opaR基因缺失对AHPND致病菌生物学特性及毒力质粒接合转移的影响  

Effects of opaR Mutation on the Biological Characteristics and Conjugative Transfer of the Virulence Plasmid of Acute Hepatopancreatic Necrosis Disease-Causing Vibrio parahaemolyticus

作　　者：王利莹 董宣 王国浩[2] 王德浩 谢景媚 罗婧斐 王显伟 黄倢 WANG Liying;DONG Xuan;WANG Guohao;WANG Dehao;XIE Jingmei;LUO Jingfei;WANG Xianwei;HUANG Jie(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;State Key Laboratory of Mariculture Biobreeding and Sustainable Goods,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao Marine Science and Technology Center,Key Laboratory of Maricultural Organism Disease Control,Ministry of Agriculture and Rural Affairs,Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity,Qingdao 266071,China;Fisheries School,Tianjin Agriculture College,Tianjin 300392,China;College of Life Science,Shandong University,Qingdao 266237,China;Network of Aquaculture Centres in Asia-Pacific,Bangkok 10900,Thailand)

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]海水养殖育种与可持续产出全国重点实验室(中国水产科学研究院黄海水产研究所)青岛海洋科技中心海洋渔业科学与食物产出过程功能实验室农业农村部海水养殖病害防治重点实验室青岛市海水养殖流行病学与生物安保重点实验室,山东青岛266071 [3]天津农学院水产学院,天津300392 [4]山东大学生命科学学院,山东青岛266237 [5]亚太水产养殖中心网,泰国曼谷10900

出　　处：《渔业科学进展》2024年第4期155-165,共11页Progress in Fishery Sciences

基　　金：国家自然科学基金(31802342);中国水产科学研究院黄海水产研究所基本科研业务费(20603022022005);中国水产科学研究院基本科研业务费(2020TD39)共同资助。

摘　　要：急性肝胰腺坏死病(acute hepatopancreatic necrosis disease, AHPND)严重影响了我国乃至全球对虾养殖业的发展。近期研究发现,致病菌毒力质粒携带trb型Ⅳ型分泌系统(typeⅣsecretion system, T4SS)可在高菌体密度下介导毒力质粒发生接合转移,从而导致AHPND致病菌多样性。为探究群体感应与T4SS表达以及毒力质粒接合转移的关系,本研究以副溶血弧菌(Vibrio parahaemolyticus)群体感应系统的高密度调控子OpaR为研究对象,在致AHPND副溶血弧菌20130629002S01::cat(Vp2S01::cat)菌株基础上,利用同源重组技术构建opaR基因缺失株Vp2S01::catΔopaR。比较了出发菌株和缺失株在生长性能、运动性和生物被膜形成能力等方面的差异,分析了opaR基因对T4SS基因表达量以及质粒接合转移效率的影响。结果显示,opaR基因缺失不影响细菌的生长特性和群集运动,但泳动能力显著增加,生物被膜形成能力显著下降;接合转移实验显示,opaR基因缺失显著提高T4SS表达水平和接合转移效率。本研究为解析群体感应系统调控AHPND致病菌T4SS表达以及毒力质粒接合转移机制提供了基础数据,可为控制AHPND致病菌毒力质粒传播提供技术支撑。Acute hepatopancreatic necrosis(AHPND)is a bacterial disease caused by Vibrio bacteria that severely affects the Pacific white shrimp farming industry.Since AHPND was first identified in China and Vietnam in 2010,it was subsequently identified in Malaysia(2011),Thailand(2012),Mexico(2013),the Philippines(2014),South Korea(2016),Bangladesh(2017),the United States(US)(2017),and Japan(2020).AHPND causes significant economic losses of more than 7 billion US$annually to the global shrimp farming industry.It has severely restricted the development of the global shrimp farming industry.Many studies have shown that the pathogens of AHPND are Vibrio spp.,including V.parahaemolyticus,V.owensii,V.campbellii,V.harveyi,V.punensis,and V.anguillarum.The virulence of these pathogenic strains was derived from a 70 kb virulent plasmid(pVA1-type plasmid).The pVA1-type plasmid carries pirAB genes encoding the binary toxin pirAB homologous to the insecticidal protein of Photobacterium phosphoreum.The pVA1-type plasmid has been confirmed to carry a novel trb typeⅣsecretion system(T4SS).Trb-T4SS can mediate the conjugative transfer of the pVA1-type plasmid at high cell density for interspecific horizontal transfer.The quorum sensing(QS)system is also known as the density-sensing system.In the surrounding environment, the QS system allows bacteria to regulate the expression of multiple genes by sensing changes in the concentration of autoinducer signaling molecules. At high cell density, high concentrations of autoinducer signaling molecules bind to receptor proteins to inhibit phosphorylation cascades and high cell density master regulator OpaR is normally expressed. OpaR is involved in the regulation of various biological processes, such as biofilm-forming ability, motility, and the expression of the type Ⅲ and type Ⅳ secretion systems. Reportedly, high concentrations of signaling molecules in the QS system regulate the expression of key genes of the T4SS to increase the conjugative transfer efficiency of drug-resistant plasmids

关 键 词：副溶血弧菌 opaR基因 生物学特性 Ⅳ型分泌系统 水平转移 

分 类 号：S917.1[农业科学—水产科学]