粘质沙雷氏菌FZSF02中转录调控因子OmpR的生物学功能  

Biological Functions of Transcription Factor OmpR in Serratia marcescens FZSF02

作　　者：贾宪波[1] 刘方晨 赵恪 林俊杰 方宇[1] 陈济琛[1] JIA Xianbo;LIU Fangchen;ZHAO Ke;LIN Junjie;FANG Yu;CHEN Jichen(Institute of Soil and Fertilizer,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China;College of Resources and Environment,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350001,China;College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350001,China)

机构地区：[1]福建省农业科学院土壤肥料研究所,福建福州350003 [2]福建农林大学资源与环境学院,福建福州350001 [3]福建农林大学生命科学学院,福建福州350001

出　　处：《福建农业学报》2021年第12期1491-1498,共8页Fujian Journal of Agricultural Sciences

基　　金：国家自然科学基金青年基金项目(31800068);福建省农业科学院科技创新团队建设项目(CXTD2021002-3)。

摘　　要：【目的】探索EnvZ/OmpR双组分调控系统的效应蛋白OmpR对粘质沙雷氏菌FZSF02灵菌红素合成及其他生物学特性的影响。【方法】同源重组法构建ompR敲除菌株,琼脂平板产色试验和qPCR检测OmpR对菌株灵菌红素合成的影响,结晶紫染色法和琼脂平板法等研究基因敲除菌株生物被膜形成能力,运动性和对不同环境胁迫因素的耐受性。【结果】序列分析显示OmpR为序列高度保守的蛋白。PCR验证证明ompR基因敲除成功;与野生型菌株相比ΔompR失去灵菌红素合成能力,qPCR试验显示灵菌红素合成基因簇中3个关键基因pigA、pigF和pigN转录水平分别降低为野生型菌株的3.8%,2.0%和2.1%;ΔompR菌株生物被膜生成能力较野生型降低37.5%(37℃)和15.1%(27℃);OmpR对该菌的生长能力、运动能力和响应环境胁迫的能力无明显影响。【结论】ompR为一新报道的特异性调控粘质沙雷氏菌灵菌红素合成的基因,且该基因对该菌生物被膜的形成有重要影响。【Objective】Biological functions of the regulatory protein ompR in the two component EnvZ/OmpR system,including prodigiosin-producing ability and other biological characteristics,of Serratia marcescens FZSF02 were studied.【Methods】Homologous recombination was used to construct ompR-knockout S.marcescens FZSF02.Effect of OmpR on the prodigiosin-producing ability was examined by LB agar plate incubation and qPCR.Methods of crystal violet staining,agar plate incubation,and others were applied to determine the biofilm-forming,mobility,and stress adaptation abilities of the transcription factor protein under various environmental stresses.【Results】OmpR was a protein with high conserved amino acid sequences.An ompR-deleted strain,FZSF02ΔompR,was successfully obtained by homologous recombination and confirmed by PCR.As a result,FZSF02ΔompR lost prodigiosin-producing ability that possessed by the wild strain.The transcriptional levels of pigA,pigF,and pigN of the prodigiosin biosynthesis gene cluster in FZSF02ΔompR were respectively 3.8%,2.0%and 2.1%of the wild type strain.The biofilm formation of FZSF02ΔompR declined 37.5%(at 37℃)and 15.1%(at 27℃)from its wild counterpart.On the other hand,OmpR exhibited no significant effect on the growth,mobility,or response to the environmental stress.【Conclusion】OmpR was a newly reported gene that specifically regulated the prodigiosin biosynthesis in S.marcescens.It also significantly affected the biofilm formation but not on the growth,mobility,or stress response.

关 键 词：粘质沙雷氏菌 OMPR 灵菌红素 生物学特性 

分 类 号：Q939[生物学—微生物学]