Xu Zhihui, Zhang Ruifu, Wang Dandan, Qiu Meihua, Feng Haichao, Zhang Nan, Shen Qirong
Key Laboratory of Plant Nutrition and Fertilization in Low-Middle Reaches of the Yangtze River, Ministry of Agriculture, Nanjing, China, and National Engineering Research Center for Organic-based Fertilizers and Jiangsu Collaborative Innovation Center for Solid Organic Waste Resource Utilization, Nanjing Agricultural University, Nanjing, China.
Appl Environ Microbiol. 2014 May;80(9):2941-50. doi: 10.1128/AEM.03943-13. Epub 2014 Feb 28.
Bacillus amyloliquefaciens strain SQR9, isolated from the cucumber rhizosphere, suppresses the growth of Fusarium oxysporum in the cucumber rhizosphere and protects the host plant from pathogen invasion through efficient root colonization. In the Gram-positive bacterium Bacillus, the response regulator DegU regulates genetic competence, swarming motility, biofilm formation, complex colony architecture, and protease production. In this study, we report that stepwise phosphorylation of DegU in B. amyloliquefaciens SQR9 can influence biocontrol activity by coordinating multicellular behavior and regulating the synthesis of antibiotics. Results from in vitro and in situ experiments and quantitative PCR (qPCR) studies demonstrate the following: (i) that the lowest level of phosphorylated DegU (DegU∼P) (the degQ mutation) impairs complex colony architecture, biofilm formation, colonization activities, and biocontrol efficiency of Fusarium wilt disease but increases the production of macrolactin and bacillaene, and (ii) that increasing the level of DegU∼P by degQ and degSU overexpression significantly improves complex colony architecture, biofilm formation, colonization activities, production of the antibiotics bacillomycin D and difficidin, and efficiency of biocontrol of Fusarium wilt disease. The results offer a new strategy to enhance the biocontrol efficacy of Bacillus amyloliquefaciens SQR9.
解淀粉芽孢杆菌菌株SQR9从黄瓜根际分离得到,它能抑制黄瓜根际尖孢镰刀菌的生长,并通过高效的根部定殖保护宿主植物免受病原菌入侵。在革兰氏阳性菌芽孢杆菌中,应答调节因子DegU调控遗传感受态、群体运动性、生物膜形成、复杂菌落结构以及蛋白酶产生。在本研究中,我们报道了解淀粉芽孢杆菌SQR9中DegU的逐步磷酸化可通过协调多细胞行为和调节抗生素合成来影响生物防治活性。体外和原位实验以及定量PCR(qPCR)研究结果表明:(i)最低水平的磷酸化DegU(DegU∼P)(degQ突变)损害复杂菌落结构、生物膜形成、定殖活性以及对枯萎病的生物防治效率,但增加了大环内酯菌素和芽孢杆菌烯的产生;(ii)通过degQ和degSU过表达增加DegU∼P水平可显著改善复杂菌落结构、生物膜形成、定殖活性、抗生素杆菌霉素D和艰难梭菌素的产生以及对枯萎病的生物防治效率。这些结果为提高解淀粉芽孢杆菌SQR9的生物防治效果提供了一种新策略。