Inthaisong Sukanya, Boonchuen Pakpoom, Jaichopsanthia Tana, Songwattana Pongpan, Khairum Apinya, Chueakhunthod Witsarut, Tharapreuksapong Akkawat, Tittabutr Panlada, Teaumroong Neung, Tantasawat Piyada Alisha
School of Crop Production Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.
School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.
Sci Rep. 2025 Jan 8;15(1):1334. doi: 10.1038/s41598-024-84787-1.
Several mungbean (Vigna radiata (L.) Wilczek) cultivars are susceptible to Cercospora leaf spot (CLS) caused by Cercospora canescens Ellis & Martin, and it is necessary to explore resistance sources and understand resistance mechanisms. However, the CLS resistance mechanisms have not yet been explored. The response to CLS revealed significantly different disease severity scores in both mungbean genotypes. Hypersensitive response (HR) started to appear at 2 days after inoculation (DAI) in SUPER5 but was never observed in CN84-1. SUPER5 exhibited fewer and smaller lesions than CN84-1 during CLS infection, resulting in SUPER5 being resistant while CN84-1 was susceptible to CLS. In this study, RNA sequencing (RNA-seq) analysis was used to unravel the mechanisms of resistance to CLS in a resistant line (SUPER5) and a susceptible variety (CN84-1) upon CLS infection. A total of 9510 DEGs including 4615 up-regulated and 4895 down-regulated genes were revealed. Of these 3242 and 1027 genes were uniquely up-regulated only in the SUPER5 and CN84-1, respectively, while 2902 and 734 genes were down-regulated only in SUPER5 and CN84-1, respectively. The 843 DEGs were enriched in biological processes mainly associated with plant defense responses, defense response to fungus, protein phosphorylation and response to chitin in Gene Ontology (GO) terms analysis. KEGG pathway analysis showed that these genes were represented in plant-pathogen interaction, the MAPK signaling pathway, plant hormone signal transduction, and cell wall component biosynthesis in response to the CLS infection specifically in SUPER5. In addition, the qRT-PCR was used to analyze the expression pattern of 22 candidate DEGs belonging to pathogenesis related (PR) proteins, resistance (R) proteins, transcription factors, hypersensitive response (HR), and the essential genes involved in cell wall activity during CLS-infected V. radiata. It was found that the expression of these genes was consistent with the RNA-seq analysis, showing a highly significant correlation with a coefficient of 0.7163 (p < 0.01). The co-expression network illustrated the interactions among these genes, which were involved in multiple functions related to the defense response. Interestingly, the ones encoding PR-2, thaumatin, peroxidase, defensin, RPM1, pectinesterase, chalcone synthase, auxin efflux carrier, and transcription factors (Pti1, Pti5, Pti6 and WRKY40) were highly significantly up-regulated in SUPER5 but not in CN84-1 upon CLS infection, suggesting that they might be involved in the CLS resistance mechanisms. Moreover, SUPER5 was found to have higher β-1,3-glucanase and chitinase activity levels than CN84-1. Our findings contribute to an understanding of the CLS resistance mechanisms and may advocate the development of more effective disease management approaches.
几种绿豆(Vigna radiata (L.) Wilczek)品种对尾孢叶斑病菌(Cercospora canescens Ellis & Martin)引起的尾孢叶斑病(CLS)敏感,因此有必要探索抗性来源并了解抗性机制。然而,CLS的抗性机制尚未得到探索。对CLS的反应显示,两种绿豆基因型的病情严重程度评分存在显著差异。超敏反应(HR)在接种后2天(DAI)开始出现在SUPER5中,但在CN84-1中从未观察到。在CLS感染期间,SUPER5的病斑比CN84-1更少、更小,导致SUPER5具有抗性而CN84-1对CLS敏感。在本研究中,利用RNA测序(RNA-seq)分析来揭示抗性品系(SUPER5)和敏感品种(CN84-1)在CLS感染后对CLS的抗性机制。共鉴定出9510个差异表达基因(DEG),其中4615个上调基因和4895个下调基因。其中,分别有3242个和1027个基因仅在SUPER5和CN84-1中上调,而分别有2902个和734个基因仅在SUPER5和CN84-1中下调。在基因本体(GO)术语分析中,843个DEG富集在主要与植物防御反应、对真菌的防御反应、蛋白质磷酸化和对几丁质的反应相关的生物学过程中。KEGG通路分析表明,这些基因在植物-病原体相互作用、丝裂原活化蛋白激酶(MAPK)信号通路、植物激素信号转导以及细胞壁成分生物合成中均有体现,特别是在SUPER5中对CLS感染的反应。此外,采用实时定量聚合酶链反应(qRT-PCR)分析了22个候选DEG在CLS感染的绿豆中的表达模式,这些DEG属于病程相关(PR)蛋白、抗性(R)蛋白、转录因子、超敏反应(HR)以及参与细胞壁活性的关键基因。结果发现,这些基因的表达与RNA-seq分析一致,相关性系数为0.7163(p < 0.01),具有高度显著性。共表达网络展示了这些基因之间的相互作用,它们参与了与防御反应相关的多种功能。有趣的是,编码PR-2、奇异果甜蛋白、过氧化物酶、防御素、RPM1、果胶酯酶、查尔酮合酶、生长素外排载体和转录因子(Pti1、Pti5、Pti6和WRKY40)的基因在CLS感染后在SUPER5中显著上调,但在CN84-1中未上调,这表明它们可能参与了CLS抗性机制。此外,发现SUPER5的β-1,3-葡聚糖酶和几丁质酶活性水平高于CN84-1。我们的数据有助于理解CLS抗性机制,并可能为开发更有效的病害管理方法提供依据。