CAREER: Using Systems Biology to Study MAP Kinase Cascades in Arabidopsis

研究小组将使用系统生物学方法来了解拟南芥中MAP激酶（MAPK）信号转导的过程。全基因组测序表明，拟南芥中有20个基因编码MAPK蛋白。这些蛋白质之间的功能冗余和串扰使得确定特定蛋白质控制哪些信号通路变得困难。为了克服这种复杂性，研究小组将使用一种新的高通量基因分型管道来创建所有20个MAPK基因突变的杂合单株植物。这20对杂合子的后代应该分离所有可能的MAPK突变组合。通过对选定的后代进行基因分型，应该可以确定哪些突变组合导致了特定的表型。研究小组不会在整个人群中进行搜索，试图分离出所有1140种不同的三突变体。相反，他们将让植物的生物学表明哪些突变组合是有用的。这项工作的结果将是对拟南芥中由MAPK级联控制的许多信号转导途径的更好理解。还将开发强调系统生物学和高通量技术概念的教育材料。Krysan博士将把高通量基因分型系统整合到他的植物功能基因组学课程中，以便为学生提供这些方法的实践经验。在高中阶段，一名当地教师将与研究小组合作，开发适合在高中环境中使用的实用练习，捕捉高通量基因分型的本质。这些材料将通过将举办的夏季讲习班分发给其他高中教师。

The research team will use a systems biology approach to understand the process of MAP kinase (MAPK) signal transduction in Arabidopsis. Whole-genome sequencing has indicated that there are 20 genes that encode MAPK proteins in Arabidopsis. Functional redundancy and cross-talk between these proteins make it difficult to determine which signaling pathways are controlled by a given protein. In order to overcome this complexity, the research team will use a novel high-throughput genotyping pipeline to create single plants that are heterozygous for mutations in all twenty MAPK genes. The progeny of these 20-tuple heterozygotes should segregate all of the possible combinations of MAPK mutations. By genotyping selected sets of progeny, it should be possible to determine which mutant combinations are causing particular phenotypes. The research team will not search through the population to try and isolate examples of all 1,140 different triple-mutants. Instead they will let the biology of the plants indicate which mutant combinations are informative. The outcome of this work will be an improved understanding of the many signal transduction pathways controlled by MAPK cascades in Arabidopsis. Educational materials will also be developed that highlight the concepts of systems biology and high-throughput technology. Dr. Krysan will incorporate the high-throughput genotyping system into his Plant Functional Genomics course in order to give students hands-on experience with these methods. At the high school level, a local teacher will collaborate with the research team to develop practical exercises suitable for use in a high school setting that capture the essence of high-throughput genotyping. These materials will be disseminated to other high school teachers through a summer workshop that will be offered.