绿僵菌系统发育及防治松褐天牛成虫的菌株筛选  

Phylogenesis of Metarhizium Strains and Their Screening for Biocontrol of Monochamus alternatus Adults(Coleoptera:Cerambycidae)

作　　者：刘程林 应玥 王瑞珍 刘云朋 王青华 孔德治 曲良建[1] Liu Chenglin;Ying Yue;Wang Ruizhen;Liu Yunpeng;Wang Qinghua;Kong Dezhi;Qu Liangjian(Key Laboratory of Forest Protection of National Forestry and Grassland Administration Ecology and Nature Conservation Institute,Chinese Academy of Forestry,Beijing 100091;Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Hangzhou 311400;Beijing Floriculture Engineering Technology Research Centre China National Botanical Garden(North Garden),Beijing 100093)

机构地区：[1]中国林业科学研究院森林生态环境与自然保护研究所、国家林业和草原局森林保护学重点实验室,北京100091 [2]中国林业科学研究院亚热带林业研究所,杭州311400 [3]国家植物园(北园)北京市花卉园艺工程技术研究中心,北京100093

出　　处：《林业科学》2025年第2期152-162,共11页Scientia Silvae Sinicae

基　　金：中国林业科学研究院基本科研业务费(CAFYBB2021ZG001,CAFYBB2020SY022);国家林业和草原局重大应急科技项目(ZD202001)。

摘　　要：【目的】研究国内外80株绿僵菌的系统发育关系及其对松褐天牛成虫的毒力,为筛选用于防治松褐天牛成虫的高毒力菌株提供参考。【方法】提取80株绿僵菌的DNA,使用引物ITS1/ITS4和T1/T22分别扩增真菌的ITS区域和β-tubulin基因序列并测序。采用MAFFT比对序列,通过IQ-TREE和Mrbayes 3.2.7软件分别采用最大似然法和贝叶斯法构建系统发育树。使用羽化3天内的健壮松褐天牛成虫测定绿僵菌的毒力。在PDA培养基上培养绿僵菌,将收集获得的孢子用0.05%的吐温-80溶液分别配置成浓度为10~7个·mL–1的孢子悬浮液,0.05%的吐温80溶液作为空白对照。采用浸渍法处理松褐天牛成虫,以累计校正死亡率为指标评价菌株的致病力。【结果】从国内外菌库中收集的123株菌株中80株为绿僵菌,分为6个主要类群,其中57株属于金龟子绿僵菌复合种,11株属于黄绿绿僵菌复合种,6株属于莱氏绿僵菌,4株属于M.viride,1株属于柱孢绿僵菌,1株属于新西兰绿僵菌。高毒力菌株主要分布在金龟子绿僵菌复合种和黄绿绿僵菌复合种;蝗绿僵菌、柱孢绿僵菌、新西兰绿僵菌、莱氏绿僵菌和M.viride对松褐天牛的成虫未表现出致病力。14株绿僵菌菌株毒力较高(LT50<15天,15天校正死亡率>80%),可作为防治松褐天牛的备选菌株。【结论】基于国内外80株绿僵菌菌株的系统发育和毒力测试结果,筛选出对松褐天牛成虫具有高毒力的菌株,其中14株具有较大应用潜力,建立了绿僵菌的系统发育关系,确认了高毒力菌株的分布。【Objective】The phylogenetic relationship of 80 Metarhizium strains isolated from domestic and abroad and their virulence to Monochamus alternatus adult were investigated in order to screen highly virulent strains for the control of M.alternatus adult.【Method】The ITS1/ITS4 and T1/T22 primer pairs were used to amplify the ITS region andβ-tubulin gene sequences of 80 Metarhizium strains,respectively.MAFFT was used to align the sequences,and the phylogenetic tree was constructed using the maximum likelihood method and the Bayesian method with IQ-TREE and Mrbayes 3.2.7 software.The virulence of Metarhizium strains was determined using M.alternatus adults in 1-3 days after emergence.Metarhizium strains were cultured on PDA medium,and the spores were collected and prepared into a spore suspension with a concentration of 107·mL−1 using 0.05%Tween 80 solution,and 0.05%Tween 80 solution was used as a blank control.The impregnation method was used to treat M.alternatus adults,and the pathogenicity of the strains was evaluated by using the cumulative corrected mortality rate of M.alternatus adults.【Result】Among the 123 strains collected from domestic and international libraries,there are 80 strains of Metarhizium,which are divided into 6 major taxa,including 57 strains belonging to M.anisopliae species complex,11 strains belonging to M.flavoviride species complex,6 strains belonging to M.rileyi,4 strains belonging to M.viride,one strain belonging to M.cylindrosporum,and one strain belonging to M.novozealandicum.The highly virulent strains were mainly found in M.anisopliae species complex and M.flavoviride species complex.M.acridum,M.cylindrosporum,M.novozealandicum,M.rileyi,and M.viride did not show pathogenicity against M.alternatus adults.A total of 14 strains of Metarhizium were highly virulence(LT50<15 d,15-day adjusted mortality>80%),which can be used as alternative strains for the control of M.alternatus.【Conclusion】Based on the phylogenetic and virulence test results of 80 strains of Metarhizium fr

关 键 词：绿僵菌 松褐天牛 菌株鉴定筛选 生物防治 

分 类 号：S763.3[农业科学—森林保护学]