防治葡萄灰霉病的贝莱斯芽胞杆菌HMB28023筛选及其抑菌物质  

Screening of Bacillus velezensis HMB28023 for Controlling Grape Gray Mold and Identification of Its Antifungal Active Compounds

作　　者：张晓云[1] 王雪美 丛蓉 陈秀叶 鹿秀云[1] 李社增[1] 郭庆港[1] 马平[1] ZHANG Xiaoyun;WANG Xuemei;CONG Rong;CHEN Xiuye;LU Xiuyun;LI Shezeng;GUO Qinggang;MA Ping(Plant Protection Institute,Hebei Academy of Agriculture and Forestry Sciences/Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China,Ministry of Agriculture and Rural Affairs,P.R.China/IPM Innovation Center of Hebei Province/International Science and Technology Joint Research Center on IPM of Hebei Province,Baoding 071000,China;Langfang Muyuan Engineering Project Management Co.,Ltd,Langfang 065000,China)

机构地区：[1]河北省农林科学院植物保护研究所,农业农村部华北北部作物有害生物综合治理重点实验室,河北省农业有害生物综合防治技术创新中心,河北省作物有害生物综合防治国际科技联合研究中心,河北保定071000 [2]廊坊市沐源工程项目管理有限公司,河北廊坊065000

出　　处：《中国生物防治学报》2025年第1期122-131,共10页Chinese Journal of Biological Control

基　　金：河北省农林科学院科技创新专项(2022KJCXZX-ZBS-1)。

摘　　要：葡萄灰霉病是由灰葡萄孢引起的一种危害严重的病害,但目前缺乏安全有效的微生物杀菌剂。本研究采用对峙培养法,测定了对源于番茄的灰葡萄孢菌表现较高拮抗活性的27株细菌的拮抗活性,发现同一细菌菌株对源于两种作物的灰葡萄孢菌的抑菌带和抑菌率有不同程度的变化;进一步通过离体叶片试验、盆栽防病试验及田间防病试验筛选获得高效防治葡萄灰霉病的生防细菌HMB28023,田间防效达到74.3%。通过形态学、16S rDNA结合gyr A、gyr B、rpo B和rpo C多基因序列分析将其鉴定为贝莱斯芽胞杆菌Bacillus velezensis。离体叶片试验结果表明该菌株发酵液、无菌体上清液和菌体悬浮液对葡萄灰霉病的防效均较好,分别为81.9%、75.1%和64.8%。通过色谱分离结合质谱分析,证明菌株HMB28023可以产生伊枯草菌素A(C_(14)—C_(15))、丰产素(丰产素A(C_(14)—C_(17))和丰产素B(C16—C_(17)))及表面活性素(C12—C16)。抑菌活性测定结果表明,伊枯草菌素A和丰产素能够显著抑制葡萄灰霉菌的生长,并造成菌丝畸形。Grape gray mold caused by Botrytis cinerea which significantly reduces the yield and quality of grape,but there is a lack of safe and effective microbial fungicides for its control.In this study,the antagonistic activity of 27 bacteria with high antagonistic activity against B.cinerea derived from tomato,was determined by the confrontation culture method.It was found that the inhibition zone and inhibition rate of the same bacterial strain against B.cinerea derived from tomato and grape exhibited variable changes.A bacterial strain HMB28023,was further obtained based on detached leaves method,potting test and field test,which field control efficacy on grape gray mold reached 74.3%.Strain HMB28023 was identified as Bacillus velezensis through morphological characteristics,16S rDNA as well as phylogenetic tree constructed by using multiple gene sequences of gyrA,gyrB,rpoB and rpoC.The results of detached leaves test indicated that the fermentation broth,cell free supernatant and bacterial suspension of strain HMB28023 all exhibited high biocontrol efficacy against grape gray mold,reaching 81.9%,75.1%and 64.8%,respectively.The lipopeptide produced by strain HMB28023 was isolated through fast protein liquid chromatography(FPLC)and identified by UPLC-Triple TOF-MS/MS.The results showed that the lipopeptide comprised iturin A(C_(14)—C_(15)),fengycin(fengycin A(C_(14)—C_(17))and fengycin B(C16—C_(17))),and surfactin(C12—C16).Both iturin A and fengycin showed strong inhibitory activities against the growth of B.cinerea,and also caused abnormal hyphal growth in vitro.

关 键 词：贝莱斯芽胞杆菌 灰葡萄孢菌 脂肽 生物防治 

分 类 号：S476[农业科学—农业昆虫与害虫防治]