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用霞草胚性悬浮细胞分离原生质体,以含0.2%琼脂糖的KM 8p培养基薄层漂浮培养,原生质体培养密度6×10~3-1×10~4/ml。培养3天再生细胞开始分裂,7天统计分裂频率最高达25.4%,10天形成小细胞团,并加降低渗透压的稀释培养基,每周一次。20—25天形成肉眼可见的小愈伤组织,植板率达3.5%。原生质体衍生的愈伤组织在增殖培养时加入0.3%-0.4%活性炭有利于生长及分化。在含6-BA 3.5 mg/L,IBA 0.8 mg/L的培养基上,再生芽的分化频率可达85%。再生芽在添加NAA 0.5 mg/L,6-BA 0.05 mg/L的1/2 MS生根培养基中2周内形成具根的再生小植株。
The protoplasts were isolated from the embryogenic suspension cells of the aspergillus oryzae and cultured in thin layer with KM 8p medium containing 0.2% agarose. The density of the protoplasts was 6 × 10 ~ 3-1 × 10 ~ 4 / ml. After 3 days of regeneration, the cells began to divide, the frequency of statistical division was up to 25.4% on the 7th day, the small cell mass was formed on the 10th day, and the culture medium was diluted to reduce the osmotic pressure once a week. 20-25 days to form a macroscopic small callus, planting rate of 3.5%. Protoplast-derived callus in the proliferation of culture by adding 0.3% -0.4% activated carbon is conducive to growth and differentiation. On the medium containing 6-BA 3.5 mg / L and IBA 0.8 mg / L, the regeneration buds could reach 85% frequency. The regenerated shoots formed rooted regenerated plantlets in 1/2 MS rooting medium supplemented with NAA 0.5 mg / L and 6-BA 0.05 mg / L for 2 weeks.