Study on Protoplast Isolation of Cymbidium faberi

Factors affecting protoplast isolation of Cymbidium faberi, including isolation material, isolation time, and concentrations of cellulase and mannitol, were studied. The results were as follows. 1. The highest yield of protoplasts from pollens was 4.56×10 6/g FW when the protoplasts were isolated for 6.5 hours in an enzymatic solution containing 1.5% cellulase, 0.3% pectinase, and 11% mannitol. But in the same conditions, the highest yield of protoplast from petal was 2.12×10 6/g FW. 2. The highest yield of protoplasts from young leaves was 3.36×10 6/g FW when the protoplasts were isolated for 20 hours in an enzymatic solution containing 1.0% cellulase, 0.3% pectinase, and 13% mannitol. 3. When the protoplasts were isolated from young protocorms for 16 hours in an enzymatic solution containing 1.0% cellulase, 0.3% pectinase, and 11% mannitol, the yield of protoplasts was 1.87×10 5/g FW. In addition, the protoplasts isolated from pollens and young leaves were easy to be purified due to little impurities in the solution, and the protoplasts from petals and protocorms were difficult to be purified because of high contents of fragments of raphides and starch pellets. So pollen and young leaf were better materials for protoplast isolation of Cymbidium faberi.