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題 名 | 番茄‘花蓮亞蔬五號’以農桿菌法轉殖Bt基因之研究=Studies on the Bt Genes Transformation by Agrobacterium Mediated Transformation for Tomato 'Hualien ADRVC No.5' |
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作 者 | 王啟正; 林立; 林學詩; | 書刊名 | 花蓮區農業改良場研究彙報 |
卷 期 | 32 2014.12[民103.12] |
頁 次 | 頁15-27 |
分類號 | 435.272 |
關鍵詞 | 番茄; Bt基因; 基因轉殖; Tomato; Bt genes; Gene transformation; |
語 文 | 中文(Chinese) |
中文摘要 | 為了要以農桿菌法轉殖 Bt 基因至番茄‘花蓮亞蔬五號’植株,取8-10 天苗齡之子葉做為培植體,感染 含有Cry1AC 及Cry1IC 載體的農桿菌,以O.D.600=0.3-0.4 或O.D.600=0.6-0.8 兩種感染菌液濃度處理, 並進行共培養及再生篩選,結果低濃度菌液感染轉殖處理之再生率(20.4-34.5%)較高濃度菌液感染轉 殖處理(13.5-28.2%)為高。經篩選後僅以含Cry1IC 轉殖再生具有Bt 基因之培植體,低菌液濃度感染 處理的Bt 基因再生植株也較多。經南方雜合檢定顯示3、22 及22-2 號轉殖株之Bt 基因(Cry1IC)有3 個複製數(copy number);而9 及20 號轉殖株則為4 個複製數。此5 株轉殖株經北方雜合檢定皆有Bt 基因表達。餵食斜紋夜蛾幼蟲生物檢定結果顯示3、20 及22 號轉殖株抗蟲力較佳,斜紋夜蛾幼蟲對其葉 片之取食率分別為14.2%、14.6%及19.8%,小於對照組(92.4%),未來可作為抗蟲品種的材料。 |
英文摘要 | To transfer Bt genes into tomato cultivar ‘Hualien AVRDC No. 5’ mediated by Agrobacterium, the 8-10 days old cotyledons of tomato were used as explants and infected with Agrobacterium carring Cry1AC or Cry1IC. The explants were infected with two bacterical densities (O.D.600=0.3-0.4 or O.D.600=0.6-0.8) and then cocutured and regenerated. The regeneration efficiency in the lower bacteria density treatment was higher (20.4-34.5%) than the higher bacteria density treatment (13.5-28.2%), and Bt genes positive plantlets were obtained if the explants infected with lower bacteria density. Transgenic plants 3, 22, and 22-2 contain three Bt (Cry1IC) copies and transgenic plants 9 and 20 with four Bt copies as confirmed by Southern bolt analysis. Bt genes were expressed in five transgenic plants proofed by northern blot analysis. Transgenic plants 3, 20 and 22 had higher resistance for armyworm according to bioassay data. The leaf consumption rates (14.2%, 14.6% and 19.8%) were lower than wild types. Those transgenic plants with insect resistance may be served as the breeding materials in the future. |
本系統中英文摘要資訊取自各篇刊載內容。