Arabidopsis 2010 - Collaborative Research: Discovering Transporters for Essential Minerals and Toxic Ions in Plants

植物的生长、发育和生存依赖于15种必需营养素和无数代谢物对特定器官、细胞和细胞内区室的吸收、转运和分类。然而，植物如何调节和协调远距离和细胞运输模式的复杂网络尚不清楚。第一个完整的植物基因组序列、大量的突变体收集和广泛的数据库为发现拟南芥中约800个转运体的功能提供了新的资源和工具。大部分未知转运蛋白是次生活性转运蛋白。该项目的主要目标是确定56种H+偶联阳离子转运体的离子特异性、组织和亚细胞分布以及生物学作用，包括CaCA、CPA1和CPA2家族。随着基因组序列的可用性和通过生物信息学分析的转运同源物的鉴定，酵母中的异源表达将用于快速描述H+偶联阳离子转运体的整个补体的功能。调节机制将由假定的自抑制结构域的突变决定。在分离膜中的运输将用于验证代表性转运体的离子特异性。组织和亚细胞分布将通过转基因植物中的启动子报告蛋白表达和原生质体模型中gfp标记的转运蛋白来确定。将使用插入敲除突变体分析转运体的生物学和生理功能。这项研究产生的新方法和信息将通过一个网址http://www.cbs.umn.edu/2010分发，每年更新几次。为了确保结果和资源可供社区使用，数据将存储在TAIR中，结果确认后将通过ABRC提供突变拟南芥系。转运蛋白影响所有生命过程，包括繁殖、新陈代谢、运动以及对激素和环境刺激的反应，因此本项目产生的资源和信息将直接促进2010年项目目标的实现。为吸引学生，包括代表性不足的少数民族学生，并为他们提供机会，利用实验和生物信息学方法解决卫生、农业和环境方面的问题，制定了一些项目。

Plant growth, development, and survival depend on the uptake, translocation, and sorting of 15 essential nutrients and countless metabolites to specific organs, cells and intracellular compartments. However, it is not known how plants regulate and coordinate the complex network of long-distance and cellular transport patterns. The first complete genome sequence of a plant, large mutant collections, and extensive databases provide opportunities to generate new resources and tools to discover the functions of ~800 transporters in Arabidopsis. The bulk of unknown transporters are secondary active transporters. The major objectives of this project are to determine the ion specificity, the tissue and subcellular distribution, and the biological roles of 56 H+-coupled cation transporters, including the CaCA, CPA1, and CPA2 families. With the availability of genomic sequence and the identification of transport homologs through bioinformatic analyses, heterologous expression in yeast will be used to quickly delineate the function of the entire complement of H+-coupled cation transporters. Regulatory mechanisms will be determined by mutation of putative autoinhibitory domains. Transport in isolated membranes will be used to verify ion specificity of representative transporters. Tissue and subcellular distribution will be determined using promoter-reporter proteins expression in transgenic plants, and GFP-tagged transporters in a protoplast model. Biological and physiological functions of transporters will be analyzed using insertional knockout mutants. New methods and information resulting from the study will be distributed through a site at http://www.cbs.umn.edu/2010 with several updates each year. To assure that results and resources are available to the community, data will be depositied at TAIR and mutant Arabidopsis lines will be available through the ABRC after results are confirmed.Transporters impact all life processes, including reproduction, metabolism, movement, and responses to hormonal and environmental stimuli, thus resources and information generated from this project will directly facilitate progress towards the goals of the 2010 project. Programs are in place to attract and provide opportunities for students, including under-represented minorities, to solve problems in health, agriculture, and the environment using experimental and bioinformatic approaches.