双组分因子LiaRS调控短短芽胞杆菌X23伊短菌素生物合成的研究  

Effects of Two-Component System LiaRS on Biosynthesis of Edeine from Brevibacillus brevis X23

作　　者：邱丽婷 张亮 刘天波 巢进 蔡海林 易克 刘清术 陈武[1] QIU Liting;ZHANG Liang;LIU Tianbo;CHAO Jin;CAI Hailin;YI Ke;LIU Qingshu;CHEN Wu(College of Plant Protection,Hunan Agricultural University,Changsha 410128,China;College of Agronomy,Hunan Agricultural University,Changsha 410128,China;Hunan Tobacco Science Research Institute,Changsha 410004,China;Hunan Xiangxi autonomous prefecture tobacco company technology center,Xiangxi 416000,China;Hunan Provincial Tobacco Company Technology Center,Changsha 410019,China;Hunan Tobacco Industry Co.,Ltd.Raw Material Procurement Center,Changsha 410007,China;Hunan Provincial Microbiology Research Institute,Changsha 410009,China)

机构地区：[1]湖南农业大学植物保护学院,长沙410128 [2]湖南农业大学农学院,长沙410128 [3]湖南省烟草科学研究所,长沙410004 [4]湖南省湘西自治州烟草公司技术中心,湘西416000 [5]湖南省长沙市烟草公司技术中心,长沙410019 [6]湖南中烟工业有限责任公司原料采购中心,长沙410007 [7]湖南省微生物研究院,长沙410009

出　　处：《中国生物防治学报》2024年第5期1036-1044,共9页Chinese Journal of Biological Control

基　　金：国家自然科学基金(32472641);中国烟草总公司科技项目(110220001019);湖南省技术攻关“揭榜挂帅”(2021NK1040);湖南中烟工业有限责任公司科技项目(KY2023YC0009);长沙市烟草公司科技项目(CS2022KJ02)。

摘　　要：双组分系统(two-component system,TCS)参与多种芽胞杆菌次级代谢产物生物合成的调控,但调控伊短菌素生物合成的TCS鲜见报道。前期转录组测序分析发现,在短短芽胞杆菌X23中,双组分因子LiaRS与伊短菌素生物合成基因簇(edeBGC)的表达趋势一致,推测其可能参与伊短菌素生物合成的调控,通过Red/ET同源重组法敲除野生型X23的liaS和liaR基因,构建了突变体菌株,平皿对峙培养、qRT-PCR定量分析和HPLC-MS靶向代谢分析等结果表明,在相同培养条件下,突变菌株的抑菌圈直径较野生型显著增加,伊短菌素生物合成基因簇的表达量显著上调,且伊短菌素的峰面积是野生型的1.3倍。本研究发现LiaRS是伊短菌素生物合成的负调控因子,为伊短菌素的代谢工程改造提供了候选基因元件。Two-component systems(TCS)perform a regulatory function in the biosynthesis of various secondary metabolites in Bacillus.However,the TCS controlling edeine biosynthesis have seldom been reported.This research,employing bioinformatics analysis,has uncovered the potential regulatory role of the two-component factor LiaRS in the biosynthesis of edeine.By employing the Red/ET homologous recombination technique,the liaS and liaR genes of the wild-type X23 were effectively eliminated,leading to the development of mutant strains.The results from plate culture,qRT-PCR quantitative analysis,and HPLC-MS targeted metabolic profiling demonstrated a significant increase in the diameter of the inhibitory zone for the mutant strains and a notable up-regulation of the expression of the edeine biosynthesis gene clusters.Moreover,the peak area of edeine was found to be 1.3 times that of the wild type.The findings of this research indicate that LiaRS functions as a suppressor of edeine biosynthesis,thereby offering potential gene elements for the metabolic engineering of edeine.

关 键 词：短短芽胞杆菌 伊短菌素 双组分系统 LiaRS Red/ET同源重组 

分 类 号：S476[农业科学—农业昆虫与害虫防治]