Aim. Optimization of conditions for genetic transformation of new perspective winter wheat genotypes. Methods. Agrobacterium-mediated transformation in culture in vitro. Results. The influence of the optical density of cells of the agrobacterial suspension, the concentration of the antibiotic cefotaxime, the duration of coculture on the frequency of obtaining kanamycin-resistant regenerants of new winter wheat genotypes by genetic transformation of callus cultures were investigated using LBA4404 and AGL0 strains. It is shown that depending on the strain the most optimal is the concentration of agrobacteria 0.2-03 OD, duration of coculture for 2-3 days and the use of cefotaxime at a concentration of 250-500 mg/L. Conclusions. The optimal parameters for conducting Agrobacterium-mediated transformation of callus cultures of new perspective winter wheat genotypes were selected. Keywords: Triticum aestivum, Agrobacterium-mediated transformation, callus cultures.

Aim. Optimization of conditions for genetic transformation of new perspective winter wheat genotypes and production of transgenic plants. Methods. Agrobacterium-mediated transformation in vitro culture using callus cultures. Results. The influence of the optical density of cells of agrobacterial suspension, the concentration of the antibiotic cefotaxime, the duration of coculture on the frequency of obtaining kanamycin-resistant regenerants of new winter wheat genotypes by genetic transformation of callus cultures were investigated. By Agrobacterium-mediated transformation of morphogenic calluses of new perspective winter wheat genotypes were obtained plant-regenerants in the genome of which revealed the complete incorporation of a genetic construct containing oat and nptII transgenes. Conclusions. Agrobacterium-mediated transformation of callus cultures of new perspective winter wheat genotypes was optimized, and transgenic plants with the target gene of ornithine-δ-aminotransferase were obtained. Keywords: Triticum aestivum, Agrobacterium-mediated transformation, callus cultures, ornithine-δ-aminotransferase gene.

No abstract

Aim. To carry out physiological and biochemical analysis of genetically modified plants of bread wheat of seed generation T2 with the double-stranded RNA suppressor of the proline dehydrogenase gene. Methods. Biochemical determination of free L-proline content and activity of the enzyme proline dehydrogenase; physiological examination of plant growth in vitro and in vivo. Results. It has been shown that transgenic plants, in contrast to control groups, grow more intensively in a selective medium with mannitol, maintaining a green color. It has been established that under normal conditions and under conditions of water deficit, plants of the T2 have an elevated level of free proline in the leaves, compared with the control genotypes. It was found that transformants are characterized by reduced activity of the enzyme of proline dehydrogenase, which manifests itself when the norm-stress-norm conditions change. Transgenic T2 plants had a higher resistance to aqueous deficiency compared to baseline, which was reflected in the nature of their growth. In conditions of soil moisture shortage, the yield of the most of the transformed lines was significantly higher than non-transformed plants. Conclusions. The obtained results allow us to conclude that the use of the pBi2E vector construct with the double-stranded RNA suppressor of the pdh gene is effective for the production of transgenic bread wheat plants with a high level of resistance to water deficiency. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation, RNA suppressor of the proline dehydrogenase gene, T2 plants, physiological-biochemical analysis.

Aim. To analyze the physiological, biochemical and economic characteristics of genetically modified plants of new promising genotypes of winter bread wheat of seed generation T2 with a double-stranded RNA suppressor of the proline dehydrogenase gene. Methods. Agrobacterium-mediated transformation in vitro; biochemical determination of proline dehydrogenase enzyme activity and free proline content; morphometric indicators and elements of crop structure; mathematical statistics. Results. It is shown that transgenic plants, in contrast to control, grow on a medium with mannitol more intensely, retaining a green color. It was found that both under normal conditions and under conditions of water deficiency, plants of seed generation T2 have an increased level of free Proline in the leaves compared to control genotypes. It was found that transformants are characterized by reduced activity of the enzyme proline dehydrogenase, which is manifested by changes in normal – stress – normal conditions. Transgenic T2 plants had a higher tolerance to water deficiency compared to the original, which was reflected in the nature of their growth. In conditions of soil moisture deficiency, the yield of most transformed lines was higher compared to untransformed plants. Conclusions. The results suggest that the use of a vector construct with a double-stranded RNA suppressor of the ProDH gene is effective for creating transgenic winter bread wheat plants with increased tolerance to water deficiency.