Sodium hypochlorite sterilization of culture medium in micropropagation of Gerbera hybrida cv. Essandre.

Micropropagation requires controlling contamination that might compromise the success of the process. Thermal sterilization is traditionally used; however, costs deriving from equipment acquisition and maintenance render this technique costly. With the purpose of finding an alternative to thermal sterilization, this research aimed at assessing the efficiency and ideal concentration of sodium hypochlorite for sterilization of culture media and glassware used during rooting of micropropagated Gerbera hybrida cv. Essandre. Two experiments were carried out. In the first one, treatments consisted of control I (no sterilization), control II (thermal sterilization), and total active chlorine concentrations of 0.0005, 0.001, 0.002 and 0.003%. In the second experiment, based on the results observed in the first experiment, treatments consisted of control I (thermal sterilization) and II (chemical sterilization), and total active chlorine concentrations of 0.002, 0.0025 and 0.003%. Plant behavior was assessed based on the length of aerial part and roots, number of roots, and dry biomass of plants. Results showed that the addition of an active chlorine concentration of 0.003% to culture media provided total control of contaminants, and there were no significant differences regarding the variables analyzed between plants obtained with thermal sterilization and with sodium hypochlorite sterilization. Thus, chemical sterilization can be used as a replacement for thermal sterilization of nutrition media for rooting of gerbera in vitro.

Glutathione and abscisic acid supplementation influences somatic embryo maturation and hormone endogenous levels during somatic embryogenesis in Podocarpus lambertii Klotzsch ex Endl.

Here we propose a protocol for embryogenic cultures induction, proliferation and maturation for the Brazilian conifer Podocarpus lambertii, and investigated the effect of abscisic acid (ABA) and glutathione (GSH) supplementation on the maturation phase. ABA, zeatin (Z) and salicylic acid (SA) endogenous levels were quantified. Number of somatic embryos obtained in ABA-supplemented treatment was signifi- cant higher than in ABA-free treatment, showing the relevance of ABA supplementation during somatic embryos maturation. Histological analysis showed the stereotyped sequence of developmental stages in conifer somatic embryos, reaching the late torpedo-staged embryo. GSH supplementation in maturation culture medium improved the somatic embryos number and morphological features. GSH 0 mM and GSH 0.1 mM treatments correlated with a decreased ABA endogenous level during maturation, while GSH 0.5 mM treatment showed constantlevels. Alltreatments resulted in decreased Z endogenous levels, supporting the concept that cytokinins are important during the initial cell division but not for the later stages of embryo development. The lowest SA levels found in GSH 0.5 mM treatment were coincident with early embryonic development, and this treatment resulted in the highest development of somatic embryos. Thus, a correlation between lower SA levels and improved somatic embryo formation can be hypothesized