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題 名 | Multiple Shoot Induction from Seedling Epicotyls and Transgenic Citrus Plant Regeneration Containing the Green Fluorescent Protein Gene=柑橘實生苗上胚軸多叢芽誘導及轉綠色螢光蛋白基因植株之再生 |
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作 者 | 段豔欣; 劉歆; 范淨; 李鼎立; 吳礽超; 郭文武; | 書刊名 | Botanical Studies |
卷 期 | 48:2 2007.04[民96.04] |
頁 次 | 頁165-171 |
分類號 | 435.33 |
關鍵詞 | 生長素; 柑橘; 遺傳轉化; 綠色螢光蛋白; 斜切方式; 芽的形成; Auxin; Citrus; Genetic transformation; Green fluorescent protein; Oblique cut; Shoot formation; |
語 文 | 英文(English) |
中文摘要 | 本研究的目的是對冰糖橙上胚軸再生芽的器官發生條件進行優化並且有效地獲得轉基因柑橘植株。採用2周苗齡的實生苗上胚軸進行器官誘導再生。通過對影響離體器官發生的兩個重要因素,即激素組合和切割方式進行分析,表明IAA在添加 BA2.0mg/l時才促進芽的形成,低於或高於此濃度對芽的形成有抑制作用。在IAA0.2和BA2.0mg/l組合下,再生芽數達到最多,平均每個外植體8.9個。在切割方式中,斜切方式在再生芽數和再生芽品質上都顯示了極好的效果。採用上面的激素組合結合斜切的方式,將綠色螢光蛋白(GFP)基因轉入冰糖橙上胚軸並獲得了12個株系的純合轉化植株。對轉基因植株進行Southern雜交鑒定,表明GFP已經穩定整合到柑橘基因組中。該轉化的成功表明將具有經濟性狀的基因引入該品種是可行的,並且獲得的帶有GFP螢光的轉基因植株可以用於柑橘體細胞融合和有性雜交等基礎研究。 |
英文摘要 | This research aimed to optimize the organogenesis of epicotyl segments and to efficiently obtain transgenic plants of 'Bingtang' sweet orange (Citrus sinensis L. Osb.), an elite citrus cultivar in China. Organogenesis induction was induced in epicotyl segments of 2 weeks old seedlings of this cultivar. Two important factors influencing organogenesis in vitro viz, hormone combination (IAA and BA) and cut modes were characterized. IAA had a positive effect on bud formation only when BA was used at the concentration of 2.0 mg/l, and an inhibitive effect was observed with higher or lower concentration of BA. The number of regenerated buds reached up to 8.9 per explant with the combination of IAA 0.2 mg/l and BA 2.0 mg/l. Among cut modes, oblique cut performed the best for its effect on the number and quality of regenerated shoots and its convenience to manipulate. With the optimized hormone combination and oblique cut mode. citrus transformation with green fluorescent protein (GFP) gene was performed and twelve independently transformed plant lines were achieved. Southern blot hybridization confirmed the stable integration of GFP gene into the citrus genome. The successful transformation of this cultivar revealed that it is possible to introduce other genes with agronomic traits into it. Furthermore, these GFP expressing transgenic plants could serve as a visual marker material for citrus somatic fusion and sexual hybridization. |
本系統中英文摘要資訊取自各篇刊載內容。