Abstract:
Fusarium oxysporum f. sp. cubense (FOC4) is a pathogen of banana fusarium wilt, which is a serious problem that has plagued the tropical banana industry for many years. The pathogenic mechanism is complex and unclear, so the prevention and control in agricultural production applications is ineffective. SNP-D4, an artificial peptide aptamer, was identified and specifically inhibited FOC4. To evaluate the efficacy of SNP-D4, FoC4 spores were treated with purified SNP-D4 to calculate the germination and fungicide rates. Damage of FOC4 spores was observed by staining with propidium iodide (PI). Eight proteins of FOC4 were identified to have high affinity for SNP-D4 by a pull-down method combined with Q-Exactive mass spectrometry. Of these eight proteins, A0A5C6SPC6, the aldehyde dehydrogenase of FOC4, was selected as an example to scrutinize the interaction sites with SNP-D4. Molecular docking revealed that Thr66 on the peptide loop of SNP-D4 bound with Tyr437 near the catalytic center of A0A5C6SPC6. Subsequently 42 spore proteins which exhibited associations with the eight proteins were retrieved for protein-protein interaction analysis, demonstrating that SNP-D4 interfered with pathways including \'translation\', \'folding, sorting and degradation\', \'transcription\', \'signal transduction\' and \'cell growth and death\', eventually causing the inhibition of growth of FOC4. This study not only investigated the possible pathogenic mechanism of FOC4, but also provided a potential antifungal agent SNP-D4 for use in the control of banana wilt disease.
摘要:
尖孢镰刀菌f.sp.古巴(FOC4)是香蕉枯萎病的病原体,这是困扰热带香蕉产业多年的严重问题。致病机制复杂且不明确,因此,农业生产应用中的预防和控制是无效的。SNP-D4,一种人工肽适体,被鉴定并特异性抑制FOC4。为了评估SNP-D4的功效,用纯化的SNP-D4处理FoC4孢子以计算发芽和杀真菌剂速率。通过用碘化丙啶(PI)染色观察到FOC4孢子的损伤。通过下拉方法结合Q-Exactive质谱法鉴定出FOC4的八种蛋白质对SNP-D4具有高亲和力。在这八种蛋白质中,选择FOC4的醛脱氢酶A0A5C6SPC6作为实例来检查与SNP-D4的相互作用位点。分子对接显示SNP-D4的肽环上的Thr66在A0A5C6SPC6的催化中心附近与Tyr437结合。随后,检索到与八种蛋白质相关的42种孢子蛋白质,用于蛋白质-蛋白质相互作用分析。证明SNP-D4干扰了包括“翻译”在内的途径,\'折叠,排序和退化,\'转录\',“信号转导”和“细胞生长和死亡”,最终导致FOC4生长的抑制。本研究不仅探讨了FOC4可能的致病机制,而且为控制香蕉枯萎病提供了潜在的抗真菌药物SNP-D4。