Direct shoot regeneration from mature zygotic embryos of Pinus elliottii x P. caribaea hybrids and plantlets production assisted by temporary immersion bioreactors.

FORTES, N.; DUARTE, E.; GONZÁLEZ, A.; SANSBERRO, P.; LUNA, C.

Vitoria-Gasteiz

Congreso; IUFRO Working Party 2.09.02 - Somatic Embryogenesis and Other Vegetative Propagation Technologies.; 2014

A protocol for large-scale propagation of Balfourodendron riedelianum Engl. (Rutaceae) by adventitious bud formation and plant regeneration was established. Vegetative buds were induced from both explants cultured on Murashige and Skoog (MS) semisolid medium (plus sucrose 30 g·L-1) containing different combinations of 6-benzyladenine (BA), thidiazuron (TDZ), and -naphtalenacetic acid (NAA). The cultures were incubated at 272ºC and photoperiod 14 h (116 mol·m-2·s-1 PPFD) for 30 days. Therefore, the regenerative explants were transferred to bioreactors containing 100 mL MS plus gibberellic acid (GA3, 0.2, 0.4 mg∙L-1), BA (0.2, 0.4 mg∙L-1) for elongation. Shoots were rooted in MS semisolid (agar 0.65%) medium with either indole-3- butyric acid (IBA, 0.5-1.5 mg∙L-1) or NAA (0.1-0.5 mg∙L-1).The regeneration frequency (40±10%) and the number of buds formed per responsive explants (8±6.15) was greater with cotyledons cultured in MS plus NAA 0.01 mg∙L-1 and TDZ 0.01 mg∙L-1. All plantlets raised in vitro were phenotypically normal and successfully hardened to ex vitro conditions.