Histological Analysis of Adventitious Bud Formation in Cultured Douglas Fir Cotyledon

Initiation of adventitious bud formation in vitro from Douglas fir cotyledons required both cytokinin and auxin at concentrations of 5 μM BAP and 5 nM NAA. Histological observations showed that these adventitious buds arose de novo from cells residing in hypodermal layers. Development of adventitious buds in culture was characterized by the sequential appearance of four anatomically distinguishable structures: 1) meristemoid, 2) bud primordium, 3) shoot apex with needle primordia, and 4) adventitious bud. The anatomical structure of tissue culture-produced buds was similar to that of vegetative buds produced on intact plants. Cultured cotyledons capable of producing adventitious buds (bud culture) were compared with bud-callus and callus cultures initiated by 5 μM BAP plus 5 μM NAA and 5μM NAA alone without BAP, respectively. Results showed that, during early stages of the culture period (i.e., prior to the appearance of meristemoid structure), cell division of bud culture was mainly located in hypodermal layers, whereas for the other culture types, bud-callus and callus cultures, cell division occurred randomly in all tissues.