中国树花可培养内生真菌分离、鉴定及系统发育分析

Identification and phylogenetic analysis of endophytic fungi in Ramalina sinensis

[目的]了解中国树花内生真菌的多样性及其系统发育关系。[方法]通过常规组织分离法,采用马铃薯葡萄糖琼脂培养基(PDA)分离培养中国树花内生真菌,对内生真菌的菌落、菌丝和孢子进行观察并结合r DNA-ITS序列的PCR扩增测序,对获得的ITS序列通过Gen Bank数据库搜索同源序列并进行分析确定其分类地位并构建系统发育树。[结果]从中国树花中分离得到可培养内生真菌51株,分属4纲、5目、7科、12属,其中青霉属(Penicillium sp.)和曲霉属(Aspergillus sp.)总相对分离率分别为47%、18%,为中国树花内生真菌优势属。[结论]首次对中国树花进行内生真菌的分离,并确定其分类地位,对分离得到的51株内生真菌进行合并相同菌落形态和显微结构,共获得18株内生真菌,通过系统发育分析发现这18株聚为5大类。

[Objective]To understand the diversity and phylogenetic relationship of endophytic fungi in Ramalina sinensis.[Methods]As research materials,Ramalina sinensis was separated by conventional organizations,using potato dextrose agar（PDA） isolated from endophytic fungi,the endophytic fungi colony,mycelium and spores were observed with r DNA-ITS sequence of PCR amplified and sequenced by Gen Bank database search and homology analysis to determine the taxonomic status and the phylogenetic tree of ITS sequence.[Results]Cultured 51 strains of endophytic fungi isolated from Ramalina sinensis,belonging to 4 classes,5 orders,7 families and 12 genera,Penicillium and Aspergillus（Penicillium sp.）（Aspergillus sp.） the total relative separation rate were 47%,18%,for Ramalina sinensis endophytic fungi the advantage is.[Conclusion] This is the first time of Ramalina sinensis to isolate endophytic fungi,and determine the classification status of 51 strains of endophytic fungi were isolated with the same colony morphology and microscopic structure,a total of 18 strains of endophytic fungi by phylogenetic analysis found that 18 strains clustered into 5 categories.