【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(C₁₄—C₁₅)、丰产素(丰产素A(C₁₄—C₁₇)和丰产素B(C₁₆—C₁₇))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。更多还原

【Abstract】 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 o27 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,wa 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 morphologica characteristics,16S rDNA as well as phylogenetic tree constructed by using multiple gene sequences of gyrA,gyr B 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,reaching81.9%,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 (C₁₄—C₁₅),fengycin (fengycin A (C₁₄—C₁₇) and fengycin B (C₁₆—C₁₇)),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.更多还原