杉木高效再生与基因转化的初步研究

High Efficiency Regeneration and Agrobacterium-mediated GeneTransformation in Cunninghamia lanceolata Hook

用取自60d龄无菌苗上的杉木茎段为外植体,在MS+6 BA0.75mg L+IBA0.25mg L培养基中诱导芽分化,丛生芽的诱导率达到95.8%,每个外植体平均有5.2个芽.切取诱导芽单苗,转至MS+NAA1.0mg L培养基上诱导根再生,外植体出根率为91.4%.在此基础上,将杉木茎段与农杆菌AGL1共培养2d,通过在含200mg L卡那霉素的再生培养基中培养,初步筛选出卡那霉素抗性苗,转化频率为1.1%.

Segments of stem from 60dayold aseptic seedlings of Chinese fir(Cunninghamia lanceolata)produced adventitious shoots on MS medium with 0.75?mg/L 6BA and 0.25?mg/L IBA.There were 5.2 shoots per explants on average and the rate of organogenesis was 95.8%.The shoots were isolated and the single shoot was cultured on root induction medium with 1.0?mg/L NAA.The rooting rate of explants was 91.4%.Segments of stem from 60dayold seedlings of Chinese fir were cocultivated with Agrobacteriums AGL1 for 2 days and cultured on regeneration medium with 200 mg/L kanamycin.3 kanamycinresistant plants have been regenerated.The transforming rate was 1.1%.