摘要： 为了分析不同地区黄瓜棒孢叶斑病菌的种内分化,能够准确地鉴定该病菌,以服务于黄瓜抗病育种,对2007-2014年采集的黄瓜棒孢叶斑病菌多主棒孢经单孢纯化获得34个菌株。利用公开发表的特异性引物CCF/CCR对其进行了PCR分子鉴定,结果表明:34个菌株均扩增得到了预期272 bp的特异片段,说明均为多主棒孢菌。应用真菌核糖体rDNA区通用引物ITS1和ITS4分别扩增各个菌株的rDNA内转录间隔区序列,经测序并比对分析,34个菌株均扩增得到了559 bp的特异片段,其中32个从黄瓜上分离的多主棒孢病菌菌株碱基序列完全一致,2个从番茄上分离的菌株碱基序列完全一致,二者的差异在于2个SNP位点T-C、G-A的突变,二者相似度为99.64%。说明,多主棒孢菌属种间的ITS区序列高度保守,利用ITS序列分析方法可以为多主棒孢菌的分类鉴定及系统发育研究提供依据,可以区分出黄瓜和番茄寄主上的多主棒孢菌。

关键词: 黄瓜, 多主棒孢菌, 分子鉴定, rDNA-ITS

Abstract: In order to analyze the differentiation of different areas Corynespora cassiicola and identify the pathogen accurately so as to serve the cucumber disease resistance breeding,34 corynespora cassiicola purified strains was collected from 2007 to 2014.Molecular identification results showed that all the isolates were Corynespora cassiicola using published specific primers CCF/CCR,all the 34 strains were amplified the expected 272 bp specific fragments.559 bp fragments of ribosomal DNA internal transcribed spacer sequences of corynespora cassiicola were obtained through PCR amplification by using universal primers ITS1 and ITS4.Sequence analysis result showed that there was no difference among the sequences of ribosomal DNA ITS of 32 strains isolated from cucumber,only 2 nucleotides mutation comparing with the sequence of the strains isolated from tomato,the similarity was 99.64%.The above results showed that the ITS sequences were highly conserved in Corynespora Güssow,the ITS sequence might be used for differential identification of species of Corynespora,the ITS sequence can distinguish different isolates of Corynespora cassiicola fromcucumber and tomato host.

Key words: Cucumber, Corynespora cassiicola (Berk.&Curt.)Wei, Molecular identification, rDNA-ITS

中图分类号: 

• S642.2

• Q78