COLONY FORMATION FROM PROTOPLASTS DERIVED FROM DOUGLAS-FIR COTYLEDONS

Proliferation of Douglas fir cells regenerated from protoplasts has been accomplished using cotyledons derived from 2-4 week old seedlings. Yield of protoplasts was significantly increased by subjecting cotyledons, prior to protoplast isolation, to a preconditioning treatment for 8 days in the presence of 15 mu;M N6-benzylaminopurine (BAP) and 500 nM 2-naphthaleneacetic acid (NAA). Protoplasts cultured on a fabric tissue support saturated with a liquid nutrient medium containing a high concentration (10 mM) of glutamine were capable of undergoing rapid cell division leading to colony formation. © 1979.