rDNA-ITS序列分析法和传统分类法对真菌进行分类鉴定

The Application of rDNA-ITS Sequence Analysis Method and Traditional Classification Method in the Classification of Fungi

通过对四川马尔康地区采集到的三株菌株的子实体进行测定,分别以三株菌种的DNA为模板,利用特异性引物做PCR扩增并测序,测序结果在Gene Bank中进行BLAST比对,找到与之同源性较高的已知菌种,通过运用DNAMAN、MEGA7.0和Clustal X2软件选择N-J法和ML法构建进化树,确定三株菌株种属。再辅以传统的形态学方法结合《中国大型真菌》图谱对菌株进行鉴定。结果表明:F1为皮革黄丝膜菌(Cortinariusmalachius(Fr.:Fr.)Fr.),F2为大丝膜菌(Cortinariuslargus Fr.),F3为黄斑蘑菇(Agaricusxanthodermus Quèl.)。rDNA-ITS序列分析法的应用虽然受核酸序列数据库完善程度的影响,但同时也丰富了数据库信息,提高了真菌分类鉴定模糊性状的区分度。相较于传统的分类法,形态学分类法和rDNA-ITS序列分析法共同对菌株进行鉴定的结果更精准,操作也更省时、快捷。

In this paper,the fruiting body of three strains collected from barkam region of Sichuan province is identified.The DNAs of three strains are respectively used as templates,and polymerase chain reaction(PCR)amplification with DNA specific primers is carried out.Amplified products are sequenced and its results are compared by BLAST in GeneBank to identify the strain with high homology to this sequence.By applying DNAMAN,MEGA7.0and ClustalX2software,Phylogenetic tree is constructed by N J method and ML method,and the genus is preliminarily identified accordingly.And then,the species are identified in accordance with the traditional morphological classification and the atlas of The Macrofungi in China.The results show that the F1is Cortinariusmalachius(Fr.:Fr.)Fr.,the F2is Cortinariuslargus Fr.and the F3is Agaricusxanthodermus Quèl.Although the application of rDNA ITS sequence analysis is affected by the perfection of nucleotide sequence database,it also enriches the database information and improves the division of fuzzy characters in fungi classification and identification.Compared with the traditional classification methods,the combination of rDNA ITS sequence analysis method and morphological classification method is more accurate,time saving and speedy in the identification of fungi.