Organelle-targeted biosensors reveal distinct oxidative events during pattern-triggered immune responses.
Organelle-targeted biosensors reveal distinct oxidative events during pattern-triggered immune responses.
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DOI：
10.1093/plphys/kiac603
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发表时间：
2023-04-03
期刊：
影响因子：
7.4
通讯作者：
Smirnoff, Nicholas
中科院分区：
文献类型：
--
作者：
Arnaud, Dominique;Deeks, Michael J.;Smirnoff, Nicholas
活性氧的产生是对病原体和病原体相关分子模式的响应，如拟南芥（拟南芥）依赖于NADPH氧化酶异构体呼吸爆发氧化酶同源物D （RBOHD）的快速细胞外氧化爆发。我们使用H2O2生物传感器roGFP2-Orp1和谷胱甘肽氧化还原状态生物传感器GRX1-roGFP2靶向各种细胞器，以揭示在鞭毛蛋白（flg22）模式触发免疫反应期间和接种紫花假单胞菌后未预料到的氧化事件。roGFP2-Orp1在处理后1和6小时以双相方式氧化，与叶绿体、线粒体和过氧化物酶体相比，在细胞质中有更强烈和更快的反应。过氧化物酶体和细胞质GRX1-roGFP2也以双相方式氧化。有趣的是，我们的研究结果表明细菌效应物部分抑制细胞质中roGFP2-Orp1氧化的第二阶段。药理学和遗传学分析表明，病原体相关分子模式诱导的胞质氧化需要参与免疫应答的BRI1-ASSOCIATED RECEPTOR KINASE （BAK1）和botryis -induced KINASE 1 （BIK1）信号传导成分，但在很大程度上不依赖于NADPH氧化酶RBOHD和呼吸爆发氧化酶同源物F （RBOHF）以及脂质过氧化物酶过氧化物酶33 （PRX33）和过氧化物酶34 （PRX34）。用鲁米诺测量的初始外胞体氧化爆发之后是第二次氧化爆发，这两次氧化爆发都发生在两波细胞质氧化之前。与胞质氧化相反，这些爆发依赖于rbohd。我们的研究结果揭示了在模式触发的免疫反应过程中复杂的氧化来源和动力学，包括细胞质氧化在很大程度上独立于先前的细胞外氧化事件。
Reactive oxygen species are produced in response to pathogens and pathogen-associated molecular patterns, as exemplified by the rapid extracellular oxidative burst dependent on the NADPH oxidase isoform RESPIRATORY BURST OXIDASE HOMOLOG D (RBOHD) in Arabidopsis (Arabidopsis thaliana). We used the H2O2 biosensor roGFP2-Orp1 and the glutathione redox state biosensor GRX1-roGFP2 targeted to various organelles to reveal unsuspected oxidative events during the pattern-triggered immune response to flagellin (flg22) and after inoculation with Pseudomonas syringae. roGFP2-Orp1 was oxidized in a biphasic manner 1 and 6 h after treatment, with a more intense and faster response in the cytosol compared to chloroplasts, mitochondria, and peroxisomes. Peroxisomal and cytosolic GRX1-roGFP2 were also oxidized in a biphasic manner. Interestingly, our results suggested that bacterial effectors partially suppress the second phase of roGFP2-Orp1 oxidation in the cytosol. Pharmacological and genetic analyses indicated that the pathogen-associated molecular pattern-induced cytosolic oxidation required the BRI1-ASSOCIATED RECEPTOR KINASE (BAK1) and BOTRYTIS-INDUCED KINASE 1 (BIK1) signaling components involved in the immune response but was largely independent of NADPH oxidases RBOHD and RESPIRATORY BURST OXIDASE HOMOLOG F (RBOHF) and apoplastic peroxidases peroxidase 33 (PRX33) and peroxidase 34 (PRX34). The initial apoplastic oxidative burst measured with luminol was followed by a second oxidation burst, both of which preceded the two waves of cytosolic oxidation. In contrast to the cytosolic oxidation, these bursts were RBOHD-dependent. Our results reveal complex oxidative sources and dynamics during the pattern-triggered immune response, including that cytosolic oxidation is largely independent of the preceding extracellular oxidation events.
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