Crude Fermented Extract Containing Gibberellic Acid Produced by Fusarium moniliforme is an Alternative to Cost Reduction in Biofactories

ABSTRACT Nidularium procerum and Nidularium innocentii (Bromeliaceae) were cultivated in vitro on media supplemented with different sources and levels of GA3 (gibberellic acid). These sources were the commercial powder (analytical degree) and fermented extract obtained by Fusarium moniliforme via solid state fermentation. The in vitro elongation and rooting of these plants were evaluated after 50 days of cultivation. The GA3 present in the fermented extract possess the same effect of purified GA3 (analytical degree) for the increase of the height of aerial part of shoots of N. innocentii, but not for the N. procerum being the GA3 fermented extract in a lesser degree. The GA3 fermented extract influences negatively the rooting in N. innocentii, while GA3 analytical degree practically does not interfere in the rooting. On the other hand, in N. procerum, both the GA3 sources reduce the root number and do not interfere in rooting percentage. GA3 crude fermented extract is an alternative to reduce costs, however, its results can vary depending on the species and parameter evaluated. The fermented extract was stored at temperature during 260 days and its shelf life presented a suitable stability, maintaining 92% of its initial GA3 amount.

Expressão transiente do gene uidA em explantes foliares de Eucalyptus saligna Sm. transformado via Agrobacterium tumefaciens / Transient expression of uidA gene in leaf explants of Eucalyptus saligna Sm. transformed via Agrobacterium tumefaciens

O objetivo desse trabalho foi avaliar a influência do pré-cultivo de explantes foliares e do meio de cultura na ressuspensão de Agrobacterium tumefaciens para infecção dos explantes. Os meios MS/2 (50% da concentração de sais) e MS N/2 (50% da concentração de NH4NO3 e KNO3) + PGR (1,0µM de TDZ (thidiazuron) + 0,1 µM de ANA (ácido naftalenoacético)) foram testados na ressuspensão da bactéria para infecção dos explantes. O pré-cultivo consistiu da manutenção dos explantes em meio de cultura para formação de calos (MS N/2 + PGR) durante um dia, sendo o tratamento sem pré-cultivo consistituído dos explantes após a excissão dos mesmos. Os explantes foram mantidos no escuro a 25 ± 2ºC mediante a utilização de plástico preto. O delineamento usado foi o inteiramente casualisado com 20 explantes. Os experimentos foram repetidos duas vezes. O meio MS/2 promoveu resultados superiores (22,4%) comparado ao meio MS N/2 + PGR (14,5%) para a percentagem de área com expressão do gene uidA. Aos 7 dias de cultivo em meio seletivo, a percentagem de área expressando o gene uidA foi 1,6 no MS/2 e 0% para o MS N/2 + PGR. O pré-cultivo produziu resultados superiores aos encontrados sem pré-cultivo, atingindo 31,4% de expressão transiente e no tratamento sem pré-cultivo 2,1%. Após 7 dias de cultivo em meio seletivo, a percentagem de área de expressão dos explantes do tratamento com pré-cultivo permaneceu 4,8% e 0% para o tratamento sem pré-cultivo. Os resultados indicam que o précultivo e ressuspensão da bactéria em meio MS/2 aumentaram a eficiência da expressão transiente do gene uidA em explantes foliares de E. saligna.

The aim of this research was to evaluate the effect of the pre-culture of leaf explants and the effect of the culture medium for the Agrobacterium tumefaciens resuspension to the explant infection. The media, MS/2 (half strength) and MS N/2 (10.3 mM NH4NO3 and 9.4 mM KNO3) + PGR (1.0 µM TDZ (thidiazuron) and 0.1 µM NAA (1-Naphthaleneacetic acid)) were tested for the bacteria resuspension. The pre-culture consisted of the maintenance of the explants on culture medium for callus formation (MS N/2+PGR) during one day and the treatment without pre-culture consisted of the use of the explants after the excision of the same ones. At the end of the co-culture, the MS/2 promoted results superiors to the MS N/2+PGR, and the area percentage that presented expression of the gene uidA was of 22.4% compared at 14.5%. To the 7 days of culture on a medium with kanamycin, the area percentage expressing the gene uidA was 1.6 in MS/2 and 0% for the MS N/2+PGR. At the end of the co-culture, the pre-culture produced results superiors to the found in the treatment without pre-culture, reaching 31.4% of expression and in the treatment without pre-culture 2.1%. After 7 days of culture on a medium with kanamycin, the area percentage of explant expression of the treatment with pre-culture stayed 4.8% and 0% for the treatment without pre-culture. The results indicate that the pre-culture and the bacteria resuspension in MS/2 increase the efficiency of the transient expression of the gene uidA in leaf explants of E. saligna.