Absract-To estimate the efficiency of proline dehydrogenase gene suppression aimed at increasing sun flower (Helianthus annuus L.) tolerance level to water deficiency and salinity, we employed the LBA4404 strain harboring pBi2E with double stranded RNA suppressor, produced based on the ProDH1 gene of Arabidop sis. Techniques for Agrobacterium mediated transformation in vitro and in planta during fertilization of sun flower have been proposed. The genotype dependent integration of T DNA in the sunflower genome was demonstrated. PCR analysis showed that ProDH1 is present in the genomes of inbred lines transformed in planta, as well as in T1 and T2 generations. Significantly, increased levels of accumulation of free L proline during early stages of in vitro cultivation under normal conditions were demonstrated in transgenic regenerants. The application of lethal doses of stressors (0.4 M mannitol and 2.0% sea salts) caused increase of L proline level in transgenic regenerants and its decline during the recovery period. This data indicates the effectiveness of suppression of the sunflower ProDH1 gene for increased osmotolerance.Keywords: sunflower (Helianthus annuus L.), Agrobacterium mediated transformation in vitro and in planta, dsRNA suppressor of proline dehydrogenase
Aim. To optimize the agrobacterium-mediated method of winter wheat transformation (Triticum aestivum L.); to select the conditions and period of inoculation to effectively transfer the genes during pollination. Methods. Agrobacterium-mediated in planta genetic transformation of winter wheat (Triticum aestivum L.) during pollination. Results. The conditions for agrobacterium-mediated transformation method of winter wheat during natural (frequency pollination was 1 %) and non-natural (frequency pollination was 4 %) pollination were defined. Conclusions. The possibility of integrating transgenes into the genome of winter wheat plants by the method of Agrobacterium-mediated transformation in planta in the process of forced and natural pollination is demonstrated. It is found that the transformation efficiency to a large extent depends on the plant genotype and the method of carrying out the transformation procedure. The selection of transgenic plants under water deficit conditions allowed to identify the plants with functional transgene. The signs of functioning transgene have been remaining in the next generation of genetically modified winter wheat.
Keywords: Triticum aestivum L., Agrobacterium-mediated transformation in planta, transgenic plants, seeds.
Aim. The evaluation of the productivity components of wheat biotech plants with the additional copy of the ornithine-δ-aminotransferase (oat) gene under water stress conditions. Methods. Field and laboratory approaches for studying the parameters of the crop structure. Results. A comparative analysis of productivity components of control plants and forms, obtained via Agrobacterium-mediated transformation were made. During plants cultivation under water deficit the genotype differences among variants were detected. Conclusions. It was showed that under osmotic stress pressure biotech plants (T2) of wheat winter genotypes Favoritka, Dostatok, Volodarka demonstrated better indices of crop structure compared with control plants. The differences between T2 Zolotocolosa and control plants were not essential. Biotechnological plants with the additional copy of the oat gene under poor water supply conditions were higher, had better developed root systems and formed the increased number of additional (lateral) shoots.
Keywords: Triticum aestivum L., biotechnological plants, ornithine-δ-aminotransferase gene, productivity components.
Aim. To investigate inheritance of transgenes in the first generation (T1) of winter wheat biotechnological plants (Triticum aestivum L.). Analyze the performance of T1 genetically modified plants with a double stranded RNA suppressor of the proline dehydrogenase (pdh) gene under normal growing conditions. Methods. PCR analysis, DNA electrophoresis; determination of indicators of the structure of the crop. Results. Molecular genetic analysis was performed and the performance indicators of control and T1 biotechnological plants were investigated. Conclusions. The first generation of genetically modified winter wheat plants resulting from Agrobacterium-mediated transformation in planta confirmed the inheritance of integrated genes. Among the transgenic variants identified plants that lack some fragments of the target gene required for partial suppression of the gene of proline dehydrogenase wheat. It is shown that at the optimal terms of growing biotechnological plants of wheat winter-annual of UK 322/17 and UK 209 h was characterized by the best indexes of structure of harvest as compared to an initial form, while the genetically changed plants of genotypes of UK 95/17 and UK 065 after the elements of the productivity did not differ from control plants.
Keywords: Triticum aestivum L., biotechnological plants, T-DNA, proline dehydrogenase gene, structural analysis indicators.
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