本研究目的為建構大型海藻裂片石蓴 (Ulva fasciata Delile) GR (glutathione reductase) 基因 UfGr 後接GFP基因 (標誌蛋白) 於農桿菌Ti vector 及轉殖水稻台農67號 (Oryza sativa L. cv. Tainung 67, TNG67) 以測試大量表現UfGr是否可以提高水稻耐低溫及高鹽逆境之能力。經 GFP 綠螢光及GR活性篩選，獲得OE-UfGR-1, OE-UfGR-3, and OE-UfGR-16 轉殖株。利用4葉齡幼苗進行逆境篩選，發現OE-UfGR-1耐低溫 (4oC) 及高鹽 (200 mM NaCl) 逆境，OE-UfGR-3 可耐高鹽逆境，OE-UfGR-16 則不耐低溫及高鹽逆境。分析低溫對於 OE-UfGR-1 抗氧化酵素superoxide dismutase (SOD)、GR、ascorbate peroxidase (APX) 及 catalase (CAT) 活性及水溶性抗氧化物含量之影響，發現 OE-UfGR-1 GR活性在低溫仍能維持較高狀態，glutathione (GSH)/oxidized glutathione (GSSG) 比值也保持較高，malondialdehyde (MDA，脂質氧化指標) 含量明顯低於 wild type。推測 OE-UfGR-1表現較高 GR 活性造成 GSSG 被有效地還原為 GSH，提高細胞還原狀態而提高水稻幼苗耐冷性，此一推論待未來實驗證明。研究結果證明大型海藻裂片石蓴UfGR基因可以幫助水稻幼苗抵抗低溫逆境。

The purpose of this study is to construct the gene of glutathione reductase (UfGr) of marine macroalga Ulva fascitat Delile following with GFP gene in Agrobacterium Ti vector and transformed to Oryza sativa L. cv. Tainung 67 (TNG67) to test whether overexpression of UfGr could enhance stress tolerance (low temperature and salt stress) in rice seedlings. The GFP green fluorescence and GR activity showed there are 3 transgenic lines, OE-UfGR-1, OE-UfGR-3, and OE-UfGR-16. Using 4-leaf age seedlings, OE-UfGR-1 is tolerant to low temperature (4oC) and salt stress (200 mM NaCl), OE-UfGR-3 is tolerant to salt stress, but OE-UfGR-16 did not tolerate low temperature and NaCl stress. The physiological analysis of OE-UfGR-1 to low temperature stress showed that OE-UfGR-1 seedlings maintained higher GR activity and glutathione (GSH)/oxidized glutathione (GSSG) ratio while malondialdehyde (MDA as lipid peroxidation marker) contents are low as compared to wild type. The activity of superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) of OE-UfGR-1 seedlings was not affected after low temperature stress.It is expected that OE-UfGR-1 seedlings effectively reduce GSSG to GSH and in turn, results in a higher reducing potential for the enhancement of low temperature tolerance.It is needed to be proven in the future. In conclusion, this study shows that overexpression of UfGr could enhance the tolerance of rice seedlings to low temperature stress.

論文審定書…………………………………………………………………i
誌謝…………………………………………………………………………ii
中文摘要……………………………………………………………………iii
英文摘要……………………………………………………………………iv
目錄…………………………………………………………………………v
圖目錄………………………………………………………………………vi
表目錄………………………………………………………………………ix
壹、前言……………………………………………………………………1
貳、材料與方法……………………………………………………………4
參、結果……………………………………………………………………11
肆、討論……………………………………………………………………13
參考文獻……………………………………………………………………15
附錄…………………………………………………………………………36

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