基于RAPD和EST-SSR标记的秀珍菇菌株聚类分析

Cluster Analysis of Ten Pleurotus geesteranus Strains Based on Random Amplification of Polymorphic DNA (RAPD) and Expressed Sequence Tag- Single Sequence Repeats (EST-SSR) Markers

本研究利用RAPD标记和EST-SSR标记对10个不同来源的秀珍菇菌株进行聚类分析。200条RAPD引物中127条有扩增产物。引物可用率为63.5%。从NCBI数据库中下载秀珍菇及相近侧耳属食用菌EST序列2167条,聚类比对后得到全长为713.541 kb的非冗余EST 1442条,搜寻后得到的29个EST-SSR全部设计引物,其中26对引物(89.7%)显示多态性。根据17条RAPD核心引物和10对EST-SSR核心引物的扩增结果,对10个秀珍菇菌株进行了RAPD标记、EST-SSR标记及二者相结合的聚类分析。3种分析结果相近,且与菌丝、子实体生长特性分析结果相统一—10个供试菌株区分为5组:1～4号菌株为一组,6号和7号菌株为一组,8号和9号菌株为一组,5号和10号菌株各自为一组。

Amplification products were obtained from ten strains of Pleurotus geesteranus using 127 （63.5%） of 200 RAPD primers tested. A total of 2167 Expressed Sequence Tags （ESTs） from P. geesteranus and related Pleurotus species were downloaded from the NCBI database and, after clustering, 1442 nonredundant ESTs with a total length of approximately 713,541 bp were obtained. Twenty-nine Expressed Sequence Tag-Single Sequence Repeats （EST-SSRs） were selected from these non-redundant ESTs and used to design EST-SSR primers. All 29 primer pairs were used for PCR amplification of genomic DNA extracted from the mycelia of ten P. geesteranus strains, and amplification products generated using 26 primer pairs （89.7%） exhibited polymorphisms. A dendrogram based on UPGMA （unweighted pair group method with arithmetic average） cluster analysis using combined data obtained using 17 RAPD core primers and 10 EST-SSR core primer pairs revealed that the 10 strains were clustered into five groups.