11份蜜环菌种质rDNA-ITS序列分析  

Sequence Analysis of rDNA-ITS of 11 Armillaria mellea Strains

作　　者：莫盛龙 夏宗元 李继 李园园 刘红昌[1,2,4] 黄明进 MO Sheng-long;XIA Zong-yuan;LI Ji;LI Yuan-yuan;LIU Hong-chang;HUANG Ming-jin(College of Agriculture,Guizhou University,Guiyang 550025,China;Dendrobium Research Institute,Guizhou University,Guiyang 550025,China;Huayuan Ecological Agriculture Development Co.Ltd in Fuquan city,Fuquan 550506,China;Guizhou Key Laboratory of Propagation and Cultivation on Medicinal Plants,Guiyang 550025,China)

机构地区：[1]贵州大学农学院,贵州贵阳550025 [2]贵州大学石斛研究院,贵州贵阳550025 [3]福泉市华源生态农业发展有限公司,贵州福泉550506 [4]贵州省药用植物繁育与种植重点实验室,贵州贵阳550025

出　　处：《特产研究》2022年第2期48-54,63,共8页Special Wild Economic Animal and Plant Research

基　　金：福泉市1000亩天麻规模化标准化生产基地建设项目([2020]59);农村产业革命——贵州省天麻专家团队建设项目(黔财农[2019]175号);天麻绿色生态化种植技术研究与示范推广(黔财农[2020]101号)。

摘　　要：本研究以收集自贵州省(贵州大学农学院M-01~M-07、都匀市M-08、凯里市M-09和福泉市M-11)和湖北省宜昌市(M-10)的11份蜜环菌种质为研究对象,提取其基因组DNA,采用通用引物ITS4/ITS5对核糖体转录间隔区(rDNA-ITS)进行扩增,利用DNAMAN软件对11份蜜环菌种质进行构建同源树并分析其序列特征,采用MEGA软件最近邻距法进行系统发育分析及遗传距离的计算。结果表明,11份蜜环菌种质可分为两个大类4个亚类,第Ⅰ-Ⅰ类包括M-01、M-02、M-04、M-05和M-08-M-10,第Ⅰ-Ⅱ类为M-03,第Ⅱ-Ⅰ类包括M-06和M-07,第Ⅱ-Ⅱ类为M-11,具有较强的同源性(93%);两个大类样品的特征序列均位于ITS1和ITS2区表现为碱基的转换或颠换;共有序列的相似度为88.71%在ITS区存在丰富的变异表现为碱基的转换、颠换和缺失,第Ⅰ类的GC含量(44.2%)高于第Ⅱ类(42.6%);种内遗传距离均为0,第Ⅰ-Ⅱ类和第Ⅱ-Ⅱ类的种间遗传距离最小(0.002),第Ⅱ-Ⅰ类和第Ⅱ-Ⅱ类的种间遗传距离最大(0.035);经NCBI数据库BLAST对比及系统发育分析这4个亚类的样品均不能与下载的蜜环菌归为一支。因此,11份密环菌种质可能是一个较为庞大的复合群,采用通用引物ITS4/ITS5扩增的核糖体基因可以对此类蜜环菌进行分类但不适合作为鉴定的持家基因。In this study,the 11 Armillaria mellea strains were collected from Guizhou Province(M-01~M-07 of Agricultural College of Guizhou University,M-08 of Duyun City,M-09 of Kaili City and M-11 of Fuquan City)and Yichang city of Hubei Province(M-10)were used as the object of study.Their genome DNA was extracted as templates of PCR amplification and gene segments of ribosomal DNA ITS were amplified using PCR amplifying technology.DNAMAN was used to construct homologous trees and analyze the sequence characteristics of 11 Armillaria mellea strains,MEGA was used to analyze the phylogeny and calculate the genetic distance.The results showed that the 11 Armillaria mellea strains could be divided into two groups and four subclass,the classⅠ-Ⅰincludes M-01,M-02,M-04,M-05 and M-08-M-10,the classⅠ-Ⅱwas M-03,the classⅡ-Ⅰincludes M-06 and M-07,the classⅡ-Ⅱwas M-11,which had strong homology(93%);The characteristic sequences of the two types of samples were located in ITS1 and ITS2 regions,showing the conversion or transformation of bases;The similarity of the common sequences was 88.71%,and there were abundant variations in ITS region,showing the conversion,transformation and absence of bases,and the GC content of the classⅠ(44.2%)higher than that of the classⅡ(42.6%);The intraspecific genetic distance were 0,the genetic distance between species of the classⅠ-ⅡandⅡ-Ⅱwas minimal(0.002),and the classⅡ-ⅠandⅡ-Ⅱwas the largest(0.035);According to NCBI database BLAST comparison and analyze the phylogeny,These four subclass of samples could not be classified with the downloaded Armillaria mellea.So the 11 Armillaria mellea strains be a relatively large composite group,the ribosomal genes amplified by universal primers ITS4/ITS5 could be used to classify Armillaria mellea but were not suitable for identification as housekeeping genes.

关 键 词：蜜环菌 核糖体基因 ITS 序列分析 

分 类 号：S567.3[农业科学—中草药栽培]