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題名 | 轉殖順義ACC合成酶基因延緩苦瓜果實後熟之研究=Studies on Delaying Fruit Ripening by Transforming Sense ACC Synthase Gene into Momordica Charantia L. |
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作者 | 林生華; 杜宜殷; Lin, Shung-hua; Do, Yi-yin; |
期刊 | 臺灣園藝 |
出版日期 | 20080600 |
卷期 | 54:2 2008.06[民97.06] |
頁次 | 頁115-125 |
分類號 | 435.266 |
語文 | chi |
關鍵詞 | 基因默化; 花粉電穿孔法; 南方氏雜交分析; 乙烯生合成; Gene silencing; Pollen electroporation; Southern hybridization; Ethylene biosynthesis; |
中文摘要 | 本研究運用苦瓜花粉電穿孔法,將含順義苦瓜ACC合成自(ACCsynthase, ACS) cDNA之質體DNA導入苦瓜基因組中後,經以100μ.mL-1抗生素Kanamycin 篩選轉殖植株,存活的轉殖植棟繼續進行GUS活性組織化學染色分析,進一步抽取呈現藍色正反應的轉殖植株之基因組DNA,以ACS cDNA、GUS及NPT II基因片段等3個探針,進行基因組DNA南方氏雜交分析。花粉電穿孔法雖可取得其轉殖基因之植株,但僅有部份外來轉殖基因整合入苦瓜基因組內。轉殖植株(GS2063)仟插繁殖'結果成熟後,測定其乙烯生成。苦瓜順義ACS基因轉殖株之果實乙烯生成高峰,較非轉殖對照果實還2天出現,且高峰值低16%。轉殖順義ACS基因有可能延緩苦瓜果實後熟。 |
英文摘要 | A plasmid containing sense ACC synthase (ACS) gene was introduced to bitter gourd (Momordica charantia L.)genome by pollen electroporation method. Some putative transgenic seeds germinated and were selected by spraying 100μg.mL-l Kanamycin. Leaves of the survived seedlings were analyzed by histochemical staining of ß-glucuronidase (GUS) activity. Genomic DNA of putative transgenic plants was extracted individually and subjected to Southern ana1ysis using ACS cDNA, GUS gene and NPTII gene as probes. Only partial fragment of transgene was integrated into the genome of bitter gourd via pollen electroporation. Transformant line GS2063 was chosen for further propagation by cutting. Mature fruits of the transgenic bitter gourd were harvested and the rate of ethy1ene production of each fruit was measured daily. Whenkept at 25℃, fruits harvested from sense ACS gene transformants took 2 days longer to reach ethylene production peak and 16% lower peak value than fruits from selfing progeny of untransformed p1ants. Transformation of sense ACC synthase gene may generate a slower ripening bitter gourd. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。