【摘要】 以四季桔上胚轴为外植体,研究农杆菌介导转化过程中的影响因素。结果显示,转化研究所用抑菌剂抗菌素中,替门的汀杀菌效果较好,且对外植体出芽无明显抑制作用。转化芽选择初期,2~3 d的预培养及选择培养初期3~5 d的暗培养有利于卡那霉素抗性芽的发生。PCR扩增检测结果显示,AFT基因已转化至四季桔,转基因植株-4℃处理6 d时,SOD酶与脯氨酸含量均高于对照。该试验建立了四季桔上胚轴遗传转化再生系统,其再生条件为:MT基本培养基附加TDZ0.5~1.0 mg/L,替门汀300 mg/L,卡那霉素100 mg/L;预培养时间为2~3 d,共培养后3~5 d的暗培养有利于转化率的提高。更多还原

【Abstract】 Epicotyls from Citrus mitis seedling were used as explants to investigate the factors which had an influence on differentiation and regeneration of anti-Kan adventitious buds in transformation mediated by Agrobacterium.Among antibiotics,Timentin was the best for differentiation of adventitious buds and growth control of agrobacterium.Growing in darkness for 3~5 days after explants co-culture with agrobacterium was help to anti-Kan buds developing.It was better for anti-Kan adventitious buds developing when explants were cultured in differentiation medium for 2~3 days before infected by agrobacterium.Transgenic plants were obtained by PCR assay,and SOD enzyme activity and content of proline were raised in transgenic plant treated by-4℃ for 4~6 days.The result showed that transformation system was established for epicotyls in Citrus mitis.The transgenic plant could be obtain in medium with TDZ 0.5~1.0 mg/L,Timentin 300 mg/L,Km 100 mg/L.Frequency of transformation would be increased as explants precultured for 2~3 d and dark culture for 3~5 d after co-culture.更多还原