Genetic transformation of promising genotypes of winter bread wheat by in planta method

  • L. V. Slivka Institute of Plant Physiology and Genetics of the National Academy of Sciences of Ukraine
  • O. V. Dubrovna Institute of Plant Physiology and Genetics of the National Academy of Sciences of Ukraine
DOI: https://doi.org/10.7124/FEEO.v28.1384

Abstract

Aim. Optimization of conditions and genetic transformation of new promising genotypes of winter bread wheat (Triticum aestivum L.) by in planta method. Methods. Agrobacterium-mediated transformation by the in planta method using strain AGL0 and vector construct pBi-OAT. Results. The influence of air temperature, optical density of cells of agrobacterial suspension, inoculation day and composition of inoculation medium on the frequency of obtaining transgenic plants of new promising genotypes of winter wheat was studied. The dependence of the frequency of obtaining transgenic plants on environmental conditions, in particular temperature, has been established. It was found that the temperature regime of 20-22 °C provided the largest number (4.4%) of wheat transformants, and when the temperature is reduced to 16-18 °C there is a decrease in the efficiency of T-DNA transfer into the plant genome and the lowest frequency of transformation is observed. Conclusions. The largest number of transformants was obtained using a inoculation medium without sucrose, the optical density of cells of the agrobacterial suspension of 0.4 op.od. and inoculation on the third day after castration of ears.

Keywords: Triticum aestivum L., Agrobacterium-mediated transformation, ornithine-δ-aminotransferase gene.

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