The advance of biotechnology in forestry has favored techniques of in vitro propagation of species that present difficulties in reproduction by sexual seeds. Luehea divaricata Mart. & Zucc, a member of the Malvaceae family, is a species that has suffered in recent decades, a large reduction in the areas occupied by natural populations, because of its use in the manufacture of furniture. Additionally, it presents slow and irregular germination and seed viability is very uneven, characteristics that contribute to a reduced ability to recover natural populations. Considering the potential for the application of techniques of tissue culture in the mass propagation of this species, this study aimed to: develop a methodology to promote an efficient surface disinfestation of seed germination in order to in vitro evaluate the ability of two types of explants of seminal in performing the in vitro establishment; evaluate the nodal segments isolated from of plants grown in vitro germination depending on the addition of BAP and NAA in different concentrations, in the WPM nutrient medium, and, finally, analyze the calli seetings responses of leave fragments to different concentrations of BAP and NAA in WPM. The project was developed at the Laboratory of Tissue Culture at the Center for Biotechnology and Breeding of the Federal University of Santa Maria. The seeds used were collected by Fepagro/Forests and the plants obtained were used as source of explants for the experiments in tissue culture. We tested the effect of different concentrations and immersion times of seeds in mercuric chloride, and also the influence of fungicides Dicarboximida and Benzimidazole in the surface disinfestations. To in vitro establish were evaluated apical and nodal segments isolated from plants after 60 days in vitro culture. The surface disinfestation of L. divaricata seeds with mercuric chloride was efficient, promoting a satisfactory germination and lack of phytotoxicity. Moreover, the use of fungicides, after disinfestation procedure with mercuric chloride, showed toxic effect on plants, failed to control contamination by microorganisms. The L. divaricata in vitro establishment was successful from both the apical and nodal segments, especially those that result in higher leaf development. Concentrations of BAP and NAA tested dont not result in a hormonal balance appropriate to promote optimal responses in vitro multiplication of L. divaricata from the culture of nodes segments and the presence of BAP influence significantly the calli induction, unlike NAA which dont not affect the calli formation on leaf fragments. Further studies should be performed to optimize the in vitro surface desinfestation, multiplication and callus induction capable of producing a larger number of aseptic cultures to continue to other steps in tissue culture. Finally, studies that aim to contribute to the L. divaricata micropropagation should continue, since the species presents barriers to the spread by seeds that can be overcome by tissue culture.
