JIA-2021-1477 轮枝镰刀菌过氧化物酶体基质受体Pex5影响致病性及FB1合成

doi:10.1016/j.jia.2022.07.044

摘要/Abstract

摘要：

过氧化物酶体内基质主要包括氧化酶、过氧化氢酶和过氧化物酶等，它们调节细胞氧化稳态和功能，绝大部分的基质含过氧化物酶体靶向信号（PTS）并由PTS受体运输入内。本文研究发现过氧化物酶体靶向信号类型1（PTS1) 受体Pex5的缺失影响了病原真菌轮枝镰刀菌多种生物学功能，结果将有助于阐明 Pex5致病及产毒的分子机制，并为控制病害和减少伏马菌素 B1（FB1）的毒害提供理论依据。同源重组的方法构建FvPEX5敲除突变体（ΔFvpex5），继而构建敲除突变体的互补菌株。通过比较野生型，敲除突变体和互补菌株三者的表型，我们探究并验证FvPex5在轮枝镰刀菌中的生物学功能；进一步通过RNA-Seq 分析ΔFvpex5中表达差异的PTS蛋白，并结合GO和KEEG注释进而解析FvPex5影响生物学功能的原因。研究发现轮枝镰刀菌 PTS1受体 FvPex5 参与 PTS1 的定位、碳源和脂质的利用、消除 ROS、细胞壁应激反应、分生孢子的形成、FB1产生以及致病性。在ΔFvpex5 突变体，RNA-Seq 分析发现差异表达的PTS1、PTS2、 PTS 相关途径中的过氧化物酶体相关基因（PEX）和 FB1 毒素相关基因，并进一步通过RT-PCR 证实这些基因的差异表达。此外，结合GO和KEEG注释，发现ΔFvpex5突变体中差异表达的PTS1、PTS2基因在碳代谢、氮代谢、脂质代谢和氧化平衡等多种生化途径中富集。FvPex5 参与了 PTS 相关基因的调控，从而影响了轮枝镰刀菌的氧化平衡、FB1产量和致病性。本研究首次发现了FvPex5对伏马毒素FB1合成起调节作用，并首次对轮枝镰刀菌中的过氧化物酶体靶向信号(PTS)蛋白进行了预测及功能分析。

Abstract:

The peroxisomal matrix oxidase, catalase and peroxidase are imported peroxisomes through the shuttling receptors, which regulates the cellular oxidative homeostasis and function. Here, we report that PTS1 shuttling receptor FvPex5 is involved in the localization of PTS1, utilization of carbon sources and lipids, elimination ROS, cell wall stress, conidiation, fumonisin B₁ (FB₁) production, and virulence in maize pathogen Fusarium verticillioides. Significantly, differential expression of PTS1-, PTS2-, PEX- and FB₁ toxin-related genes in wild type and ΔFvpex5 mutant were examined by RNA-Seq analyses and confirmed by RT-PCR assay. In addition, different expression of PTS1 and PTS2 genes of the ΔFvpex5 mutant were enriched in diverse biochemical pathways, such as carbon metabolism, nitrogen metabolism, lipid metabolism and the oxidation balance by combining GO and KEGG annotations. Overall, we showed that FvPex5 is involved in the regulation of genes associated with PTS, thereby affecting the oxidation balance, FB₁ and virulence in F. verticillioides. The results help to clarify the functional divergence of Pex5 orthologs, and may provide a possible target for controlling F. verticillioides infections and FB₁ biosynthesis.

Key words: Fusarium verticillioides , FvPex5 , PTS , fumonisin B₁ , virulence

. JIA-2021-1477 轮枝镰刀菌过氧化物酶体基质受体Pex5影响致病性及FB1合成[J]. Journal of Integrative Agriculture, 2022, 21(10): 2957-2972.

YU Wen-ying, LIN Mei, YAN Hui-juan, WANG Jia-jia, ZHANG Sheng-min, LU Guo-dong, WANG Zong-hua, Won-Bo SHIM. The peroxisomal matrix shuttling receptor Pex5 plays a role of FB₁ production and virulence in Fusarium verticillioides[J]. Journal of Integrative Agriculture, 2022, 21(10): 2957-2972.

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