黑穗醋栗组织培养和遗传转化

Study of tissue culture and genetic transformation on blackcurrant

通过对不同激素质量浓度配比实验,建立黑穗醋栗茎尖高频再生体系。结果表明,MS+6-BA1.0mg·L-1+NAA0.1mg·L-1培养基可高效诱导黑穗醋栗茎尖的分化,1/2MS培养基可快速诱导根的生成,形成再生植株。25mg·L-1卡那霉素可以抑制茎尖的分化,20mg·L-1卡那霉素可以抑制分化小苗的生根。利用农杆菌介导法将osMAPK4基因转化黑穗醋栗,获得了抗卡那霉素的再生植株。提取转基因植株基因组DNA,进行PCR检测,结果表明osMAPK4基因已经被整合到黑穗醋栗基因组中。

High-efficient regeneration system of blackcurrant from shoot meristem was established by tissue culture technique and the different phytohormone combination in this experiment. The result showed that the medium of MS＋6-BA1.0 mg· L^-1＋NAA 0.1 mg· L^-1 could efficiently induce the differentiation of bud of shoot meristem, the medium of 1/2MS quickly induced the regeneration of roots and regenerated plant. Kanamycin at 25 mg· L^-1 in the medium inhibited the regeneration of shoots from the meristem of explants. Kanamycin at 20 mg· L^-1 in the medium showed obvious inhibition to the regeneration of roots from the shoots. The osMAPK4 gene were transformed into blackcurrant by Agrobacterium method and some plants resistant to Kanamycin were obtained. The genomic DNA of transgenic blackcurrant was extracted and analyzed by PCR. The results indicated that the osMAPK4 gene had been transformed into the blackcurrant.