双孢蘑菇SCAR标记的建立及在菌株群鉴定中的应用

Establishment of SCAR Markers and its Aplication for Identification of Strain-Groups in Agaricus Bisporus

根据15条具有菌株群特异性的SRAP、ISSR、RAPD扩增产物DNA条带测序结果,设计64对SCAR引物,其中6对引物能成功扩增出目的条带,即有6条特异片段(A、C、E、G、H、J)被成功转化为SCAR标记。通过这6个SCAR标记的不同组合,将40个双孢蘑菇菌株分为9个类群和6个类群,与基于SRAP、ISSR、RAPD资料的聚类分析结果的组间距离值取13和16时的分类结果完全吻合。对A片段的SCAR-PCR检测结果是:可从40个双孢蘑菇菌株中鉴别出双孢蘑菇333号菌株(编号01)。

Fifteen strain or strain groups-specific DNA bands amplificated from SRAP,ISSR,RAPD PCR were selected to be sequenced. According to the sequence data,64 pairs of SCAR primer were designed for SCAR-PCR test. Six pairs of SCAR primer could be used to amplify 6 objective bands successfully. It indicated that these 6 specific fragments marked as A,C,E,G,H,J respectively,were converted into SCAR marker successfully,by their relevant SCAR primer pairs A1,C4,E5,G5,H2,J3 respectively. Through combination among A1,C4,E5,G5,J3 fragments,40 strains could be divided into 6 and 9 strain groups. This result is agreement completely with the result that 40 varieties were classified into 6 and 9 major variety groups at 16 and 13 of distance value in the genetic relationship dendrogram. Additionally,sample 333（No.01） could be identified by PCR with primer pair A1,directly.