A simple and efficient method for isolation of pineapple protoplasts

Protoplasts have showed great value for new germplasms generation. A method was described for the isolation of large numbers of pineapple protoplasts. The procedure utilized 5% mannitol solution as the primary osmoticum and an enzyme mixture of 1.5% macerozyme R-10, 1.5% cellulase R-10 and 5% mannitol solution, pH 5.5, followed by differential and gradient centrifugations. Overall, the method was found to be a simplified and effective alternative to those previously described for pineapple protoplast isolation, obtaining the highest protoplast yield (3.6 × 106/g FW) and the highest protoplast viability (88.9%).