fliY基因失活减弱沙福芽孢杆菌ST7菌株的锰氧化能力  

Inactivation of fliY attenuates the manganese-oxidizing capacity of Bacillus safensis ST7

作　　者：朱忠梅 吴莉丹 张心仪 方争林 牛熙[1] 黄世会 王嘉福[1] 冉雪琴[2] ZHU Zhongmei;WU Lidan;ZHANG Xinyi;FANG Zhenglin;NIU Xi;HUANG Shihui;WANG Jiafu;RAN Xueqin(The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),College of Life Sciences,Institute of Agro-Bioengineering,Guizhou University,Guiyang 550025,Guizhou,China;College of Animal Science,Guizhou University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州大学生命科学学院贵州大学农业生物工程研究院山地植物资源保护与种质创新省部共建教育部重点实验室,贵州贵阳550025 [2]贵州大学动物科学学院,贵州贵阳550025

出　　处：《微生物学通报》2023年第1期107-117,共11页Microbiology China

基　　金：国家自然科学基金(31960641);贵州省科技创新人才团队项目(黔科合平台人才[2019]5615);贵州省“百”层次创新型人才培养项目(黔科合人才[2016]4012)。

摘　　要：【背景】沙福芽孢杆菌ST7菌株具有较强的锰氧化能力,但其分子机制不清楚。【目的】着重研究鞭毛马达开关蛋白基因(fliY)对沙福芽孢杆菌锰氧化能力的影响。【方法】根据同源重组原理,以沙福芽孢杆菌ST7菌株为起始菌株,构建fliY基因敲除的突变株ΔfliY,测定菌落迁徙、细菌生物膜和锰氧化率等,研究fliY基因突变后菌株的运动能力、生物膜生成和锰氧化能力是否发生变化。【结果】经克隆测序,证实突变株ΔfliY中fliY基因的后半段被卡纳霉素抗性基因取代,fliY基因失活;与野生型菌株ST7相比,突变株ΔfliY在全营养的LB培养基中生长变化不大,但在含锰的PYCM培养基中,突变株的生长速度减慢、菌落较小、生物膜生成量显著下降,运动性和锰氧化能力分别下降65%和20%。【结论】fliY基因不仅影响菌株的生长和运动,而且参与细菌的趋化和锰氧化等生物学过程。[Background]Bacillus safensis ST7 has strong ability of oxidizing manganese,the molecular mechanism of which,however,remains unclear.[Objective]The aim is to study the effect of the flagellar motor switch protein FliY on the manganese-oxidizing ability of B.safensis.[Methods]Based on the principle of homologous recombination,a fliY-deleted mutant(ΔfliY)was constructed using B.safensis ST7 as the starting strain.The changes in mobility,biofilm formation,and manganese-oxidizing capacity ofΔfliY were evaluated based on colony migration,biofilm production,and Mn(II)-oxidizing rate.[Results]After cloning and sequencing,it was confirmed that fliY was inactivated with the last part replaced by kanamycin resistance gene inΔfliY.The growth ofΔfliY in LB medium was similar to that of the starting strain.However,ΔfliY showed retarded logarithmic growth,small colonies,and reduced biofilm production in the PYCM medium containing Mn(II).The motility and Mn(II)-oxidizing rate ofΔfliY decreased by about 65%and 20%,respectively.[Conclusion]The gene fliY not only affects the growth and motility but also participates in biological processes such as chemotaxis and manganese oxidation of B.safensis.

关 键 词：沙福芽孢杆菌 鞭毛马达开关蛋白基因 生物膜 鞭毛 锰氧化 

分 类 号：X172[环境科学与工程—环境科学]