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
王彩云 1侯俊 2王永 1成忠均 1柳敏 1周茂嫦 1李恒谦 1张翔宇 1阮培均1
作者信息
- 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
- 折叠
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Armillar-ia mellea、Armillaria gallica.
Abstract
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 bio-logical 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 isola-ted 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 bio-logical characters.The complete rDNA-ITS,rDNA-EF1-α sequences of obtained 19 strains by rDNA-ITS and rDNA-EF1-α amplification were divided into 3 species by GC content analysis,multiple sequence alignment,BLAST align-ment and phylogenetic analysis.By analyzing the biological characteristics and rDNA-ITS,rDNA-EF1-α sequence 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Key words
Armillaria/Gastrorlia elata/biological trait/rDNA-ITS/elongation factor引用本文复制引用
基金项目
"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)
出版年
2024
NETL
NSTL
万方数据