四株马铃薯甲虫白僵菌菌株的鉴定  

Identification of Four Beauveria bassiana Strains from Colorado Potato Beetle

作　　者：马金奉[1] 邓春生[1] 张燕荣[1] 王妮珊[1] 耿兵[1] 

机构地区：[1]中国农业科学院农业环境与可持续发展研究所农业清洁流域团队,北京100081

出　　处：《新疆农业科学》2014年第5期876-884,共9页Xinjiang Agricultural Sciences

基　　金：国家公益性行业(农业)科研专项(201103026-2)

摘　　要：【目的】通过形态学特征明确四株马铃薯甲虫致病菌分类地位,探讨利用18S及ITS核酸序列鉴定四株致病菌的可行性。【方法】通过传统形态学特征及18S、ITS序列分别对四株马铃薯甲虫白僵菌1572、1573、1576和1577进行鉴定。【结果】孢子形态、产孢结构、菌落等形态学特征表明四菌株均为球孢白僵菌。四菌株18S rDNA序列分析获得1 665个共同碱基,在GenBank比对,从结果中选择相似度大于98%的序列与四菌株一起构建系统发育树Ⅰ,发现白僵菌属归于一个分支,四菌株均归属于白僵菌属球孢白僵菌。剔除遗传关系比较远的、冗余的属外菌重新构建18S系统发育树Ⅱ,结果四菌株都与Beauveria bassiana(登录号为EU334676.1)分到一个枝上。ITS核酸序列分析获得513个共同碱基,在GenBank比对,显现的序列均属白僵菌属,下载这些序列与四菌株构建ITS系统发育树Ⅰ,发现球孢与布氏白僵菌归属在一个分支。选取Stephen A.Rehner注册的白僵菌属六个分支的代表种及数据库中部分球孢和布氏白僵菌与四菌株一起构建ITS系统发育树Ⅱ,四菌株与代表种Cordyceps bassiana(AY532041.1)分到一个枝上,但在该枝上有三株布氏白僵菌,球孢与布氏白僵菌不能完全区分开。【结论】利用18S和ITS核酸序列均可将白僵菌鉴定到属,根据ITS序列可以确定四株白僵菌归属于球孢白僵菌的分支。[ Objective] In order to clarify species of four fungi strains which can infect Colorado potato beetle effectively and discuss feasibility of identifying them by 18S and ITS rDNA sequence. [ Method] In the research, species of four strains were identified by method of traditional morphology, and at the same time, 18S and ITS rDNA sequence were used to classify four strains named as 1572, 1573, 1576 and 1577. [ Resuit] Four strains were all classified as Beauveria bassiana by their morphological features such as spore mor- phology, spore structure and colonies. The four strains had 1 665 bp same conserved domain of 18S rDNA, phylogenetic tree I was constructed based on 18S rDNA sequence and the strains whose maximum likelihood were more than 98%. It showed that genus Beauveria constituted a branch, and strains in this research all be- longed to Beauveria bassiana. Then phylogenetic tree II was constructed based on 18S rDNA and the se- quences similar to four strains, which showed that the four strains were close to Beauveria bassiana （EU334676.1） and they were assigned to the same branch. At the same time, the four strains had 513 bp common sequences of ITS. When the common sequence was submitted to GenBank, similar sequence ap- peared, but they all belonged to genus Beauveria. All the sequences appeared in the database were downloaded to construct phylogenetic tree I with the four strains, and the result was that this tree could be divided into sev- eral branches, but 56 Beauveria bassiana and 13 Beauveria brongniartii could not be separated, because they were divided into a common branch. Six different species of Beauveria from Stephen A. Rehner submitted were chosen to construct phylogenetic tree II with some strains in the database and the four strains, the result was that the four strains were close to Cordyceps bassiana （ AY532041.1 ）, and they were assigned to the same branch, but there were three Beauveria brongniartii in this branch, too, Beauveria bassiana and Beauveria brongniartii still could n

关 键 词：马铃薯甲虫 产孢结构 系统发育学分析 球孢白僵菌 鉴定 

分 类 号：Q93-331[生物学—微生物学]