外源添加胞外囊泡对海假交替单胞菌生物被膜形成及厚壳贻贝附着的影响  

Effects of exogenous addition of outer membrane vesicles on biofilm formation of Pseudoalteromonas marina and settlement of mussel(Mytilus coruscus)

作　　者：吴冠举 缪天音 胡晓梦 张稳 杨金龙[1,2,3] 梁箫 WU Guanju;MIAO Tianyin;HU Xiaomeng;ZHANG Wen;YANG Jinlong;LIANG Xiao(International Research Center for Marine Biosciences,Ministry of Science and Technology,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,Ministry of Education,Shanghai Ocean University,Shanghai 201306,China;Shanghai Collaborative Innovation Center for Cultivating Elite Breeds and Green-Culture of Aquaculture Animals,Shanghai 201306,China)

机构地区：[1]上海海洋大学,国家海洋生物科学国际联合研究中心,上海201306 [2]上海海洋大学,水产种质资源发掘与利用教育部重点实验室,上海201306 [3]上海市水产动物良种创制与绿色养殖协同创新中心,上海201306

出　　处：《大连海洋大学学报》2023年第6期994-1002,共9页Journal of Dalian Ocean University

基　　金：上海市优秀学术带头人项目(20XD1421800);国家自然科学基金(41876159);国家重点研发计划(2022YFE0204600)。

摘　　要：为探究胞外囊泡(outer membrane vesicles,OMVs)对海假交替单胞菌(Pseudoalteromonas marina)生物被膜形成及厚壳贻贝(Mytilus coruscus)(壳长为0.88 mm±0.16 mm,壳高为0.55 mm±0.12 mm)稚贝附着的影响,从海假交替单胞菌中提取并纯化了OMVs,并添加到海假交替单胞菌中共同孵育形成生物被膜,在OMVs终质量浓度为0.1、1、10、20μg/mL时检测形成的生物被膜对贻贝稚贝附着的诱导作用。结果表明:在海假交替单胞菌中成功提取到了OMVs,从菌体中提取获得的OMVs较上清液中更多;添加10μg/mL的OMVs时,形成的生物被膜对贻贝稚贝的附着诱导活性最高,稚贝附着率达到56.42%;随着OMVs添加量的增加,细菌密度和膜厚呈增长趋势,胞外产物中α-多糖、β-多糖、脂质和蛋白质含量呈升高趋势;相关性分析显示,生物被膜中的脂质含量与贻贝附着显著相关(P<0.05)。研究表明,OMVs的添加可促进海假交替单胞菌生物被膜形成,提高生物被膜胞外产物的含量,并推测主要通过增加脂质含量提高生物被膜对贻贝附着的诱导能力。To investigate the effects of outer membrane vesicles(OMVs)on the biofilm formation of Pseudoalteromonas marina and the settlement of mussel(Mytilus coruscus)plantigrades with shell length(0.88 mm±0.16 mm)and shell height of(0.55 mm±0.12 mm),OMVs were extracted and purified from P.marina and added into P.marina bacterial fluid to form biofilms,and the inducing capacity of biofilms on the settlement of mussel juveniles was detected at the final OMVs concentration of 0.1,1,10 and 20μg/mL.The results showed that OMVs were successfully extracted from P.marina,with the more content of OMVs extracted from the bacterium than that in the supernatant.The biofilms had the maximal inducing activity to mussel plantigrades,with the settlement rate of 56.42%at the OMVs concentration of 10μg/mL.With the increase in OMVs concentration,the bacterial density and biofilm thickness of biofilms were found to be increased,with increase inα-polysaccharides,β-polysaccharides,lipids and proteins in extracellular products.Correlation analysis showed that the lipid content in the biofilms was significantly correlated with mussel settlement(P<0.05),indicating that addition of OMVs promoted the formation of P.marina biofilms and increases the content of extracellular products of biofilms.It is speculated that the improvement in inducing ability of biofilmd to mussel settlement after adding OMVs is primarily due to the increase in lipid content.

关 键 词：胞外囊泡 生物被膜 厚壳贻贝 稚贝 附着 

分 类 号：S968.3[农业科学—水产养殖]