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定量蛋白质组学分析揭示了香蕉品种“巴西”对尖孢镰刀菌古巴专化型 1 号和 4 号生理小种抗性差异的原因。

Quantitative proteomics analysis reveals resistance differences of banana cultivar 'Brazilian' to Fusarium oxysporum f. sp. cubense races 1 and 4.

机构信息

Guangdong Province Key Laboratory of Microbial Signals and Disease Control, College of Agriculture, South China Agricultural University, Guangzhou 510642, China; State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, South China Agricultural University, Guangzhou 510642, China.

Guangdong Province Key Laboratory of Microbial Signals and Disease Control, College of Agriculture, South China Agricultural University, Guangzhou 510642, China; State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, South China Agricultural University, Guangzhou 510642, China.

出版信息

J Proteomics. 2019 Jul 15;203:103376. doi: 10.1016/j.jprot.2019.05.004. Epub 2019 May 9.

Abstract

Banana Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (Foc), is one of the most devastating diseases in banana production. Foc is classified into three physiological races. However, the resistance mechanisms of banana against different Foc races are poorly understood. In this study, we performed a comparative proteomics analysis to investigate the resistance mechanisms of 'Brazilian' against Foc1 and Foc4. The proteomes of 'Brazilian' roots inoculated with Foc1 and Foc4 and mock inoculated control at 48 h were analyzed using TMT based quantitative analysis technique. A total of 7325 unique protein species were identified, of which 689, 744, and 1222 protein species were differentially accumulated in Foc1 vs. CK, Foc4 vs. CK, and Foc1 vs. Foc4, respectively. The differential accumulations of candidate protein species were further confirmed by RT-qPCR, PRM, and physiological and biochemical assays. Bioinformatics analysis revealed that the differentially abundance protein species (DAPS) related to pattern recognition receptors, plant cell wall modification, redox homeostasis, and defense responses were differentially accumulated after Foc1 and Foc4 infection, suggesting that 'Brazilian' differed in resistance to the two Foc races. Our study lay the foundation for an in-depth understanding of the interaction between bananas and Foc at the proteome level. SIGNIFICANCE: The banana fusarium wilt disease is one of the most destructive disease of banana and is caused by Fusarium oxysporum f. sp. cubense (Foc). Foc is classified into three physiological races, namely, Foc1, Foc2, and Foc4. Among these races, Foc1 and Foc4 are widely distributed in south China and significantly lose yield. Although both physiological races (Foc1 and Foc4) can invade the Cavendish banana cultivar 'Brazilian', they have significant pathogenicity differences. Unfortunately, how the resistance differences are produced between two races is still largely unclear to date. In this study, we addressed this issue by performing TMT-based comparative quantitative proteomics analysis of 'Brazilian' roots after inoculation with Foc1 and Foc4 as well as sterile water as the control. We revealed that the series of protein species associated with pattern recognition receptors, plant cell wall modification, redox homeostasis, pathogenesis, phytohormones and signal transduction, plant secondary metabolites and programmed cell death etc. were involved in the response to Foc infection. Notably, the potential role of lipid signaling in banana defense against Foc are not reported previously but rather unveiled for the first time in this study. The current study represents the most extensive analysis of the protein profile of 'Brazilian' in response to Foc inoculation and includes for the first time the results from comparison quantitative proteomics analysis between plants inoculated with a pathogenic strain Foc4 and a nonpathogenic strain Foc1 of 'Brazilian', which will lay the foundation for an in-depth understanding of the interaction between bananas and Foc at the proteome level.

摘要

香蕉枯萎病由尖孢镰刀菌古巴专化型(Foc)引起,是香蕉生产中最具破坏性的病害之一。Foc 分为三个生理小种。然而,香蕉对不同 Foc 小种的抗性机制尚不清楚。在这项研究中,我们进行了比较蛋白质组学分析,以研究‘巴西’对 Foc1 和 Foc4 的抗性机制。使用 TMT 基于定量分析技术分析了接种 Foc1 和 Foc4 以及模拟接种对照的‘巴西’根 48 小时的蛋白质组。共鉴定到 7325 个独特的蛋白质种类,其中 Foc1 与 CK、Foc4 与 CK 和 Foc1 与 Foc4 相比,分别有 689、744 和 1222 种蛋白质差异积累。通过 RT-qPCR、PRM 和生理生化测定进一步验证了候选蛋白种类的差异积累。生物信息学分析表明,与模式识别受体、植物细胞壁修饰、氧化还原稳态和防御反应相关的差异丰度蛋白(DAPS)在 Foc1 和 Foc4 感染后差异积累,表明‘巴西’对两种 Foc 小种的抗性存在差异。我们的研究为在蛋白质组水平深入了解香蕉与 Foc 的相互作用奠定了基础。

意义

香蕉枯萎病是香蕉最具破坏性的病害之一,由尖孢镰刀菌古巴专化型(Foc)引起。Foc 分为三个生理小种,即 Foc1、Foc2 和 Foc4。其中,Foc1 和 Foc4 在中国南方广泛分布,对产量有显著影响。尽管两个生理小种(Foc1 和 Foc4)都能侵染卡文迪什香蕉品种‘巴西’,但它们的致病性差异显著。不幸的是,两种小种之间产生抗性差异的机制在很大程度上仍不清楚。在这项研究中,我们通过对接种 Foc1 和 Foc4 以及无菌水作为对照的‘巴西’根进行基于 TMT 的比较定量蛋白质组学分析来解决这个问题。我们揭示了一系列与模式识别受体、植物细胞壁修饰、氧化还原稳态、发病机制、植物激素和信号转导、植物次生代谢物和程序性细胞死亡等相关的蛋白种类参与了对 Foc 感染的反应。值得注意的是,脂质信号在香蕉防御 Foc 中的潜在作用以前没有报道过,但在本研究中首次揭示。本研究代表了对‘巴西’接种 Foc 后蛋白质谱最广泛的分析,并且首次包括了对接种致病性菌株 Foc4 和非致病性菌株 Foc1 的‘巴西’植物进行比较定量蛋白质组学分析的结果,这将为在蛋白质组水平深入了解香蕉与 Foc 的相互作用奠定基础。

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