【摘要】 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。更多还原

【Abstract】 Protoplasts of Gypsophila oldhamiana Miq. were enzymatically isolated from the suspension cultured cells. The protoplasts were cultured in the thin solid KM 8 P medium solidified by 0.2% agarose, which floating on the surface of liquid KM 8 P medium. In the media, the optimum plant growth regulators and their concentrations and combinations were 2, 4-dichloropheno-xyacetic acid (2,4-D) 0.5mg/L, naphthalene acetic acid (NAA) 0.8mg/L and zeatin 0.3mg/L, The optimum density of cultured protoplasts was ranging 6× 103/ml to 1×104/ml. The culture condition was 25× in dark. The first divisions of regenerated cells were observed at 3 days from initial culture and the division frequency reached to 25.4% at 7 days. Ten days after culture, a dilution medium which was half concentration in mannitol was added and the dilution was repeatedat one week interval thereafter. The cultures were transferred to light condition with an intensity about 800-1000 1x. Micro-calli observed with naked eyes were formed after being cultured for 20-25 days. A 3.5% plating efficiency was achieved. When transferred on modified MS differentiation medium supplemented with indol butyric acid (ISA) 0.8mg/L and 6-benzylaminopurine(6-BA) 3.5 mg/L, calli were differentiated adventitious shoots. The differentiation frequency was up to 85%.Regenerated shoots were rooted in a half strength MS medium supplemented withNAA 0.5 mg/L and 6-BA 0.05 mg/L. The plantlets with normal morphology were transplanted into peat/soil mixture and growth vigorously under green house conditions.更多还原