Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, 50 Zhongling Street, Nanjing, 210014, China,
Appl Microbiol Biotechnol. 2015 Feb;99(4):1897-910. doi: 10.1007/s00253-014-6195-4. Epub 2014 Nov 16.
The antagonistic activity of lipopeptides in Bacillus subtilis 916 has been well documented, yet relatively little is known about their mechanism in biofilm formation and environmental colonization. This study sought to examine the interaction of B. subtilis 916 on Rhizoctonia solani-infected rice sheath to elucidate the mechanism of colonization on plant leaves. Results showed that the mutants Δbac, Δsrf, and Δsrf + bac of B. subtilis 916, deficient in bacillomycin L and surfactin production, respectively, not only altered colony morphology but also changed swarming motility, reduced antagonistic activity, and decreased biofilm formation. In particular, biofilm formation in mutant Δbac, not Δsrf or Δsrf + bac, were restored with addition of surfactin and bacillomycin L at 10 and 50 μg/mL, respectively. Moreover, surfactin and bacillomycin L were able to restore or enhance swarming motility in the corresponding mutants at 10 μg/mL, respectively. With the aid of green fluorescent protein tagging, it was demonstrated that B. subtilis 916 formed a robust biofilm on the rice sheath blight lesion and colonized well on R. solani-infected rice sheath, while its corresponding mutants performed poorly. These observations also correlated with the rice cultivar pot experiments, in which B. subtilis 916 exhibited greater biocontrol than its mutants. Our results suggest that surfactin and bacillomycin L contribute differently but synergistically to the biocontrol of rice sheath blight in B. subtilis 916 through its antifungal activity, biofilm formation, and colonization.
枯草芽孢杆菌 916 中脂肽的拮抗活性已有很好的记录,但对其在生物膜形成和环境定殖中的机制知之甚少。本研究旨在研究枯草芽孢杆菌 916 对水稻纹枯病菌的相互作用,以阐明其在植物叶片定殖的机制。结果表明,枯草芽孢杆菌 916 的突变体Δbac、Δsrf 和Δsrf+bac,分别缺乏杆菌霉素 L 和表面活性素的产生,不仅改变了菌落形态,而且改变了群集运动性,降低了拮抗活性,减少了生物膜的形成。特别是,突变体Δbac 的生物膜形成,而不是Δsrf 或Δsrf+bac,在用 10 和 50μg/mL 的表面活性素和杆菌霉素 L 分别添加后得到恢复。此外,表面活性素和杆菌霉素 L 分别能够在相应的突变体中以 10μg/mL 恢复或增强群集运动性。借助绿色荧光蛋白标记,证明枯草芽孢杆菌 916 在水稻纹枯病病斑上形成了一个强大的生物膜,并很好地定殖在感染纹枯病菌的水稻叶鞘上,而其相应的突变体则表现不佳。这些观察结果也与水稻品种盆栽试验相吻合,枯草芽孢杆菌 916 的生物防治效果优于其突变体。我们的研究结果表明,表面活性素和杆菌霉素 L 通过其抗真菌活性、生物膜形成和定殖,对枯草芽孢杆菌 916 防治水稻纹枯病有不同但协同的作用。