文章摘要
美人鱼发光杆菌美人鱼亚种dly基因缺失株构建及其生物学特性研究*
Liu Dingyuan1,2, Wang Chunyuan2, Yu Yongxiang2, Wang Yingeng2, Li Jingze2, Zhang Xiaosong2, Qin Lei1, Zhang Zheng2
投稿时间:2023-02-11  修订日期:2023-02-26
DOI:
中文关键词: 美人鱼发光杆菌美人鱼亚种  缺失株  胞外产物  生物学特性  致病性
英文关键词: Photobacterium damselae subsp.damselae  mutant strain  extracellular products  biological characteristics  pathogenicity
基金项目:山东省自然科学基金项目(ZR2021MC027; ZR2020QC216)和山东省重点研发计划(科技示范工程)课题(2021SFGC0701)共同资助。
作者单位邮编
刘定远 江苏海洋大学 江苏 连云港
农业农村部海水养殖病害防治重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 
266071
王春元 农业农村部海水养殖病害防治重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 
于永翔 农业农村部海水养殖病害防治重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 
王印庚 农业农村部海水养殖病害防治重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 
李京泽 农业农村部海水养殖病害防治重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 
张晓松 农业农村部海水养殖病害防治重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 
秦蕾 江苏海洋大学 江苏 连云港 
张正* 农业农村部海水养殖病害防治重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 266071
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中文摘要:
      美人鱼发光杆菌美人鱼亚种(Photobacterium damselae subsp.damselae, PDD)是广泛分布于全球海洋环境中的一种致病菌。本文以实验室保存的一株高致病性PDD菌株(PDD1608)作为研究对象,初步探究毒力基因dly对PDD菌株的生物学特性和致病力的影响。利用λRed重组技术成功构建毒力基因dly缺失株Δdly PDD1608::Cm,比较野生株和缺失株的生长、涌动性、药物敏感性、生理生化特性、生物被膜形成能力、菌株及胞外产物(extracellular products, ECP)的溶血性和磷脂酶活性等生物学特性。选用海水青鳉鱼(Oryzias melastigma)作为实验动物,通过人工感染实验测定野生株和缺失株及其 ECP 对海水青鳉鱼的致病性。毒力基因dly缺失后导致 PDD菌株的生长变慢,缺失株的涌动性、溶血性和磷脂酶活性均降低;野生株和缺失株的药物敏感性和生理生化特性未产生变化;与野生株相比,缺失株的生物被膜形成能力有显著性差异(P<0.05);人工感染实验表明缺失株及其ECP对海水青鳉鱼的致病性降低。毒力基因dly影响PDD菌株的生长、涌动性、溶血性和磷脂酶活性等多种生物学特性,并且与PDD菌株及其ECP的致病力强弱密切相关。
英文摘要:
      As a pathogen, Photobacterium damselae subsp.damselae (PDD) distribute widely in Marine environment. In the past decade, reports on the pathogenicity of PDD to Marine animals have gradually increased, and the host species of PDD are also diverse. Therefore, it is necessary to further study the pathogenicity of PDD in order to establish targeted prevention and control technical measures to reduce its harm to Marine organisms.In this study, a high-virulence PDD strain (PDD1608)was selected to explore the effect of virulence gene dly on the biological characteristics and pathogenicity. The dly-deleted mutant strain Δdly PDD1608::Cm was successfully constructed by λRed recombination technique. The biological characteristics of wild type strain and mutant strain were compared, including growth, swarming motility , drug susceptibility, physiological and biochemical characteristics, biofilm formation ability, hemolytic and phospholipase activity of extracellular products (ECP). Meanwhile, Oryzias melastigma was used as the target host, the pathogenicity of wild type and mutant strains and their ECPs to Oryzias melastigma were detected by artificial infection test. There was a positive correlation between the results of bacterial solution and ECP challenge, indicating that the deletion of dly gene affects the virulence of PDD strain.Deletion of the virulence gene dly resulted in slower growth of PDD mutant strain and reduced swarming, hemolytic and phospholipase activities ; the drug susceptibility, physiological and biochemical characteristics of the wild type strain and mutant strain did not change; compared with the wild type strain, the biofilm formation ability of the mutant strain was significantly different (P<0.05); artificial infection test showed that the pathogenicity of the mutant strain and its ECP decreased.The results of this study showed that virulence gene dly affects many biological characteristics of PDD strains, such as growth, swarming motility , hemolytic and phospholipase activity, and it was closely related to the pathogenicity of PDD strain and ECP.
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