籼稻绿芽悬浮细胞原生质体再生成株

采用籼稻Hu-18绿芽为外植体，在N6附加2 mg/L 2,4-D的培养基上诱导愈伤组织。加20 d后转人AA 培养基进行悬浮培养。继代培养45 d左右形成了分裂旺盛的胚性细胞悬浮系。即从绿芽诱导至胚性细胞悬浮系的建立仅用了65 d左右。继代后前6 d，细胞干重几乎每2 d增加1倍，而培养液的渗透压及pH 值迅速下降。取继代后4 d的悬浮细胞游离原生质体，产率为8.74X10⁶/g鲜重。纯化后的原生质体在KPR培养基中进行琼脂糖包埋培养，原生质体植扳率为12.0％ 。将20 d后的小愈伤组织(0.1 mm)转入 N6附加0.5 mg/L 2,4- D、1 mg/ L BA、1 mg/L KT、0.3 mg/L ZT的培养基上增殖，待长成直径2～3 mm 左右时，用N₆附加1 mg/L BA，1 mg/L KT，0.3 mg/L ZT的培养基进行分化培养， 5 d后同时出现芽根生长，最终再生成绿色植株，绿苗分化率为3.5株/10000个原生质体。

Plant Regeneration from Protoplasts Derived from Cell Suspensions of Green Shoots in Indica Rice

The calli induced from green shoots of indica rice Hu-18 on N₆ medium with 2 mg/L 2,4-D were subcultured for about 45 days in AA liquid medium. A finely divided, fast growing cell suspension was established. During the first 6 days after subculture, dry weight of suspension cells doubled every 2 days, while the osmostic pressure and pH value of the culture medium were rapidly reduced. 4 days after subculture suspension cells were used for protoplast isolation. Protoplast yield reached 7. 84×10⁶ protoplasts/g cells. Protoplasts were cultured in KPR medium with 0. 6% agarose. The plating efficiency of protoplasts was 12.0%. After 20 days, colonies (0.1 mm) were transferred onto N6 medium with 0. 5 mg/L 2,4-D, 1mg/L BA, 1 mg/L KT and 0. 3 mg/L ZT The growing calli 2-3 mm in diameter, were translcrred onto N6 medium with 1 mg/L BA, 1 mg/L KT, 0. 3 mg/L ZT and 0. 5 mg/L NAA for redifferentlaiion After 5 days, shoots and roots were observed. The green plantlet frequency was about 3. 5 per 10000 protoplasts.