Characterization of the Tomato Secretome Using Integrated Functional and Computational Strategies

植物细胞壁在植物生物学的许多方面都起着核心作用，包括器官的生长和分化、对微生物病原体的防御、营养物质的代谢和运输以及对环境胁迫的反应。此外，植物细胞壁可以被认为是一个重要的隔间，作为与环境的接口，因此大量的分子信号发生在壁环境中和穿过壁环境。根据其多功能的性质，植物细胞壁含有大量复杂的蛋白质或蛋白质组，包括具有广泛催化功能的酶和结构蛋白质。然而，越来越明显的是，对细胞壁蛋白质组的范围、组成和动态性质的了解仍然非常有限，也被称为分泌组。该项目将使用一种集成的方法对番茄分泌组进行分类，将一套新开发的功能筛查与高度纯化的壁蛋白提取物、生物信息工具和计算预测相结合。这一基础设施将被用来评估在与壁生物学密切相关的两个生物过程中细胞壁蛋白质组的定性和定量动态：果实成熟和植物防御反应。为此，我们将研究一个标准的番茄实验品种的蛋白质和信使核糖核酸的表达，并与几个非软化番茄突变体的蛋白质和mRNA表达进行对比。这项研究将产生一系列番茄组织中植物分泌组的全面目录，重要的是，为深入了解在果实成熟和病原菌攻击过程中似乎有助于并在信号转导中发挥作用的细胞外蛋白。由此产生的信息应该能够让人们对如何提高水果质量和农业生产率有重要的见解。将开发一个可公开访问的番茄分泌组数据库(SecreTom)，以存储项目数据，提供对所有相关计算工具的访问，并作为植物壁蛋白质组研究的中心。SecreTOM将通过茄科基因组数据库(sgn：http://www.sgn.cornell.edu/)，)访问，该数据库也将作为项目数据的长期储存库。此外，还将举办一年一度的夏季蛋白质组学研讨会，包括研讨会系列、海报演示和以实验室为基础的实践培训。将通过现有康奈尔方案网络的支持，确保本科生和研究生中代表性不足的群体的参与。

The plant cell wall plays a central role in many diverse aspects of plant biology, including organ growth and differentiation, defence against microbial pathogens, metabolism and transport of nutrients and responses to environmental stresses. In addition, the plant cell wall can be thought of as a vital compartment that acts as the interface with the environment and so substantial molecular signaling occurs in and across the wall environment. In accordance with its multifunctional nature, the plant cell wall houses a large and complex protein population, or proteome, including enzymes with a broad range of catalytic functions as well as structural proteins. However, it is increasingly evident that there is still a very limited understanding of the extent, composition and dynamic properties of the cell wall proteome, also termed the 'secretome'. This project will use an integrated approach to catalog the tomato secretome, coupling a suite of newly developed functional screens with sequencing of highly purified wall protein extracts, bioinfomatic tools and computational prediction. This infrastructure will be used to evaluate the qualitative and quantitative dynamics of the cell wall proteome during two biological processes that are intimately associated with wall biology: fruit ripening and plant defence responses. To this end, protein and mRNA expression will be studied in a standard experimental tomato genotype and contrasted with those in several non-softening tomato mutants.This research will generate a comprehensive catalog of the plant secretome in a range of tomato tissues and importantly, provide insights into the extracellular proteins that appear to contribute to, and play a role in, signaling during fruit ripening and in response to pathogen attack. The resulting information should allow important insights into how to improve fruit quality and agricultural productivity. A publicly accessible tomato secretome database (SecreTom) will be developed to house the project data, to provide access to all associated computational tools, and to act as a hub for plant wall proteome research. SecreTom will be accessible through the Solanaceous Genomes Database (SGN: http://www.sgn.cornell.edu/), which will also serve as the long-term repository for the project data. In addition, an annual summer Proteomics Workshop will be held, comprising a seminar series, poster presentations, and lab-based practical training. The participation of under-represented groups at the undergraduate and graduate levels will be ensured through the support of a network of existing Cornell programs.