枯草芽胞杆菌对藜麦黑茎病的防治效果及其抑菌物质鉴定

doi:10.16409/j.cnki.2095-039x.2025.02.001

摘要/Abstract

摘要： 藜麦黑茎病是近年来发现的藜麦生产中普遍存在的病害之一，本研究前期从藜麦穗中分离出1株对藜麦黑茎病病原菌Ascochyta caulina具有良好抑制作用的枯草芽胞杆菌LS3，为明确其防效及抑菌物质组成，本研究采用生长速率法，对菌株LS3挥发性和非挥发性抑菌物质及其作用效果进行测定；采用随机区组设计，对菌株LS3发酵液进行田间防治效果试验；采用GC-MS对其挥发性抑菌物质进行鉴定；采用硫酸铵沉淀法和离子交换层析法，对LS3菌株的非挥发性抑菌物质进行分离和纯化，并使用HPLC-MS对其进行鉴定。结果表明，菌株LS3挥发物对藜麦黑茎病病原菌的抑制率为43.47%，发酵液的抑制率为61.45%，发酵液经高温灭菌后抑制率仅为13.08%；不同浓度LS3菌株发酵液均对藜麦黑茎病具有一定防治效果，其中2.0×10⁶ cfu/mL发酵液的防效达66.81%；对其主要挥发物成分进行鉴定，发现为2-壬醇、苯酚和十三烷醇，其中2-壬醇和苯酚对藜麦黑茎病菌的抑制效果较好，EC₅₀分别为2.57和1.24 μg/mL；将LS3菌株的抗菌粗蛋白分离、纯化后，显示只有一个活性峰，对该峰进行聚丙烯酰凝胶电泳发现1条较清晰的条带，经HPLC-MS检测后发现该蛋白是分子量大小约为32.6 kDa的鞭毛蛋白。菌株LS3表现出良好的抑菌作用，鉴定其抑菌物质为其进一步开发和利用鉴定基础。

关键词: 藜麦黑茎病, 枯草芽胞杆菌, 抑菌作用, 抗菌物质

Abstract: Quinoa black stem disease is one of the common diseases found in quinoa production in recent years. In the previous study, a strain of Bacillus subtilis LS3 with good inhibitory effect on Ascochyta caulina was isolated from quinoa ear. In order to clarify its control effect and the composition of antifungal substances, the growth rate method was used to determine the inhibitory effect of volatile and non-volatile antifungal substances of strain LS3. The field control effect of LS3 strain fermentation broth was determined by randomized block design. The volatile antifungl substances were identified by GC-MS. The non-volatile antifungl substances of LS3 strain were isolated and purified by ammonium sulfate precipitation and ion exchange chromatography, and identified by HPLC-MS. The results showed that the inhibition rate of volatiles of strain LS3 was 43.47%, the inhibition rate of fermentation broth was 61.45%, and the inhibition rate of fermentation broth after high temperature sterilization was only 13.08%. Different concentrations of LS3 strain fermentation broth had different control effects on quinoa black stem disease. When the concentration was 2.0×10⁶ cfu/mL, the control effect was 66.81%. The main volatile components of LS3 were identified as 2-nonanol, phenol and tridecanol. Among them, 2-nonanol and phenol had better inhibitory effect on the pathogen, and the EC₅₀ values were 2.57 and 1.24 μg/mL, respectively. The crude antifungal protein of LS3 strain was separated and purified, and only one active peak was found. A clear band was found by polyacrylamide gel electrophoresis. After HPLC-MS detection, it was found that the protein was a flagellin with a molecular weight of about 32.6 kDa. LS3 strain showed good antifungal activity, and identification of antifungal substances was the basis for further development and utilization of LS3 strain.

Key words: Ascochyta caulina, Bacillus subtilis, antifungal effect, antifungal substances

中图分类号:

S476

任璐, 孙江伟, 吕红, 殷辉, 秦楠, 赵晓军. 枯草芽胞杆菌对藜麦黑茎病的防治效果及其抑菌物质鉴定[J]. 中国生物防治学报, 2025, 41(1): 112-121.

REN Lu, SUN Jiangwei, Lü Hong, YIN Hui, QIN Nan, ZHAO Xiaojun. Control Effect of Bacillus subtilis on Quinoa Black Stem Disease and Identification of Its Antifungal Substances[J]. Chinese Journal of Biological Control, 2025, 41(1): 112-121.

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