Callus formation from fusion product culture between cotyledon protoplast of Amaranthus tricolor and mesophyll protoplast of Solanum tuberosum.
1988
Chung J.D. | Kim G.H.
Factors affecting isolation, fusion and culture of protoplasts originated from cotyledon of Amaranthus tricolor and from mesophyll of Solanum tuberosum were tested for more viable protoplast yield, high fusion frequency and more preferred cell division and cell colony formation. The highest healthy protoplasts were obtained when the sliced cotyledon of 12 days old which was preplasmolysed in 0.3M mannitol solution with CPW inorganic salts for 1 hour under dark was digested in 2.0% cellulase Onozuka R-10, 0.3% driselase, 0.3% macerozyme, 0.3M mannitol and CPW inorganic salts containing solution. Frequency of cell division and colony formation from protoplast cultures of Amaranthus tricolor were increased when protoplasts in 1x10** (5)/ml density were cultured in 8p-KM medium containing 0.2M glucose, 0.1M mannitol, 0.2mg/l 2,4-D, 1.0mg/l NAA and 0.5mg/l BA. On the other hand, cell division and cell colony formation from culture of protoplast in 5x10** (4)/ml density of Solanum tuberosum were stimulated in 8p-KM medium containing 0.2M sucrose, 0.025M mannitol, sorbitol, xylitol and inositol, 0.2mg/l 2,4-D, 1.0mg/l NAA and 0.5mg/l BA. Fusion frequency between two species was highly increased in 30% PEG(1540) solution at 25deg C for 10 minutes, followed by elution with 100mM CaCl2 2H2O containing solution with a pH adjusted to 10.5.
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