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Plasma membrane localization and potential endocytosis of constitutively expressed XA21 proteins in transgenic rice.

机译:组成型表达的XA21蛋白在转基因水稻中的质膜定位和潜在的内吞作用。

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The rice pattern recognition receptor (PRR) XA21 confers race-specific resistance in leaf infection by bacterial blight Xathomonas oryzae pv. oryzae (Xoo), and was shown to be primarily localized to the endoplasmic reticulum (ER) when expressed with its native promoter or overexpressed in the protoplast. However, whether the protein is still ER-localization in the intact cell when overexpressed remains to be identified. Here, we showed that XA21, its kinase-dead mutant XA21PK736EP, and the triple autophosphorylation mutant XA21PS686A/T688A/S699A GFP fusions were primarily localized to the plasma membrane (PM) when overexpressed in the intact transgenic rice cell, and also localized to the ER in the transgenic protoplast. The transgenic plants constitutively expressing the wild-type XA21 or its GFP fusion displayed race-specific resistance to Xoo at the adult and seedling stages. XA21 and XA21PK736EP could be internalized probably via the SCAMP-positive early endosomal compartment in the protoplast, suggesting that XA21 might be endocytosed to initiate resistance responses during pathogen infection. We also established a root infection system and demonstrated that XA21 also mediated race-specific resistance responses to Xoo in the root. Our current study provides an insight into the nature of the XA21-mediated resistance and a practical approach using the root cell system to further dissect the cellular signaling of the PRR during the rice-Xoo interaction.Digital Object Identifier http://dx.doi.org/10.1093/mp/ssq038
机译:水稻模式识别受体(PRR)XA21在细菌性白叶枯萎病(Xathomonas oryzae)叶片感染中赋予种族特异性抗性。 oryzae ( Xoo ),当以其天然启动子表达或在原生质体中过表达时,显示主要位于内质网(ER)。但是,该蛋白在过表达时是否仍处于完整细胞中的ER-定位尚待确定。在这里,我们发现XA21,其激酶死亡突变体XA21P K736EP 和三重自磷酸化突变体XA21P S686A / T688A / S699A GFP融合蛋白主要定位于质膜( PM)在完整的转基因水稻细胞中过表达时,也位于转基因原生质体中的ER处。组成型表达野生型XA21或其GFP融合体的转基因植物在成年和幼苗期对 Xoo 具有种族特异性的抗性。 XA21和XA21P K736EP 可能通过原生质体中SCAMP阳性的早期内体区室被内在化,这表明XA21可能被内吞并在病原体感染过程中引发耐药性。我们还建立了根部感染系统,并证明XA21还介导了根中对 Xoo 的种族特异性抗性反应。我们当前的研究提供了对XA21介导的抗性的本质的了解,并提供了一种实用的方法来利用根细胞系统进一步解析水稻与 Xoo 相互作用期间PRR的细胞信号传导。 http://dx.doi.org/10.1093/mp/ssq038

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