Repression Mediated Embryonic Paterning in Arabidopsis

拟南芥中抑制介导的胚胎模式

• 批准号: 1457381
• 负责人: Jeffrey Long
• 金额: $ 56.74万
• 依托单位: University of California-Los Angeles
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1457381&HistoricalAwards=false
• 关键词: Repression; Mediated; Embryonic; Paterning; Arabidopsis

Plants begin their life with a very basic body plan consisting of a shoot at the top and a root at the bottom. These structures form in the embryo, and proper development of the shoot and root is controlled in large part by a plant hormone called auxin. Auxin is synthesized and transported throughout the embryo and therefore each cell must make a decision whether to respond or not respond to its presence. This project seeks to uncover the mechanisms used by cells to dampen the response to auxin, and the resulting gene expression and developmental changes that occur when this process is disrupted. As auxin is used to pattern organs throughout the life of the plant, these studies will be broadly applicable to plant development and will apply to the growth and patterning of commercially available crop plants. Reagents created from this work will also be used during outreach programs for the general public at the Mildred E. Mathias Botanical Garden as well as in undergraduate classrooms at the University of California Los Angeles.This project seeks to understand the relationship between two classes of transcription factors, the AUX/IAA and AUXIN RESPONSE FACTORS, during Arabidopsis embryo patterning. Specifically, the action of three AUX/IAAs (AUX/IAA8,9,31) and three ARFs (ARF1,2,9) that work with the co-repressor TOPLESS (TPL) to repress gene expression will be explored. Genome wide targets of AUX/IAA8 and ARF2 will be determined during embryogenesis using chromatin immunoprecipitation followed by deep sequencing. These targets will then be compared to the known targets of TPL to define a shared transcriptional network. RNA-seq in both aux/iaa8 and arf2 mutant backgrounds will be used to correlate gene expression changes with genome binding. The study will also document both the protein and mRNA accumulation patterns of all six of these genes and a characterization of a hextuple mutant will be carried out using well-characterized fluorescent reporters and in situ hybridizations. Finally, the biological function of the B3 repression motif found in ARF2 will be explored and an in planta repression assay will be used to determine the transcriptional activity of ARF2 at a cellular level. Taken together, these experiments will provide the field with one of the most complete characterizations of an AUX/IAA and ARF protein to date. As these proteins are used throughout development and are found in almost all plants, the results should be applicable to many studies on how plant hormones regulate development.

植物开始他们的生活与一个非常基本的身体计划组成的一个芽在顶部和一个根在底部。这些结构在胚胎中形成，芽和根的正常发育在很大程度上由一种称为生长素的植物激素控制。生长素在整个胚胎中合成和运输，因此每个细胞必须决定是否对其存在做出反应。该项目旨在揭示细胞抑制对生长素反应的机制，以及当这一过程被破坏时所产生的基因表达和发育变化。由于生长素在植物的整个生命过程中用于器官的图案化，这些研究将广泛适用于植物发育，并将适用于商业上可获得的作物植物的生长和图案化。从这项工作中创建的试剂也将用于Mildred E.马蒂亚斯植物园以及在加州大学洛杉矶的本科生教室。本项目旨在了解两类转录因子，AUX/IAA和生长素反应因子，在拟南芥胚胎图案化过程中的关系。具体而言，将探索三种AUX/IAAs（AUX/IAA 8，9，31）和三种ARF（ARF 1，2，9）与辅阻遏物TOPLESS（TPL）一起抑制基因表达的作用。将在胚胎发生期间使用染色质免疫沉淀法，然后进行深度测序，确定AUX/IAA 8和ARF 2的全基因组靶标。然后将这些靶标与TPL的已知靶标进行比较，以定义共享的转录网络。aux/iaa 8和arf 2突变体背景中的RNA-seq将用于将基因表达变化与基因组结合相关联。该研究还将记录所有六个基因的蛋白质和mRNA积累模式，并将使用表征良好的荧光报告和原位杂交进行六重突变体的表征。最后，将探讨在ARF 2中发现的B3抑制基序的生物学功能，并使用植物内抑制试验来确定ARF 2在细胞水平上的转录活性。总之，这些实验将提供该领域的AUX/IAA和ARF蛋白质的最完整的表征之一。由于这些蛋白质在整个发育过程中使用，并且几乎在所有植物中都存在，因此这些结果应该适用于许多关于植物激素如何调节发育的研究。