Lu Junming, Huang Yanbing, Liu Rui, Liang Ying, Zhang Hongyan, Shen Naikun, Yang Dengfeng, Jiang Mingguo
Guangxi Key Laboratory for Polysaccharide Materials and Modifications, School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, China.
Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Beibu Gulf Marine Research Center, Guangxi Academy of Sciences, Nanning, China.
Front Microbiol. 2024 Sep 25;15:1456847. doi: 10.3389/fmicb.2024.1456847. eCollection 2024.
wilt of banana, also recognized as Panama disease, is caused by the soil-borne fungus f. sp. tropical race 4 (FOC TR4). In recent years, strategies utilizing biocontrol agents, comprising antifungal microorganisms and their associated bioactive compounds from various environments, have been implemented to control this destructive disease. Our previous study showed that Gxun-2 had significant antifungal effects against FOC TR4. However, there has been little scientific investigation of the antibacterial or antifungal activity. The aim of this study was to isolate, identify and evaluate the inhibition strength of active compounds in Gxun-2, so as to explain the mechanism of the strain inhibition on FOC TR4 from the perspective of compounds.
The main antibacterial compounds of strain Gxun-2 were isolated, purified and identified using by fermentation extraction, silica gel column chromatography, thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and nuclear magnetic resonance (NMR) techniques. The effect of the compounds on the mycelial growth, morphology and spore germination of strain FOC TR4 was observed by 96-well plate method and AGAR diffusion method.
Among the metabolites produced by the strain, four antifungal compounds which were identified phenazine (CHN), phenazine-1-carboxylic acid (PCA) (CHNO), 2-acetamidophenol (CHNO) and aeruginaldehyde (CHNOS) were identified through HPLC and NMR. Of these compounds, phenazine and PCA exhibited the most pronounced inhibitory effects on the spore germination and mycelial growth of FOC TR4. Phenazine demonstrated potent antifungal activity against FOC TR4 with a minimum inhibitory concentration (MIC) of 6.25 mg/L. The half-maximal effective concentration (EC) was calculated to be 26.24 mg/L using the toxicity regression equation. PCA exhibited antifungal activity against FOC TR4 with an MIC of 25 mg/L and an EC of 89.63 mg/L. Furthermore, phenazine and PCA triggered substantial morphological transformations in the mycelia of FOC TR4, encompassing folding, bending, fracturing, and diminished spore formation.
These findings indicate that strain Gxun-2 plays a crucial role in controlling FOC TR4 pathogenesis, predominantly through producing the antifungal compounds phenazine and PCA, and possesses potential as a cost-efficient and sustainable biocontrol agent against wilt of banana in forthcoming times.
香蕉枯萎病,也被称为巴拿马病,由土壤传播的真菌尖孢镰刀菌古巴专化型热带4号小种(FOC TR4)引起。近年来,已实施利用生物防治剂的策略来控制这种毁灭性病害,这些生物防治剂包括来自各种环境的抗真菌微生物及其相关生物活性化合物。我们之前的研究表明,Gxun - 2对FOC TR4具有显著的抗真菌作用。然而,关于其抗菌或抗真菌活性的科学研究较少。本研究的目的是分离、鉴定和评估Gxun - 2中活性化合物的抑制强度,以便从化合物角度解释该菌株对FOC TR4的抑制机制。
采用发酵提取、硅胶柱色谱、薄层色谱(TLC)、高效液相色谱(HPLC)和核磁共振(NMR)技术,对菌株Gxun - 2的主要抗菌化合物进行分离、纯化和鉴定。通过96孔板法和琼脂扩散法观察这些化合物对FOC TR4菌株菌丝生长、形态和孢子萌发的影响。
通过HPLC和NMR鉴定出该菌株产生的代谢产物中的四种抗真菌化合物,分别为吩嗪(CHN)、吩嗪 - 1 - 羧酸(PCA)(CHNO)、2 - 乙酰氨基酚(CHNO)和铜绿醛(CHNOS)。在这些化合物中,吩嗪和PCA对FOC TR4的孢子萌发和菌丝生长表现出最显著的抑制作用。吩嗪对FOC TR4显示出强大的抗真菌活性,最低抑菌浓度(MIC)为6.25 mg/L。使用毒性回归方程计算得出半数有效浓度(EC)为26.24 mg/L。PCA对FOC TR4表现出抗真菌活性,MIC为25 mg/L,EC为89.63 mg/L。此外,吩嗪和PCA引发了FOC TR4菌丝体的大量形态变化,包括折叠、弯曲、断裂和孢子形成减少。
这些发现表明,菌株Gxun - 2在控制FOC TR4致病过程中起着关键作用,主要通过产生抗真菌化合物吩嗪和PCA,并且在未来有潜力作为一种经济高效且可持续的香蕉枯萎病生物防治剂。