Successive Callus Induction-Protoplast Isolation Process Derived from Leaf of Sweet Violet, Viola odrata

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The most important aim of this research is to take possession of protoplasts having high intrinsic activity. Active protoplasts have an advantage of cell differentiation into roots, stem and leaves. Much more active protoplasts will be isolated from calli than from an ordinary, direct procedure of tissue cells, because the former protoplasts are derived from calli having a vigorous cell division ability but the latter protoplasts result from terminally differentiated tissues such as leaves. Finding a preferable callus induction, the treatment of leaves of sweet violet, Viola odrata, was examined by the addition of the plant growth regulators, dicamba and thidiazuron. The result shows that 1 to 2 ppm of dicamba and 1 ppm of thidiazuron are preferably better condition. The plant cell walls of the calli of V. odrata, then, were decomposed by the cellulase "Onozuka" RS to isolate protoplasts. Recommended concentration of the cellulase for protoplast isolation was found to be about 200 units/mL.
2003-01-31