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Localization of DIR1 at the tissue, cellular and subcellular levels during Systemic Acquired Resistance in Arabidopsis using DIR1:GUS and DIR1:EGFP reporters

机译:使用DIR1:GUS和DIR1:EGFP报告基因在拟南芥系统获得性抗性中将DIR1定位在组织,细胞和亚细胞水平

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Background Systemic Acquired Resistance (SAR) is an induced resistance response to pathogens, characterized by the translocation of a long-distance signal from induced leaves to distant tissues to prime them for increased resistance to future infection. DEFECTIVE in INDUCED RESISTANCE 1 (DIR1) has been hypothesized to chaperone a small signaling molecule to distant tissues during SAR in Arabidopsis. Results DIR1 promoter:DIR1-GUS/dir1-1 lines were constructed to examine DIR1 expression. DIR1 is expressed in seedlings, flowers and ubiquitously in untreated or mock-inoculated mature leaf cells, including phloem sieve elements and companion cells. Inoculation of leaves with SAR-inducing avirulent or virulent Pseudomonas syringae pv tomato (Pst) resulted in Type III Secretion System-dependent suppression of DIR1 expression in leaf cells. Transient expression of fluorescent fusion proteins in tobacco and intercellular washing fluid experiments indicated that DIR1's ER signal sequence targets it for secretion to the cell wall. However, DIR1 expressed without a signal sequence rescued the dir1-1 SAR defect, suggesting that a cytosolic pool of DIR1 is important for the SAR response. Conclusions Although expression of DIR1 decreases during SAR induction, the protein localizes to all living cell types of the vasculature, including companion cells and sieve elements, and therefore DIR1 is well situated to participate in long-distance signaling during SAR.
机译:背景技术系统获得性抗性(SAR)是一种对病原体的诱导抗性反应,其特征是长距离信号从诱导叶转移到远处组织,以引发它们以增强对未来感染的抵抗力。据推测,拟南芥SAR期间,抗性诱导的1号(DIR1)缺陷会陪伴小信号分子向远处的组织。结果构建了DIR1启动子:DIR1-GUS / dir1-1,检测DIR1的表达。 DIR1在幼苗,花朵中表达,并在未经处理或模拟接种的成熟叶细胞(包括韧皮部筛子元件和陪伴细胞)中普遍存在。用诱导SAR的无毒或强毒的丁香假单胞菌pv番茄(Pst)接种叶片会导致III型分泌系统依赖性抑制叶细胞中DIR1表达。烟草和细胞间洗涤液中荧光融合蛋白的瞬时表达表明DIR1的ER信号序列将其靶向细胞分泌到细胞壁。但是,没有信号序列表达的DIR1挽救了dir1-1 SAR缺陷,这表明DIR1的胞质池对于SAR反应很重要。结论尽管DIR1的表达在SAR诱导过程中降低,但该蛋白定位于脉管系统的所有活细胞类型,包括陪伴细胞和筛网元件,因此DIR1位置优越,可以参与SAR期间的长距离信号传导。

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