激素对油菜子叶再生及转基因瞬间表达的影响

Effects of Phytohormones on Shoot Regeneration and Transient Expression of Transgene in Cotyledon of Rapeseed(Brassica napus L.)

以甘蓝型油菜"湘油15"子叶为外植体,以MS为基本培养基,采用正交试验设计,研究了预培养基2,4-D和6-BA浓度、分化/共培养基NAA和6-BA浓度对子叶芽再生频率和转GUS基因瞬间表达率的影响.结果表明,激素对芽再生频率和转基因瞬间表达率都有重要影响,但激素对芽再生和转基因瞬间表达的影响效应具有不一致性.激素对芽分化的影响程度从大到小依次为再生培养基NAA、预培养基6-BA、预培养基2,4-D、再生培养基6-BA,推导的对于芽再生的最佳激素组合方案为预培养基2,4-D 1.0 mg/L和6-BA1.0 mg/L、再生分化培养基NAA0.2 mg/L和6-BA3.0 mg/L;激素对GUS基因瞬间表达的影响程度从大到小依次为共培养基NAA、共培养基6-BA、预培养基6-BA、预培养基2,4-D,推导的对于瞬间表达的最佳激素组合方案为预培养基2,4-D 0mg/L和6-BA0.2 mg/L,共培养基NAA0.05 mg/L和6-BA3.0 mg/L.推导的对于芽再生频率×GUS基因瞬间表达率的最佳激素组合方案为预培养基2,4-D 1.0 mg/L和6-BA1.0 mg/L,共/分化培养基NAA0.05 mg/L和6-BA3.0 mg/L.

The cotyledons of a Brassica napus variety ＂Xiangyou 15＂ were used as the explants and MS was used as the basic medium in the experiment. The effects of the concentration of 2,4-D, 6-BA in the preculture medium and that of NAA, 6-BA in the differentiation medium and/or co-culture medium on the shoot regeneration frequency and the transient expression frequency of transformed GUS gene were investigated by using the orthogonal design. The results showed that the concentration of the phytohormones influenced the shoot regeneration frequency and the transient expression frequency of transgene greatly, but the effects on the shoot regeneration frequency and the transient expression frequency of transgene had non-consistency. As for shoot regeneration frequency, the putative optimum phytohormone combination was 1.0 mg/L of 2,4-D, 1.0 mg/L of 6-BA in the pre-culture medium and 0. 2 mg/L of NAA. 3.0 mg/L of 6-BA in the differentiation medium. As for transient expression frequency of transgene, the putative optimum phytohormone combination was 0mg/L of 2,4-D, 0. 2 mg/L of 6-BA in the pre-culture medium and 0.05 mg/L of NAA, 3.0 mg/L of 6-BA in the co-culture medium. As for the shoot regeneration frequency×transient expression frequency of transgene, the putative optimum phytohormone combination was 1. 0 mg/L of 2,4-D, 1.0 mg/L of 6-BA in the pre-culture medium and 0. 05 mg/L of NAA, 3.0 mg/L of 6-BA in the co-culture and/or differentiation medium.