Douglas fir plantlets were produced in tissue culture under defined conditions from cotyledon explants obtained from 2- to 4-week-old seedlings. Tissue pieces were cultured on the surface of a fabric tissue support (100 percent polyester) saturated with liquid nutrient medium; this facilitated periodic changes of the medium to meet the requirements at successive developmental stages without transfer of cultured tissues. Plant growth regulators were needed to stimulate adventitious bud formation. Plantlets were regenerated by rooting excised shoots at 19 degrees C on agarsolidified medium containing sucrose and the auxin naphthalene-2-acetic acid. After root initiation, plant growth regulators were removed; this resulted in stimulation of root elongation and the subsequent development of plantlets, which were then established in soil.
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