短短芽孢杆菌Brevibacillus brevs HAB-5主要抑菌活性成分的分析及其特性研究 |
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引用本文: | 杨廷雅,孙亮,周婷婷,沈适存,吴小燕,缪卫国,郑服丛. 短短芽孢杆菌Brevibacillus brevs HAB-5主要抑菌活性成分的分析及其特性研究[J]. 中国生物防治, 2014, 0(2): 222-231 |
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作者姓名: | 杨廷雅 孙亮 周婷婷 沈适存 吴小燕 缪卫国 郑服丛 |
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作者单位: | 海南大学环境与植物保护学院/海南省热带生物资源可持续利用重点实验室,海口570228 |
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基金项目: | 海南大学社会服务专项基金(HDSF201305);国家自然科学基金(31160359);国家“973”计划项目(2011CB111612);国家农业产业技术体系建设(CARS-034-GW8);教育部博士点基金(20104601110004);海南大学科研启动资金(kyqd1006) |
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摘 要: | 本文采用乙醇低温沉淀法及 Sephadex G-50凝胶柱等手段分离纯化短短芽孢杆菌 Brevibacillus brevs HAB-5主要活性成分。针对芒果炭疽病菌C7-2,菌株HAB-5发酵产生抑菌活性物质的最佳培养基为改良的LB,命名为ZS,其最佳发酵条件为28℃、2 d和装液量为50 mL/250 mL。菌株HAB-5总蛋白没有拮抗活性,但HAB-5菌液的上清液能产生较强的抑菌效果;同时,在单独培养和与病原菌对峙培养下,取距HAB-5菌落1 cm处的培养基(MB)均对病原菌产生了明显的抑菌圈。菌株HAB-5上清液经硫酸铵沉淀不具有抑菌活性,但采用低温乙醇法等提取具有抑菌作用的菌株HAB-5上清液或MB,其粗提物均产生较强的抑菌效果,再经Sephadex G-50凝胶柱过柱和SDS-PAGE分析,在14.4 kD位置均具有较强信号的条带。因此,菌株HAB-5主要抑菌活性物质为肽类,其中14.4 kD大小的肽类是菌株HAB-5重要抑菌成分。
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关 键 词: | 短短芽孢杆菌 芒果炭疽病菌 抑菌活性成分 肽类 |
Analysis on the Key Antagonistic Active Compositions of Brevibacillus brevs HAB-5 and Its Characteristics |
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Abstract: | In this study the active antagonistic components from Brevibacillus brevs HAB-5 strain were isolated and purified mainly through low-temperature ethanol precipitation and Sephadex G-50 Gel column. The optimal fermentation medium ZS of HAB-5 was developed from LB to make HAB-5 produce the strongest antagonistic activity against Colletotrichum gloeosporioides Pen C7-2, and the optimal culture condition was incubation at 28 ℃for 2 d with medium liquid volume in 50 mL/250 mL. Total bacterial proteins from HAB-5 did not show antagonism against the pathogen, but its supernatant exhibited strong antifungal effect. In the meantime, the medium block (MB), taken 1 cm from HAB-5 colony from ZS medium produced a significant inhibition zone against the pathogen. The HAB-5 supernatant treated by ammonium sulfate did not have antifungal activity, however, the crudes that were extracted from the supernatant or MB through low-temperature ethanol precipitation showed strong antifungal effect against the pathogen. Furthermore, the SDS-PAGE analysis of the crudes purified with Sephadex G-50 gel column showed a single electrophoresis band at 14.4 kD. Therefore, our results indicate that the primary antagonistic activity of peptides and the peptide with 14.4 kD may play an important role in it. |
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Keywords: | Brevibacillus brevs Colletotrichum gloeosporioides antagonistic active compositions peptide |
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