Optimization of the method of Agrobacterium-mediated in planta transformation of winter wheat genotypes

DOI: https://doi.org/10.7124/FEEO.v34.1641
Keywords: Triticum aestivum, Agrobacterium-mediated transformation, optimization

Abstract

Aim. Optimization of the Agrobacterium-mediated in planta transformation of winter wheat genotypes. Methods. Genetic transformation, molecular genetic analysis; of mathematical statistics. Results. The influence of the composition of the inoculation medium and the method of application of the agrobacterium cell suspension on the frequency of seed setting and the formation of transgenic plants of new promising genotypes of winter wheat was studied. The dependence of obtaining the amount of grain and transformants on the genotypic characteristics of plants when using different inoculation conditions and transformation procedures was established. The higher efficiency of the MC-32 inoculation medium, which additionally contained MES, magnesium sulfate, and ammonium chloride, was shown compared to the MC-31 medium supplemented with sodium thiosulfate. A significant difference in the rate of seed setting was revealed when using different methods of applying A. tumefacins cell suspension. Method I proved to be the most effective, when a bacterial culture was applied to the pistil receptacle, and after the liquid in which the agrobacteria were resuspended completely dried, pollination was carried out with pollen obtained from an intact ear of the same plant. Conclusions. Selected inoculation conditions for efficient gene transfer in the pollination process for Agrobacterium-mediated in planta transformation of new winter wheat genotypes.

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