【摘要】 以罗汉果子叶为外植体,研究了影响根癌农杆菌介导的罗汉果遗传转化的若干因素,建立了罗汉果遗传转化体系。结果表明,5 d苗龄的子叶、预培养1 d、侵染20 min、共培养4 d、共培养温度22℃、AS100μmol/L、Hy浓度不定芽筛选为10 mg/L,生根筛选为20 mg/L转化率最高。抗性苗经PCR和Southern印迹分析,初步证实NPR1基因已整合至罗汉果基因组中,转化率为1.5%。更多还原

【Abstract】 Using cotyledon of Siraitia grosvenorii as explants,an efficient and reliable transformation system was established.Several factors affecting genetic transformation of Agrobacterium-mediated S.grosvenorii were studied.The results showed that 5 days aged cotyledon using for infection explants,pre-culture 1 d,bacterial infection time 20 min,co-culture 4 d with 22 ℃,AS concentrate 100 μmol/L,10 mg/L of hygromycin was suitable screening concentration for the differentiation of adventitious bud and root could get the better transformation efficiency under 20 mg/L of hygromycin.The transformants were confirmed by PCR and Southern blotting analysis with the transformation rate 1.5%.更多还原