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  

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

作　　者：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 Guo Fuqiang;Peng Zhixin;Li Qingyu;Zhang Chengsheng;Guo Yali;Zhu Di;Zhang Wei;Gou Jianyu;Zhao Fubin;Gao Gui;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;Qianxinan Prefecture Branch,Guizhou Tobacco Company,Xingyi 562400,Guizhou Province,China;Zunyi Branch of Guizhou Provincial Tobacco Company,Zunyi 563000,Guizhou Province,China;Shandong Qingdao Tobacco Co.Ltd.,Qingdao 266000,Shandong Province,China)

机构地区：[1]中国农业科学院烟草研究所,山东青岛266101 [2]国家盐碱地综合利用技术创新中心,山东东营257300 [3]贵州省烟草公司黔西南州公司,兴义562400 [4]贵州省烟草公司遵义市公司,遵义563000 [5]山东青岛烟草有限公司,青岛266000

出　　处：《植物保护学报》2024年第6期1335-1346,共12页Journal of Plant Protection

基　　金：贵州省烟草公司黔西南州公司科技项目(2022-07);中国烟草总公司重大科技项目(110202101057(LS-17));国家盐碱地综合利用技术创新中心揭榜挂帅项目(GYJ2023005)。

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。To control the severe tobacco disease caused by Phytophthora nicotianae,the preferred car-bon 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 ster-ile 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 HGl 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 transcrip-tome 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 pre-ferred six carbon sources.When dextrin was used as the carbon source,the antimicrobial rate of the co-culture extract reached a maximum of 76.45%,and its fermentation broth effectively prevented the in-fection 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 HGl,and reduced the level of oxidative stress,thus facilitated the normal progress of life activities.

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 

分 类 号：TS2[轻工技术与工程—食品科学与工程]