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从山东菏泽和泰安两地感病芍药(Paeonia lactiflora Pall.)植株的叶片上进行了红斑病病原分离纯化和致病力检测,并对致病菌进行了形态学鉴定及rDNA-ITS序列分析。研究结果表明,病样组织中可分离到两种疑似病原菌,分离比例为4.8︰1,初步鉴定为链格孢(Alternaria alternata)和细极链格孢(A.tenuissima)。进一步rDNA-ITS分析结果表明,两种病原菌5.8S rDNA及其两侧ITS区序列分别与Gen Bank中序列号为KF380822.1、HM467832.1的同源性最高,均为99%,由此确定两种病原菌分别为链格孢Alternaria alternata和细极链格孢A.tenuissima,其序列已在Gen Bank登录,登录号分别为KR912224和KR912225。因此,确定山东省芍药红斑病是由链格孢A.alternata和细极链格孢A.tenuissima两种病原菌引起的复合侵染。
The pathogen was isolated and purified from leaves of Paeonia lactiflora Pall. In Heze and Tai’an of Shandong Province. Morphological identification and rDNA-ITS sequence analysis of pathogenic bacteria were carried out. The results showed that two kinds of suspected pathogenic bacteria were isolated from diseased tissues, with a separation ratio of 4.8︰1, initially identified as Alternaria alternata and A.tenuissima. Further rDNA-ITS analysis showed that the 5.8S rDNA sequences of the two pathogenic bacteria and ITS sequences on the two sides were the highest homologies with GenBank with the sequence numbers of KF380822.1 and HM467832.1, both of which were 99% The two pathogens were Alternaria alternata and A. tenuissima, respectively. The sequences of these two strains were registered in Gen Bank with accession numbers KR912224 and KR912225, respectively. Therefore, it was confirmed that the peony erythematosus in Shandong province was caused by the complex infection caused by two pathogenic bacteria, A. alternata and A.tenuissima.