Isolation of Novel MAMP-like Activities and Identification of Cognate Pattern Recognition Receptors in Arabidopsis thaliana Using Next-Generation Sequencing (NGS)–Based Mapping

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2018-02-13 DOI:10.1002/cppb.20056
Li Fan, Eunyoung Chae, Andrea A. Gust, Thorsten Nürnberger
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引用次数: 5

Abstract

Activation of pattern-triggered plant immunity requires recognition of microbe-derived molecular patterns (MAMPs) by plant-encoded pattern recognition receptors (PRRs). Many plant PRRs are found in selected plant genera only. Transfer of single PRRs or of cassettes expressing several PRRs (PRR stacking) across plant genus boundaries offers the potential to boost disease resistance by improving pathogen recognition features in economically important crop plants. The success of such an approach is most dependent on the availability of a large number of plant PRRs. Here, an efficient method for the identification of novel PRRs in the model plant Arabidopsis thaliana (hereafter, Arabidopsis for simplicity) is described. This method takes advantage of natural variation in microbial pattern sensitivity among hundreds of Arabidopsis accessions currently available. Identification of pattern-sensitive as well as pattern-insensitive accessions facilitates next-generation sequencing (NGS)–assisted mapping of PRRs. This approach is potentially applicable to the identification of PRRs that recognize patterns of any chemical nature. © 2017 by John Wiley & Sons, Inc.

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利用基于下一代测序(NGS)的定位法分离拟南芥中新的MAMP样活性和鉴定同源模式识别受体。
激活模式触发的植物免疫需要植物编码模式识别受体(PRRs)识别微生物衍生分子模式(MAMPs)。许多植物PRRs仅在选定的植物属中发现。单个PRRs或表达多个PRRs的磁带(PRR堆叠)跨植物属边界的转移提供了通过改善经济上重要作物的病原体识别特征来增强抗病能力的潜力。这种方法的成功在很大程度上取决于能否获得大量的植物PRRs。本文描述了一种在模式植物拟南芥(以下简称拟南芥)中鉴定新型PRRs的有效方法。该方法利用了目前可获得的数百种拟南芥材料中微生物模式敏感性的自然变化。鉴定模式敏感和模式不敏感的加入有助于下一代测序(NGS)辅助的PRRs制图。这种方法可能适用于识别任何化学性质的PRRs。©2017 by John Wiley & Sons, Inc。
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Current protocols in plant biology
Current protocols in plant biology Agricultural and Biological Sciences-Plant Science
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期刊介绍: Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.
期刊最新文献
Issue Information Isolation, Library Preparation, and Bioinformatic Analysis of Historical and Ancient Plant DNA Isolation of Plant Root Nuclei for Single Cell RNA Sequencing Selective Enrichment Coupled with Proteomics to Identify S-Acylated Plasma Membrane Proteins in Arabidopsis In-Plate Quantitative Characterization of Arabidopsis thaliana Susceptibility to the Fungal Vascular Pathogen Fusarium oxysporum.
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