安徽省土壤绿僵菌分离株的分子鉴定

Molecular identification of Metarhizium strains isolated from soil in Anhui

目的进一步验证ITS序列的系统发育分析可为绿僵菌属种的鉴定提供重要的参考依据。方法对分离自安徽土壤的13株绿僵菌菌株的内转录间隔区(ITS)片段进行PCR扩增和序列测定,采用Blast方法将测序结果在GenBank中进行同源搜索,依据邻接法构建获得与其相关菌株的ITS序列系统发育树。结果供试菌株分别位于系统发育树的3个分支上,分支I包括8个菌株和金龟子绿僵菌小孢变种,1个菌株和金龟子绿僵菌鳞鳃金龟变种形成分支III,另外4个菌株和黄绿绿僵菌棉蚜变种聚为分支X。结论结合同源比较的数据,将这8个、4个和1个绿僵菌菌株分别鉴定为金龟子绿僵菌小孢变种、黄绿绿僵菌棉蚜变种和金龟子绿僵菌鳞鳃金龟变种。

Objective To further manifest that the phylogenetic analysis of ITS sequence could provide an important reference basis for genus identification of Metarhizium.Method The internal transcribed spacer（ITS） sequences of l3 Metarhizium strains isolated from Anhui soil was amplified and sequenced.We searched for homology using the BLAST programs available at the web site of the National Center for Biotechnology Information,and the phylogenetic tree for the genus Metarhizium was constructed by Neighbor-Joining method.Result The tested strains were located in 3 clades of the phylogenetic tree.8 Metarhizium strains and the Metarhizium anisopliae var.anisopliae type were classified into clade I,1 strain and the M.anisopliae var.lepidiotum type were grouped into clade III,and 4 strains and the M.flavoviride var.pemphigum type were classified into clade X.Conclusion Combined with the homologous comparative data,the eight strains,the one strain and the four strains are identified as Metarhizium anisopliae var.anisopliae,M.anisopliae var.lepidiotum and M.flavoviride var.pemphigum,respectively.