Kulesh S. Agrobacterium-mediated transformation of bread wheat using a double-stranded RNA suppressor of the proline dehydrogenase gene.

In the dissertation work, the possibility of obtaining the method of Agrobacterium-mediated transformation in vitro and in planta of biotechnological plants of bread wheat using agrobacterial strains AGL0 and LBA4404 bearing the plasmid pBi2E, which in its composition contains a double-stranded RNA suppressor of the gene of prolin dehydrogenase, has been experimentally proved and their morphological, physiological-biochemical and cytogenetic characteristics were studied. Some elements of the technology for the production of transgenic wheat plants by the Agrobacterium-mediated transformation method have been improved. Using Agrobacterium-mediated transformation in vitro and in planta, the bread wheat of the Zimoyarka variety plants with a double-stranded RNA suppressor of the proline dehydrogenase gene, characterized by increased tolerance to water deficiency, were obtained. A comprehensive analysis of the osmotolerance of biotechnological plants of generations T0−T2 in the laboratory and field experiments with pot plants was carried out. The cytogenetic analysis showed a significantly higher level of karyotypic variability in transgenic forms than non-transgenic ones. It was found that the chromosomal variability of transgenic plants was manifested by aneuploidy and structural changes in chromosomes. The features of the meiosis in genetically modified wheat plants containing the double-stranded RNA suppressor of the proline dehydrogenase gene obtained by Agrobacterium-mediated transformation using in vitro and in planta methods were investigated and their differences from non-transgenic forms were charactrerized.