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从枯草芽孢杆菌 Sneb545 中分离鉴定诱导系统抗性决定因子对大豆胞囊线虫的抗性。

Isolation and identification of induced systemic resistance determinants from Bacillus simplex Sneb545 against Heterodera glycines.

机构信息

College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning, China.

College of Biology Science and Technology, Shenyang Agricultural University, Shenyang, Liaoning, China.

出版信息

Sci Rep. 2020 Jul 14;10(1):11586. doi: 10.1038/s41598-020-68548-4.

Abstract

Heterodera glycines is one of the most destructive pathogens of soybean. Soybean seeds coated with Bacillus simplex Sneb545 have shown resistance to H. glycines as a result of induced systemic resistance (ISR) in the plants. In this study, we aimed to identify the resistance-inducing determinants from this B. simplex strain. Combining the ISR bioassay, six ISR-active compounds were isolated from a culture of B. simplex Sneb545 using organic solvent gradient extraction, silica gel column chromatography, Sephadex LH-20 column chromatography, and semi-preparative high-performance liquid chromatography (HPLC), and all systems were based on activity tracking. The compounds were determined as cyclic(Pro-Tyr), cyclic(Val-Pro), cyclic(Leu-Pro), uracil, phenylalanine, and tryptophan using H NMR and C NMR. In plants from seeds coated with Bacillus simplex Sneb545, these six ISR-active compounds delayed the development of H. glycines in soybean roots. Moreover, cyclic(Pro-Tyr), cyclic(Val-Pro), and tryptophan reduced the number of nematodes in soybean roots. The expression levels of defense-related genes with cyclic(Val-Pro), tryptophan and uracil treatment soybean analysed using Quantitative real-time PCR (qRT-PCR). The results indicate cyclic(Val-Pro), tryptophan and uracil induced the expression of defense-related genes involved in the SA- and JA-pathways to against H. glycines. Our research results provide new agents for the control of H. glycines.

摘要

大豆孢囊线虫是大豆最具破坏性的病原体之一。由于植物诱导的系统抗性(ISR),包被苏云金芽孢杆菌 Sneb545 的大豆种子对大豆孢囊线虫表现出抗性。在这项研究中,我们旨在从这种苏云金芽孢杆菌菌株中鉴定出诱导抗性的决定因素。结合 ISR 生物测定,使用有机溶剂梯度萃取、硅胶柱色谱、Sephadex LH-20 柱色谱和半制备高效液相色谱(HPLC)从苏云金芽孢杆菌 Sneb545 的培养物中分离出六种具有 ISR 活性的化合物,所有系统均基于活性跟踪。使用 H NMR 和 C NMR 确定这些化合物为环(脯氨酸-酪氨酸)、环(缬氨酸-脯氨酸)、环(亮氨酸-脯氨酸)、尿嘧啶、苯丙氨酸和色氨酸。在包被苏云金芽孢杆菌 Sneb545 的种子的植物中,这六种 ISR 活性化合物延迟了大豆孢囊线虫在大豆根系中的发育。此外,环(脯氨酸-酪氨酸)、环(缬氨酸-脯氨酸)和色氨酸减少了大豆根系中的线虫数量。使用定量实时 PCR(qRT-PCR)分析用环(缬氨酸-脯氨酸)、色氨酸和尿嘧啶处理的大豆的防御相关基因的表达水平。结果表明,环(缬氨酸-脯氨酸)、色氨酸和尿嘧啶诱导了与 SA 和 JA 途径相关的防御相关基因的表达,以对抗大豆孢囊线虫。我们的研究结果为防治大豆孢囊线虫提供了新的药剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b75a/7360772/299b95e8a5c5/41598_2020_68548_Fig1_HTML.jpg

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