벼 도열병균, pyricularia oryzae의 원형질체 형성

Formation of protoplasts from pyricularia oryzae

The optimum conditions of protoplasts formation from Pyricularia oryzae were investigated with lytic enzymes and osmotic stabilizers. The mycelia were begun to refease the protoplasts in response to the complex enzyme solution after 30-60 minutes and reached to maximum after 2-3hrs. Among the lytic enzymes tested, the mixture solution containing ${\beta}-Glucuronidase$(0.01 ml/ml), Cellulase ONOZUKA-RS(20mg/ml), Driselase (10mg/ml), and Macerozyme R-10 (10mg/ml) resulted in the highest rate of protoplasts releasing of Pyricularia oryzae. The best stabilizer was 0.6M KCl at pH 7.0. Shen the mycelia were digested with enzyme mixture, the stationary culture was better than shaking culture for higher protoplast formation.