本研究目的探討大型海藻裂片石蓴 (Ulva fasciata Delile) glutathione reductase (UfGR; NCBI number DQ286546.1) 基因轉殖水稻台農67號 (Oryza sativa L. cv. Tainung 67, TNG67) 對於水稻鹽耐受性之影響。藉由農桿菌Ti –vector 轉殖 ubiquitin:: UfGR + 35S::GFP 後獲得 homozygous gene之 T2種子。轉殖株GR活性高於野生型有OE-UfGR-1 及OE-UfGR-3，以四葉齡水耕幼苗進行鹽處理 (150 mM NaCl) 後，存活率高於野生型。150 mM NaCl 處理下，轉殖株之effective 和 Maxium PSII 活性 (Fv/Fm、Fv'/Fm') 與chlorophyll a、chlorophyll b、及carotenoids 含量降低程度低於於野生株， 轉殖株之H2O2含量及malondialdehyde ( MDA，脂質氧化指標) 含量明顯少較於野生型而proline 含量增加則高於野生型。轉殖株GR比活性在150 mM NaCl 處理下較野生型高，轉殖株ascorbate peroxidase (APX) 及 catalase (CAT) 比活性被鹽處理促進之程度也較高，尤其是OE-UfGR-3較為明顯。轉殖株之抗氧化物glutathione (GSH) 含量、GSH/oxidized glutathione (GSSG) 比率、ascorbate (AsA) 含量、及AsA/dehydroascorbate (DHA) 比率高於野生型，但是轉殖株與野生型鹽處理之 Na+ 含量增加、K+ 含量降低、及K+/Na+ 比率降低之程度相同，因此UfGR基因大量表現於水稻可有效提高ascorbate–glutathione cycle效率，降低氧化傷害，增進耐鹽性，但是不調控K+ 及Na+之含量而提高耐鹽性。

This study was to examine whether the Agrobacterium-mediated overexpression of a marine macroalgal glutathione reductase (UfGR; NCBI number DQ286546.1) of Ulva fasciata Delile in Japonica rice (Oryza sativa L. cv. Tainung 67, TNG67) improves the tolerance of rice to salt stress. Using the transformation of ubiquitin:: UfGR + 35S::GFP, the hydroponically grown seedling (4-leaf age) of transgenic line with homozygous gene expression (T2) were used to determine the GR activity and their responses to salt challenge (150 mM NaCl). Transgenic lines, OE-UfGR-1 and OE-UfGR-3 that GR specific activity is higher than wild type showed higher survival ratio after recovery to normal condition without NaCl. Under 150 mM NaCl conditions,the decrease in effective PSII activity (Fv/Fm), maximum PSII activity (Fv'/Fm') and the contents of chlorphyll a, chlorophyll b, and carotenoids in the transgenic lines is lower than those in wild type. Transgenic lines showed a lower H2O2 and malondialdehyde (MDA, lipid peroxidation indiicator) contents following NaCl treatment, but the increae in proline contents is higher in transgenic lines as compared to the wild type after NaCl treatment. The GR specific activity of transgenic lines could be induced under NaCl treatment and the induction is higher in transgenic lines, while the specific activity of ascorbate peroxidase (APX) and catalase (CAT) showed a similar trend. Glutathione (GSH) content, GSH/oxidized glutathione (GSSG) ratio, ascorbate (AsA) content, and AsA/dehydroascorbate (DHA) ratio were highly induced by NaCl treatment, particularly in the transgenic lines. But, the increae in Na+ content as well as the decrease in K+ contents and K+/Na+ ratio by NaCl treatment were same between wild type and transgenic lines. It could be concluded that overexpression of UfGR effectively enhances the tolerance of rice to slat stress via the enhancement of ascorbate–glutathione cycle for the reduction of oxidative damage under salt stress, but not the regulation of K+ and Na+ content.

目錄
論文審定書....................................................................................................................i

致謝 ii

摘要 iii

Abstract iv

目錄 v

圖目錄 vi

表目錄 xi

縮寫字對照 xv

壹、前言 1

貳、材料與方法 4

参、結果 12

肆、討論 16

參考文獻 20

附錄 66

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