产褪黑素绿色木霉工程菌的构建及生理特性分析  

Construction and characterization of melatonin-producing engineering strain of Trichoderma viride

作　　者：李哲 张豪[1] 霍雪雪 黄艳华 郝永任 LI Zhe;ZHANG Hao;HUO Xuexue;HUANG Yanhua;HAO Yongren(Biology Institute,Qilu University of Technology(Shandong Academy of Sciences),Ji’nan 250014,Shandong,China;State Key Laboratory of Environmental Chemistry and Ecotoxicology,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,Beijing 100085,China)

机构地区：[1]齐鲁工业大学(山东省科学院)生物研究所,山东济南250014 [2]中国科学院生态环境研究中心环境化学与生态毒理学国家重点实验室,北京100085

出　　处：《微生物学通报》2023年第1期148-162,共15页Microbiology China

基　　金：山东省自然科学基金(ZR202102270296);国家自然科学基金(21607169,3227010260);山东省科学院青年科学基金项目(2020QN0019);济南市创新团队项目(2019GXRC033)。

摘　　要：【背景】褪黑素(melatonin)是动植物内广泛存在的一种小分子生物胺类物质,在促进生物生长和提高环境耐受性等方面发挥重要作用。木霉(Trichoderma)既是重要的生防菌株也是高效的工业产品生产菌株,能够合成丰富的代谢产物。【目的】针对目前木霉菌株中还未发现褪黑素合成的问题,构建具有褪黑素合成能力的绿色木霉(Trichoderma viride)工程菌,并对其生理特性进行研究。【方法】在绿色木霉Tv-1511中异源表达了来源于人基因组的芳烷基胺N-乙酰转移酶(aralkylamineN-acetyltransferase,AANAT)编码基因hAANAT和乙酰复合胺-O-甲基转移酶(acetylserotonin-O-methyltransferase,ASMT)编码基因hASMT,高效液相色谱法(highperformance liquid chromatography,HPLC)检测了木霉工程菌合成褪黑素的产量,并利用生化方法检测了工程菌的生长、抗逆及对植物的促生抗病能力。【结果】获得了具有褪黑素合成能力的绿色木霉工程菌,此株工程菌具有更好的生长和产孢特性、更强的逆境胁迫耐受能力、更高的病原菌拮抗性,以及更有效的植物促生能力。【结论】褪黑素对绿色木霉的生长和抗逆具有积极的作用,通过遗传改造获得具有褪黑素合成能力的木霉菌株,对工农业生产应用具有重要意义。[Background]Melatonin is a small-molecule biogenic amine ubiquitous in animals and plants,which promotes the growth and environmental adaptation of organisms.Trichoderma act as important biocontrol strains and engineering strains of industrial products,which can produce diverse secondary metabolites.[Objective]Since the melatonin synthesis in Trichoderma strains remains unclear,we constructed a Trichoderma viride strain capable of synthesizing melatonin and characterized its physiological properties.[Methods]In this study,the genes h AANAT encoding human alkylamine N-acetyltransferase and hASMT encoding human acetylserotonin O-methyltransferase were heterologously expressed in T.viride Tv-1511.The yield of melatonin synthesized by the engineered strain was determined by high performance liquid chromatography(HPLC).Furthermore,the growth,stress resistance,and growth-promoting and biocontrol abilities of the engineered strain were determined by biochemical methods.[Results]A melatonin-producing strain of T.viride was obtained,which had better growth and sporulation characteristics,stronger stress tolerance,stronger antagonism to pathogens,and higher plant growth-promoting ability than the start strain.[Conclusion]Melatonin plays a positive role in the growth and stress resistance of T.viride.It is of great significance to construct the Trichoderma strains with melatonin-synthesizing ability by genetic modification.

关 键 词：绿色木霉 褪黑素 基因编辑 基因功能 生理特性 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]