266 Direct and indirect regeneration from Pfaffia tuberosa (Spreng.) Hicken calli and its β-ecdysone production FLORES, R.1*; MALDANER, J.2; BRONDANI, D.3; CEZAROTTO, V.4; GIACOMELLI, S.R.4; GARLET, T.M.B.5; NICOLOSO, F.T.6 1Instituto Federal Farroupilha, Campus São Vicente do Sul. RS, Brasil. 2Fundação Estadual de Pesquisa Agropecuária. Santa Maria, RS, Brasil. 3Universidade Federal de Santa Catarina, Campus Blumenau. SC, Brasil. 4Universidade Regional Integrada, Campus Frederico Westphalen. RS, Brasil 5Universidade Federal de Santa Maria (UFSM), Campus Palmeira das Missões. RS, Brasil. 6UFSM, Campus sede. Santa Maria, RS, Brasil. *E-mail:
[email protected] ABSTRACT: This study aimed to establish calli from nodal segments of P. tuberosa and to evaluate the effect of plant growth regulators (PGR) on plant regeneration and production of β-ecdysone from in vitro-grown tissues. Calli were induced from nodal segments on Murashige and Skoog (MS) basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) combined with N6-benzylaminopurine (BAP). The β-ecdysone concentrations [on a dry weight (DW) basis] in in vitro grown and field-grown tissues were measured by high performance liquid chromatography. Direct regenerants (ranging from 4.0 to 75.0%) arose from sprouts present in the explants only in the medium containing 2,4-D. Maximum frequency of indirect shoot regeneration from calli (20.0%) was observed in the medium containing 2,4-D and BAP (1.0 µM at each). The accumulation of β-ecdysone in the calli was associated with the presence of shoots and influenced by the concentrations of 2,4-D.