IngentaConnect The determination of multiple microsatellite genotypes and DNA se...

Authors: MATSUKI, Y.¹; ISAGI, Y.²; SUYAMA, Y.³

Source: Molecular Ecology Notes, Volume 7, Number 2, March 2007 , pp. 194-198(5)

Abstract:

Pollen plays important roles in the reproduction and gene flow of flowering plants, and its haploid DNA sequence provides useful information for studies of plant evolution and genealogy. We describe a new method for multiple microsatellite genotyping and DNA sequencing from a single pollen grain. The haploid DNA was extracted from a single pollen grain by using a simple DNA extraction method, and multiple microsatellite genotypes and DNA sequences of multiple chloroplast loci were determined. Using nine pairs of microsatellite primers, more than 90% of genotypes were successfully determined, and 71% and 100% of DNA sequences were determined at two chloroplast DNA loci, the trnL intron region and the trnL/trnF intergenic spacer region, respectively. This simple method of genetic analysis for a single pollen grain will facilitate detailed study of pollination, evolution and genealogy.

Keywords: chloroplast DNA; DNA sequencing; microsatellite; multiplex PCR; single pollen grain; whole genome amplification

Document Type: Research article

DOI: 10.1111/j.1471-8286.2006.01588.x

Affiliations: 1: Graduate School for International Development and Cooperation, Hiroshima University, Kagamiyama 1-5-1, Higashi-Hiroshima 739-8529, Japan, 2: Graduate School of Integrated Arts and Sciences, Hiroshima University, Kagamiyama 1-7-1, Higashi-Hiroshima 739-8521, Japan, 3: Field Science Center, Graduate School of Agricultural Science, Tohoku University, 232-3, Ohkuchi, Naruko-cho, Tamatsukuri-gun, Miyagi 989-6711, Japan

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The determination of multiple microsatellite genotypes and DNA sequences from a single pollen grain