甘蓝型黄籽油菜高油品系下胚轴再生体系研究

Studies on Hypocotyl Derived Shoot Regeneration System of Yellow-seeded Rapeseed(Brassica napus L.) with High Oil Content

为获得甘蓝型油菜高含油量双低恢复系,育成高含油量双低油菜杂交种,以2个不同甘蓝型黄籽油菜高油双低品系(GY1、GY2)下胚轴为外植体,采用正交设计,研究了预培养基中2,4-D和6-BA质量浓度、预培养时间、分化培养基中NAA和6-BA质量浓度组合以及AgNO3质量浓度5种不同因素对其下胚轴芽苗再生的影响。结果表明,除AgNO3质量浓度外,其他4种因素对2个油菜品系下胚轴芽苗再生均具有显著或极显著影响。2个油菜品系间和不同处理间的芽苗再生频率差异均达显著水平,且油菜品系与处理间存在明显互作效应。4d苗龄,在MS+6-BA 0.5mg/L+2,4-D 0.5mg/L预培养基上预培养3d后,在添加NAA 0.05mg/L+6-BA 2.0mg/L的分化培养基中芽苗再生频率最高,GY1和GY2的下胚轴芽再生频率分别为45.2%和41.7%。初步建立了甘蓝型油菜高油品系的下胚轴再生体系,为高含油量油菜遗传转化奠定了基础。

In the experiment,the hypocotyls of two rapeseed（Brassica napus L.） lines（GY1,GY2） with high oil content and double low quality chosen as research objects,some factors affecting shoot regeneration frequency were investigated using orthogonal design to set up better regeneration system of rapeseeds for establishing foundation of obtaining transgenic restorer and breeding hybrids with high oil content and double low quality through biological engineering.The results showed that 2,4-D and 6-BA concentration in the pre-culture medium and pre-culture days as well as NAA,6-BA concentration combination in differentiation medium were the main factors that affected shoot regeneration of the two lines,but AgNO3 concentration in the study had no significant effect.The differences of shoot regeneration frequency among two lines or different treatments were significant,and line×treatment interaction was also significant.It＇s found that 4 days-old seedlings of two lines cultured in the pre-culture medium containing 6-BA 0.5 mg/L and 2,4-D 0.5 mg/L for 3 d,followed by the differentiation medium NAA 0.05 mg/L ＋ 6-BA 2.0 mg/L,exhibited the highest shoot regeneration rates 45.2% and 41.7%,establishing a stable and efficient regeneration systems which laid a foundation for genetic transformation of the two lines.