In vitro propagation of seven Daphne L. species

Cultures of seven Daphne species: Daphne caucasica, D. cneorum, D. giraldii, D. retusa, D. jasminea, D. laureola and D. tangutica were established in vitro on MS/WPM based media. Five of the species responded best on MS-based media (D. tangutica, D. laureola, D. caucasica, D. retusa and D. giraldii), while the remaining two species performed best on WPM-based media (D. cneorum, and D. jasminea). Shoot proliferation was achieved from both apical and nodal explants. Shoots were sub-cultured from stock cultures, cut into nodal explants 3-5 cm long and place vertically on basal media supplemented with different concentrations and combinations of cytokinins and auxins. Individual species displayed different responses to the various cytokinins and auxins. Among species, D. jasminea produced the greatest proliferation rate with an average of 7.84 + 0.6 shoots per explant on WPM supplemented with 2.32 mu M BA + 0.0045 mu M TDZ + 0.054 mu M NAA, while the best multiplication rate for the same species grown on the same media supplemented with a single cytokinin (BA) and no auxin was 2.60 + 1.3 shoots per explant. Following multiplication, new shoots transferred to the elongation trails and then 50-100 mm Shoots used for rooting experiments. Increased rooting efficiencies were observed on in vitro-generated shoots with the two-layer medium or dipping methods over when PGRs were uniformly incorporated into the medium. Maximum rooting frequencies (average) ranged from 59% in D. tangutica to 85% in D. jasminea. Following in vitro rooting, rooted shoots immersed in 0.01% solution of humates and planted into a standard horticultural substrate composed and watered weekly with a solution containing half-strength MS salts.