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一种结合转录组学和生理学方法来理解应对……中氧化应激的适应性机制

A combined transcriptomic and physiological approach to understanding the adaptive mechanisms to cope with oxidative stress in .

作者信息

Park Jiyeun, Lee Hyun-Hee, Moon Heeji, Lee Nahyun, Kim Sieun, Kim Jung-Eun, Lee Yoonji, Min Kyunghun, Kim Hun, Choi Gyung Ja, Lee Yin-Won, Seo Young-Su, Son Hokyoung

机构信息

Department of Agricultural Biotechnology, Seoul National University , Seoul, Republic of Korea.

Department of Integrated Biological Science, Pusan National University , Busan, Republic of Korea.

出版信息

Microbiol Spectr. 2023 Sep 6;11(5):e0148523. doi: 10.1128/spectrum.01485-23.

Abstract

In plant-pathogen interactions, oxidative bursts are crucial for plants to defend themselves against pathogen infections. Rapid production and accumulation of reactive oxygen species kill pathogens directly and cause local cell death, preventing pathogens from spreading to adjacent cells. Meanwhile, the pathogens have developed several mechanisms to tolerate oxidative stress and successfully colonize plant tissues. In this study, we investigated the mechanisms responsible for resistance to oxidative stress by analyzing the transcriptomes of six oxidative stress-sensitive strains of the plant pathogenic fungus . Weighted gene co-expression network analysis identified several pathways related to oxidative stress responses, including the DNA repair system, autophagy, and ubiquitin-mediated proteolysis. We also identified hub genes with high intramodular connectivity in key modules and generated deletion or conditional suppression mutants. Phenotypic characterization of those mutants showed that the deletion of , , and caused sensitivity to oxidative stress, and further investigation on those genes revealed that transcriptional elongation and DNA damage responses play roles in oxidative stress response and pathogenicity. The suppression of also led to hypersensitivity to oxidative stress, and we demonstrated that plays a crucial role in heme biosynthesis and is essential for peroxidase activity. This study increases the understanding of the adaptive mechanisms to cope with oxidative stress in plant pathogenic fungi. IMPORTANCE Fungal pathogens have evolved various mechanisms to overcome host-derived stresses for successful infection. Oxidative stress is a representative defense system induced by the host plant, and fungi have complex response systems to cope with it. is one of the devastating plant pathogenic fungi, and understanding its pathosystem is crucial for disease control. In this study, we investigated adaptive mechanisms for coping with oxidative stress at the transcriptome level using oxidative stress-sensitive strains. In addition, by introducing genetic modification technique such as CRISPR-Cas9 and the conditional gene expression system, we identified pathways/genes required for resistance to oxidative stress and also for virulence. Overall, this study advances the understanding of the oxidative stress response and related mechanisms in plant pathogenic fungi.

摘要

在植物与病原体的相互作用中,氧化爆发对于植物抵御病原体感染至关重要。活性氧的快速产生和积累直接杀死病原体并导致局部细胞死亡,从而防止病原体扩散到相邻细胞。同时,病原体已发展出多种机制来耐受氧化应激并成功定殖于植物组织。在本研究中,我们通过分析植物病原真菌的六个氧化应激敏感菌株的转录组,研究了其对氧化应激的抗性机制。加权基因共表达网络分析确定了几个与氧化应激反应相关的途径,包括DNA修复系统、自噬和泛素介导的蛋白水解。我们还在关键模块中鉴定出具有高模块内连通性的枢纽基因,并构建了缺失或条件性抑制突变体。对这些突变体的表型特征分析表明,缺失[具体基因1]、[具体基因2]和[具体基因3]会导致对氧化应激敏感,对这些基因的进一步研究表明转录延伸和DNA损伤反应在氧化应激反应和致病性中发挥作用。抑制[具体基因4]也会导致对氧化应激超敏,并且我们证明[具体基因4]在血红素生物合成中起关键作用,对过氧化物酶活性至关重要。本研究增进了对植物病原真菌应对氧化应激的适应性机制的理解。重要性 真菌病原体已进化出多种机制来克服宿主衍生的压力以成功感染。氧化应激是宿主植物诱导的一种代表性防御系统,真菌具有复杂的反应系统来应对它。[具体真菌名称]是一种具有破坏性的植物病原真菌,了解其病理系统对于疾病控制至关重要。在本研究中,我们使用氧化应激敏感菌株在转录组水平上研究了应对氧化应激的适应性机制。此外,通过引入CRISPR-Cas9等基因编辑技术和条件性基因表达系统,我们鉴定了抗氧化应激和毒力所需的途径/基因。总体而言,本研究推进了对植物病原真菌氧化应激反应及相关机制的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/30c0/10581207/643e289b6733/spectrum.01485-23.f001.jpg

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