The obtaining of wheat plants with yeast genes of trehalose biosynthesis TPS1 and TPS2
DOI:
https://doi.org/10.15407/dopovidi2020.06.092Keywords:
genetic transformation, TPS2, trehalose, Triticum aestivum, yeast genes of trehalose TPS1Abstract
vPlants of several common wheat varieties (Triticum aestivum L.) were established in in vitro culture, and their ability of plant regeneration was studied. In order to obtain plants with enhanced drought tolerance, varieties Myronivska 67 and Myrkhad were picked up for a further transfer of yeast (Saccharomyces cerevisiae) trehalose biosynthesis genes (TPS1 and TPS2) into their genomes. For this purpose, constructs pBract214-TPS1 and pBract214-TPS2 were created using the Gateway-cloning technique. Both constructs contained TPS1 and TPS2 genes under the control of the constitutive maize ubiquitin promoter (PUbi) and hygromycin-phosphotransferase (hpt) selectable marker gene. Transformation was carried out with the use of two A. tumefaciens strains GV3101 carrying the constructions pBract214-TPS1 and pBract214-TPS2. Wheat immature embryos were used as explants for the transformation. Selection of transgenic lines was carried out on nutrient medium supplemented with 30 mg/L hygromycin (as selective agent). The presence of yeast TPS1 and TPS2 sequences in genomic DNA isolated from transgenic wheat plants was confirmed by the PCR analysis using primers specific to these genes.
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