Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii
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摘要: 以长心卡帕藻为材料,利用海螺酶和鲍酶两种海藻工具酶与纤维素酶成功制备其原生质体。确定了最适酶比例为20%鲍酶与12%纤维素酶,渗透剂为2.0 mol/L葡萄糖,最适酶解条件为30℃黑暗酶解4.0 h,原生质体密度为32.60×104 个/mL,产量为65.20×104 个/g。20℃,光强1500-2000 lx,光周期12 h/d培养,原生质体出现类愈伤组织细胞团和再生植株两种分化途径,酶解后组织块培养出现了再生新枝和类愈伤组织细胞团两种分化途径。Abstract: In this study, protoplasts were successfully isolated from Kappaphycus alvarezii using snail enzymes, abalone enzymes and cellulase. The optimum enzymic ratio was fixed to be 20% of abalone enzyme, 12% of cellulase and the osmotic stabilizer was 2.0 mol/L glucose. The optimum enzymic hydrolysis conditions were found to be dark enzymolysis at 30℃ continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×104 mL-1, 65.20×104 g-1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20℃, light intensity of 1 500-2 000 lx and photoperiod of 12 h/d, two developmental pathways were investigated: (1) callus-like cell mass and regenerated plantlet occurred on protoplast; (2) young shoots and calluslike cell mass occurred in tissue blocks after enzymolysis.
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Key words:
- Kappaphycus alvarezii /
- protoplast /
- regenerated plantlet /
- callus-like /
- young shoots
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