臺灣博碩士論文加值系統

蝴蝶蘭雖原產於熱帶或亞熱帶，比一般植物具有較高的耐熱性，但高溫時間過久，仍會導致休眠，生長遲滯，如再遭遇通風不良，或溼度太高，則容易遭致軟腐病以及其他病害的發生。因此，我們利用PCR篩選扣減式基因庫的技術，選取蝴蝶蘭在高溫逆境下的基因並探討其表現。分析篩選基因的結果，我們找到一些EST，其產物可能是植物面對高溫逆境時所產生的保護蛋白，如熱休克轉錄調節因子 (heat shock transcription factor) 和低分子量的熱休克蛋白 (small heat shock protein) 等。另外我們也找到一些與auxin訊息傳導相關的EST，如AUX1 permease和ChaC-like protein (cation transporter)。而北方墨點分析 (Northern blot) 結果意外發現，這些EST的表現似乎受生理時鐘所調控。由北方墨點分析的結果，啟動子的分析，以及相關的文獻研究，我們推測auxin訊息傳導途徑和蝴蝶蘭高溫逆境的反應可能有關。

Abstract

Phalaenopsis amabilis var. formosa Shimadzu is one of the common tropic and sub-tropic plants that possess some thermotolarance under heat stress. Despite of this capacity, however, the Phalaenopsis would easily be dormant or cease growing, and could be severely infected by pathogen during long-term high temperature. To determine how Phalaenopsis responses to high temperature, we constructed a heat-induced partial cDNA library using PCR-select subtraction. Within the 120 ESTs selected for this study, there are genes encoding the heat shock transcription factor and the small heat shock protein that are involved in known protective mechanisms at higher temperaure. In addition, we obtained some ESTs conding for proteins associated with auxin signal transduction pathway, such as AUX1 permease and Chac-like protein (cation transporter). Interestingly, Northern blot analyses indicated that the transciption levels of those ESTs seem to be regulated by circadian rhythm. Meanwhile, we analyzed the promoter region of these putative auxin-related ESTs to acquirte more information. Our results suggest that the auxin signaling pathway probably interact with heat sock response in Phalaenopsis.

目錄

中文摘要-------------------------------------------------------------------- ii
英文摘要-------------------------------------------------------------------- iii
名詞縮寫-------------------------------------------------------------------- iv
一、前言 ------------------------------------------------------------------- p. 1
二、材料與方法
1. PCR篩選扣減式基因庫的構築 ---------------------------------- p. 8
2. 點墨點法 ------------------------------------------------------------- p. 13
3. 北方式墨點法 ------------------------------------------------------- p. 18
4. 啟動子區域的分析 ------------------------------------------------- p. 23
三、結果 -------------------------------------------------------------------- p. 24
四、討論 -------------------------------------------------------------------- p. 33
參考文獻 -------------------------------------------------------------------- p. 38
表 ----------------------------------------------------------------------------- p. 42
圖 ----------------------------------------------------------------------------- p. 47
附錄 -------------------------------------------------------------------------- p. 57

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