Optimization of Agrobacterium-mediated transformation conditions of prospective genotypes of winter bread wheat by in planta method

  • O. V. Dubrovna Institute of Plant Physiology and Genetics of the National Academy of Sciences of Ukraine
  • L. V. Slivka Institute of Plant Physiology and Genetics of the National Academy of Sciences of Ukraine
DOI: https://doi.org/10.7124/FEEO.v28.1377

Abstract

Aim. Optimization of conditions for genetic transformation of new promising genotypes of winter bread wheat (T. aestivum L.) by in planta method. Methods. Agrobacterium-mediated transformation by in planta method using the strain AGL0 and vector construct pBi2E. 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 winter wheat genotypes was studied. The dependence of the frequency obtaining of transgenic plants from environmental conditions, in particular temperature, has been established. It was found that the temperature regime of 20-22°C provided the largest number (4.8%) 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 (0.7%). Conclusions. The largest number of transformants was obtained using a inoculation medium without sucrose, the optical density of cells of the agrobacterial suspension of 0.6 op.od. and inoculation on the third day after castration of ears.

Keywords: T. aestivum, Agrobacterium-mediated transformation in planta, optimization of conditions.

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