【摘要】 由于超声波处理可在外植体上产生许多微小伤口 ,从而提高农杆菌与外植体之间的接触面积 ,提高农杆菌对外植体的转化频率 .用野生型发根农杆菌 (Agrobacterium rhizogenes) A4 菌株感染黄瓜 (Cucumis sativus L.)无菌苗的子叶和下胚轴 ,建立了黄瓜的发根遗传转化体系 .同时 ,对超声波处理时间、外植体放置方向、预培养时间、外植体类型、乙酰丁香酮的添加与否等影响因素进行了研究 .结果表明 ,采用外植体在添加 0 .5 m g/ L NAA和 2 mg/ L 6 - BA的 MS培养基上预培养 4 d,超声波处理 2 s,共培养 3d,在预培养共培养期间子叶上表皮向下可以取得最佳的转化效果 ,发根诱导率为 94 .7% .乙酰丁香酮对发根诱导频率影响不明显 .冠瘿碱检测表明 ,发根确实能合成农杆碱更多还原

【Abstract】 Sonication assisted Agrobacterium mediated transformation(SAAT) tremendously improves the efficiency of Agrobacterium infection by introducing large numbers of microwounds into the target plant tissue. Cotyledon and hypocotyl segments of cucumber(Cucumis sativus L.)were transformed by Agrobacterium rhizogenes A 4. The effects of the cucumber explants transformed, duration of sonication treatment, length of explants preculture, addition of acetosyringone(AS) and the orientation of the explants during the co cultivation were studied. The highest frequency of hairy root induced was obtained when explants were pre cultured for 4 days and treated with sonication for 2 s, followed with the abaxial side of the explants in contact with the culture medium for 3 days. No significant differences were observed with acetosyringone added. Transformation was confirmed by agropine detection.更多还原