解淀粉芽孢杆菌BA-26抗菌物质分离及对灰葡萄孢抑菌作用研究

doi:10.13560/j.cnki.biotech.bull.1985.2019-0055

生物技术通报 ›› 2019, Vol. 35 ›› Issue (7): 83-89.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0055

解淀粉芽孢杆菌BA-26抗菌物质分离及对灰葡萄孢抑菌作用研究

刘超^1,2,3, 刘洪伟^2,3, 汪步青^1,2, 赵雯雅^2,3, 王雅娜^2,3, 张丽萍^2,3

1. 河北工业大学,天津 300000;
2. 河北省科学院生物研究所,石家庄 050081;
3. 河北省主要农作物病害微生物控制工程技术研究中心,石家庄 050081

收稿日期:2019-01-05 出版日期:2019-07-26 发布日期:2019-07-29
作者简介:刘超,男,硕士研究生,研究方向：应用微生物;E-mail：lch1092@126.com
基金资助:
河北省高层次人才资助项目（B2018003019）,河北省科技计划项目（17222906D）,河北省科学院科技计划项目（18302,2018G01）

Isolation of Antifungal Substances from Bacillus amyloliquefaciens BA-26 and Its Antifungal Activity against Botrytis cinerea

LIU Chao^1,2,3, LIU Hong-wei^2,3, WANG Bu-qing^1,2, ZHAO Wen-ya^2,3, WANG Ya-na^2,3, ZHANG Li-ping^2,3

1. Hebei University of Technology,Tianjin 300000;
2. Institute of Biology,Hebei Academy of Science,Shijiazhuang 050081;
3. Main Crops Disease of Microbial Control Engineering Technology Research Center in Hebei Province,Shijiazhuang 050081

Received:2019-01-05 Published:2019-07-26 Online:2019-07-29

摘要/Abstract

摘要： 分析解淀粉芽孢杆菌（Bacillus amyloliquefaciens）BA-26发酵液中抑菌物质存在情况,明确活性物质的抑菌效果,初步探索其抗植物病原真菌的作用机制。通过硫酸铵沉淀、大孔树脂吸附及反相高效液相色谱法（RP-HPLC）,从菌株BA-26的发酵液中分离纯化抗菌物质;采用平板对峙法检测抗菌物质抑菌活性;用二倍稀释法测定各活性组分的最小抑菌浓度（MIC）,并研究其对菌丝和孢子萌发的影响。从菌株BA-26发酵液中获得抗菌粗提物抑菌谱广,对立枯丝核菌等多种植物病原真菌有强抑制作用;分离纯化得到11个具有抑制真菌作用的活性组分;其中A6-19和A6-20组分对灰葡萄孢（Botrytis cinerea）抑菌活性最强,最小抑菌浓度为7.81 μg/mL;菌株BA-26产生的抗真菌活性物质能使灰葡萄孢菌丝生长受阻、膨大变粗、细胞膜破坏并能抑制其孢子萌发。明确了解淀粉芽孢杆菌BA-26菌株产抗菌物质情况,初步探究其抑菌作用机制,为抗菌物质在防治病原真菌等方面的应用奠定基础。

关键词: 解淀粉芽孢杆菌, 抗菌化合物, 灰葡萄孢, 抑菌作用

Abstract: The objective of this study are to analyze the antifungal substances produced by Bacillus amyloliquefaciens BA-26,to clarify the antifungal effects of these active substances,and to explore the mechanisms of their inhibition to plant pathogenic fungi. Antifungal substances produced by B. amyloliquefaciens BA-26 were isolated and purified by ammonium sulfate deposition,macroporous resin and RP-HPLC methods. Using the plate diffusion method,the inhibitory spectrum and the antifungal activities of these components were detected. The minimum inhibitory concentration of each purified component was determined by double dilution method,and their antifungal mechanisms to hyphae and spore germination were studied. The crude extract from strain BA-26 fermentation presented a broad inhibitory spectrum,and had significant inhibitory effect on various plant pathogenic fungi,such as Rhizoctonia solani and so on. Eleven antifungal components were isolated from the antifungal crude extract. The antifungal activity of A6-19 and A6-20 components showed the strongest antifungal activity against Botrytis cinerea,and the minimum inhibitory concentration was 7.81 μg/mL. These components inhibited the growth of B. cinerea mycelium,and resulted in them enlarged and thickened;in addition,they destroyed cell membrane,and inhibited spore germination. In this study,antifungal substances of B. amyloliquefaciens BA-26 were isolated and their mechanisms were preliminarily explored. These results laid the foundation for the application of these antimicrobial substances,especially in the field of anti-pathogenic fungi.

Key words: Bacillus amyloliquefaciens, antibacterialcompounds, Botrytis cinerea, bacteriostatic effects

刘超, 刘洪伟, 汪步青, 赵雯雅, 王雅娜, 张丽萍. 解淀粉芽孢杆菌BA-26抗菌物质分离及对灰葡萄孢抑菌作用研究[J]. 生物技术通报, 2019, 35(7): 83-89.

LIU Chao, LIU Hong-wei, WANG Bu-qing, ZHAO Wen-ya, WANG Ya-na, ZHANG Li-ping. Isolation of Antifungal Substances from Bacillus amyloliquefaciens BA-26 and Its Antifungal Activity against Botrytis cinerea[J]. Biotechnology Bulletin, 2019, 35(7): 83-89.

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解淀粉芽孢杆菌BA-26抗菌物质分离及对灰葡萄孢抑菌作用研究

Isolation of Antifungal Substances from Bacillus amyloliquefaciens BA-26 and Its Antifungal Activity against Botrytis cinerea

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