Development of methods of wheat Agrobacterium-mediated transformation

Abstract

Aims. Agrobacterium – mediated wheat transformation is marked by a number of limitations. Low frequencies of regeneration, genetic transformation, obtain of fertile plants are among them. The optimization of protocols of Agrobacterium – mediated transformation in vitro and in planta can increase the effectiveness of the whole method. Methods. Agrobacterium – mediated transformation of Triticum aestivum L. was established. Varions explants of bread wheat were used. There are apical meristems from 3-day aseptic seedlings, upper, midle, low parts of 5–6 –days seedlings. The selection of transformants was carried out on cultural media with the 50 mg/l of kanamicine sulfate. Results. The procedures of Agrobacterium – mediated transformation in vitro and in planta were improved. Using LBA4404 strain harboring binary vector pBi2E with dsRNA-suppressor of proline dehydrogenase gene transgenic wheat plants were obtained. PCR-analysis showed that exson as well as intron of ProDH of Arabidopsis were introducted into wheat genome. Conlusions. Protocols of wheat Agrobacterium – mediated transformation may be using for wheat genetic improvement

Keywords: Triticum aestivum L., Agrobacterium – mediated transformation in vitro and in planta, dsRNA-suppressor of proline dehydrogenase gene.