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Obtaining of wheat (Triticum aestivum L.) lines with yeast genes of trehalose biosynthesis
SUMMARY. Yeast (Saccharomyces cerevisiae) trehalose biosynthesis genes (TPS1 and TPS2) were transferred into genomes of several common wheat varieties using two methods of Agrobacterium-mediated transformation (in vitro and in planta) to enhance their drought tolerance. For this purpose, vectors pBract214-TPS1 and pBract214-TPS2 were constructed using Gateway-cloning technique. Both vectors contained TPS1 and TPS2 genes under the control of the constitutive maize ubiquitin promoter (PUbi) and hygromycin-phosphotransferase (hpt) selec-table marker gene. Three-five days callus obtained from wheat immature embryos was used as explants for the transformation in vitro. Selection of transgenic plants was carried out on nutrient medium supplemented with 30 mg/L hygromycin (as selective agent). Seeds of wheat (transgenic generation T1) were obtained after in planta method of transformation. Integration and presence of yeast genes in wheat genomic DNA isolated from transgenic plants were confirmed by PCR analysis using primers specific to TPS1 and TPS2 genes.
Key words: trehalose, yeast genes of trehalose TPS1, TPS2, genetic transformation, Agrobacterium tumefaciens, in vitro, in planta, Triticum aestivum
E-mail: kvasko.anna ukr.net, yemets.alla nas.gov.ua
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|Coded & Designed by Volodymyr Duplij||Modified 24.09.21|