Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing, China.
Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Mississippi State, Mississippi, USA.
Appl Environ Microbiol. 2021 Mar 11;87(7). doi: 10.1128/AEM.02840-20.
YL-1 has extensive antimicrobial activities against phytopathogens, and its genome harbors a pyoverdine (PVD) biosynthesis gene cluster. The alternative sigma factor PvdS in PAO1 acts as a critical regulator in response to iron starvation. The assembly of the PVD backbone starts with peptide synthetase enzyme PvdL. PvdF catalyzes formylation of l-OH-Orn to produce l--hydroxyornithine. Here, we describe the characterization of PVD production in YL-1 and its antimicrobial activity in comparison with that of its PVD-deficient , , and mutants, which were obtained using a -based site-specific mutagenesis strategy. Using methods, we examined the effect of exogenous iron under low-iron conditions and an iron-chelating agent under iron-sufficient conditions on PVD production, antibacterial activity, and the relative expression of the PVD transcription factor gene in YL-1. We found that strain YL-1, the mutant, and the (pUCP26-) complemented strain produced visible PVDs and demonstrated a wide range of inhibitory effects against Gram-negative and Gram-positive bacteria under low-iron conditions and that with the increase of iron, its PVD production and antibacterial activity were reduced. The antibacterial compounds produced by strain YL-1 under low-iron conditions were PVDs based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Moreover, the antibacterial activity observed was correlated with control efficacies of strain YL-1 against rice bacterial leaf blight (BLB) disease caused by pv. oryzae. Collectively, PVDs are responsible for the antibacterial activities of strain YL-1 under both natural and induced low-iron conditions. The results demonstrated that PVDs are essential for the broad-spectrum antibacterial activities of strain YL-1 against both Gram-positive and Gram-negative bacteria under low-iron conditions. Our findings also highlight the effect of exogenous iron on the production of PVD and the importance of this bacterial product in bacterial interactions. As a biocontrol agent, PVDs can directly inhibit the proliferation of the tested bacteria in addition to participating in iron competition.
YL-1 对植物病原体具有广泛的抗菌活性,其基因组中含有一个绿脓菌素(PVD)生物合成基因簇。PAO1 中的替代 sigma 因子 PvdS 作为对铁饥饿的反应的关键调节剂。PVD 骨干的组装始于肽合成酶酶 PvdL。PvdF 催化 l-OH-Orn 的甲酰化,生成 l--羟基鸟氨酸。在这里,我们描述了 YL-1 中 PVD 产生的特征及其与 PVD 缺陷的比较, 和 突变体的抗菌活性,这些突变体是使用基于 的位点特异性诱变策略获得的。使用 方法,我们研究了在低铁条件下外源性铁和在铁充足条件下铁螯合剂对 PVD 产生、抗菌活性和 YL-1 中 PVD 转录因子基因 的相对表达的影响。我们发现,菌株 YL-1、 突变体和(pUCP26-)互补菌株在低铁条件下产生可见的 PVDs,并表现出对革兰氏阴性和革兰氏阳性细菌的广泛抑制作用,并且随着铁的增加,其 PVD 产生和抗菌活性降低。根据液相色谱-串联质谱(LC-MS/MS)分析,菌株 YL-1 在低铁条件下产生的抗菌化合物是 PVDs。此外,观察到的抗菌活性与菌株 YL-1 对 pv. oryzae 引起的水稻细菌性叶斑病(BLB)的控制效果相关。总的来说,PVDs 负责菌株 YL-1 在自然和诱导低铁条件下的抗菌活性。结果表明,PVDs 是菌株 YL-1 在低铁条件下对革兰氏阳性和革兰氏阴性细菌具有广谱抗菌活性的必要条件。我们的研究结果还强调了外源性铁对 PVD 产生的影响以及该细菌产物在细菌相互作用中的重要性。作为一种生物防治剂,PVDs 可以直接抑制测试细菌的增殖,除了参与铁竞争之外。