Molecular markers for Nassella trichotoma (Poaceae) to study genetic variation in New Zealand

利用分子标记研究新西兰三裂针茅(禾本科)的遗传变异

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Abstract

PREMISE OF THE STUDY: Simple sequence repeat (SSR) markers were developed for the study of genetic diversity of New Zealand Nassella trichotoma (Poaceae) and to support future studies in its native range. METHODS AND RESULTS: Genomic DNA was extracted from N. trichotoma leaf material and subjected to Roche 454 sequencing. From a total of 745 putative SSRs, 48 with di- to pentanucleotide repeats were screened, 32 primer pairs were designed, and 15 polymorphic markers were optimized for multiplex PCR on 105 N. trichotoma samples from four New Zealand regions. Each locus resulted in two to six alleles per locus, and four of the loci cross-amplified in N. tenuissima. The mean observed and expected heterozygosity ranged from 0.00 to 0.90 and 0.00 to 0.50 per locus, respectively. CONCLUSIONS: The novel SSR markers are valuable for the study of genetic diversity of N. trichotoma and might also be useful for closely related species.

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