黄瓜‘新津春四号’遗传转化体系的建立

Establishment of Genetic Transformation System of Cucumber ‘New Jinchun 4’

摘要:

对根癌农杆菌介导‘新津春四号’黄瓜(Cucumbis sativus L.)遗传转化的影响因素进行了研究。结果表明，黄瓜叶片适宜的潮霉素(Hygromycin, Hyg)筛选浓度为40 mg L-1；500 mg L-1羧苄青霉素(Carbenicillin, Carb)可有效抑菌。预培养2 d有利于转化；共培养2 d有利于提高转化频率并避免农杆菌的过度生长；添加150 μmol/L乙酰丁香酮(Acetosyringone, AS),农杆菌浸染液pH 5.7、浓度0.8，浸染时间8 min为最佳遗传转化条件。再生植株经gus基因的瞬时表达检测及PCR检测证明hpt基因已成功转化。

Abstract:

The genetic transformation system of cucumber ‘New Jinchun 4’ mediated by Agrobacterium tumefaciens were studied. The cucumber leaves as explants were pre-cultured on MS + 1.0 mg L-1 6-BA + 0.15 mg L-1 IAA for 2 days. After infected by Agrobacterium (OD600=0.8, pH=5.7) for 8 min supplemented with 150 μmol/L acetosyringone (AS), the explants were co-cultured on MS + 1.0 mg L-1 6-BA + 0.15 mg L-1 IAA + 500 mg L-1 Carb + 40 mg L-1 Hyg for 2 days. The hpt gene were transformed successfully to regenerated plantlets by gus instantaneous expression and PCR test.