贝莱斯芽孢杆菌SF18-3的鉴定及对辣椒细菌性斑点病的防效评价  

Identification and Evaluation of Bacillus subtilis SF18-3 Against Bacterial Spot Disease of Pepper

作　　者：王辉[1] 刘丽[1] 于舒怡[1] 关天舒[1] 邹春蕾[2] 李柏宏[1] 郑丽娇 WANG Hui;LIU Li;YU Shuyi;GUAN Tianshu;ZOU Chunlei;LI Baihong;ZHENG Lijiao(Institute of Plant Protection,,Liaoning Academy of Agricultural Sciences,Shenyang 110161,China;Institute of Vegetable Research,Liaoning Academy of Agricultural Sciences,Shenyang 110161,China;Xinmin Rural Revitalization and Development Center,Xinmin Liaoning110300,China)

机构地区：[1]辽宁省农业科学院植物保护研究所,沈阳110161 [2]辽宁省农业科学院蔬菜研究所,沈阳110161 [3]新民市乡村振兴发展中心,辽宁新民110300

出　　处：《沈阳农业大学学报》2024年第4期417-425,共9页Journal of Shenyang Agricultural University

基　　金：沈阳市中青年科技创新人才支持计划项目(RC210404);沈阳市农村科技特派团专项项目(23-411-2-19)。

摘　　要：近年辽宁地区辣椒细菌性斑点病[Xanthomonas campestris pv.vesicatoria(Doidge)Dye]发生严重,为挖掘新的优质高效生防资源,以辣椒细菌性斑点病菌BWB3为靶标,采用稀释平板涂布法从动物粪肥混合物中分离获得1株高效生防细菌SF18-3。为进一步明确SF18-3的生防潜力,利用抑菌圈法优化最佳发酵条件后,采用双抗体夹心法测定其发酵液中6种胞外酶活性,测定其室内抑菌活性并评价田间盆栽防治效果,并通过形态特征观察、生理生化试验与16S rDNA序列分析确定其分类地位。结果表明:以NB为基础培养基140 r·min^(-1)恒温30℃发酵84 h后SF18-3抑菌效果最好,发酵液中蛋白酶活性与木聚糖酶活性较高,分别为536.13 U·L^(-1)与564.22 U·L^(-1);最佳发酵条件下平板抑菌圈直径为3.40 cm,盆栽防治效果达70.17%;显微镜观察SF18-3菌体呈杆状、有芽孢;革兰氏染色为阳性。16Sr DNA序列与已知贝莱斯芽孢杆菌(Bacillus velezensis)OK169608及(B.velezensis)OK147645聚为一类,结合其菌落形态与生理生化特征,确定菌株SF18-3为贝莱斯芽孢杆菌(Bacillus velezensis)。In recent years,bacterial spot disease of pepper[Xanthomonas campestris pv.vesicatoria(Doidge)Dye]has occurred seriously in Liaoning province.In this study,a strain of biocontrol bacterium SF18-3,known for its high efficacy,was obtained from a mixture of animal manure using the dilution plate coating method.The target organism for this isolation was bacterial mottle blotch BWB3 in pepper plants.To further clarify the biocontrol effect of SF18-3,the optimal fermentation conditions were optimized by antibacterial zone method,and the 6 extracellular enzyme activities were determined by double antibody sandwich method.The antibacterial activities were determined in laboratory test and the control efficiency were evaluated in pod experiment.The taxonomic status of SF18-3 was determined by morphological characteristics observation,physiological and biochemical tests and 16S rDNA sequence analysis.The results showed that SF18-3 had the best bacterial inhibition effect after fermentation for 84 h at 30℃with NB as the basal medium at 140 r·min^(-1).The protease activity and xylanase activity in the fermentation broth were higher,which were 536.13 U·L^(-1) and 564.22 U·L^(-1),respectively.Under optimal fermentation conditions,the diameter of the transparent circle was 3.40 cm on the plate and a 70.17%control efficiency on potting.SF18-3 was rod-shaped,spore,Gram-positive bacterium,and 16Sr DNA sequence was clustered with Bacillus velezensis OK169608 and B.velezensis OK147645.According to colony morphology and physiological and biochemical characteristics,strain SF18-3 was identified as B.velezensis.

关 键 词：辣椒细菌性斑点病 贝莱斯芽孢杆菌 胞外酶 防治效果 鉴定 

分 类 号：S476[农业科学—农业昆虫与害虫防治]