Isolation and culture of protoplasts from embryonic cotyledons of Pinus densiflora S. et Z.
1985
Lee, K.J. (Seoul National Univ., Suwon (Korea R.)) | Lee, J.S. | Youn, Y. | Lee, S.K. (Forest Genetics Inst., Suwon (Korea R.))
Protoplasts were isolated from cotyledons of germinating Pinus densiflora S. et Z. seeds. The seeds were germinated for nine days, and excised embryonic cotyledons about 3-4mm long were incubated with Cellulase Onozuka R-10 and Macerozyme. After 8 hrs of incubation, large number of viable protoplasts were isolated. Isolated protoplasts were cultured in a medium containing basal salts of B5 medium, vitamines, amino acids, organic acid, sugars and growth hormones. The first evidence of protoplast budding was observed after twelve hrs in culture, and it suggested that high potential of the embryonic cotyledons for rapid cell division affected the early budding, rather than effect of culture medium was shown in twelve hrs
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