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標題: | TDR4與PIF1a在果實紅熟過程中調節茄紅素累積之功能性研究 Functional studies of TDR4 and PIF1a involved in the regulation of lycopene accumulation in the ripening of tomato fruit |
作者: | Guan-Han Li 李觀漢 |
指導教授: | 謝旭亮 |
關鍵字: | 茄紅素,藍光,TDR4,PIF1a,PSY1, Lycopene,Blue light,TDR4,PIF1a,PSY1, |
出版年 : | 2018 |
學位: | 碩士 |
摘要: | 番茄紅熟過程受環境與植物賀爾蒙所影響,紅熟過程中由綠轉紅是透過胡蘿蔔素生合成路徑所產生的茄紅素所致,而開啟此路徑之重要酵素為PSY (Phytoene synthase),它受環境與賀爾蒙所調控之轉錄因子所影響,透由PSY的啟動而產生茄紅素累積;在先前研究中,TDR4為調控PSY1的轉錄因子之一且受到乙烯調節;但我們研究中也發現藍光可誘導TDR4的表現,進而影響茄紅素的大量累積,另一轉錄因子PIF1a也可與PSY1的啟動子結合而抑制其表現,來降低茄紅素產生,且PIF1a為參與在光訊息傳遞之中的轉錄因子;為了瞭解這二個基因如何參與在茄紅素生合成路徑,我們透過基因表現與蛋白質交互作用來瞭解。基因表現結果中顯示TDR4可以促進PIF1a的表現,並於PIF1a的啟動子區域發現可供TDR4結合的CArG序列。由Electrophoretic mobility shift assay (EMSA)結果顯示TDR4對該序列有結合專一性,利用重組蛋白質進行pull-down assay與Bimolecular Fluorescence Complementation (BiFC)分析中發現TDR4可與PIF1a結合且位於細胞核中。因此,兩者的交互作用可能會影響茄紅素累積,詳細機制仍有待進一步研究。 The ripening process in tomato fruit is regulated by environmental factors and plant hormones. Tomato fruit turns from green into red color that is due to lycopene accumulation. The lycopene is synthesized from the carotenoid pathway and this pathway is mainly controlled by an enzyme called phytoene synthase (PSY). Environmental factors and plant hormones trigger transcription factors that can induce or reduce PSY gene expression to regulate tomato ripening. In previous studies, the transcription factor TDR4 participated in the regulation of carotenoid biosynthetic pathway. TDR4 triggered by ethylene is well known. But we found that TDR4 could be induced by blue light. However, the mechanisms that integrate light and ethylene signaling are unknown. Another transcription factor regulated by light is PIF1a. PIF1a can reduce PSY1 gene expression and lycopene content. To further understand how these two genes regulate PSY1 expression, resulting in lycopene accumulation and fruit ripening, we perform gene expression and protein-protein interaction studies. The results derived from qPCR data indicated that TDR4 might up-regulate PIF1a gene expression. We found that the CArG-motif bound by TDR4 is also present in the promoter region of PIF1a. The Electrophoretic mobility shift assay (EMSA) confirmed that TDR4 could specifically bind to the PIF1a promoter. Pull-down assays and Bimolecular Fluorescence Complementation (BiFC) studies indicated that TDR4 interacted with PIF1a in the nucleus. Thus, it may be interesting to understand the molecular mechanisms underlying TDR4 and PIF1a interaction in regulating lycopene accumulation in tomato fruit in the near future. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70604 |
DOI: | 10.6342/NTU201802960 |
全文授權: | 有償授權 |
顯示於系所單位: | 植物科學研究所 |
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