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产褪黑素绿色木霉工程菌的构建及生理特性分析
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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)


Construction and characterization of melatonin-producing engineering strain of Trichoderma viride
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    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(aralkylamine N-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(high performance 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。

    Abstract:

    [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 hAANAT 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.

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李哲,张豪,霍雪雪,黄艳华,郝永任. 产褪黑素绿色木霉工程菌的构建及生理特性分析[J]. 微生物学通报, 2023, 50(1): 148-162

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  • 收稿日期:2022-04-19
  • 最后修改日期:2022-05-12
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  • 在线发布日期: 2023-01-03
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