基于HPCE和多基因联合序列鉴定卵孢长根菇的分类研究  

Study on Classification of Oudemansiella raphanipes Based on HPCE and Multi Gene Combined Sequence Identification

作　　者：龚娜[1] 马晓颖[1] 陈珣[1] 刘国丽[1] 肇莹[1] 肖军[1] GONG Na;MA Xiaoying;CHEN Xun;LIU Guoli;ZHAO Ying;XIAO Jun(Institute of Edible Fungi,Liaoning Academy of Agricultural Sciences,Shenyang,Liaoning 110161)

机构地区：[1]辽宁省农业科学院食用菌研究所,辽宁沈阳110161

出　　处：《北方园艺》2022年第18期117-123,共7页Northern Horticulture

基　　金：揭榜挂帅科技攻关专项资助项目(2021JH1/10400035);辽宁省农业科学院院长基金资助项目(2022QN2303);辽宁省科技特派活动专项计划资助项目(2021JH5/10400035);沈阳市科技特团专项资助项目(21-116-3-23);国家食用菌产业技术体系资助项目(CARS-20)。

摘　　要：以5株卵孢长根菇菌株为试材,采用ITS和nLSU序列的测定并综合分析,结合GenBank中相关物种序列及菌株的形态学特征,进行卵孢长根菇的分子生物学鉴定,并且采用高效毛细管电泳检测,研究了卵孢长根菇的高效毛细管指纹图谱,通过主成分分析、相似度分析和聚类分析,对其进行鉴定和评价,以期探究新的快速准确的鉴定方法解决卵孢长根菇种类繁多,种内分类难度大,命名混乱等问题。结果表明:参试菌株总体亲缘关系较近,均为卵孢长根菇(Oudemansiella raphanipes),Or2、Or5和Or1亲缘关系较近的菌株均不同程度的聚为一支,Or2、Or5和Or1相对于Or4、Or6亲缘关系更近,基于指纹图谱鉴别卵孢长根菇子实体形态的多样性,通过“W”形的三联差异峰作为鉴别卵孢长根菇不同子实体出菇表型的关键指示峰。卵孢长根菇具有丰富的物种多样性,采用HPCE和多基因联合序列的鉴定方法可有效区分不同形态种,为快速筛选优质卵孢长根菇菌株提供参考依据。Five isolates of Oudemansiella raphanipes were used as the experimental materials.The molecular biological identification of Oudemansiella raphanipes was determined and comprehensive analysised by using ITS and nLSU sequence, combined with the sequences of related species in GenBank and morphological characteristics of the strains. In addition, high performance capillary electrophoresis was used to study the high performance capillary fingerprinting of Oudemansiella raphanipes,and the identification and evaluation were carried out by principal component analysis, similarity analysis and cluster analysis, in order to explore a new rapid and accurate identification method and to solve the problem of confusing nomenclature problems due to the large number of species of Oudemansiella raphanipes.The results showed that all test strains had close relationships, were all Oudemansiella raphanipes.Strains with close genetic relationships of Or2,Or5 and Or1 all clustered into one branch to varying degrees, Or2,Or5 and Or1 had closer genetic relationships than Or4 and Or6.The ‘W’ shaped triad differential peak was used as the key indicator peak for the identification of different fruiting bodies of Oudemansiella raphanipes.There was rich species diversity in Oudemansiella raphanipes.The identification method of HPCE and multi gene joint sequence could effectively distinguish different morphological species, which provided reference for rapid screening of high-quality Oudemansiella raphanipes strains.

关 键 词：卵孢长根菇 毛细管电泳 多基因联合分析 

分 类 号：S646.9[农业科学—蔬菜学]