中国烟草科学2023，Vol.44Issue(6)：36-43,8.DOI:10.13496/j.issn.1007-5119.2023.06.006
固态共培养木霉和芽孢杆菌抗烟草黑胫病及其活性成分研究
Study on Co-culture of Trichoderma asperellum and Bacillus subtilis by Solid-state Fermentation against Tobacco Black Shank Disease and Their Bioactive Secondary Metabolites
摘要
Abstract
In order to improve the biomass of Trichoderma asperellum HG1 and Bacillus subtilis tpb55 in the co-culture system,and increase their anti-oomycetes activities against Phytophthora nicotianae,and to explore the bioactive ingredients,the solid-state fermentation system was constructed.The anti-oomycetes activities were evaluated by the mycelium growth rate method.The solid-state fermentation conditions were determined by the anti-oomycetes activities and biomass of HG1 and tpb55.The bioactivity guided separation and isolation of fermentation extracts by silica gel and octadecylsilyl silica gel(ODS)column chromatography(CC),as well as high performance liquid chromatography(HPLC),together with NMR and ESI mass spectrometry were applied to clarify and identify the bioactive ingredients.Finally,pot experiments were applied to verify the control effect of solid-state co-culture of HG1 and tpb55 on tobacco black shank.The results showed that the optimal solid-state fermentation condition was as follows:the inoculation sequence was first B.subtilis(106 CFU/mL,3％)12 h and then inoculated with T.asperellum(106 CFU/mL,1.5％).After 14 d of fermentation,under the optimal condition,the bacterial amounts of tpb55 were 1.58x1010 CFU/g and the spore amounts of HG1 were 6.75× 1010 CFU/g,with the strongest anti-oomycete activity towards P.nicotianae(inhibition rate 73.25％).A pure compound was isolated from the bioactive fraction and identified as harziandione.The results of pot experiments showed that the control effect of solid co-culture strains on tobacco black shank was significantly better than that of single culture,reached 61.94％.The co-cultured strains could also improve soil physical and chemical properties.This study provided a new idea and method for the research and development of co-culture biocontrol agents for tobacco black shank disease.关键词
固态发酵/共培养/抗卵菌活性/harziandioneKey words
solid state fermentation/co-culture/anti-oomycetes activity/harziandione分类
农业科技引用本文复制引用
彭治鑫,赵栋霖,林伟,高贵,管志坤,王耀斌,张成省,李伟..固态共培养木霉和芽孢杆菌抗烟草黑胫病及其活性成分研究[J].中国烟草科学,2023,44(6):36-43,8.基金项目
福建省烟草公司南平市公司科技项目(NYK2021-01-03) （NYK2021-01-03）
中国烟草总公司黔西南州公司科技项目(2022-07) （2022-07）
山东省烟草公司科技项目(201837020027209) （201837020027209）
青岛市科技惠民示范专项(23-3-8-xdny-2-nsh) （23-3-8-xdny-2-nsh）
