CAREER: Regulatory Mechanisms of Pathogen-Mediated Cellular Stress Signaling in Arabidopsis: Taking Plant Molecular Biology to the Urban Garden

非技术摘要：该项目将建立一个创新和连贯的平台，以显著提高我们对植物中病原体介导的内质网（ER）胁迫的理解，培训下一代少数民族科学家，并使城市社区参与公民科学。该项目的研究范围涉及细胞应激和未折叠蛋白质积累与植物免疫系统之间的独特联系。在病原体攻击时，植物细胞增加新肽的产生，从而对ER中的蛋白质折叠机制造成负担并引起细胞应激。通过这个项目，研究人员将了解拟南芥中有效缓解这种应激反应的分子机制。从这项研究中获得的见解将影响农业科学，阐明机制，以开发作物植物的能力，在增加细胞压力下发挥作用-一个更可持续的未来的必要性。位于伯明翰，AL，一个城市，拥有全国最多样化和贫困人口之一（74%的非洲裔美国人，25%低于贫困线），研究人员处于独特的地位，通过为代表性不足和社会经济地位低下的少数民族开展外展计划，促进植物生物学相关教育。研究人员将实施一个综合服务和发现为基础的学习推广平台“OUTPACE”（拓展植物病理学临床教育），该平台针对本科生和公民科学家：当地社区花园的低收入城市园艺家庭。OUTPACE将使学生获得植物免疫的实验室和实地知识，并协助社区种植者诊断植物疾病。这些努力与当前旨在加强公民参与和可持续生活以及消除城市营养不良和饥饿的区域举措保持一致。技术摘要：肌醇需要酶1（IRE 1）是一种高度保守的真核内质网（ER）应激传感器。虽然慢性ER应激与后生动物免疫紊乱有关，但IRE 1a在植物免疫中的作用仍然难以捉摸。最近，主要研究人员在拟南芥免疫信号传导中建立了IRE 1a及其bZIP 60转录因子的客户mRNA之间的第一个已知联系。基于这些发现，研究人员将把他们的研究问题扩展到植物免疫中ER信号传导的其他关键领域，并解决以下目标：1）了解振荡ER胁迫条件下生物胁迫依赖的IRE 1a激活和失活的分子机制; 2）阐明IRE 1a在细胞适应或病原体介导的细胞死亡中的调控机制; 3）解读IRE 1a和水杨酸受体NPR 1之间的反馈机制; 4）全基因组范围内发现和表征新型IRE 1a底物。为了实现这些目标，研究人员将采用成熟的和新开发的方法，包括：突变体/转基因植物研究，基于植物荧光标记的显微镜，病原体感染和细胞死亡测定，蛋白质-蛋白质相互作用和酶测定，一种新的基于酵母标记的系统，转录谱分析和生物信息学辅助分析。

NON-TECHNICAL ABSTRACT:This project will establish an innovative and coherent platform to significantly enhance our understanding of pathogen-mediated endoplasmic reticulum (ER) stress in plants, train next-generation minority scientists, and engage an urban community in citizen science. The research scope of the project addresses a unique connection between the cellular stress and accumulation of unfolded proteins, and the plant immune system. Upon pathogen attack, plant cells increase production of new peptides, which consequently puts a burden on the protein folding machinery in the ER and causes cellular stress. Through this project, the investigators will learn about the molecular mechanisms underlying the efficient alleviation of this stress response in Arabidopsis. The insights gained from this research will impact the agricultural sciences by elucidating mechanisms to develop crop plants with the capacity to function under increased cellular stress - a necessity for a more sustainable future.Situated in Birmingham, AL, a city that has one of the most diverse and impoverished populations nationwide (74% African American, 25% below the poverty line), the investigators are in a unique position to foster plant biology-related education through outreach programs for underrepresented and socioeconomically underprivileged minorities. The investigators will implement an integrative service- and discovery-based learning outreach platform "OUTPACE" (OUTreach plant PAthology Clinic & Education) that is directed towards undergraduate students and citizen scientists: low-income urban gardening families of the local Community Gardens. OUTPACE will enable the students to acquire laboratory- and field-based knowledge of plant immunity and assist community growers in diagnosing plant diseases. These efforts are well aligned with current regional initiatives to enhance civic engagement and sustainable living, and combat urban malnutrition and hunger. TECHNICAL ABSTRACT:Inositol-Requiring Enzyme 1 (IRE1) is a highly conserved eukaryotic endoplasmic reticulum (ER) stress sensor. While chronic ER stress is linked with metazoan immune disorders, the role of IRE1a in plant immunity remains elusive. Recently, the principal investigator established the first known connection between the IRE1a and its client mRNA for the bZIP60 transcription factor in Arabidopsis immune signaling. Based on these findings, the investigators will expand their research questions to other key areas of ER signaling in plant immunity and address the following objectives: 1) Understanding the molecular mechanisms of biotic stress-dependent IRE1a activation and deactivation under oscillating ER stress conditions; 2) Elucidating the regulatory mechanisms of IRE1a functions in cellular adaptation or pathogen-mediated cell death; 3) Deciphering the feedback mechanisms between IRE1a and the salicylic acid receptor NPR1; 4) Genome-wide discovery and characterization of novel IRE1a substrates. To achieve these goals, the investigators will employ well-established and newly developed methodologies, including: mutant/transgenic plants studies, in planta fluorescent reporter-based microscopy, pathogen infections and cell death assays, protein-protein interaction and enzymatic assays, a novel yeast reporter-based system, transcriptional profiling, and bioinformatics-aided analyses.