文心蘭SPLs (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE) 參與高室溫誘導開花之調控機制

SPLs (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE) are involved in the regulatory mechanism of high ambient temperature-induced flowering in Oncidesa.

杜建勳(Jian-Syun Du)

指導教授 ： 葉開溫

國立臺灣大學/生命科學院/植物科學研究所/碩士(2015年)

https://doi.org/10.6342/NTU.2015.00223

摘要

文心蘭開花機制至今仍不甚明瞭。前人研究發現三個文心蘭SPL (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE) 基因參與在營養期及生殖期間的轉換，也參與在高環境溫度 (即高室溫) 誘導開花之中。該基因可能扮演調控者之角色，其基因表現與下游開花基因FLOWERING LOCUS T (FT) 及APETALA1 (AP1) 具正相關性。為了探究文心蘭OgSPL與開花基因間之調控關係，我們進一步釣取文心蘭OgFT、OgLEAFY (OgLFY)、OgAP1以及OgSPLs等基因之啟動子進行研究。結果顯示文心蘭SPL蛋白於細胞核中表現，並可調控OgFT及OgAP1啟動子，但無法調控OgLFY啟動子。此外，文心蘭OgFT蛋白於細胞核以及細胞質中表現，並可與阿拉伯芥AtFD蛋白交互作用於細胞核中，其蛋白複合體可進一步調控三個OgSPL啟動子。進一步將釣取之相關基因啟動子並接上GUS報導基因，進而轉殖到阿拉伯芥中。從GUS組織染色結果中，OgSPLs與開花基因的啟動子具有部分相同之表現位置，顯示OgSPL與相關開花基因之間存在調控之可能性。而經由GUS活性測定分析，說明文心蘭OgAP1啟動子可以受高環境溫度所誘導。總結上述結果，OgSPLs在高環境溫度誘導文心蘭開花路徑中為重要之調控者，並影響開花基因之表現。

關鍵字

開花 ； 高環境溫度 ； 文心蘭 ； SPLs基因 ； FT基因

並列摘要

The mechanism of flowering and phase transition from vegetative to reproductive stage is still not well-known in Oncidesa. Our previous study has shown that three SPL (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE) genes in Oncidesa (OgSPLs) are identified as key regulators in the phase transition of reproductive process and high ambient temperature-induced flowering, which relate to induction of the down-stream floral genes, such as FLOWERING LOCUS T (FT), APETALA1 (AP1). To comprehend the regulatory mechanism between OgSPLs and the floral genes, we cloned OgFT, OgLEAFY (OgLFY), OgAP1, and OgSPLs promoter. Our results showed that OgSPLs localizes in nucleus and could regulate OgFT, OgAP1 promoter but not OgLFY promoter. Additionally, OgFT localizes in nucleus and cytoplasm which could interact with AtFD and further regulate OgSPLs promoter. Moreover, we generated Arabidopsis transgenic plants which has GUS reporter gene driven by the target genes’ promoter of Oncidesa. GUS staining data revealed the overlapped expression of OgSPLs and floral genes, indicating the possibility of regulation network between OgSPLs and floral regulation genes. GUS activity analysis showed that OgAP1 promoter could be induced by high ambient temperature. Our results suggested that OgSPLs are the master-regulator to coordinate the floral genes on high ambient temperature-induced flowering and phase transition in Oncidesa.

並列關鍵字

flowering ； high ambient temperature ； Oncidesa ； SPLs ； FT

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