Diferentes métodos de transformação genética de Nicotiana tabacum L. para expressão de proteínas

Abstract

Plants are a promising expression system for the production of heterologous proteins. These are an attractive alternative because they have some advantages such as large-scale production, low growth cost, production of complex proteins and lower risk of contamination with human pathogens. The production of proteins in plants can be transiently or stable, where in the transient form there is no integration of the transgene in the plant genome, while in the stable form there is an integration and the modifications are transferred to the next generations. Thus, the aim of this work was to evaluate the transient and stable transformation of Nicotiana tabacum plants. For the transient transformation, we used the agroinfiltration technique, containing Agrobacterium and the pJL TRBO-G vector, in tobacco leaves. An evaluation was carried out 4,5 and 6 days after the infection and the presence of GFP was not observed. For the transformation, the co-cultivation technique with Agrobacterium was used, containing the vector pH7WGD2::bvl, and in vitro tobacco leaf discs. Plant regeneration was carried out through organogenesis. A PCR analysis revealed a transformation efficiency of 0.55% relative to the total buds and, from the Western Blot, it was confirmed that 2 plants were expressing a BVL lectin. According to the obtained results, it was observed the need to optimize the techniques to improve the efficiency of both transformations. As for transformed plants, further analysis must be performed to elucidate the role of the BVL lectin in plant physiology and tobacco plant defense.