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对分离出的链格孢菌株用ISSR和RAPD分子标记技术分析研究其遗传多样性,比较2种分子生物学方法在链格孢遗传分析中的优劣,为研究烟草赤星病菌遗传多样性及烟草抗病品种的培育奠定基础。采用ISSR和RAPD分子标记方法对来自不同地区的28份烟草赤星病菌进行遗传多样性分析,筛选出10个ISSR引物和10个RAPD引物;ISSR扩增出多态性条带112条,多态性条带比率为86.82%,菌株间相似性系数为0.53～0.97;RAPD引物扩增出多态性条带70条,多态性条带比率为81.39%,菌株间相似性系数为0.57～0.94。用SPSS17.0软件对2种标记遗传距离进行相关性分析,发现2种分子标记结果呈显著正相关,表明2种分子标记方法都适合于烟草赤星病菌遗传多样性研究,ISSR是一种多态性优于RAPD的标记技术。根据2种标记的结果,利用NTSYS软件按UPGMA方法进行聚类分析,发现烟草赤星病菌遗传多样性与地理差异没有显著相关性。
The isolates of Alternaria were analyzed by ISSR and RAPD molecular markers to study the genetic diversity of Alternaria alternata. The advantages and disadvantages of two molecular biological methods in genetic analysis of Alternaria alternata were compared. Genetic diversity of Tobacco Alternaria alternata and tobacco The cultivation of resistant varieties laid the foundation. ISSR and RAPD markers were used to analyze the genetic diversity of 28 tobaccos isolates from different regions. Ten ISSR primers and 10 RAPD primers were screened out. ISSR amplified 112 polymorphic bands, The percentage of bands was 86.82%, and the similarity coefficient was 0.53 ~ 0.97. The number of polymorphic bands amplified by RAPD primers was 70, the percentage of polymorphic bands was 81.39%, and the similarity coefficient was 0.57 ~ 0.94. SPSS17.0 software was used to analyze the genetic distance of the two markers and found that there was a significant positive correlation between the two molecular markers, indicating that both of the two molecular markers are suitable for the genetic diversity of R. solani, ISSR is a polymorphism Sex better than RAPD labeling technology. According to the results of two markers, using UPGMA clustering analysis by NTSYS software, we found there is no significant correlation between genetic diversity and geographic differences.