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

植物的生长、发育和存活依赖于15种必需营养物质和无数代谢产物到特定器官、细胞和细胞内隔间的吸收、运输和分选。然而，植物如何调节和协调长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、大量的突变体集合和广泛的数据库提供了产生新的资源和工具来发现~800转运蛋白在拟南芥中功能的机会。大部分未知的转运蛋白是次要的活性转运蛋白。该项目的主要目标是确定56个H偶联阳离子转运蛋白的离子特异性、组织和亚细胞分布以及生物学作用，包括cacA、CPA1和CPA2家族。随着基因组序列的获得和生物信息学分析对运输同源物的鉴定，酵母中异源表达将被用来快速描述整个H偶联阳离子转运蛋白的功能。调控机制将通过假定的自抑制结构域的突变来确定。分离膜中的转运将用于验证典型转运体的离子专一性。组织和亚细胞的分布将通过转基因植物中启动子-报告蛋白的表达和原生质体模型中GFP标记的转运蛋白来确定。转运蛋白的生物学和生理功能将使用插入基因敲除突变体进行分析。这项研究产生的新方法和信息将通过http://www.cbs.umn.edu/2010的一个网站分发，每年更新几次。为了确保社区可以获得结果和资源，将在TAIR储存数据，并在结果得到确认后通过ABRC获得突变的拟南芥品系。转运体影响所有生命过程，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，该项目产生的资源和信息将直接促进实现2010年项目目标的进展。已经制定了一些计划，以吸引学生，包括代表人数不足的少数群体，利用实验和生物信息学方法解决健康、农业和环境方面的问题。

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.