利用農桿菌將不轉式與反轉式之顫楊cinnamate 4-hydroxylase(C4H)基因,轉殖到經過傳統選育之優良赤桉品系,並已獲得轉殖之細胞團、癒合組織、芽體及苗木。基因轉殖已經利用聚合酶連鎖反應(polymerase chain reaction,PCR)檢測確認。轉殖C4H基因之芽體經比較其芽體增殖及抽長速率,皆顯著優於轉殖β-glucouronidase(GUS)報告基因之芽體或非轉殖之芽體。目前已經生產出轉殖不轉式與反轉式C4H基因之苗木各2個轉殖系。自扦插繁殖之轉基因樹苗採取葉片萃取總量DNA,進行南方式雜交分析,結果進一步確認本研究轉殖之外源基因已經嵌入赤桉之染色體基因組。依據不同轉殖系,利用扦插技術加以複製繁殖出上百株的扦插苗,並用以進行造林試驗以及未來生長性狀的測定。
The gene encoding cinnamate 4-hydroxylase (C4H) from Populus tremuloides (quaking aspen) was transferred in a sense or antisense orientation into a superior clone of Eucalyptus camaldulensis via Agrobacterium tumefaciens. Transformed cells, calli, shoots, and plants were obtained, and transformation was verified by polymerase chain reaction (PCR). Transgenic shoots carrying the C4H gene proliferated and elongated at a much greater rate than both transgenic control shoots harboring the β-glucouronidase (GUS) gene and untransformed control shoots. Two independent lines of each of the transgenic plants carrying sense or antisense C4H were produced. Genomic DNA analyses confirmed that the aspen C4H gene was successfully transferred and integrated into the chromosomal genome of these E. camaldulensis transgenic plants. More than 100 rooted cuttings derived from these transgenic plants were generated and have been out-planted for further characterization.
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