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[目的]分离纯化枯草芽孢杆菌(Bacillus subtilis)菌株XG-1的抗西瓜枯萎病菌(Fusarium oxysporum f.sp niveum FON)的拮抗蛋白。[方法]通过硫酸铵分级沉淀获得粗蛋白,采用DEAE-Sepharose FF离子交换柱和Sephacryl S-100凝胶柱层析进行分离纯化,对具抗西瓜枯萎病菌的蛋白纯进行MALDI-TOF-MS结构分析。[结果]该拮抗蛋白与来源于枯草芽孢杆菌(B.subtilis)的鞭毛蛋白(gi|14278900)同源性较高,蛋白分值为248,覆盖率为63%,推测该拮抗蛋白可能是菌株XG-1的一种鞭毛蛋白,分子量约为30.6kD。[结论]该研究为阐明生防菌XG-1的作用机理提供理论依据,为西瓜枯萎病的防治提供重要的参考。
[Objective] The research aimed to isolate and purify the antagonistic protein of Fusarium oxysporum f.sp niveum FON against Bacillus subtilis strain XG-1. [Method] The crude protein was fractionated by ammonium sulfate fractionation and purified by DEAE-Sepharose FF ion exchange column and Sephacryl S-100 gel column chromatography. The pure proteins with Fusarium oxysporum f.sp. tumefaciens were separated by MALDI-TOF-MS analysis. [Result] The antagonistic protein had high homology with flagellin from B. subtilis (gi | 14278900) with a score of 248 and a coverage rate of 63%. It is presumed that this antagonist may be a strain A flagellin of XG-1 with a molecular weight of about 30.6 kD. [Conclusion] The study provided theoretical basis for elucidating the mechanism of biocontrol fungi XG-1 and provided important reference for the prevention and control of watermelon wilt.