Rutin hydrate induces autophagic cell death and oxidative stress response in phytopathogenic fungus Fusarium graminearum

IF 0.8 4区农林科学 Q3 AGRICULTURE, MULTIDISCIPLINARY

Zemdirbyste-agriculture Pub Date : 2023-12-20 DOI:10.13080/z-a.2023.110.042

Emre Yörük

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引用次数: 0

Abstract

Fusarium graminearum infects cereal crops of high commercial and nutritional value such as barley, wheat, and maize, and causes various diseases. Due to the increasing world population and changing climatic conditions, the importance of controlling this pathogen is increasing. The aim of this study was to investigate the potential antifungal effect of the flavonoid compound rutin hydrate (RH) on the F. graminearum PH-1 reference strain at different levels. The EC 50 (half effective concentration) value was determined by the agar dilution method on the PDA (potato dextrose agar) medium containing different concentrations of rutin hydrate. To examine the effect of rutin hydrate on cell viability, the WST-1 toxicity test was performed. To determine the effect of rutin hydrate on genomic stability and methylation profiles, the CRED-RA (coupled restriction enzyme digestion-random amplification) analysis was performed. To determine the expression levels of atg5 , cat , kmet1 , sporulation ( mgv1 ), and apoptosis ( mst20 ) related genes, the qRT-PCR (quantitative real-time polymerase chain reaction) was performed. To detect the presence of oxidative stress and autophagy, the DCF-DA (2’,7’-dichlorofluorescein) and MDC (monodansylcadaverine) staining tests were used. The EC 50 value of rutin hydrate was determined as 580 µg mL −1 . The WST-1 analysis revealed that rutin hydrate caused cytotoxicity (58.53%) in F. graminearum ( p < 0.05). In the CRED-RA analysis, genomic template stability (GTS) decreased by 9.53%, and the methylation levels for Hap II and Msp I enzymes were 11.6% and 5.24%, respectively. There was a significant difference in the atg5 (2.66 ± 0.48; p < 0.01), cat (3.00 ± 0.46; p < 0.01), and kmet1 (6.29 ± 2.24; p < 0.05) expression in the rutin hydrate-treated samples. Autophagy and oxidative stress response with a significant difference ( p < 0.05) in the rutin hydrate-treated F. graminearum was confirmed by DCF-DA and MDC tests. This is the first report demonstrating the antifungal effect of rutin hydrate on F. graminearum at physiological, transcript, and epigenetic levels.

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水合芦丁诱导植物病原真菌禾谷镰刀菌的自噬细胞死亡和氧化应激反应

禾谷镰刀菌（Fusarium graminearum）会侵染大麦、小麦和玉米等具有较高商业和营养价值的谷类作物，并引发各种病害。由于世界人口的增加和气候条件的变化，控制这种病原体的重要性与日俱增。本研究旨在探讨黄酮类化合物水合芦丁（RH）在不同浓度下对禾谷镰孢属 PH-1 参考菌株的潜在抗真菌作用。在含有不同浓度水合芦丁的 PDA（马铃薯葡萄糖琼脂）培养基上，采用琼脂稀释法测定 EC 50（半数有效浓度）值。为检测芦丁水合物对细胞活力的影响，进行了 WST-1 毒性试验。为确定水合芦丁对基因组稳定性和甲基化图谱的影响，进行了 CRED-RA（耦合限制性酶消化-随机扩增）分析。为确定 atg5、cat、kmet1、孢子（mgv1）和凋亡（mst20）相关基因的表达水平，进行了 qRT-PCR（定量实时聚合酶链反应）分析。为了检测氧化应激和自噬的存在，使用了 DCF-DA（2',7'-二氯荧光素）和 MDC（单丹参素）染色试验。芦丁水合物的 EC 50 值被确定为 580 µg mL -1 。WST-1 分析表明，水合芦丁对禾谷镰刀菌具有细胞毒性（58.53%）（p < 0.05）。在 CRED-RA 分析中，基因组模板稳定性（GTS）降低了 9.53%，Hap II 和 Msp I 酶的甲基化水平分别为 11.6% 和 5.24%。在水合芦丁处理的样本中，atg5（2.66 ± 0.48；p < 0.01）、cat（3.00 ± 0.46；p < 0.01）和 kmet1（6.29 ± 2.24；p < 0.05）的表达量有明显差异。经 DCF-DA 和 MDC 测试证实，自噬和氧化应激反应在水合芦丁处理的禾谷镰刀菌中有显著差异（p < 0.05）。这是第一份在生理、转录本和表观遗传水平上证明水合芦丁对禾谷镰刀菌有抗真菌作用的报告。

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来源期刊

Zemdirbyste-agriculture AGRICULTURE, MULTIDISCIPLINARY-

CiteScore

1.80

自引率

11.10%

发文量

审稿时长

>12 weeks

期刊介绍： Zemdirbyste-Agriculture is a quarterly scientific journal which covers a wide range of topics in the field of agricultural sciences, agronomy. It publishes articles of original research findings in the English language in the field of agronomy (soil and crop management, crop production, plant protection, plant breeding and genetics, biotechnology, plant nutrition, agrochemistry, soil science, microbiology etc.) and related areas. Articles are peer-reviewed. Review, debating papers as well as those of a methodological nature will also be considered.