Figueroa Ana Cristina, Díaz María Soledad, Turco Mauricio, Fernández Trotta Andrea, Marino Bibiana, Soria Néstor Walter, Beltramo Dante Miguel, Alasino Roxana Valeria
Centro de Excelencia en Productos y Procesos de Córdoba-CEPROCOR- Complejo Hospitalario, Santa María de Punilla, X6154 Córdoba, Argentina.
Cátedra de Biotecnología, Facultad de Ciencias de la salud, Unidad Asociada al CONICET: Área de Cs. Agrarias, Ingeniería, Cs. Biológicas, Universidad Católica de Córdoba, Avenida Armada Argentina 3555, X5016DHK Córdoba, Argentina.
J Appl Microbiol. 2024 Dec 2;135(12). doi: 10.1093/jambio/lxae306.
Thecaphora frezzii, the causal agent of peanut smut, causes significant grain losses in Argentina. Current control strategies are insufficient to manage this pathogen. We investigate the effect of antioxidants on the in vitro development of T. frezzii hyphae, to identify compounds with antifungal activity, also evaluate protein and lipid profiles as potential targets for these compounds.
The antifungal activity was evaluated in both, solid and liquid media, and minimum inhibitory concentration and minimum fungicidal concentration were calculated. The protein profile was assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis, while lipids were analyzed by thin-layer chromatography (TLC) and gas chromatography with flame ionization detection, both before and after hyphal treatment. Lipase activity was analyzed using agar Tween 20 and TLC, while lipid peroxidation was evaluated by the thiobarbituric acid-reactive substance (TBARS)assay. Microscopy was used to observe morphological and metabolic changes. Butylated hydroxyanisole, methylparaben, and lipoic acid showed inhibitory effects on T. frezzii. Lipoic acid was chosen for further study due to its lack of environmental toxicity. Lipoic acid induced the loss of cytosolic proteins, hydrolysis of triglycerides, and increased levels of free fatty acids, monoacylglycerols, and diacylglycerols. It also caused a decrease in ergosterol levels and alterations in the fungal cell wall and membrane, ultimately leading to cell death.
This study demonstrates the efficacy of lipoic acid in inhibiting the in vitro development of T. frezzii.
弗氏内孢堆黑粉菌是花生黑粉病的病原体，在阿根廷导致了严重的谷物损失。目前的控制策略不足以管理这种病原体。我们研究抗氧化剂对弗氏内孢堆黑粉菌菌丝体外发育的影响，以鉴定具有抗真菌活性的化合物，并评估蛋白质和脂质谱作为这些化合物的潜在靶点。
在固体和液体培养基中评估抗真菌活性，并计算最低抑菌浓度和最低杀菌浓度。使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳评估蛋白质谱，同时在菌丝处理前后通过薄层色谱（TLC）和火焰离子化检测气相色谱分析脂质。使用琼脂吐温20和TLC分析脂肪酶活性，同时通过硫代巴比妥酸反应性物质（TBARS）测定评估脂质过氧化。使用显微镜观察形态和代谢变化。丁基羟基茴香醚（BHA）、对羟基苯甲酸甲酯和硫辛酸对弗氏内孢堆黑粉菌有抑制作用。由于硫辛酸缺乏环境毒性，因此选择其进行进一步研究。硫辛酸导致胞质蛋白丢失、甘油三酯水解以及游离脂肪酸、单酰甘油和二酰甘油水平升高。它还导致麦角固醇水平降低以及真菌细胞壁和细胞膜发生改变，最终导致细胞死亡。
本研究证明了硫辛酸在抑制弗氏内孢堆黑粉菌体外发育方面的有效性。
