10株桑黄菌基于rDNA ITS序列的分子鉴定

Molecular Identification of 10 Phellinus Strains Based on rDNA ITS Sequences

建立桑黄菌的分类学基本框架,可为桑黄菌的开发利用奠定基础。利用rDNAITS序列分析技术,在分子水平上对采自秦巴山区的9株野生桑黄菌和1株实用桑黄菌进行分类鉴定。以真菌rDNAITS序列分析中的通用引物ITS1和ITS4,分别对10个菌株的基因组DNA模板PCR扩增出目的片段,将扩增产物割胶纯化、克隆并测序,获得10个菌株的rDNAITS序列,并结合GenBank中已登录桑黄菌的rDNAITS同源序列,应用MEGA4.1软件和Neighbor-Joining法,分别计算遗传距离及构建系统发育进化树分析亲缘关系。结果表明10株桑黄菌均为针层孔菌属(Phellinus),其中:S1、S4、S6、Rh、Sc菌株初步鉴定为鲍氏针层孔菌(Phellinus baumii);S2、S3、S5、S7、Gy菌株初步鉴定为裂蹄木层孔菌(Phellinus linteus)。

The establishment of basic taxonomic features for Phellinus would facilitate utilization of this fungus.By rDNA ITS sequence analysis,9 wild Phellinus strains collected from the Qinling Mountains and 1 Phellinus strain at practical use were classified and identified at the molecular level.Two universal primers ITS1 and ITS4 which are generally used in the analysis of fungal rDNA ITS sequences were used to amplify genomic DNAs of the 10 Phellinus strains respectively.The amplified products were isolated,purified,cloned and sequenced,yielding rDNA ITS sequences of the 10 strains.MEGA 4.1 software and Neighbor-Joining algorithm were employed to calculate genetic distances and construct phylo-genetic trees to analyze their lineage relationships with the known homologous sequences registered in GenBank.The results showed that all 10 strains belonged to genus Phellinus sp.（Phellinus）,where strains S1,S4,S6,Rh,and Sc were preliminarily identified as Powell Phellinidium （Phellinus baumii）,and strains S2,S3,S5,S7,and Gy were preliminarily identified as Fungus Phellinidium （Phellinus linteus）.