Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii

作者：Zhang Si; Liu Cui; Jin Yuemei; Chi Shan; Tang Xianming; Chen Fuxiao; Fang Xu; Liu Tao^*

来源：Acta Oceanologica Sinica, 2014, 33(10): 114-123.

DOI：10.1007/s13131-014-0546-y

• Kappaphycus alvarezii
• protoplast
• regenerated plantlet
• callus-like
• young shoots

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 30A degrees C continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60x10(4) mL(-1), 65.20x10(4) g(-1) tissue for Kappaphycus alvarezii. Finally, under the temperature of 20A degrees 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 callus-like cell mass occurred in tissue blocks after enzymolysis.

• 出版日期2014-10
• 单位山东大学; 国家海洋环境预报中心; 中国海洋大学