cbpD基因对溶藻弧菌毒力及相关生物学特性的影响  

Effects of cbpD gene on virulence and related biological characteristics of Vibrio alginolyticus

作　　者：苏雯晓 邓益琴 臧树军 王茜 林梓阳[2,4] 冯娟 SU Wenxiao;DENG Yiqin;ZANG Shujun;WANG Qian;LIN Ziyang;FENG Juan(Shanghai Ocean University/National Demonstration Center for Experimental Fisheries Science Education,Shanghai 201306,China;South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences/Key Laboratory of South China Sea Fishery Resource Exploitation&Utilization,Ministry of Agriculture and Rural Affairs,Guangzhou 510300,China;Tianjin Agricultural University,Tianjin 300392,China;Ocean Collage,Hainan University,Haikou 570228,China)

机构地区：[1]上海海洋大学/水产科学国家级实验教学示范中心,上海201306 [2]中国水产科学研究院南海水产研究所/农业农村部南海渔业资源开发利用重点实验室,广东广州510300 [3]天津农学院,天津300392 [4]海南大学海洋学院,海南海口570228

出　　处：《南方水产科学》2022年第5期81-90,共10页South China Fisheries Science

基　　金：中国水产科学研究院基本科研业务费资助(2022GH03);国家自然科学基金项目(31902415);广东省自然科学基金项目(2019A1515011833);中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助(2021SD15);广州市科技计划资助项目(202201010162)。

摘　　要：为研究几丁质结合蛋白(Chitin-binding protein D,CbpD)对溶藻弧菌(Vibrio alginolyticus)毒力和相关生物学特性的影响,通过同源重组技术构建溶藻弧菌ZJ-T的cbpD基因缺失突变株ZJ-T-ΔcbpD,比较突变株与野生株对斑马鱼(Danio rerio)的毒性,以及毒力相关生理过程,包括生长能力、运动性、胞外蛋白酶分泌活性、溶血活性、抗生素敏感性、生物膜形成、过氧化氢(H_(2)O_(2))和铜离子(Cu^(2+))抗性以及铁离子(Fe^(3+))吸收能力的差异。研究发现,cbpD缺失后,溶藻弧菌对斑马鱼的毒力显著减弱,且细菌游动能力、涌动能力和胞外蛋白酶活性均降低,突变株对Cu2+的抗性提高;cbpD的突变不影响溶藻弧菌在富营养培养基中的生长、生物膜的形成、溶血活性、对大部分抗生素敏感性、对H_(2)O_(2)的抗性和Fe3+的获取能力。结果表明,cbpD可能通过正调控溶藻弧菌的运动能力和胞外蛋白酶分泌活性,而促进溶藻弧菌的毒力。In order to study the effect of the chitin binding protein D(CbpD)on the virulence and related biological characteristics of Vibrio alginolyticus,we constructed the cbpD gene deletion mutant strain of V.alginolyticus ZJ-T by homologous recombination technique.Then we compared the virulence to Danio rerio and other related physiological processes between the wild type strain and the cbpD gene mutant strain,including the growth ability,motility,extracellular protease secretion activity,hemolytic activity and antibiotic sensitivity,biofilm formation,hydrogen peroxide and copper ion resistance as well as ion absorption capacity.The results show that due to the absence of cbpD gene,the swimming motility,swarming motility and extracellular protease activity all reduced,while the resistance to copper ion of the mutant strain strengthened,and its virulence to D.rerio was weakened.However,the deletion of cbpD gene did not affect the growth of the strain in nutrient-rich medium,biofilm formation,hemolytic activity,stress response to H_(2)O_(2),uptake and utilization of iron and sensitivity to most antibiotics.In conclusion,the results indicate that cbpD may promote the virulence by positively regulating the motility,extracellular protease secretion activity of V.alginolyticus.

关 键 词：溶藻弧菌 cbpD基因 基因敲除 毒力调控 生物学特性 

分 类 号：R917.1[医药卫生—药物分析学]