2019-03-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/683219摘要：植物在面對自然界中的逆境時,會透過訊號傳遞來啟動防禦反應。植物需要藉由其細胞膜上的受器pattern recognition receptors (PRRs)來辨識由病原菌來的pathogen/microbe-associated molecular patterns (PAMPs/MAMPs)或辨識因植物本身受傷而產生的damage-associated molecular patterns (DAMPs) 。然而，目前對於DAMPs在植物中的PRRs研究甚少。因此，本計劃將探討由植物本身因受傷而釋出的化合物，硫代葡萄糖&#33527;(glucosinolates)水解的產物—&#33096;化合物(3-butenenitrile; 3BN)，在植物中的反應及訊號傳遞途徑。首先，通過不同訊號傳遞的阿拉伯芥T-DNA插入突變株來分析其對3BN的反應，以定量即時聚合&#37238;鏈鎖反應(qRT-PCR)來分析訊號傳遞相關基因的表現量，並以不同的生理生化實驗分析植物對於3BN的反應，接著利用代謝體學來分析植物體內的代謝物的代謝變化。在第二年的計劃中，分別收集在不同時間點(0,1,3,6,12,24小時)以3BN處理的植株材料進行RNA-seq分析，從中挑選出參與3BN反應之PRRs及轉錄因子，並將這些後選基因進行基因選殖，以農桿菌GV3101轉殖於阿拉伯芥中。在第三年的計劃中，我們將PRRs及轉錄因子的阿拉伯芥基因轉殖株進行一系列的抗病及抗蟲分析。本計劃預期可以了解3BN在植物體的訊號傳遞途徑及代謝反應，並找出參與其中非常上游的PRRs及最下游的轉錄因子。PRRs接受外源訊息並傳至下游的轉錄因子，從而調控發育/抗病蟲害基因表達。因此，本計劃有助於了解植物對於抗病及抗蟲之受傷調控機制。<br> Abstract: Plants are sessile organisms and constantly exposed to biotic and abiotic stresses. Membrane-anchored pattern recognition receptors (PRRs) are the key regulators to bridge extracellular environmental cues to intercellular signaling molecules. Plants can recognize pathogen/microbe-associated molecular patterns (PAMPs/MAMPs) from pathogens and recognize damage-associated molecular patterns (DAMPs) from themselves through PRRs. Glucosinolate-derived DAMPs such as 3-butenenitrile (3BN) is released upon disruption of plant tissue. However, little is known about the PRRs of DAMPs in plants. Therefore, the proposed project aims to decipher how Arabidopsis response to 3BN and signaling pathways in plants. First, the T-DNA insertion mutants in different signal pathways will be used to study their response after 3BN treatment, the expression level of signal transduction-related genes will be analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), different physiological and biochemical experiments will be performed to analyze how plant response to 3BN. Then, metabolomics study will be used to analyze the metabolic process of the metabolites in plants. In the second year, plant materials treated with 3BN at different time points (0, 1, 3, 6, 12, 24 hours) will be collected for RNA-seq analysis to identify the PRRs and transcription factors (TFs) involved in the 3BN signaling pathway. Candidate PRRs and TFs will be cloned and transformed into Arabidopsis by Agrobacterium tumefaciens GV3101. In the third year, we will conduct a series of disease and insect resistance assays for the PRRs and TFs Arabidopsis transgenic plants. This project is expected to decipher the metabolic process upon 3BN treatment, and to identify the upstream PRRs and downstream TFs for understanding the recognition/perception and signaling of the 3BN in Arabidopsis. Therefore, this project will gain an insight of wound regulatory mechanism for pathogen and insect resistance in plants.訊號傳遞PRR受器轉錄因子硫代葡萄糖&#33527水解產物&#33096化合物signal transductionpattern recognition receptorstranscription factorsglucosinolate hydrolysis productsnitrile