基于生物学特性及分子标记的贵州蜜环菌遗传多样性分析  

Genetic Diversity Analysis of Halimasch(Armillaria mellea)Resources in Guizhou Based on Biological Traits and Molecular Marker

作　　者：王彩云 侯俊 王永 成忠均 柳敏 周茂嫦 李恒谦 张翔宇 阮培均 WANG Cai-yun;HOU Jun;WANG Yong;CHENG Zhong-jun;LIU Min;ZHOU Mao-chang;LI Heng-qian;ZHANG Xiang-yu;RUAN Pei-jun(Bijie Inst.of TCM,Bijie 551700;Rural Rejuvenation Bureau of Dafang County,Bijie 551700)

机构地区：[1]毕节市中药研究所,贵州毕节551700 [2]大方县乡村振兴局,贵州毕节551700

出　　处：《微生物学杂志》2024年第3期49-59,共11页Journal of Microbiology

基　　金：“十二五”国家科技支撑计划项目(2015BAI05B03);贵州省科研机构创新能力建设专项(黔科合服企[2020]4012);贵州省人才基地建设项目(RCJD2020-21);贵州省科技计划项目(黔科合支撑[2020]4Y098号);广州市科技计划项目(201903010008);毕节市创新平台建设项目(毕科合[2022]10号)。

摘　　要：为研究贵州天麻主产区蜜环菌的生物种及其遗传多样性,在贵州天麻主产区采集蜜环菌样本,利用双抗培养基分离菌索或子实体、菌材,定期观察并统计蜜环菌母种及原种的生物学特性;挑取菌索提取总DNA,利用真菌通用引物ITS和延伸因子EF1-α基因扩增rDNA-ITS、rDNA-EF1-α片段并送测序及鉴定;用DNAStar软件进行GC含量分析及多序列比对,用MEGA软件构建进化树。从贵州毕节各县区、黔东南州雷山县等分离到17株蜜环菌,并与生产上普遍用的A9、京234做比较,通过生物学特性初步分析各蜜环菌的形态差异;通过rDNA-ITS、rDNA-EF1-α扩增,得到19株菌株的rDNA-ITS、rDNA-EF1-α完整序列,经GC含量分析、多序列比对、BLAST比对鉴定以及系统发育分析,将19株菌株分为3个种。通过对蜜环菌菌株生物学特性及rDNA-ITS、rDNA-EF1-α序列分析,发现贵州天麻主产区蜜环菌野生资源主要为Armillaria sp.,其次为Armillaria mellea、Armillaria gallica。Biological species and genetic diversity of halimasch(Armillaria mellea)were studied in main producing areas of Guizhou rhizoma gastrodiae(Gastrodia elata).The samples were collected in representative main producing areas,whose cordules or fruiting bodies and bacterial materials were isolated by double antibody medium,and the biological characteristics of the mother species and original species of Armillaria were observed and counted regularly.Total DNA was extracted from the fungicolor,and the rDNA-ITS,rDNA-EF1-α fragments were amplified by fungal universal primers ITS and EF1-α,that were sequenced.The Contig Express software and the DNAStar software were used to concatenate the sequences and used for GC content analysis and multiple sequence alignment,respectively,and the MEGA software was used to construct the evolutionary tree.The total seventeen Armillaria strains were isolated from samples of each County of Bijie Guizhou Province and Leishan County of southeast Qian,compared with A9 and Jing234 common used in production,the whose differences of the morphology were preliminarily analyzed by biological characters.The complete rDNA-ITS,rDNA-EF1-αsequences of obtained 19 strains by rDNA-ITS and rDNA-EF1-αamplification were divided into 3 species by GC content analysis,multiple sequence alignment,BLAST alignment and phylogenetic analysis.By analyzing the biological characteristics and rDNA-ITS,rDNA-EF1-αsequence of Armillaria strains,it is found that the resources in Guizhou Province was Armillaria sp.,followed by A.mellea and A.gallica,the most widely distributed resources in Guizhou Province was Armillaria sp..

关 键 词：蜜环菌 天麻 生物学特性 RDNA-ITS 延伸因子 

分 类 号：Q939.5[生物学—微生物学]