Culture media, growth regulators and nitrogen sources in callus formation regulation of Brazilwood (Caesalpinia echinata Lam.)

Abstract

The regulation of callus formation is the first step to indirect somatic embryogenesis in vitro. Therefore, we investigated hormonal and nutrient factors on development of callus in leaf tissues of Caesalpinia echinata (Brazilwood). Explants were cultivated in culture media MS, B5, WPM, and White. The effect of nitrogen sources (NH4NO3, KNO3and glutamine)and the interaction between auxins (2,4-D, IAA and IBA) and cytokinins (BAP and KIN) were also evaluated. Media MS, B5 and White were not significantly different and provided better results. MS medium containing 2.4 g L-1 NH4NO3, 1.35 g L-1glutamine, and 4.11 g L-1 of KNO3 in the presence of 1 mg.L-1 2,4-D and 5 mg.L-1 BAP increased cellular proliferation as determined by fresh weight of callus. The combination of plant growth regulators at 0.5 mg L-1 2,4-D + 5.0 mg L-1 BAP and 0.5 mg L-1 IBA + 5.0 mg L-1 BAP supplemented with 2.4 g L-1 NH4NO3 in MS medium stimulated growth of callus. In general, callus showed friable, non-embryogenic characteristics, with accumulation of phenolics and the presence of meristematic areas (meristems).