2016-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/691930摘要：植物受到傷害與昆蟲咬傷後，茉莉酸訊息傳導的調控對於植物啟動防禦反應尤為重要，在近年來獲得 深入的研究。許多研究指出，茉莉酸是植物受傷反應與防禦昆蟲咬傷機制中最重要的植物荷爾蒙。在 初步的研究中，我們發現有一個 AP2/ ERF 基因家族中的轉錄子叫 ORA47，會結合在ORA47 的下 游基因之啟動子上新的順式元件，並且活化眾多茉莉酸生合成基因，參與了茉莉酸的生合成。所以我 們假設，ORA47 在茉莉酸訊息中是個主要的調控者。此外，我們找到 MYC2 與WRKY26 是 ORA47 的直接調控上游轉錄因子。這些成果拓展了阿拉伯芥受傷訊息傳中未知的領域。本計畫中我們將利用 遺傳方法、ChIP、EMSA 與酵母菌雜合技術進行更多的實驗去驗證WRKY26 在ORA47 表達中所扮 演的色。此外本計畫中也擬搜尋與WRKY26 物理交互作用的上游調控蛋白與其直接上游的調控轉錄 子。此外，在ORA47 啟動子中含有快速反應逆境元件(RSRE)。當植物受傷時，此區域可能會被CAMTA3 轉錄因子辨認並啟動ORA47 的表現。本計畫內我們將決定MYC2, WRKY26 與 CAMTA 對ORA47 時間序列與表現程度上的影響。預期最終將揭示在阿拉伯芥的受傷反應裡，介與MAPK 與茉莉酸生 合成訊息傳導路徑中的成員。<br> Abstract: The regulation of jasmonate (JA) signal transduction after wounding or herbivory is particularly important for plants’ ability to initiate defense reactions and received intensive study in recent years. Many studies have demonstrated that JA is the most important hormone that response to mechanical wounding and defense against herbivores. In our preliminary study we discovered that the ORA47 protein, an AP2/ERF transcription factor, binds to the novel cis-element in the promoters of many JA biosynthetic genes. We propose that ORA47 is a master regulator of JA signaling. Furthermore we found that MYC2 and WRKY26 are the direct upstream transcription factors to regulate ORA47 gene. These expand an area has not been known in the Arabidopsis wounding signal transduction pathway. In this project we will carry out experiments to confirm that WRKY26 plays direct role of ORA47 gene expression using genetics, chromatin immunoprecipitation, electrophiretic mobility shift assay, and yeast one and two hybrid methods. In addition, searching for upstream regulatory proteins showing physical interaction with WRKY26 protein and activating the WRKY26 gene expression will be also investigated. In addition, ORA47 contains a Rapid Stress Response Element (RSRE) in the promoter region. It could be the ORA47 lies downstream of the CAMTA3 transcription factor which binds to RSRE under wounding condition. In this project we would like to know the time course and amplitude of ORA47 regulated by MYC2, WRKY26, and CAMTA under wound stress. Consequently, we anticipate to reveal the signal transduction components between MAPK and JA biosynthesis in response to wounding of Arabidopsis thaliana.茉莉酸訊息傳導WRKY26MYC2CAMTA3ORA47wound signal transductionWRKY26MYC2CAMTA3ORA47WRKY26，MYC2 與 CAMTA3轉錄因子直接調控ORA47基因表現並參予阿拉伯芥茉莉酸的生合成