双孢蘑菇SSR分子标记开发及其在遗传多样性分析中的应用

采用Primer 5.0软件设计引物，共设计有效扩增SSR引物112对，经筛选，其中97对在39份双孢蘑菇基因组中扩增出明显差异条带，25对在As2796单孢分离子代群体中表现出多态性，19对引物在单孢分离子代中遵循孟德尔自由分离定律。进一步研究表明共检测到67个等位基因，平均每个位点2.68个，平均Neis基因多样性指数为0.502 3。8份双孢蘑菇品种的遗传相似系数变化范围为0.336 3～0.864 1，平均值为0.538 5；基于Nei-s 遗传相似系数的UPGMA聚类分析，在遗传距离0.46处，8个双孢蘑菇品种（系）分为2个类群，亲缘关系的远近与其种质来源有一定的相关性。这些潜在的多态性SSR 的开发为双孢蘑菇遗传多样性分析、图谱构建提供了更丰富的候选分子标记基础，为进一步进行双孢蘑菇新品种的选育和种质资源的分析评价及管理提供了遗传依据。

Development and application of SSR markers in Agaricus bisporus

A total of 112 primer pairs were successfully designed， of which 97 primers produced stable and specific bands in 39 Agaricus bisporus genotypes （including 31 single spore isolates and 8 other strains）. Twenty five primer pairs showed polymorphism among single spore isolates and 19 primer pairs followed the law of segregation in single spore isolates population. Further study showed that 67 alleles were detected with the average of 268.The average of Neis genetic diversity index （H） was 0.502 3. The genetic similarity coefficient ranged from 0.336 3 to 0.864 1 with an average of 0538 5. The UPGMA cluster analysis based on SSR data showed that these parental lines could be classified into two groups at the threshold of 0.46， which was related to their pedigree analysis. The results indicated that molecular markers was usable in map structure， analysis of genetic polymorphism in A.bisporus. It also provided the genetic basis for the breeding of new varieties of A.bisporus or the collection and management of germplasm resources.