松材线虫侵染前后马尾松树体内微生物多样性分析  

Microbial Diversity Analysis of Pinus massoniana Before and After Infected by Pine Wood Nematode

作　　者：温晓健 巫建军 李永先 王璇 理永霞[1] 张星耀[1] WEN Xiao-jian;WU Jian-jun;LI Yong-xian;WANG Xuan;LI Yong-xia;ZHANG Xing-yao(Key Laboratory of Forest Protection of National Forestry and Grassland Administrstion,Institute of Forest Ecology,Environment and Nature Conservation,Chinese Academy of Forestry,Beijing l00091,China;Suichang County Ecological Forestry Development Center,Suichang 323399,Zhejiang,China)

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]浙江省遂昌县生态林业发展中心,浙江遂昌323399

出　　处：《林业科学研究》2022年第1期48-58,共11页Forest Research

基　　金：国家林业和草原局重大应急科技项目(ZD202001)。

摘　　要：[目的]以浙江省松材线虫病疫区健康和枯死马尾松(Pinus massoniana Lamb.)为研究对象,探究松材线虫(Bursaphelenchus xylophilus)侵染前后马尾松树体内微生物群落结构的差异,同时为利用马尾松内生微生物防治松材线虫病积累微生物种质资源。[方法]采用组织分离法、形态学和分子生物学相结合的方法,对健康和枯死马尾松不同组织内的(根部、干部和针叶)真菌和细菌进行分离鉴定。对分离的木霉菌株构建系统发育树,并分析了松材线虫取食木霉对其种群数量的影响。[结果]从健康和枯死马尾松树体内分别分离得到真菌19个属共106个菌株和10个属共86个菌株,优势类群均为木霉属真菌。不同组织间真菌多样性水平存在差异,干部和针叶的多样性指数高于根部。健康马尾松真菌的Shannon-Wiener指数(H)、Simpson指数(D)和均匀度指数(J)依次为2.37、0.87和0.51,高于枯死马尾松的1.72、0.76和0.39,二者的相似性指数小于0.5。从健康和枯死马尾松树体内分别分离得到细菌13个属共59个菌株和12个属共56个菌株,优势类群均为伯克氏菌属、肠杆菌属和沙雷氏菌属细菌。健康和枯死马尾松中均为干部的细菌多样性最高,针叶的细菌多样性最低,二者的相似性指数大于0.5。木霉作为马尾松树体内的优势类群,本次共分离到6个种,通过取食实验发现木霉FXY7显著抑制了松材线虫的种群数量。[结论]健康马尾松树体内的真菌多样性高于枯死马尾松,二者的真菌菌群相似性较低,而且不同组织间真菌菌群相似性也较低,但健康和枯死马尾松的细菌群落具有一定的相似性。这表明松材线虫入侵会影响马尾松体内的真菌多样性,但对细菌的影响较小。本研究中还分离到多株可能对松材线虫病具有生防潜力的内生菌,为探索松材线虫病的新型防控技术奠定了基础。[Objective]Taking healthy and dead Pinus massoniana Lamb.in Bursaphelenchus xylophilus infected area in Zhejiang province as research object to examine the differences in the microbial communities structure of P.massoniana trees before and after infected by B.xylophilus,and to accumulate the microbial resources for controlling pine wilt disease by using endophytic microorganisms of P.massoniana.[Method]The methods of tissue isolation,morphology and molecular biology were used to isolate and identify the fungi and bacteria in different tissues(root,stem and needle)of healthy and dead P.massoniana.Phylogenetic tree was constructed for the isolated Trichoderma strains,and the effect of Trichoderma spp.on B.xylophilus population was analyzed by feeding experiments.[Result]106 fungal strains belonging to 19 genera were isolated from healthy P.massoniana,and 86 strains belonging to 10 genera were isolated from the dead pine,the dominant genera were both Trichoderma.There were differences in fungal diversity among different tissues,the diversity indexes of stem and needle were higher than those of root.The Shannon-Wiener index,Simpson index and evenness index of fungal community from healthy pine were 2.37,0.87 and 0.51,respectively,they were all higher than those of dead pine,which were 1.72,0.76 and 0.39.The similarity indexes of fungal community in healthy and dead P.massoniana were less than 0.5.59 bacterial strains belonging to 13 genera were isolated from healthy P.massoniana,and 56 strains belonging to 12 genera were isolated from the dead pine,the dominant groups were Burkholderia,Enterobacter and Serratia.The bacterial diversity of stem was the highest in both the healthy and dead pine,and that of needle was the lowest.The similarity indexes of bacterial community in healthy and dead P.massoniana were more than 0.5.Trichoderma was the dominant group in P.massoniana,six species were isolated in this study.The feeding experiment showed that the sample FXY7 of Trichoderma significantly inhibited the population of

关 键 词：松材线虫 马尾松 真菌 细菌 多样性 

分 类 号：S763.1[农业科学—森林保护学]