贝莱斯芽孢杆菌TCS001产脂肽类物质抑菌活性及发酵条件优化  

Antifungal activity and fermentation optimization of lipopeptides produced by Bacillus velezensis TCS001

作　　者：杨然迪 杨怡妍 曹灏 金京 陈杰 YANG Randi;YANG Yiyan;CAO Hao;JIN Jing;CHEN Jie(National Joint Local Engineering Laboratory for High-Efficient Preparation of Biopesticide,College of Forestry and Biotechnology,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学林业与生物技术学院生物农药高效制备技术国家地方联合工程实验室,杭州311300

出　　处：《农药学学报》2024年第3期504-516,共13页Chinese Journal of Pesticide Science

基　　金：浙江省重点研发计划——生态安全高效果蔬病害生物防治制剂研发(2021C02046)。

摘　　要：贝莱斯芽孢杆菌Bacillus velezensis TCS001是一株新型海洋生境生防细菌。为研究TCS001发酵液中脂肽类物质对植物病原菌的抑制作用并提高其产量,通过菌丝生长速率法测定了TCS001脂肽类物质对17种植物病原菌的抑制作用,并以灰葡萄孢菌Botrytis cinerea为靶标探究了其抑菌机理;利用单因素法和响应面分析法,对TCS001脂肽类物质的发酵条件进行了优化,以提高脂肽类物质的产量。结果表明:TCS001脂肽类物质对17种植物病原菌的抑制率为56.45%~97.15%,其中对山核桃干腐病菌Botryosphaeria dothidea、苹果轮纹病菌Macrophoma kawatsukai、油菜菌核病菌Sclerotinia sclerotiorum、小麦根腐病菌Bipolaris sorokiniana和番茄灰霉病菌Botrytis cinerea的抑制率较高,分别为97.15%、94.87%、92.64%、91.04%和89.55%。活体盆栽试验和光学显微镜观察表明,TCS001脂肽类物质对灰葡萄孢菌具有较好的抑制效果,并对菌丝有明显致畸作用。单因素试验结合响应面分析表明,影响TCS001脂肽类物质产量的3个关键因子为培养基氮源、碳源和发酵温度,最佳发酵条件为:可溶性淀粉10.5 g/L,花生饼粉18.5 g/L,NaCl 3.0 g/L,装液量32%,接种量3%,发酵液pH值为6.0,发酵温度31℃,种龄16 h,转速164 r/min,发酵培养时间48 h。响应面预测值与实际测定值之间具有较好的拟合性。最佳发酵条件下,TCS001脂肽类物质产量为0.653 g/L,是优化前产量0.41 g/L的1.59倍,该结果可为TCS001菌株的后续开发奠定基础,并为相关研究提供理论依据。Bacillus velezensis TCS001 is a novel biocontrol bacterium from marine habitat.This study investigated the inhibitory effects of lipopeptides produced by TCS001 on 17 plant pathogens through mycelial growth-rate assay,and the underlying anti-fungal mechanisms were preliminarily explored with Botrytis cinerea.Results showed that the inhibitory rates of TCS001 lipopeptides against 17 plant pathogens ranged from 56.45%to 97.15%.The inhibitory rates to Botryosphaeria dothidea(causing Carya cathayensis canker disease),Macrophoma kawatsukai,Sclerotinia sclerotiorum,Bipolaris sorokiniana,and Botrytis cinerea,were 97.15%,94.87%,92.64%,91.04%,and 89.55%,respectively.Pot test and optical microscope observation demonstrated that lipopeptides could limit B.cinerea growth in vivo and have teratogenic effect on hypha.The fermentation conditions for optimal lipopeptides production of TCS001 were determined by a single-factor experiment combined with response surface methodology.The three key factors that affected lipopeptide production most were the nitrogen source,carbon source,and fermentation temperature.The optimized fermentation conditions were as follows:soluble starch 10.5 g/L,peanut cake powder 18.5 g/L,NaCl 3.0 g/L,liquid volume 32%,inoculum volume 3%,fermentation pH 6.0,fermentation temperature 31°C,seed age 16 h,rotation speed 164 r/min,fermentation time 48 h.There is a good fit between the predicted values and the actual values of the response surface.The actual yield of lipopeptides increased to 0.653 g/L,which was 1.59 times that of 0.41 g/L before optimization.The results lay a foundation for the subsequent development of TCS001 and provide a theoretical basis.

关 键 词：贝莱斯芽孢杆菌 脂肽类物质 抑菌活性 发酵 优化 

分 类 号：S476[农业科学—农业昆虫与害虫防治]