Xu Gaoge, Zhao Yuxin, Du Liangcheng, Qian Guoliang, Liu Fengquan
College of Plant Protection, Nanjing Agricultural University, China/Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education, Nanjing, 210095, China.
Microb Biotechnol. 2015 May;8(3):499-509. doi: 10.1111/1751-7915.12246. Epub 2015 Feb 13.
Lysobacter enzymogenes is an important biocontrol agent with the ability to produce a variety of lytic enzymes and novel antibiotics. Little is known about their regulatory mechanisms. Understanding these will be helpful for improving biocontrol of crop diseases and potential medical application. In the present study, we generated an hfq (encoding a putative ribonucleic acid chaperone) deletion mutant, and then utilized a new genomic marker-free method to construct an hfq-complemented strain. We showed for the first time that Hfq played a pleiotropic role in regulating the antibacterial antibiotic biosynthesis and extracellular lytic enzyme activity in L. enzymogenes. Mutation of hfq significantly increased the yield of WAP-8294A2 (an antibacterial antibiotic) as well as the transcription of its key biosynthetic gene, waps1. However, inactivation of hfq almost abolished the extracellular chitinase activity and remarkably decreased the activity of both extracellular protease and cellulase in L. enzymogenes. We further showed that the regulation of hfq in extracellular chitinase production was in part through the impairment of the secretion of chitinase A. Collectively, our results reveal the regulatory roles of hfq in antibiotic metabolite and extracellular lytic enzymes in the underexplored genus of Lysobacter.
解淀粉芽孢杆菌是一种重要的生防菌,能够产生多种裂解酶和新型抗生素。人们对其调控机制知之甚少。了解这些机制将有助于改善作物病害的生物防治及潜在的医学应用。在本研究中,我们构建了一个hfq(编码一种假定的核糖核酸伴侣)缺失突变体,然后利用一种新的无基因组标记方法构建了一个hfq互补菌株。我们首次表明,Hfq在调控解淀粉芽孢杆菌的抗菌抗生素生物合成和胞外裂解酶活性方面发挥了多效性作用。hfq突变显著提高了WAP-8294A2(一种抗菌抗生素)的产量及其关键生物合成基因waps1的转录水平。然而,hfq失活几乎消除了解淀粉芽孢杆菌的胞外几丁质酶活性,并显著降低了胞外蛋白酶和纤维素酶的活性。我们进一步表明,hfq对胞外几丁质酶产生的调控部分是通过损害几丁质酶A的分泌实现的。总的来说,我们的结果揭示了hfq在解淀粉芽孢杆菌这一研究较少的属中对抗生素代谢物和胞外裂解酶的调控作用。