Produção de plantas e metabólitos de Cleome dendroides Schult. & Schult. f. (Cleomaceae) utilizando diferentes sistemas de cultivo in vitro

Abstract

Cleome dendroides is an endemic species of the Atlantic Forest of Rio de Janeiro and Espírito Santo states, biome changed by intense human activities that threaten its populations. There are no physiological, biotechnological, phytochemical or pharmacological studies of this specie. Considering the phytochemical profile and the medical potential of the genus, it becomes relevant to establish protocols for the production of plants and metabolites of C. dendroides using different in vitro culture systems. In this work, studies were performed on the in vivo germination of the species evaluating the influence of the substrate, temperature and light. There was no dormancy, and germination was achieved at 20, 25 and 20-30°C, on sand or vermiculite substrates. An efficient methodology of germination under in vitro conditions was defined by comparing the different stages of post-seminal development with seedlings from germination under in vivo conditions. The plantlets obtained from the in vitro germination were used as a source of explants for in vitro propagation system. The morphogenic response was evaluated considering the source and type of explant, types and concentrations of growth regulators and number of subcultures. The methodology was efficient for the production of shoots and maintenance of in vitro plants. The best condition for in vitro propagation was using solidified medium supplemented with BA, regardless of the tested explants. The shoots were efficiently elongated, rooted and acclimatized. It was also established an efficient protocol for root culture and shoots regeneration from these cultures. The effect of source of explant, the types and growth regulator concentrations on the proliferation of roots and shoot regeneration was evaluated. IBA provided higher multiplication rates of the roots, whereas BA was effective in the regeneration of shoots from the newly formed roots. A protocol of callus culture and cell suspension culture was established. The effect of the origin and type of explant, the types and concentrations of growth regulators on callus formation was evaluated. PIC + KIN was the most efficient combination for callus induction from explants derived from in vitro germinated seedlings. Callus cultures were maitained for at least two years. Cell suspension cultures were established with the same combination of growth regulators (PIC + KIN), maintaining a biomass of about five times the initial fresh weight of three successive subcultures. Histochemical and phytochemical analysis revealed the presence of alkaloids in callus and suspension cell cultures. Phytochemical analyses of field plants, acclimatization in vitro plant and culture roots were performed, which indicated the presence of glucosinolate derivatives. The results demonstrated the possibility of producing medicinal important metabolites from C. dendroides by biotechnological methods