库尔勒香梨的EST-SSR 标记开发及遗传多样性分析
Development of EST-SSR marker and genetic diversity analysis of Korla Fragrant Pear
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摘要: 利用已公开的梨EST 序列(2 398 条)开发SSR 引物,分析梨EST 序列中SSR 分布及特征,针对库尔勒香梨进行引物筛选,并分析受香梨优斑螟为害程度差异较大香梨18 株样本的遗传多样性。结果表明:EST 序列中有1 813 条无冗余序列,141 条含162 个SSR,占全部EST 的7.78%,出现频率8.94%,平均分布距离为4.93 kb,但不同微卫星的重复类型间差异很大(9.51~79.87 kb),二核苷酸重复最为常见,占SSR 比例达到51.85%。从设计的123 对引物中,选取41 对SSR 引物进行筛选,发现17 对引物扩增效果和多态性较好,共检测出111 条多态性条带,等位基因Na 为5.53,有效等位基因数Ne 为3.74,Nei’s 基因多样性指数He 为0.69,Shannon’s 多样性指数I 为1.40,受害较重的G 组香梨个体主要集中在UPGMA 聚类的0.78 分支处。Abstract: The SSR primers were developed by reference to the public 2 398 strips of EST sequence in pear, and by analyzing their distribution and characteristics of SSR markers,then the suitable primers specific to Kuerle Pear were sorted out,to clarify the genetic diversity of 18 pear trees which showed great variation in damage level by disoperation of Euzophera pyriella. The results showed that there were 1 813 strips of the non-redundant sequence in total of 2 398 EST sequence,of which 141 strips contained 162 SSR,amounting to 7.78% of the whole EST sequence. Their appearance rate was 8.94%,and the average distribution distance was 4.93 kb which showed extensively difference among microsatellite sequences(from 9.51 to 79.87 kb). The most common sequence was the dinucleotide repeat,which accounted for 51.85% of the whole. Choosing 41 pairs from the devised 123 pairs of primers to conduct primer screening,it was found that there were 111 strips of sequence with polymorphism bands,among which 17 strips showed favorable amplifying results and polymorphism. The allele Na was 5.53, and the number of valid allele gene Ne was 3.74,the H value of Nei’sgene diversity(H)was 0.69,and I value of Shannon’s information index(I)was 40. The individuals suffering serious damage were concentrated on the branch of 0.78 in the cluster of UPGMA.