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为明确毒莴苣EST序列中SSR的特点,并开发EST-SSR分子标记用于毒莴苣及其近似种的分类鉴定及遗传多样性分析,以筛选的7 438条毒莴苣EST序列为基础,利用SSRIT软件结合人工搜索查找SSR位点,并采用Primer Premier 5.0软件设计EST-SSR引物,以毒莴苣及其近似种为材料对所合成引物进行多态性检测,应用UPGMA法构建11个毒莴苣及其近似种的系统发育树。结果表明,共搜索到SSR位点1 465个,发现重复基元598种,其中二、三、六核苷酸重复是主要的重复类型。合成的24对引物中有9对表现出多态性,利用这9对EST-SSR引物分析毒莴苣等11份材料的亲缘关系,其相似性系数在0.432~1.000之间;于0.513处可被划分为3大类:Ⅰ类包括翅果菊和北山莴苣,Ⅱ类只有宿根莴苣1种,Ⅲ类包括毒莴苣和莴苣。
In order to clarify the characteristics of SSR in toxic lettuce EST sequences and develop EST-SSR molecular markers for the classification and genetic diversity analysis of toxic lettuce and its approximate species, based on the 7 438 EST sequences screened from Lettuce, SSRIT The software combined with artificial search to find SSR loci, and Primer Premier 5.0 software designed EST-SSR primers to toxic lettuce and its approximate species as the material to detect the polymorphism of the primers synthesized by UPGMA method to construct 11 poisonous lettuce and its Approximate species of phylogenetic tree. The results showed that a total of 1 465 SSR loci were found and 598 repeat motifs were found. Among them, two, three and six nucleotide repeats were the major repeat types. Nine of the 24 pairs of primers showed polymorphism. The 9 pairs of EST-SSR primers were used to analyze the genetic relationship of 11 materials such as poison lettuce with the similarity coefficient of 0.432-1.000; Divided into three categories: Ⅰ category, including plum-bergamot and Kitayama lettuce, Ⅱ only perennial lettuce 1, Ⅲ category, including lettuce and lettuce.