甘蓝型油菜下胚轴离体培养再生植株研究

目的]为进一步通过基因转化获得预期优良性状的甘蓝型油菜提供借鉴。方法]以甘蓝型油菜的下胚轴为外植体,研究不同苗龄、不同基因型、不同预培养条件以及各种生长调节剂组合对油菜外植体高频率再生的影响。结果]试验中,7 d苗龄的甘蓝型油菜下胚轴再生频率可达35.83%;经3 d预培养处理的油菜下胚轴芽再生频率可提高至57.78%,油菜品种史力丰下胚轴的芽分化率较高,达35.00%,其次是N481-1,而N370-1的芽分化率最低;其最高分化率的激素组合为6-BA 3.0 mg/L+NAA 0.05 mg/L+TDZ 1.0 mg/L,达38.89%。史力丰品种6~8 d无菌种子实生苗的下胚轴在预培养基MS+2,4-D 1.0 mg/L+蔗糖30 g/L+琼脂4.5 g/L上预培养3 d,在最适分化培养基MS+6-BA 3.0 mg/L+NAA 0.05 mg/L+TDZ 1.0 mg/L+AgNO35.0 mg/L+蔗糖30 g/L+琼脂4.5 g/L中培养,再生频率最高。结论]建立了甘蓝型油菜下胚轴离体培养高频率再生植株模式。

Research on the Plant Regeneration from Hypocotyl of Brassica napus L

Objective] The breeding method for the elite character of Brassica napus L.based on the plant gene transformation was researched in the experiment.Method] In the experiment,the hypocotyl of Brassica napus L.was taken as explants.And the effect of the different seedling ages,different genotypes,different culture conditions and various growth regulator combinations on the rape regeneration with high-frequency was analyzed.Results] The regeneration frequency of plantlet from the hypocotyl of Brassica napus L.at 7 day-age reached to 35.83% and the regeneration frequency of plantlet from the hypocotyl of Brassica napus L.pre-cultured for 3 days was as high as 57.78%.The ability of rape variety-Shilifeng in bud differentiation from of hypocotyl was strong(35.00%);N481-1,the second and N370-1,lowest.The highest frequency(38.89%) of plantlet regeneration was from the hormone combination: 6-BA 3.0 mg/L+NAA 0.05 mg/L+TDZ 1.0 mg/L.the highest frequency of plant regeneration of variety-Shilifeng could be obtained after the hypocotyl of its seedling at 6-8 day-age was pre-cultured on the medium MS+2,4-D 1.0 mg/L+30 g/L sugar+4.5 g/L agar for 3 days and then transferred on the differentiation medium with MS+6-BA 3.0 mg/L+NAA 0.05 mg/L+TDZ 1.0 mg/L+AgNO3 5.0 mg/L+30 g/L sugar+4.5 g/L agar. Conclusion] The model of high-frequency plantlet regeneration from the hypocotyl of Brassica napus L.was established.