基于拮抗及ITS序列分析的河南香菇主产区种质资源鉴定及遗传多样性分析

来源:用户上传      作者:崔筱　刘芹　孔维丽　张玉亭　胡素娟　王彦坡　康源春　孔维威　袁瑞奇

　　摘 要：用核糖体基因转录间隔区域（Internal Transcribed Spacer，ITS）序列及拮抗法对河南食用菌主产区香菇栽培菌株进行鉴定，并分析遗传多样性。以收集的41个香菇栽培菌株为样本，两两之间进行拮抗测试分析，提取DNA并进行PCR扩增、回收、测序，采用比对邻接法（neighbor-joinging，NJ）构建系统进化树。结果表明，16个差异菌株拮抗现象明显，其余菌株无拮抗或者拮抗不显著，41个香菇菌株初步分为16个不同菌株及7个不同亚种;ITS序列比对及系统发育分析将41个香菇品种聚为独立进化的4个大类，即3个不同的菌株和8个不同的亚种，种内遗传距离为0.000 0～0.006 5，与Pleurotus ostreatus（KY962509）种间遗传距离为0.374 0。综上所述，“ITS+拮抗法”结合可以明确区分菌株间拮抗、无拮抗及拮抗不明显现象基因序列的相似度、遗传距离的差异及亲缘关系的远近，为香菇品种的鉴定提供理论支撑。
　　关键词：香菇;ITS;拮抗鉴定;遗传多样性分析
　　中图分类号：S646 文献标志码：A 文章编号：1673-2871（2022）07-031-08
　　Identification of germplasms and genetic diversity of Lentinus edodes from main production areas in Henan province based on antagonism and ITS sequence analysis
　　CUI Xiao LIU Qin KONG Weili ZHANG Yuting HU Sujuan WANG Yanpo KANG Yuanchun KONG Weiwei YUAN Ruiqi
　　（1. Institute of Plant Nutrition， Agricultural Resources and Environmental Science， Henan Academy of Agricultural Sciences， Zhengzhou 450002， Henan， China; 2. Xixia Edible Fungus Production Office， Xixia 474599， Henan， China）
　　Abstract：Internal Transcribed Spacer （ITS） sequence and antagonism were used to identify and analyze the genetic diversity of Lentinus edodes isolated from the main producing area of edible fungi in Henan. The 41 cultivated L. edodes strains from the main producing areas of edible fungi in the province were collected as samples， and the antagonism test was conducted between pairs， and DNA of the samples were extracted， amplified by PCR， recovered and sequenced， and a phylogenetic tree was constructed by neighbor-joining （NJ） method. The results showed that： The antagonism was obvious in 16 different strains， while the rest showed no or insignificant antagonism， and the 41 L. edodes strains were divided into 16 different strains and 7 different subspecies; ITS sequence alignment and phylogenetic analysis clustered 41 L. edodes strains into independently evolved 4 broad classes， 3 different strains and 8 different subspecies and the intraspecific genetic distances were between 0.000 0 and 0.006 5， of which the interspecific genetic distance with Pleurotus ostreatus（KY962509） is 0.374. The combination of “ITS + antagonism” method could clearly distinguish the similarity of gene sequence of the phenomenon of antagonism， no antagonism and antagonism of which is notobvious， genetic distance and relationship between strains， which provide theoretical support for the identification of L. edodes varieties.
　　Key words： Lentinus edodes; ITS; Antagonistic identification; Genetic diversity analysis

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