详细的蛋白质组学和代谢组学研究揭示了转基因豌豆对枯萎病胁迫的抗性响应机制。

IF 6 1区 生物学 Q1 PLANT SCIENCES Plant, Cell & Environment Pub Date : 2024-11-05 DOI:10.1111/pce.15261
Subhasis Karmakar, Sabarinathan Selvaraj, Dipak Gayen, Mirza J Baig
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引用次数: 0

摘要

豌豆对半干旱地区的农民来说至关重要,但它却因镰刀菌引起的枯萎病而减产。本研究表明,引入水稻草酸氧化酶 4(Osoxo4)基因可显著提高抗枯萎病能力。通过基因表达分析、酶活性测定、生化评估、组织化学染色和体内外生物测定(包括孢子萌发试验),证实了转基因品系的抗性增强。我们进行了蛋白质组学和代谢组学分析,以研究抗性增强的机制。基于 LC-MS/MS 的无标记蛋白质组学对野生感染的转基因豌豆和野生型豌豆叶片进行了鉴定,共鉴定出 2386 个蛋白质,其中 1048 个蛋白质的丰度发生了显著变化--转基因植物中 738 个蛋白质上调,310 个蛋白质下调。值得注意的是,HMG1/2-like 蛋白、Putative nucleosome assembly protein C364.06、DEAD-box ATP-dependent RNA helicase 3、Lipoxygenase 1、Annexin D1 和 Annexin-like protein RJ4 等蛋白质显著上调,这表明它们在培育抗枯萎品种中可能发挥作用。代谢组分析表明,转基因鸽子豆中氨基酸、糖、草酸、糖醇和肌醇的水平升高,糖和淀粉代谢以及肌醇磷酸代谢的通路上调,表明其对枯萎病胁迫的恢复能力增强。这项研究强调了独特的调控蛋白和代谢物,有助于深入了解胁迫适应性,并为培育抗病鸽子豆品种的遗传干预提供指导。
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A Detailed Proteomics and Metabolomics Landscape Sheds Light on the Mechanistic Insights Into the Resistance Response of Transgenic Pigeon Pea Against Wilt Stress.

Pigeon pea, vital for farmers in semi-arid regions, suffers yield losses from Fusarium wilt caused by Fusarium udum. This study demonstrates that introducing the rice oxalate oxidase 4 (Osoxo4) gene significantly boosts wilt resistance. Enhanced resistance in transgenic lines was confirmed through gene expression analysis, enzyme activity assays, biochemical assessments, histochemical staining and in vitro and in vivo bioassays, including spore germination tests. We performed proteomics and metabolomics analyses to investigate mechanisms of enhanced resistance. LC-MS/MS-based label-free proteomics of wilt-infected transgenic and wild-type pigeon pea leaves identified 2386 proteins, with 1048 showing significant abundance changes-738 upregulated and 310 downregulated-in transgenic plants. Notably, proteins such as HMG1/2-like protein, Putative nucleosome assembly protein C364.06, DEAD-box ATP-dependent RNA helicase 3, Lipoxygenase 1, Annexin D1 and Annexin-like protein RJ4 were significantly upregulated, indicating their potential role in developing wilt-resistant cultivars. Metabolomic analysis showed elevated levels of amino acids, sugars, oxalic acid, sugar alcohols and myo-inositol in transgenic pigeon pea, with upregulated pathways in Sugar and Starch Metabolism and Inositol Phosphate Metabolism, indicating enhanced resilience to wilt stress. This study highlights unique regulatory proteins and metabolites, offering insights into stress adaptation and guiding genetic interventions for breeding disease-resistant pigeon pea varieties.

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来源期刊
Plant, Cell & Environment
Plant, Cell & Environment 生物-植物科学
CiteScore
13.30
自引率
4.10%
发文量
253
审稿时长
1.8 months
期刊介绍: Plant, Cell & Environment is a premier plant science journal, offering valuable insights into plant responses to their environment. Committed to publishing high-quality theoretical and experimental research, the journal covers a broad spectrum of factors, spanning from molecular to community levels. Researchers exploring various aspects of plant biology, physiology, and ecology contribute to the journal's comprehensive understanding of plant-environment interactions.
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