Kvasko A. Creation of drought tolerant wheat lines with yeast trehalose biosynthesis genes

The thesis is devoted to the creation of drought tolerant bread wheat lines of Ukrainian varieties by Agrobacterium-mediated transformation methods in vitro and in planta using genetic vectors containing yeast trehalose biosynthesis genes TPS1 and TPS2. Genetic vectors containing yeast trehalose biosynthesis genes TPS1 and TPS2: pGWB2-TPS1, pGWB2-TPS2 and pBract214-TPS1, pBract214-TPS2 were created using the Gateway-cloning method. Agrobacterium-mediated transformation of tobacco, as a model plant, with vectors pGWB2-TPS1, pGWB2-TPS2 under control of the cauliflower mosaic virus promoter (P35S) was conducted. The vectors pBract214-TPS1 and pBract214-TPS2 with target genes TPS1 and TPS2 under control of a strong constitutive maize ubiquitin promoter (PUbi) were used for genetic transformation of bread wheat. In order to determine the highest morphogenetic potential in vitro culture of seven varieties of Ukrainian bread wheat: Vykhovanka, Zymoyarka, Myronivska 67, Shchedrist, Zhuravka Odesa, Kesaria Poliska and Mirhad were established. Agrobacterium-mediated transformation of these selected wheat varieties was conducted. For the first time, five varieties of Ukrainian bread wheat were transformated using in planta method. Using molecular genetic analysis presence of transgenes in tobacco and wheat plants was confirmed. PCR analysis with specific primers for TPS1 and TPS2 genes resulted in fragments of 640 bp and 758 bp corresponding to positive control (vector constructs with TPS1 and TPS2 genes, respectively). Analysis of the obtained wheat lines showed increased drought tolerance of the plants.