Ashkenazi V, Chani E, Lavi U, Levy D, Hillel J, Veilleux R E
Department of Genetics, The Hebrew University of Jerusalem, Rehovot, Israel.
Genome. 2001 Feb;44(1):50-62. doi: 10.1139/gen-44-1-50.
Three genomic libraries were constructed using a mixture of DNA from Solanum phureja Juz. & Buk., and S. chacoense Bitt. Two of the libraries were enriched for ATT and GT repeats (a 27-fold enrichment was achieved). In total, 3500 clones of the conventional library, 1,000 of the library enriched for ATT, and 12,000 of the one enriched for GT were screened with five different repeat motifs, and a total of 18 primer pairs was obtained. Another group of 12 primer pairs was obtained from the SSR-containing sequences in the public databases (18 SSR-containing sequences were utilized). From among 30 newly developed primer pairs, 12 previously published ones, and 12 pairs developed for tomato, 7 were used to identify 12 different potato cultivars and introductions, and 12 were used to study phylogenetic distance among seven wild and cultivated potato species. Two SSR markers were sufficient to discriminate the 12 cultivars. The mean number of alleles per polymorphic locus was 5 for the 12 cultivars and 4.5 for the seven species. The results obtained in this study confirm those achieved in similar studies in other plant species regarding the abundance and use of SSR markers in identifying species and cultivars.
使用来自秘鲁茄（Solanum phureja Juz. & Buk.）和查科茄（S. chacoense Bitt.）的DNA混合物构建了三个基因组文库。其中两个文库富含ATT和GT重复序列（实现了27倍的富集）。总共用五种不同的重复基序筛选了常规文库的3500个克隆、富含ATT的文库的1000个克隆以及富含GT的文库的12000个克隆，共获得18对引物。另外12对引物是从公共数据库中含简单序列重复（SSR）的序列获得的（利用了18个含SSR的序列）。从新开发的30对引物、12对先前发表的引物以及为番茄开发的12对引物中，7对用于鉴定12个不同的马铃薯品种和引种材料，12对用于研究7个野生和栽培马铃薯物种之间的系统发育距离。两个SSR标记足以区分这12个品种。12个品种中每个多态性位点的等位基因平均数为5，7个物种为4.5。本研究获得的结果证实了在其他植物物种的类似研究中关于SSR标记在鉴定物种和品种中的丰度及用途所取得的成果。
