Autoria: MELO, B. P. de; LOURENCO-TESSUTTI, I. T.; MORGANTE, C. V.; SANTOS, N. C.; PINHEIRO, L. B.; LINS, C. B. de J.; SILVA, M. C. M.; MACEDO, L. L. P.; FONTES, E. P. B.; GROSSI-DE-SA, M. F.

Resumo: The first successful attempt to generate genetically modified plants expressing a transgene was preformed via T-DNA-based gene transfer employing Agrobacterium tumefaciens-mediated genetic transformation. Limitations over infectivity and in vitro tissue culture led to the development of other DNA delivery systems, such as the biolistic method. Herein, we developed a new one-step protocol for transgenic soybean recovery by combining the two different transformation methods. This protocol comprises the following steps: agrobacterial preparation, seed sterilization, soybean embryo excision, shoot-cell injury by tungsten-microparticle bombardment, A. tumefaciens-mediated transformation, embryo co-cultivation in vitro, and selection of transgenic plants. This protocol can be completed in approximately 30?40 weeks. The average efficiency of producing transgenic soybean germlines using this protocol was 9.84%, similar to other previously described protocols. However, we introduced a more cost-effective, more straightforward and shorter methodology for transgenic plant recovery, which allows co-cultivation and plant regeneration in a single step, decreasing the chances of contamination and making the manipulation easier. Finally, as a hallmark, our protocol does not generate plant chimeras, in contrast to traditional plant regeneration protocols applied in other Agrobacterium-mediated transformation methods. Therefore, this new approach of plant transformation is applicable for studies of gene function and the production of transgenic cultivars carrying different traits for precision-breeding programs.

Ano de publicação: 2020

Tipo de publicação: Artigo de periódico

Unidade: Embrapa Recursos Genéticos e Biotecnologia

Palavras-chave: Agrobacterium-mediated transformation, Cultura de Tecido, Embryonic axis, Glycine Max, High-efficiency plant transformation, Particle bombardment, Planta geneticamente modificada, Recuperação trangênica de soja, Soja