芹菜胚性细胞悬浮系原生质体分离及再生植株

利用芹菜胚性细胞悬浮系成功分离得到大量原生质体, 获得芹菜大量原生质体的最佳反应体系为: 酶液组成为3.0%纤维素酶Onozuka R210、1.0%离析酶R210、11%甘露醇、0.5% CaCl₂ ·2H₂O和0.1% MES; 摇床转速为80 r/min, 温度(25 ±2) ℃, 酶解时间5～6 h; 原生质体产量为25.00 ×10⁶ /g, 原生质体活力83.41%。原生质体浅层培养, 培养基为1 /2 MS + 1 mg/L 2, 4-D + 0.5 mg/L KT + 11%甘露醇+ 500 mg/L水解络蛋白, 两天后, 重新再生细胞壁之后进行第1次分裂, 逐步降低渗透压至甘露醇3% ,大约30 d形成小细胞团。小愈伤组织经增殖培养后在1 /2MS + 500 mg/L CH + 0.25 mg/L KT固体分化培养基诱导出不定芽, 30 d后再转入MS基本培养基, 获得完整的再生植株。

Protoplast Isolation and Plant Regeneration from Somatic Embryogenic Cell Suspension of Celery(Apium graveolens L.)

Protop lastswere successfully isolated from embryogenic suspensions incubating in enzyme solution containing 3.0% cellulose Onozuka R210, 1.0% macerozyme R210, 0.5% CaCl₂ ·2H₂O, 0.1%MES and 11% mannitol. The enzyme mixture was shaken (80 r /min) for 6 h at (25 ±2) ℃. The yield andviability were 25.00 ×10⁶ /g and 83.41%. Purified protop lasts were cultured in mediem 1 /2 MS + 1 mg/L2, 4-D + 0.5 mg/L KT + 11% mannitol + 500 mg/L CH initially with shallow liquid layers. Two days later,cell wall was regenerated and simultaneously intiated the first division, through reducing manntol, mini-culliwere observed in 30 days. Shoots regenerated in medium 1 /2MS + 0.25 mg/L KT + 500 mg/L CH.