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 an...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°C continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×10^4 mL-1, 65.20×10^4 g-1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20°C, 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.展开更多
基金The National Science Foundation Project under contract No.2007FY210500the National Department Public Benefit Research Foundation of China under contract No.200805075+2 种基金the Province Science and Technology in the Guangdong Project under contract Nos 2010B060200010 and 2010B020201015the Science Expenditure in the Hainan Project under contract No.11-20410-0015the National Natural Science Foundation of China under contract Nos 41206106 and 41222038
文摘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°C continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×10^4 mL-1, 65.20×10^4 g-1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20°C, 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.
