Arabidopsis 2010: Integrating Gene Expression With Biochemical Phenotype in Plant-Insect Interactions

尽管食草动物对生态、进化和经济产生了巨大影响，但对植物对敌人反应的分子基础的研究更多地集中在微生物上，而不是昆虫上。植物对昆虫反应的研究主要局限于表型（化学）防御。很少有人尝试将分子和化学反应与任何一种害虫联系起来，以了解植物防御的机制。该项目将采用约2500个应激反应基因的微阵列（由以前的NSF植物基因组项目开发）以及硫代葡萄糖苷生物合成途径中的关键基因。基因表达和硫代葡萄糖苷谱将用于植物对昆虫和病原体的反应，这些昆虫和病原体代表了咀嚼和吸吮进食类型以及十字花科的一系列专业化。这个有限的概念验证项目的主要目标是证明基因功能可以通过使用多元统计处理来推断，将阵列结果与防御代谢物反应谱和生态结果联系起来。防御方面的“基因功能”将根据与已知功能相关的表达模式（例如，在信号传导途径，代谢物合成中）和生态结果的背景下（例如，对昆虫的影响，对食草动物的抗性），使用标准的多变量聚类技术，一致性分析和Mantel协变量统计来推断。统计鉴定的植物防御反应关键成分的身份和功能最终将在不同环境条件下与特定昆虫和微生物进行生物测定时使用突变体和敲除来确认。表达模式的图像、代谢物谱数据、综合结果、方案更新和获取材料的说明将每月发布在以下tair链接的URL: http://schultzlab.cas.psu.edu/。代谢组学数据将存档于基因组拟南芥资源网络（http://www.york.ac.uk/res/garnet/beale.htm），功能基因组学数据将存档于TAIR （http://www.arabidopsis.org/info/2010_projects/）。拟南芥2010项目的目标包括确定植物防御背景下基因的功能，生产胁迫微阵列和化学分析协议，作为广泛的研究人员有用的工具，以及开发分子生态学家可以用来连接基因型和表型反应的统计协议。此外，该项目将与现有的2个拟南芥2010项目(Wurtele et al.；“视觉信息学工具”和Lewis等人。“苯丙类代谢物网络”)，并与马克斯·普朗克化学生态学研究所（德国耶拿）建立国际合作。该有限项目的广泛影响将包括培养3名研究生，利用PSU农业科学与化学系现有的K-12，教师和本科生研究经验联系，以及开发将拟南芥反应作为K-8环境中教学工具的平台。

Studies of the molecular bases of plant responses to enemies have focused more on microbes than on insects, despite the great ecological, evolutionary, and economic impacts of herbivores. Research on plant responses to insects has been limited mainly to phenotypic (chemical) defenses. Little attempt has been made to link molecular and chemical responses to either type of pest to understand the mechanisms underlying plant defense. This project will employ microarrays of approximately 2500 stress-responsive genes (developed by a previous NSF Plant Genome project) plus critical genes in the glucosinolate biosynthetic pathway. Gene expression and glucosinolate profiles will be developed for plants responding to insect species and pathogens representing chewing and sucking feeding types and a range of specialization on Brassicaceae. The major goal of this limited proof-of-concept project is to demonstrate that gene function can be inferred by using multivariate statistical treatments to link array results to defense metabolite response profiles and to ecological outcomes. "Gene function" in terms of defense will be inferred in terms of expression pattern in relation to known functions (e.g., in signaling pathways, metabolite synthesis) and in the context of the ecological outcome (e.g., impact on insect, resistance to herbivory) using standard multivariate clustering techniques, concordance analysis, and Mantel covariate statistics. Identities and functions of critical components of plant defense responses identified statistically will eventually be confirmed using mutants and knockouts in bioassays with specific insects and microbes under varying environmental conditions. Images of expression patterns, metabolite profile data, integrative results, protocol updates, and instructions for obtaining material will be posted monthly at the following TAIR-linked URL: http://schultzlab.cas.psu.edu/ . Metabolomics data will be archived at the Genomic Arabidopsis Resource Network (http://www.york.ac.uk/res/garnet/beale.htm) and functional genomics data at TAIR (http://www.arabidopsis.org/info/2010_projects/ ). Arabidopsis 2010 objectives addressed by this project include determining the functions of genes in the context of plant defense, producing a stress microarray and chemical analytical protocols as tools useful to a wide range of investigators, and developing statistical protocols molecular ecologists can use to link genotypic and phenotypic responses. In addition, the project will be coordinated with 2 existing Arabidopsis 2010 programs (Wurtele et al. "Visual informatics tools" and Lewis et al. "Phenylpropanoid metabolite networks") and establishes an international collaboration with the Max Planck Institute for Chemical Ecology (Jena, Germany). Broader impacts of the limited project will include training 3 graduate students, exploiting existing K-12, teacher, and undergraduate research experience links that already exist in the PSU College of Agricultural Sciences and Chemistry Department, and developing platforms for using Arabidopsis responses as a teaching tool in the K-8 environment.