering of P. sativum accessions, using the neighbour joining method and based on simple matching distances, was distinct and distant to P. fulvum. Secondly, PCR with a higher annealing temperature and fluorescent labeling identified simple and allelic loci markers useful for creating a genotype/fingerprint database for P. sativum cultivars. This is the first report to demonstrate the use of Pisum specific STMS sequences for both diversity analysis and genotype identification.

Keywords: diversity; fingerprinting; Pisum; RAMS; STMS

Language: English

Document Type: Regular paper

Affiliations: 1: Molecular Plant Genetics and Germplasm Development Group, Department of Crop Production, Joint Centre for Crop Improvement, The University of Melbourne, Victoria 3010, Australia 2: Victorian Institute for Dryland Agriculture, Department of Natural Resources and Environment, Private Bag 260, Horsham, Victoria 3401, Australia

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