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棘孢木霉菌与枯草芽胞杆菌共培养的最适碳源及其促进机理

引用本文：郭富强,彭治鑫,李清钰,张成省,郭亚利,朱迪,张维,芶剑渝,赵福彬,高贵,赵栋霖.棘孢木霉菌与枯草芽胞杆菌共培养的最适碳源及其促进机理.植物保护学报,2024,51(6):1335-1346

DOI：10.13802/j.cnki.zwbhxb.2024.2024057

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作者	单位	E-mail
郭富强	中国农业科学院烟草研究所, 山东青岛 266101
彭治鑫	中国农业科学院烟草研究所, 山东青岛 266101
李清钰	中国农业科学院烟草研究所, 山东青岛 266101
张成省	中国农业科学院烟草研究所, 山东青岛 266101 国家盐碱地综合利用技术创新中心, 山东东营 257300
郭亚利	贵州省烟草公司黔西南州公司, 兴义 562400
朱迪	贵州省烟草公司黔西南州公司, 兴义 562400
张维	贵州省烟草公司黔西南州公司, 兴义 562400
芶剑渝	贵州省烟草公司遵义市公司, 遵义 563000
赵福彬	山东青岛烟草有限公司, 青岛 266000
高贵	贵州省烟草公司黔西南州公司, 兴义 562400	gwufei@163.com
赵栋霖	中国农业科学院烟草研究所, 山东青岛 266101 国家盐碱地综合利用技术创新中心, 山东东营 257300	zhaodonglin@caas.cn

中文摘要:为有效防治烟草疫霉病，筛选棘孢木霉菌Trichoderma asperellum HG1和枯草芽胞杆菌Bacillus subtilis Tpb55单独培养的碳源及两者共培养的碳源，测定不同碳源下2种菌共培养发酵液和发酵液提取物对烟草疫霉菌Phytophthora nicotianae的抑制效果及最适碳源条件下共培养发酵液的盆栽防效，分离鉴定了2种菌共培养提取物中的抑菌活性物质，通过转录组学方法分析添加3.75 g/L糊精和未添加糊精条件下棘孢木霉菌HG1的基因表达差异，并采用实时荧光定量PCR（real-time quantitative PCR，RT-qPCR）技术对转录组测序结果进行验证。结果表明：棘孢木霉菌HG1单独偏好的碳源有4种，枯草芽胞杆菌Tpb55单独偏好的碳源有3种，2种菌共同偏好的碳源有6种；以糊精为碳源时，棘孢木霉菌HG1和枯草芽胞杆菌Tpb55共培养提取物对烟草疫霉菌的抑制率最高可达76.45%，其发酵液对烟草疫霉病的室内盆栽防效为100.0%，且对烟株的生长发育无负作用；以糊精作为碳源的2种菌共培养提取物中分离鉴定出的（E）-7，9-二烯-11-羰基十八酸甲酯化合物对烟草疫霉菌的抑制率为72.93%；添加糊精能激发棘孢木霉菌HG1中谷胱甘肽代谢系统，降低其氧化应激的程度，从而有利于生命活动的正常进行。

中文关键词:棘孢木霉菌枯草芽胞杆菌碳源共培养发酵优化转录组分析

Optimal carbon source for co-culture of Trichoderma asperellum HG1 and Bacillus subtilis Tpb55 and its promoting mechanisms

Author Name	Affiliation	E-mail
Guo Fuqiang	Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, Shandong Province, China
Peng Zhixin	Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, Shandong Province, China
Li Qingyu	Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, Shandong Province, China
Zhang Chengsheng	Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, Shandong Province, China National Center of Technology Innovation for Comprehensive Utilization of Saline-Alkali Land, Dongying 257300, Shandong Province, China
Guo Yali	Qianxinan Prefecture Branch, Guizhou Tobacco Company, Xingyi 562400, Guizhou Province, China
Zhu Di	Qianxinan Prefecture Branch, Guizhou Tobacco Company, Xingyi 562400, Guizhou Province, China
Zhang Wei	Qianxinan Prefecture Branch, Guizhou Tobacco Company, Xingyi 562400, Guizhou Province, China
Gou Jianyu	Zunyi Branch of Guizhou Provincial Tobacco Company, Zunyi 563000, Guizhou Province, China
Zhao Fubin	Shandong Qingdao Tobacco Co. Ltd., Qingdao 266000, Shandong Province, China
Gao Gui	Qianxinan Prefecture Branch, Guizhou Tobacco Company, Xingyi 562400, Guizhou Province, China	gwufei@163.com
Zhao Donglin	Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, Shandong Province, China National Center of Technology Innovation for Comprehensive Utilization of Saline-Alkali Land, Dongying 257300, Shandong Province, China	zhaodonglin@caas.cn

Abstract:To control the severe tobacco disease caused by Phytophthora nicotianae, the preferred carbon sources of Trichoderma asperellum HG1 and Bacillus subtilis Tpb55 individually, as well as those preferred by two strains in combination were screened in this study. The antimicrobial effects of the sterile filtrates and co-culture extracts from the two strains under different carbon sources and combinations on P. nicotianae were determined. The efficacy of control of potted plant disease with the co-cultivation fermentation broth under the optimal conditions was also explored. The antimicrobial substance in the co-culture extracts were separated and identified. The gene expressions of T. asperellum HG1 with 3.75 g/L or without the optimal carbon source dextrin were analyzed by transcriptomics. Furthermore, real-time quantitative PCR (RT-qPCR) technology was employed to validate the results of the transcriptome sequencing. The results indicated that T. asperellum HG1 individually preferred four carbon sources, B. subtilis Tpb55 individually preferred three carbon sources, and both microbes jointly preferred six carbon sources. When dextrin was used as the carbon source, the antimicrobial rate of the coculture extract reached a maximum of 76.45%, and its fermentation broth effectively prevented the infection of P. nicotianae, with a prevention effect of 100.0%, and without any negative impact on the growth and development of tobacco. (E)-7,9-diene-11-carbonyl-octadecanoic acid methyl ester, isolated and identified from the co-cultured extract with dextrin as the carbon source, exhibited an antimicrobial rate of 72.93% against P. nicotianae at a concentration of 0.3 mg/mL. The addition of dextrin stimulated the glutathione metabolic system in T. asperellum HG1, and reduced the level of oxidative stress, thus facilitated the normal progress of life activities.

keywords:Trichoderma asperellum Bacillus subtilis carbon source co-culture fermentation optimization transcriptome analysis

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