文章摘要
解淀粉芽孢杆菌F11抗真菌活性研究
Study on antifungal activity of Bacillus amyloliquefaciens F11
投稿时间:2020-08-23  
DOI:10.13254/j.jare.2020.0450
中文关键词: 解淀粉芽孢杆菌,生防菌,胞外代谢物粗提液,脂肽抗生素,抑菌机理
英文关键词: Bacillus amyloliquefaciens, biocontrol bacteria, extracellular metabolites, lipopeptide antibiotics, bacteriostatic mechanism
基金项目:贵州省科技计划项目(黔科合基础[2017]0106,黔科合平台人才[2017]5788号);国家自然科学基金项目(31760133);贵州大学人才引进项目([2015]10号);贵州省一流学科建设项目(GNYL[2017]009);教育部2019年度省级一流本科专业建设点项目
作者单位E-mail
冯蓉 贵州大学农学院, 贵阳 550025
贵州大学新型肥料资源与技术研究所, 贵阳 550025 
 
刘丽 贵州大学农学院, 贵阳 550025
贵州大学新型肥料资源与技术研究所, 贵阳 550025 
liuliz706@sina.com 
陈海念 贵州大学农学院, 贵阳 550025
贵州大学新型肥料资源与技术研究所, 贵阳 550025 
 
杨胜竹 贵州大学农学院, 贵阳 550025
贵州大学新型肥料资源与技术研究所, 贵阳 550025 
 
雷钦卿 贵州大学农学院, 贵阳 550025  
丁志峰 贵州大学农学院, 贵阳 550025  
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中文摘要:
      解淀粉芽孢杆菌F11对核盘病菌、玉米大斑病菌、猕猴桃软腐病菌、烟草炭疽病菌等12种作物病原菌具有高效、广谱的抑菌效果,研究其生物防治机理可为该菌株应用于植物病害防治提供理论依据。本研究采用混菌法、牛津杯法及显微镜观察法探究了菌株F11发酵液不同稀释倍数、发酵液3种分离液(菌株发酵液离心分离获得上清液A,上清液A继而进行酸沉淀和甲醇提取分别获得发酵上清液B和胞外代谢物粗提液)对病原菌的抑制效果及抑菌特性。混菌法结果表明,菌株F11的抑菌效果随稀释倍数的增加而逐渐下降,但1 000倍稀释液对樟树炭疽病菌(Colletotrichum gloeosporioides)的相对抑制率和10倍稀释液与发酵原液对烟草黑胫病菌(Phytophthora parasitica var.nicotianae)、核盘病菌(Sclerotinia sclerotiorum)、玉米大斑病菌(Setosphaeria turcica)、猕猴桃软腐病菌(Botryosphaeria dothidea)的相对抑制率均达100%,且所列稀释液与发酵原液间无显著差异。牛津杯法结果表明,上清液B中不含抑菌活性物质,而上清液A与胞外代谢物粗提液中均含有抑菌活性物质。显微镜观察结果显示,胞外代谢物粗提液中的抑菌活性物质可使病原菌菌丝消融、断裂、扭曲、缠绕变形、原生质体凝集渗漏、形成泡囊结构等。研究表明,菌株F11病原菌的抑菌效果随发酵液稀释倍数增加而下降,其对樟树炭疽病菌(Colletotrichum gloeosporioides)的防治效果最好,发酵液经1 000倍稀释后相对抑制率仍达100%;对烟草黑胫病菌、核盘病菌、玉米大斑病菌、猕猴桃软腐病菌四种病原菌则可采用10倍稀释液代替发酵原液进行防治。其抑菌活性物质存在于胞外代谢物粗提液中,该物质的抑菌特性和脂肽抗生素的作用基本一致,推测为一类脂肽抗生素。
英文摘要:
      Bacillus amyloliquefaciens F11 inhibits the growth and reproduction of 12 crop pathogens with a high inhibition rate, which shows that this strain possesses a high efficiency and broad-spectrum antibacterial effect. This provides a theoretical basis for the application of this strain in plant disease control. In this study, the mixed bacteria method, the Oxford cup method, and a microscope were used to investigate the inhibitory effect and antibacterial characteristics of the fermentation broth of strain F11 with different dilution ratios and three separate liquids. Supernatant A was separated from the fermentation broth of the strain by centrifugation, and supernatant B and the crude extract of the extracellular metabolites were separated and extracted from supernatant A by acid precipitation and methanol extraction with centrifugation, respectively. The results of the mixed bacteria method showed that the antibacterial effect of strain F11 decreased with an increase in the dilution ratio, but the relative inhibition rates of 1 000×diluent on Colletotrichum gloeosporioides and 10×diluent and fermentation broth on Phytophthora parasitica var. nicotianae, Sclerotinia sclerotiorum, Setosphaeria turcica, and Botryosphaeria dothidea were all 100%. There was no significant difference between the fermentation broth and the diluent. The results of the Oxford cup method showed that there were no antibacterial substances in supernatant B, while both supernatant A and the crude extract of extracellular metabolites contained these substances. The microscopic observations showed that the antibacterial substances in the crude extract could cause the hyphae to melt, break, twist, and intertwine to deform, leak protoplast, and form vesicle structures. The bacteriostatic effect of strain F11 decreased as the dilution ratio of the fermentation broth increased. The results showed that it had the best control effect on Colletotrichum gloeosporioides as the relative inhibition rate of the 1 000×diluent of the fermentation broth was 100%, and the 10×diluent could be used to substitute the fermentation broth to control the pathogens Phytophthora parasitica var. nicotianae, Sclerotinia sclerotiorum, Setosphaeria turcica, and Botryosphaeria dothidea. The inhibitory activity was found in the crude extract of the extracellular metabolites. Its properties were the same as those of lipopeptide antibiotics, which implied that it is probably a lipopeptide antibiotic.
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