Preparation and regeneration of protoplasts from Nannochloropsis oculata
微拟球藻原生质体制备与再生

The influence of enzyme digestion time and initial algal density on algal cells were considered.Protoplasts of Nannochloropsis oculata,a marine microalga,were prepareded by digestion with the mixture of 4% Hemicellulase and 2% Driselase.The protoplasts were examined after the treatment of Calcofluor White strain.The results shown that the highest preparation efficiency occurred at the initial cells density of 2×107~3×107 cells/mL and a digestion of 1～ 3h digestion.The resulting protoplasts could regenerate on regeneration medium.The growth of protoplasts was consistent with wild algal cells.Considered the influence of regeneration,we selected the optimal treatment time of 1h and initial algal density of 2×107 cells/mL.