Propagação in vitro, viabilidade das sementes e análise fitoquímica dos extratos de calos de Himatanthus sucuuba Wood

Abstract

Studies of the viability and vigor of stored seed, in vitro propagation and induction of friable calli from H. sucuuba foliar and stem explants were the objectives of this work. For the seed evaluation was carried out the preconditioning removing the tegument and endosperm of 50 seeds. The tetrazolium at 0.075% concentration allowed to evaluate 70% of viability in stored seeds and germination resulted in 78% being 47% developed normal seedlings and 31% abnormal seedlings. Both tests showed seed viability after 14 months of storage. The most efficient culture medium for seed growth was MS supplemented with AIA (5.0 mg.L-1) resulting in 80% germination and seedlings with 5.97 cm in height and 4.2 nodal segments. The combination of BAP + AIA (0.1 + 8.0 mg.L-1) stimulated 100% sprouting and 30% rooting of the nodal segments and differed from the other treatments. The micropropagated and acclimated seedlings ex vitro in the substrate plant soil showed growth in height (8.41 cm) and stem diameter (2.2 mm) after 90 days in the nursery. Friable calli from foliar and stem explants those were formed during cultivation in MS medium with 2,4-D + TDZ addition (1.0 + 1.5 mg.L-1) induced 73.33% of friable calli from foliar explants in the dark, while 2,4-D + TDZ (4.0 + 2.0 mg.L-1) induced 76.67% of friable calli from stem explants in the presence of light. Methanolic extracts obtained from friable calli were analyzed by comparative thin layer chromatography (CCDC) and by nuclear magnetic resonance of hydrogen (1H NMR). The analysis allowed to observe the presence of phenolic substances (by intensifying the spots when the plate was revealed with NP-PEG and the presence of signals between 6 and 8 ppm in the 1H NMR spectrum), sugars (by the presence of signals between 3 and 4 ppm in the 1 H NMR spectrum and the anomeric signals in 4.09 ppm (d, J = 8 Hz, sugar with beta configuration) and 5.16 ppm (d, J = 3.5 Hz, sugar with alpha configuration). There are also evidence of terpenoids (purple spots when plaque was developed with anisaldehyde and signals at 5.3 ppm and singlets between 0.6 and 1 ppm present in the 1H-NMR spectrum). Seeds of H. sucuuba can be stored and maintain viability. Regarding micropropagation, studies with growth regulators that optimize in vitro rooting are still necessary. It has been proven that friable calli can be produced by foliar and stem explants and both present secondary metabolite production, although there is larger quantities from friable calli of foliar explants. Keywords: Medicinal species, tetrazolium, phytoregulators, secondary metabolites, Amazon.