Isolation and molecular analysis of the pepper Bs4C resistance gene that mediates recognition of the cognate Xanthomonas TAL effector protein AvrBs4

介导同源黄单胞菌 TAL 效应蛋白 AvrBs4 识别的辣椒 Bs4C 抗性基因的分离和分子分析

• 批准号: 215184390
• 负责人: Professor Dr. Thomas Lahaye
• 金额: --
• 依托单位: Abteilung für Allgemeine Genetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/215184390?language=en
• 关键词: Isolation; molecular; analysis; pepper; Bs4C

Microbial diseases are a continuous threat to cultivated crops that provide the basis of our global food supply. Wild relatives of cultivated crops represent a rich resource of antimicrobial immune systems that however need to be characterized genetically before their potential can be exploited in breeding programs. Due to technical limitations, plant immune systems were studied in the past primarily in model plants with small genomes. Yet, next-generation sequencing (NGS) technologies facilitate now direct isolation of valuable traits from complex plant genomes and transcriptomes. In the framework of this proposal we want to isolate and characterize the pepper resistance (R) gene Bs4C that confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv), the causal agent of bacterial spot disease on pepper and tomato. NGS-based transcriptome profiling was used to identify Bs4C candidate genes (2.2.1.5). We will apply reverse transcription (RT)-PCR, genetic mapping and functional analysis to clarify which of the identified candidate genes represents Bs4C (3.2.1.1 - 3.2.1.8). If we fail to identify Bs4C within the identified transcripts we will apply NGS-based genome re-sequencing, genetic mapping and functional analysis to identify Bs4C (3.2.1.12). Upon isolation we will characterize the Bs4C gene at the molecular level to clarify how it mediates recognition of Xcv and execution of an immune response (3.2.1.9 - 3.2.1.11). At the technical level we aim to establish a novel R gene cloning strategy that is applicable to complex crop genomes and that implements NGS technology.

微生物疾病对作为全球粮食供应基础的栽培作物构成持续威胁。栽培作物的野生近缘种代表着丰富的抗菌免疫系统资源，然而，在育种计划中利用其潜力之前，需要对其进行遗传表征。由于技术限制，过去主要在基因组较小的模型植物中研究植物免疫系统。然而，新一代测序（NGS）技术现在有利于从复杂的植物基因组和转录组中直接分离有价值的性状。在此提案的框架内，我们想要分离并表征辣椒抗性 (R) 基因 Bs4C，该基因赋予对野油菜黄单胞菌 pv 的抗性。 vesicatoria (Xcv)，辣椒和番茄细菌性斑点病的病原体。基于 NGS 的转录组分析用于识别 Bs4C 候选基因 (2.2.1.5)。我们将应用逆转录 (RT)-PCR、遗传图谱和功能分析来阐明哪些已识别的候选基因代表 Bs4C (3.2.1.1 - 3.2.1.8)。如果我们未能在已识别的转录本中识别 Bs4C，我们将应用基于 NGS 的基因组重测序、遗传图谱和功能分析来识别 Bs4C (3.2.1.12)。分离后，我们将在分子水平上表征 Bs4C 基因，以阐明它如何介导 Xcv 的识别和免疫反应的执行 (3.2.1.9 - 3.2.1.11)。在技​​术层面，我们的目标是建立一种适用于复杂作物基因组并实施NGS技术的新型R基因克隆策略。

项目成果

RNA-seq pinpoints a Xanthomonas TAL-effector activated resistance gene in a large-crop genome

• DOI: 10.1073/pnas.1212415109
• 发表时间: 2012-11-20
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Strauss, Tina;van Poecke, Remco M. P.;Lahaye, Thomas
• 通讯作者: Lahaye, Thomas