An Arabidopsis and Tomato Mesophyll Protoplast System for Fast Identification of Early MAMP-Triggered Immunity-Suppressing Effectors
An Arabidopsis and Tomato Mesophyll Protoplast System for Fast Identification of Early MAMP-Triggered Immunity-Suppressing Effectors
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DOI：
10.1007/978-1-62703-986-4_17
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Brunner, Frederic
中科院分区：
文献类型：
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作者：
Fraiture, Malou;Zheng, Xiangzi;Brunner, Frederic
植物叶肉原生质体中的瞬时表达允许以简化且同步的方式快速表征体内基因功能，而不会由于使用基于细菌的基因或蛋白质递送系统而产生偏差。它提供了测试微生物效应子是否可以颠覆植物免疫信号的早期事件的可能性，这些信号在识别微生物相关分子模式 (MAMP) 后被激活，即所谓的 MAMP 触发免疫 (MTI)。在这里，我们描述了拟南芥和番茄叶肉原生质体效应基因的分离和转染，使用非侵入性荧光素酶报告基因测定和检测激活的丝裂原激活蛋白激酶（MAPK）的简单方法，以中等通量的方式识别和研究抑制 MTI 早期信号转导事件的新效应子。
Transient expression in plant mesophyll protoplasts allows rapid characterisation of gene functions in vivo in a simplified and synchronized manner without bias due to the use of bacteria-based gene or protein delivery systems. It offers the possibility to test whether microbial effectors can subvert early events of plant immune signaling that are activated upon recognition of Microbe-Associated Molecular Patterns (MAMPs), the so-called MAMP-triggered immunity (MTI). Here, we describe the isolation and transfection with effector genes of Arabidopsis thaliana and Solanum lycopersicum mesophyll protoplasts, the use of a non-invasive luciferase reporter assay and a simple method to detect activated Mitogen-Activated Protein Kinases (MAPKs) to identify and study, in a medium-throughput manner, new effectors suppressing early signal transduction events of MTI.