ecp基因簇在路德维希肠杆菌AA4定殖松材线虫中的功能研究  

Study on the Function of ecp Gene Cluster in Colonization of Pine Wood Nematode by Enterobacter Ludwigii AA4

作　　者：王明越 何杉 张丽霞 王思佳 苑志博 吴迪[1,2] 王传珍[4] 潘佳亮 包盛波 王琦 牛犇[1,2] WANG Mingyue;HE Shan;ZHANG Lixia;WANG Sijia;YUAN Zhibo;WU Di;WANG Chuanzhen;PAN Jialiang;BAO Shengbo;WANG Qi;NIU Ben(State Key Laboratory of Tree Genetics and Breeding,Northeast Forestry University,Harbin 150040,China;College of Life Science,Northeast Forestry University,Harbin 150040,China;Sino Green Agri-Biotech Co.,Ltd.,Beijing 102101,China;Yantai Service Center of Forest Resources Monitoring and Protection,Yantai 264000,China;General Station of Forest and Grassland Pest Management,National Forestry and Grassland Administration,Key Laboratory of National Forestry and Grassland Administration on Forest Pest Monitoring and Warning,Shenyang 110034,China;Department of Plant Pathology,China Agricultural University,Beijing 100193,China)

机构地区：[1]东北林业大学林木遗传育种全国重点实验室,哈尔滨150040 [2]东北林业大学生命科学学院,哈尔滨150040 [3]中农绿康(北京)生物技术有限公司,北京102101 [4]烟台市森林资源监测保护服务中心,烟台264000 [5]国家林业和草原局森林和草原病虫害防治总站,林草有害生物监测预警国家林业和草原局重点实验室,沈阳110034 [6]中国农业大学植物病理学系,北京100193

出　　处：《中国生物防治学报》2024年第5期1022-1035,共14页Chinese Journal of Biological Control

基　　金：国家重点研发计划(2022YFD1401900);国家自然科学基金(32071741)。

摘　　要：松材线虫病生防细菌路德维希肠杆菌EnterobacterludwigiiAA4能够有效地定殖并杀死松材线虫。大肠杆菌共有型菌毛(Escherichia coli common pilus,ECP)在动植物共生与病原细菌中普遍存在,能够通过增强细菌与宿主细胞之间及生物膜群落中细菌之间的相互作用,提高共生与病原细菌的定殖能力。大肠杆菌共有型菌毛由ecp基因簇编码。为了探究ecp基因簇在菌株AA4定殖松材线虫中的功能,本研究通过BLAST比对分析,鉴定了菌株AA4中的ecp基因簇;利用Red重组技术构建了菌株AA4的ecp基因簇缺失突变体,并对上述突变体定殖松材线虫的能力与定殖相关表型进行了分析。研究结果表明,ecp基因簇缺失突变体菌毛装配能力显著减弱,运动性、生物膜形成能力以及在松材线虫上的定殖能力显著降低。因此,AA4的ecp基因簇可能通过调节菌毛装配,影响其运动性与生物膜形成能力,进而实现对菌株AA4定殖松材线虫的调控。上述研究结果将为揭示松材线虫病生防路德维希肠杆菌AA4定殖松材线虫的分子机制奠定理论基础,为松材线虫病生防微生物制剂的研发与高效应用提供科学依据。Enterobacter ludwigii AA4,a biocontrol bacterial strain,can efficiently colonize and kill pine wood nematode(PWN)to prevent the occurrence of pine wilt disease(PWD).Escherichia coli common pilus(ECP)can promote bacterial colonization by enhancing the bacteria-host interactions and bacterial interspecies interplay within the biofilms.The ECP is encoded by the ecp gene cluster.In order to explore the function of ECP in colonization of PWN by strain AA4,we identified the ecp gene cluster in AA4 by using BLAST alignment.By employing the Red(λ,β,exo)-mediated DNA homologous recombination technology,we further constructed the ecp gene cluster knock-out deletion mutant and analyzed its ability to colonize the PWN plus the colonization-related phenotypes.The outcomes showed that the pilus assembly,motility,biofilm formation of the knock-out mutant and its residence on the PWN was significantly reduced.Thus,ecp gene cluster may regulate the colonization of the PWN by E.ludwigii AA4 through affecting its motility and biofilm formation via modulation of its pilus synthesis.Altogether,our results will serve as theoretical bases for revealing the molecular mechanisms underlying the PWN colonization by strain AA4 and will provide scientific evidence guiding the development of biopesticides for controlling the PWD and their efficient application.

关 键 词：ecp基因簇 菌毛 路德维希肠杆菌 松材线虫 定殖 

分 类 号：S476.2[农业科学—农业昆虫与害虫防治]