Analysis of Antifungal Substances Produced by Bacillus subtilis Isolate Against Fusarium oxysporum
QIAO Xin-Lei, FAN Xue-Rui, HUO Xiao-Yi, ZHANG Xue-Chao, XIAO Jia-Wen, ZHANG Dong-Dong*
College of Life Science, Hebei Agricultural University/Hebei Provincial Engineering Research Center of Agricultural Wastes Resource Utilization, Baoding 071000, China
Abstract：Bacillus subtilis Z-5 has significant antagonistic effect against many plant pathogenic fungi. In this study, antagonistic effect of strain Z-5 on Fusarium oxysporum was detected by confrontation culture. The activities of protease, amylase, cellulase and siderophore in the sterile filtrate were detected by the method of transparent circle. The antifungal substances were extracted by acid precipitation, and the antagonistic effect of the extract against F. oxysporum was detected by inhibition zone method. The extract was separated by high performance liquid chromatography(HPLC) and the activities of the separated components were detected by inhibition zone method. The structures of antifungal substances were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS) and mass spectrometry/mass spectrometry(MS/MS). The results showed that strain Z-5 presented significant antagonistic activity against F. oxysporum, and the sterile filtrate had significant protease, amylase and cellulase activities as well as the siderophore. The extracts of the sterile filtrate showed significant antagonistic effect against F. oxysporum and three antifungal components were obtained, which were identified as lipopeptide antibiotics C14 iturin A, C15 iturin A and C14-C18 fengycin A. The detection of antifungal substances from B. subtilis strain Z-5 laid the foundation for the study of its antifungal mechanism and the effective use of biocontrol agents.
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