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題名 | 開發青蔥之高效率試管內再生系統=Development of a Highly Efficient in vitro Plant Regeneration System in Welsh Onion |
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作 者 | 葉賢舜; 尤進欽; | 書刊名 | 宜蘭大學生物資源學刊 |
卷期 | 5:2 2009.12[民98.12] |
頁次 | 頁65-75 |
分類號 | 435.221 |
關鍵詞 | 青蔥; 組織培養; 抗生素篩選; 發根; Welsh onion; Tissue culture; Antibiotics selection; Rooting; |
語文 | 中文(Chinese) |
中文摘要 | 組織培養再生系統是農桿菌媒介基因轉殖法的重要操作平台。本試驗以“黑葉”品種青蔥為材料,評估三種組織培養再生流程誘得之癒合組織,經農桿菌感染及低濃度抗生素(10 mg L^(-1) G418)篩選7週後的擬轉殖類芽鞘組織分化能力。結果顯示,以MS培養基含1 mg L^(-1) 2,4-D 及1 mgL^(-1) BA培養莖盤所誘導出的緊寶、球狀型癒合組織可轉綠並分化出類芽鞘組織,為最佳的農桿菌感染目標組織。為提高再生率,在農桿菌感染前切除癒合組織上一半長度的類芽鞘組織,可提高擬轉殖枝梢再生率至約34.3%。感染後,在癒合組織轉綠階段剖切培植體可維持正常增殖;延至形成叢生枝梢團後施以剖切,28.6-47.3%的培植體則會誘導絨狀根發生。枝梢根系形成階段,置於含有50或80 mg L^(-1) G418 和500 mg L^(-1) carbenicillin的發根篩選培養基上皆無法發根。為克服此問題,藉由移除培養基中的G418及減半carbenicillin濃度,發根率最高可達88.9%。本試驗證明青蔥經農桿菌轉殖法,透過組織培養系統可成功再生擬轉殖株。 |
英文摘要 | Tissue culture regeneration is a key platform for Agrobacterium tumefaciens -mediated gene transformation. In this study, the Welsh onion (Allium fistulosum L.) variety ”Hak-ip” was used to evaluate callus proliferation after transformation. After inoculation with A. tumefaciens and a selected low-concentration antibiotic (G418 at 10 mg L^(-1)) for 7 weeks, the coleoptile-like tissue differentiation of the putative transformant plants was studied. The results show that MS medium containing 1 mg L^(-1) 2, 4-D and 1 mg L^(-1) BA induced the stem discs of the Welsh onion to form compact and nodular calli, which turned green and regenerated coleoptile-like tissue, indicating that tissue cultured under these conditions is an excellent material for use in A. tumefaciens transformation. Removal of at least half the length of the coleoptile-like tissue increased the regeneration rate to 34.3% in the putative transformant shoots. After transformation, dissection of the calli until they turned green maintained normal proliferation; while upon later dissection, when a shoot clump had formed, a fluffy rooting formation was induced in 28.6-47.3% of the explants. During root formation, rooting was completely inhibited when the shoots were rooted in medium containing 50 or 80 mg L^(-1) G418 and 500 mg L^(-1) carbenicillin, and was boosted up to 88.9% by elimination of G418 and reduction of the carbenicillin concentration by half in the medium. In conclusion, this study demonstrated that A. tumefaciens-transformed putative Welsh onion plants can be successfully regenerated using a tissue culture regeneration system. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。