芽胞杆菌与嘧菌酯协同应用对葡萄叶部微生物群落及霜霉菌线粒体功能的影响  

Synergistic effects of Bacillus subtilis and azoxystrobin on microbial community on grapevine leaves and mitochondrial function of downy mildew pathogen Plasmopara viticola

作　　者：毕秋艳[1] 张诗琪 赵建江[1] 路粉[1] 吴杰 韩秀英[1] Bi Qiuyan;Zhang Shiqi;Zhao Jianjiang;Lu Fen;Wu Jie;Han Xiuying(Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China,Ministry of Agriculture,Integrated Pest Management Center of Hebei Province,Institute of Plant Protection,Hebei Academy of Agricultural and Forestry Sciences,Baoding 071000,Hebei Province,China)

机构地区：[1]河北省农林科学院植物保护研究所,河北省农业有害生物综合防治工程技术研究中心,农业部华北北部作物有害生物综合治理重点实验室,保定071000

出　　处：《植物保护学报》2022年第4期1252-1262,共11页Journal of Plant Protection

基　　金：河北省自然科学基金(C2019301093);国家重点研发计划(2016YFD0200505)。

摘　　要：为明确生防菌剂枯草芽胞杆菌Bacillus subtilis HMB20428与化学杀菌剂嘧菌酯协同应用对葡萄霜霉菌Plasmopara viticola的抑制机制,结合生物学测定与显微表型观察方法分析其对葡萄霜霉病菌及活孢子囊的抑制作用,采用高通量测序技术分析其对葡萄叶部微生物种群多样性的影响,通过测定生理生化指标研究其对葡萄霜霉菌线粒体功能的作用。结果显示,枯草芽胞杆菌HMB20428与嘧菌酯协同应用对葡萄霜霉病菌的抑制作用显著,对活孢子囊的抑制率为99.05%;两者协同应用对微生物细菌种群有显著影响,枯草芽胞杆菌的优势占位作用明显;但对微生物真菌种群无显著影响;两者协同应用造成葡萄霜霉菌线粒体膜通透性转运孔开放度增加至90.59%,对跨膜电位的抑制率为94.29%,呼吸链酶复合物Ⅰ~Ⅴ的活性分别显著下降56.39%、22.47%、47.90%、60.52%和90.91%,说明葡萄霜霉菌线粒体膜在受到明显损伤和出现代谢功能障碍的同时,呼吸链酶活性明显被抑制,从而导致葡萄霜霉菌的能量代谢和生长受阻。表明枯草芽胞杆菌HMB20428与嘧菌酯协同应用时主要通过促进枯草芽胞杆菌竞争占位和增强对霜霉菌线粒体功能的破坏作用达到预防与治疗葡萄霜霉病的目的。In order to clarify the inhibitory mechanism of the biocontrol agent Bacillus subtilis HMB20428 and the chemical fungicide azoxystrobin against plant-pathogenic oomycete Plasmopara viticola,bioassay and microscopic phenotyping combination were used to study the inhibitory effect on live sporangia of P.viticola,high-throughput sequencing was used to analyze the influence on the bacterial and fungal diversity of microbial populations,and physiological and biochemical index determination were used to study the effect on mitochondrial function of P.viticola.The results showed that B.subtilis HMB20428 and azoxystrobin had significant synergistic effects on P.viticola,and the inhibition rate of live sporangia was 99.05%;the synergistic application had significant impact on the microbial bacterial population,improving the dominant position of B.subtilis,however there was no significant impact on the microbial fungal population;the opening of mitochondrial permeability transition pore increased to 90.59%,the inhibition rate to mitochondrial transmembrane potential was 94.29%,and the mitochondrial respiratory chain complex Ⅰ-Ⅴ activities were significantly reduced by 56.39%,22.47%,47.90%,60.52% and 90.91%,respectively,indicating that the mitochondrial membrane of P.viticola was damaged and metabolic dysfunction occurred,while the enzyme activity of mitochondrial respiratory chain was inhibited,resulting in energy metabolism and growth of P.viticola hindered.In conclusion,the synergistic application of B.subtilis HMB20428 and azoxystrobin achieves the purpose of preventing and curing grape downy mildew mainly through improving the competitive occupation of grapevine leaf niche by B.subtilis and enhancing the damage to mitochondrial function of P.viticola.

关 键 词：枯草芽胞杆菌 葡萄霜霉菌 协同作用 微生物种群 线粒体功能 机制 

分 类 号：S436.631.1[农业科学—农业昆虫与害虫防治]