Center for Research on Plant Transporters (CROPS) - A Plant Community Resource for the Structure and Function of Plant Membrane Transporters Underlying Important Crop Traits

PI: Geoffrey Chang（加州大学圣地亚哥分校）co -PI: Erin Connolly（南卡罗莱纳大学），Leon Kochian和Miguel Piñeros （USDA-ARS/康奈尔大学），Michael Stowell（科罗拉多大学博尔德分校），Julian Schroeder和Milton Saier（加州大学圣地亚哥分校）植物需要从土壤中吸收养分，并将这些养分输送到整个植物的专门细胞中。这些营养物质在每个植物细胞内通过植物细胞膜和细胞器膜进行运输。转运蛋白是介导几乎所有分子和离子通过植物细胞膜的蛋白质。许多转运蛋白的自然变异已被证明对与植物生长和产量以及对非生物和生物胁迫的耐受性相关的关键农艺性状很重要。然而，这些蛋白质的基本生化、功能和结构特征在很大程度上是未知的。为了解决这些问题以及在这一研究领域对新工具的迫切需求，我们正在建立一个新的中心，称为植物转运体研究中心（crop），致力于研究植物转运体的功能和结构特征，这些转运体按其农艺重要性优先排序。该中心将利用现有的先进研究基础设施，大规模生产纯化的植物转运蛋白，并快速发现/生成新的单域抗体，这些抗体可作为有价值的工具，并可能为更大的植物科学界带来变革和支持资源。此外，作物将决定关键植物膜转运蛋白及其自然变异的分子结构，为了解其在植物中的作用提供一个新的框架，为不同作物物种的重要农业性状提供基础。在过去的四分之一世纪里，我们对植物转运体的了解急剧增加。这使得人们越来越认识到植物转运体在许多农艺性状中发挥着重要作用，例如水和养分的有效获取/利用以及作物对由盐度、酸度和重金属引起的不利土壤环境的耐受性。自然遗传变异是培育各种农业相关性状的基础。了解遗传变异如何转化为转运体结构和功能的变化，对于在作物改良中更有效地利用这种变异至关重要。作物将侧重于与边际条件下的粮食安全和可持续性以及人类健康/营养和食品安全有关的农艺性状的关键问题。具体而言，通过外部合作和内部专业知识，作物将针对关键转运体，包括涉及微量营养素获取，铝耐受性，磷获取效率，氮获取效率，耐盐性和病原体和昆虫抗性的转运体。作为第一步，庄稼将建立一个高效的管道，基于成熟的技术，为植物科学界生产植物膜运输资源。这些资源包括大量纯化的植物转运蛋白、强大的纳米体体外生产平台、新型合成单域抗体和最先进的结构测定技术。CROPS致力于为更广泛的植物膜运输界提供所有数据和生物资源。对蛋白质、抗体试剂和蛋白质结构信息的获取将通过项目网站/数据库和通过长期知识库的传播提供。具体来说，所有结构x射线和EM结构数据（GC和MS）将存储在蛋白质数据库（http://www.pdb.org）和EMDataBank （http://www.emdatabank.org）中。所有植物和合成纳米体DNA序列将被注释并存入GenBank和Gramene。质粒构建体将储存在Addgene （https://www.addgene.org）中，这是一个非营利性质粒储存库。在外联方面，CROPS将提供机制，邀请更广泛的社区利用它的管道，作为它们自己的研究的基础。研究人员将被邀请为感兴趣的特定植物转运体提交提名。研究生和博士后将接受培训，并获得一些最先进的技术，如快速抗体（纳米体）进化，膜蛋白结构/功能生物学，以及基于分子的特性分析，以促进独立的职业发展。农作物将非常重视为来自弱势群体的高中生和本科生提供研究培训和实验室经验，重点是发展写作和演讲技能，并指导学生承担个人责任，获得科学教育和项目管理技能。

PI: Geoffrey Chang (University of California-San Diego)Co-PIs: Erin Connolly (University of South Carolina), Leon Kochian and Miguel Piñeros (USDA-ARS/Cornell University), Michael Stowell (University of Colorado-Boulder), Julian Schroeder and Milton Saier (University of California, San Diego)Plants need to take up nutrients from the soil and transport these nutrients to specialized cells throughout the whole plant. These nutrients are transported across plant cell membranes and organelle membranes inside of each plant cell. Transporters are proteins that mediate the passage of virtually every molecule and ion across plant cell membranes. Many natural variants of transport proteins have been shown to be important for key agronomic traits associated with plant growth and yield, and tolerance to abiotic and biotic stresses. However, the fundamental biochemical, functional, and structural characteristics of these proteins are largely unknown. To address these questions and the glaring need for new tools in this research area, we are establishing a new center called the Center for Research on Plant TransporterS (CROPS), devoted to the functional and structural characterization of plant transporters prioritized by their agronomic importance. This center will leverage an existing advanced research infrastructure to produce purified plant transporters on a large-scale and rapidly discover/generate novel single-domain antibodies that can be used as valuable tools and potentially transformative and enabling resources for the greater plant science community. In addition, CROPS will determine the molecular structures of key plant membrane transporters and their natural variants, providing a new framework for understanding their functions in the context of their in planta roles, underpinning agriculturally important traits in diverse crop species.Our understanding of plant transporters has increased dramatically over the past quarter century. This has led to a growing awareness that plant transporters play important roles in many agronomic traits, such as efficient acquisition/use of water and nutrients and crop tolerance to adverse soil environments arising from salinity, acidity, and the presence of heavy metals. Natural genetic variation is the foundation for breeding a wide variety of agriculturally relevant traits. Understanding how genetic variation translates into transporter structural and functional changes is critical to provide for a more efficient exploitation of this variation in crop improvement. CROPS will focus on key questions regarding agronomic traits that relate to food security and sustainability under marginal conditions and human health/nutrition and food safety. Specifically, through external collaborations and internal expertise, CROPS will target key transporters including those involved in micronutrient acquisition, aluminum tolerance, phosphorous acquisition efficiency, nitrogen acquisition efficiency, salt tolerance and pathogen and insect resistance. As a first step, CROPS will establish an efficient pipeline based on proven technologies for the production of plant membrane transport resources for the plant scientific community. These resources include large quantities of purified plant transporter proteins, a powerful in vitro production platform for nanobodies, novel synthetic single-domain antibodies, and state-of-the-art structural determination techniques. CROPS has a strong commitment to providing access to all data and biological resources to the broader plant membrane transport community. Access to proteins, antibody reagents, and information about protein structure will be provided through a project website/database and by dissemination through long-term repositories. Specifically, all structural x-ray and EM structural data (GC and MS) will be deposited in the Protein Data Bank (http://www.pdb.org) and the EMDataBank (http://www.emdatabank.org). All plant and synthetic nanobody DNA sequences will be annotated and deposited in GenBank and Gramene. Plasmid constructs will be deposited in Addgene (https://www.addgene.org), a not-for-profit plasmid repository. With regard to outreach, CROPS will provide mechanisms for inviting the wider community to tap into its pipeline as a basis for their own research. Researchers will be invited to submit nominations for specific plant transporters of interest. Graduate students and postdocs will receive training and access to some of the most state-of-the-art techniques in rapid antibody (nanobody) evolution, membrane protein structural/functional biology, and molecular-based trait analysis for advancing independent careers. CROPS will have a strong emphasis on providing research training and laboratory experiences to high school and undergraduate students from underrepresented groups, with a focus on development of writing and presentation skills as well as mentoring students to take personal responsibility and gain scientific educational and project management skills.