2020-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/661172摘要：在動物&#32454;胞中，麩氨酸受體ionotropic glutamate receptor (iGluR)在神經訊號傳導 中扮演著重要作用。在阿拉伯芥中(Arabidopsis thaliana) 被發現至少20 個麩氨酸 受體glutamate receptor-like(GLR)的基因。這些基因大多&#25968;在根部有表現，然而 ，生理功能卻鮮少知道。早先研究表示，阿拉伯芥第三群(clade III) 麩氨酸受體 GLR3.2/GLR3.4 及GLR3.6影響根的生長發育。本研究中我們將使用阿拉伯芥為模 型系&#32479;。我們將建立麩氨酸受體GLR3.6突變株和過分表達轉殖株，並觀察其根部 的外表型。我們將尋找麩氨酸受體GLR3.6之磷酸化位點，之後，我們將建立麩氨 酸受體GLR3.6磷酸化位點點突變之過分表達轉殖株，並觀察其根部的外表型。我 們將使用鈣離子呈像工具和通道阻礙劑，調查GLR3.6磷酸化位點點突變過分表達 轉殖株中鈣離子之訊號傳導。為探究植物麩氨酸受體磷酸化如何透過生長素影響根 的生長發育，以DR5 為啟動子之報導基因β-glucuronidase (GUS)將被引進使用 ，以監測在麩氨酸受體GLR3.6磷酸化位點點突變之過分表達轉殖株的植物生長素 分布。此外，我們將分析麩氨酸受體GLR3.6下游受調控之基因(如與細胞周期有關 之KRP4)及蛋白質。我們最終希望闡明麩氨酸受體3.6磷酸化其調控根生長發育之 分子機制, 並應用至經濟作物(如水稻)以改進產率。 <br> Abstract: In animal cells, glutamate receptors play important role in the neutral signal transduction. At least 20 type of ionotropic glutamate receptor (iGluR)-like channels have been identified in Arabidopsis thaliana. Most of them express in root tissues, however, the functions are little known. Previous studies showed that clade III GLR (GLR3.2/GLR3.4) and GLR3.6 are involved in root growth in Arabidopsis. Arabidopsis thaliana will be used as our model system. The T-DNA insertion mutants and overexpression lines of GLR3.6 will be isolated and generated. These will be subjected to phenotyping for root development. We will try to identify the phosphorylation sites of GLR3.6. Overexpression lines of GLR3.6 phosphorylation site point-mutated lines will be generated followed by phenotyping for root development. The calcium signaling will be investigated using calcium imaging tools and channel blockers. To determine how auxin involves in the GLR-mediated regulation of root development, DR5 promoter&#65534;driven β-glucuronidase (GUS) reporter will be utilized to monitor the auxin amount in the mutant lines. Downstream target genes (i.e. cell cycle gene KRP4) and proteins will be identified. In particular, calcium-binding protein will be our top focus. Once the target genes and proteins are identified, we will further try to elucidate the role of GLR3.6 phosphorylation in regulating root development. In the future, we hope to apply to crops to improve crop (i.e. rice) yield.麩氨酸受體生長素根鈣離子磷酸化Glutamate receptorauxinrootcalciumphosphorylation.