Wen Ying, Lu Xiao-Ping, Ren Rui, Mi Fu-Gui, Han Ping-An, Xue Chun-Lei
College of Agronomy, Inner Mongolia Agricultural University, Huhhot, China.
Yi Chuan. 2013 Feb;35(2):225-32. doi: 10.3724/sp.j.1005.2013.00225.
A total of 57 498 non-redundant ESTs were identified from 210 878 ESTs of Sorghum in NCBI by sequence analysis. In all, 3 338 SSRs were distributed in 3 116 ESTs with an average frequency of one SSR per 11.28 kb, which included 215 SSR motifs. Analysis of SSR motifs revealed that the trinucleotides were major motifs, accounting for 68.33%.The dinucleotides motifs accounted for 17.97%. There were 1 694 sequences from 3 338 EST-SSR sequences could be designed into primers and the proportion was 50.75%. Fourteen primers were selected to amplify EST-SSR loci with 50 collections of Sorghum bicolor × S. sudanenes, 7 collections of S. bicolor and 3 collections of S. sudanenes. Seventy-two allele variations were detected and the frequency was 5.14 gene loci per primer. The polymorphism index of each primer was in the range of 0.54-0.93. The genetic distance ranged from 0.1646 to 0.6398. This showed abundant genetic diversity in the materials. The materials were divided into 5 groups with clustering analysis of EST-SSR data. Each group included the varieties with similar parents or similar regional distribution. Meanwhile, 4 specific molecular markers were found. Primer D1763 was specific in the registered variety GB-4-2 which was the progeny of S. bicolor 314A × S. sudanenes White Skull. The marker was specific in justification of the germ difference. These results showed that the EST-SSR was an effective marker for genetic diversity analysis and specificity studies on S. bicolor × S. sudanenes.
通过序列分析,从NCBI中高粱的210878条EST中鉴定出总共57498条非冗余EST。共有3338个SSR分布在3116条EST中,平均频率为每11.28kb有一个SSR,其中包括215个SSR基序。对SSR基序的分析表明,三核苷酸是主要基序,占68.33%。二核苷酸基序占17.97%。3338条EST-SSR序列中有1694条序列可以设计成引物,比例为50.75%。选择14对引物对50份双色高粱×苏丹草、7份双色高粱和3份苏丹草进行EST-SSR位点扩增。共检测到72个等位基因变异,每个引物的频率为5.14个基因位点。每个引物的多态性指数在0.54-0.93范围内。遗传距离在0.1646至0.6398之间。这表明材料中存在丰富的遗传多样性。利用EST-SSR数据进行聚类分析,将材料分为5组。每组包括亲本相似或区域分布相似的品种。同时,发现了4个特异性分子标记。引物D1763在登记品种GB-4-2中具有特异性,该品种是双色高粱314A×苏丹草白骷髅的后代。该标记在鉴别种质差异方面具有特异性。这些结果表明,EST-SSR是分析双色高粱×苏丹草遗传多样性和特异性研究的有效标记。