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Isolation and Culture of Mesophyll Protoplasts from in vitro Cultured Populus alba × P . glandulosa
박용구,한경환 ( Young Goo Park,Kyung Hwan Han ) 한국산림과학회 1986 한국산림과학회지 Vol.73 No.1
N/A This study was carried out to investigate the optimum conditions for isolation and culture of mesophyll protoplasts from Populus alba × P. glandulosa. The results obtained from the experiments are as follows; 1) The suitable concentration of BAP for shoot multiplication was 0.4 ㎎/ℓ. 2) High yield and viability of isolated protoplasts were obtained by our high enzyme-short time incubation method. 3) Optimum enzyme concentrations for mesophyll protoplast isolation were Cellulase 2%, Macerozyme 0.8%, Hemicellulase 1.2%, Driselase 2%, and Pectolyase Y-23 0.05%. 4) 0.6M mannitol in enzyme solution was the most effective for protoplast isolation and viability. 5) The most adequate pH level of enzyme solution was pH 5.6. 6) The effect of DTT and MES buffer was significant. 7) For protoplast purification, 0.6M sucrose was the most proper concentration. 8) The adding effect of Dextran T40 in floating solution was important. 9) The mesophyll protoplasts isolated through our high enzyme-short time incubation method revealed successful response to culture condition over 3 weeks of culture.