Rapid Identification of Phytotoxins Produced by Glomerella cingulata Using High‐Resolution Mass Spectrometry‐Based Qualification, Targeted Structural Confirmation and Their Characteristics Investigation

IF 3.5 4区 生物学 Q2 MICROBIOLOGY
Xin Yu, Zhiyang Liu, Huidi Zhang, Caixia Wang, Sen Lian, Xiangli Dong, Baohua Li, Pingliang Li
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引用次数: 0

Abstract

Glomerella cingulata is a pathogenic fungus that can cause apple Glomerella leaf spot (GLS), a new and destructive apple disease in China. Phytotoxins are important factors closely related to the disease process, but there is still no report on the phytotoxins of G. cingulata. The aim of this study was to rapidly identify the phytotoxins of this pathogen using a strategy of HRMS‐based preliminary qualification, followed by targeted structure confirmation and also investigation of phytotoxicity characteristics. First, the crude toxin sample was directly analyzed by the UPLC‐HRMS and GC‐MS, and the data were processed to screen for possible phytotoxic compounds using MS library and the phytotoxicity‐related literature. The reference standards of credible phytotoxic compounds were then subjected to targeted structure validation (signal comparison between standards and compounds in crude toxin via HPLC‐DAD, UPLC‐MS/MS, and GC‐MS), and also the phytotoxicity assay. The results confirmed six phytotoxins produced by G. cingulata, namely 5‐hydroxymethyl‐2‐furancarboxylic acid (HMFCA), 2,5‐bis(hydroxymethyl)furan (BHMF), 2‐furoic acid (FA), 2,3‐butanediol, trans‐aconitic acid (TAA), and cis‐aconitic acid (CAA). Of these, HMFCA and TAA exhibited greater phytotoxicity. Main characteristics: All of them were non‐host‐selective toxins, and toxins were synergistically phytotoxic to the host when mixed. BHMF, HMFCA, FA, TAA, and CAA could be commonly produced by all tested strains, and their phytotoxicity can be significantly inhibited or even eliminated at high temperatures or high pH. The elucidation of the phytotoxins of G. cingulata in this work could provide information on the pathogenesis and control of apple GLS.
利用基于高分辨率质谱的定性、定向结构确证及其特性调查快速鉴定茵陈格氏菌产生的植物毒素
苹果疫霉菌(Glomerella cingulata)是一种致病真菌,可引起苹果疫霉菌叶斑病(GLS),这是我国苹果病害中一种新的毁灭性病害。植物毒素是与病害发生过程密切相关的重要因素,但目前还没有关于颖枯丝核菌植物毒素的报道。本研究的目的是采用基于 HRMS 的初步定性、定向结构确认和植物毒性特征研究的策略,快速鉴定该病原菌的植物毒素。首先,用 UPLC-HRMS 和 GC-MS 直接分析粗毒素样品,然后利用质谱库和植物毒性相关文献处理数据,筛选可能的植物毒性化合物。然后,对可信的植物毒性化合物参考标准进行了有针对性的结构验证(通过 HPLC-DAD、UPLC-MS/MS 和 GC-MS 对标准品和粗毒素中的化合物进行信号比较),并进行了植物毒性检测。结果确认了 G. cingulata 产生的六种植物毒素,即 5-羟甲基-2-呋喃羧酸(HMFCA)、2,5-双(羟甲基)呋喃(BHMF)、2-糠酸(FA)、2,3-丁二醇、反式乌头酸(TAA)和顺式乌头酸(CAA)。其中,HMFCA 和 TAA 的植物毒性更大。主要特征:它们都是非寄主选择性毒素,混合使用时对寄主具有协同植物毒性。所有受试菌株都能产生 BHMF、HMFCA、FA、TAA 和 CAA,在高温或高 pH 条件下,它们的植物毒性会被显著抑制甚至消除。这项工作对 G. cingulata 植物毒素的阐明可为苹果 GLS 的致病机理和防治提供信息。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Basic Microbiology
Journal of Basic Microbiology 生物-微生物学
CiteScore
6.10
自引率
0.00%
发文量
134
审稿时长
1.8 months
期刊介绍: The Journal of Basic Microbiology (JBM) publishes primary research papers on both procaryotic and eucaryotic microorganisms, including bacteria, archaea, fungi, algae, protozoans, phages, viruses, viroids and prions. Papers published deal with: microbial interactions (pathogenic, mutualistic, environmental), ecology, physiology, genetics and cell biology/development, new methodologies, i.e., new imaging technologies (e.g. video-fluorescence microscopy, modern TEM applications) novel molecular biology methods (e.g. PCR-based gene targeting or cassettes for cloning of GFP constructs).
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