Bringing Plant Immunity to Light: A Genetically Encoded, Bioluminescent Reporter of Pattern-Triggered Immunity in Nicotiana benthamiana
Bringing Plant Immunity to Light: A Genetically Encoded, Bioluminescent Reporter of Pattern-Triggered Immunity in Nicotiana benthamiana
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DOI：
10.1094/mpmi-07-22-0160-ta
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发表时间：
2023-02-07
影响因子：
3.5
通讯作者：
Steinbrenner，Adam D.
中科院分区：
文献类型：
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作者：
Garcia，Anthony G. K.;Steinbrenner，Adam D.
植物依靠先天免疫系统来抵御各种生物攻击者。先天免疫的关键成分包括细胞表面模式识别受体（PRR），其识别害虫和病原体相关的分子模式（PAMP）。与其他类型的受体不同，这些受体在激活时通常具有可见的细胞死亡免疫输出，PRR通常缺乏用于评估功能的快速方法。在这里，我们描述了一个基因编码的生物发光报告免疫激活异源表达PRRs的模式organismNicotianabenthamiana。我们刻画了N.本氏菌的转录组的变化响应于根癌农杆菌和随后的PAMP处理，以鉴定模式触发免疫（PTI）相关的标记基因，然后将其用于产生启动子-荧光素酶融合真菌生物发光途径（FBP）构建体。一种报告基因构建体pFBP_2xNbLYS1：：LUZ允许通过异源表达的PRR稳健地检测PTI活化。与已知的PTI信号传导途径一致，受体样蛋白（RLP）PRR的报告基因活化依赖于RLP PRR的已知衔接子，即，SOBIR1. FBP报告基因最大限度地减少了测定PRR功能所需的劳动力、试剂和时间，并在生物相关PAMP浓度下显示出稳健的灵敏度，使其成为高通量筛选的理想选择。本文所描述的工具将是强有力的研究PRR功能和表征植物细胞表面受体的结构-功能。作者已经在知识共享CC 0“无权利保留”许可下将作品奉献给公共领域，在法律允许的范围内放弃他或她在全球范围内对作品的所有权利，包括所有相关和相邻权利，2023.
Plants rely on innate immune systems to defend against a wide variety of biotic attackers. Key components of innate immunity include cell-surface pattern-recognition receptors (PRRs), which recognize pest- and pathogen-associated molecular patterns (PAMPs). Unlike other classes of receptors that often have visible cell-death immune outputs upon activation, PRRs generally lack rapid methods for assessing function. Here, we describe a genetically encoded bioluminescent reporter of immune activation by heterologously expressed PRRs in the model organismNicotiana benthamiana. We characterizedN. benthamianatranscriptome changes in response toAgrobacterium tumefaciensand subsequent PAMP treatment to identify pattern-triggered immunity (PTI)-associated marker genes, which were then used to generate promoter-luciferase fusion fungal bioluminescence pathway (FBP) constructs. A reporter construct termedpFBP_2xNbLYS1::LUZallows for robust detection of PTI activation by heterologously expressed PRRs. Consistent with known PTI signaling pathways, reporter activation by receptor-like protein (RLP) PRRs is dependent on the known adaptor of RLP PRRs, i.e., SOBIR1. The FBP reporter minimizes the amount of labor, reagents, and time needed to assay function of PRRs and displays robust sensitivity at biologically relevant PAMP concentrations, making it ideal for high throughput screens. The tools described in this paper will be powerful for investigations of PRR function and characterization of the structure-function of plant cell-surface receptors.The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2023.