Abstract
Genetic engineering has amplified the possibilities of crop breeding and supported sustainability ideals. Agrobacterium tumefaciens is the preferred transformation system, since it produces transgenic plants with more stable transgene expression and inheritance. The agrobacteria and plant tissue must be co-cultivated in conditions that allow gene transfer. This study aimed to evaluate how co-cultivation time and temperature affect the transformation of immature maize embryos of the Hi-II hybrid (model genotype) and the Brazilian BR 451 variety with A. tumefaciens EHA101:pTF102. The pTF102 plasmid carries a uidA reporter gene that enables transient transformation to be quickly verified by GUS histochemical assays. Increasing the co-cultivation period from three to five days at 20°C resulted in a higher number of GUS positive embryos and blue spots per embryo in the BR 451 Brazilian variety, indicating better bacterial T-DNA transfer into the target explant cells. This condition raised the BR 451 response level to match the response level of the Hi-II control genotype, indicating that this Brazilian variety is suitable for genetic transformation.
Highlights
Maize (Zea mays L.) is a highly important crop for food production, occupying 13% of the agricultural area in the world (USDA, 2014) and being extensively used for human and animal nutrition
The blue spots represent the location where the reporter gene was inserted in the embryo and the number of blue spots per embryo represents the intensity of the transient genetic transformation
Differences among maize genotypes in the capacity to be infected have been widely reported in the literature (Huang & Wei, 2005; Frame et al, 2006; Omer et al, 2013; Ombori et al, 2014; Souza et al, 2017), with genotype dependence being the main limitation in maize genetic transformation by this method (Hiei, Ishida, & Komari, 2014)
Summary
Maize (Zea mays L.) is a highly important crop for food production, occupying 13% of the agricultural area in the world (USDA, 2014) and being extensively used for human and animal nutrition. Maize has been a target species for biotechnological innovation, being the first genetically modified cereal released in the world market. In Brazil, 88.4% of maize is genetically modified (ISAAA, 2016), but there is no Brazilian transgenic maize commercially released in the current market. Despite the unmatched commercial success of maize transformation and significant progress made in the molecular techniques for this species, the effective rate of genetic transformation of maize is still insufficient (Yadava et al, 2016). The inefficiency of genetic transformation is attributable to various limitations of the available maize tissue culture and transformation protocols.
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