Aim. To study the influence of antibiotics Ceftriaxone and Timentin on the morphogenic calli and roots formation, shoots regeneration in two bread wheat cultivars Triticum aestivum L., to study effect of Ceftriaxone on the growth dynamics of plants. Methods. In vitro plant tissue culture, analysis of Variance, correlation and regression analysis. Results. The influence of β-lactam antibiotics Ceftriaxone and Timentin on morphogenetic processes (morphogenic calli formation, shoots regeneration and rhizogenesis), was studied in apical 18-day-old wheat calli. Two wheat genotypes of different growth habits, winter and winter-spring, were used. Conclusions. Timentin and Ceftriaxone stimulate morphogenic calli formation in bread wheat apical calli. As compared to Timentin, Ceftriaxone has strong concentration-dependent and genotype-dependent influence on shoots regeneration. The roots formation depended primarily on the wheat genotype, independently from an antibiotic applied. The presence of Ceftriaxone in culture medium stimulates rooting and growth of regenerating plants as well as their biomass increment.

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Aim. To create a genetic construct carrying the bacterial protein Cas9 gene, the reporter β-glucuronidase gus gene, as well as the marker phosphinotricin-N-acetyltransferase bar gene for plant genome editing. Methods. Molecular-biological, biotechnological, microbiological and bioinformatic methods were used in the study; Golden Gate molecular cloning method was used to create genetic constructs. Results. The genetic construct pSPE2053 which carries the Cas9 endonuclease gene, the gus and bar genes was created; the assembly correctness of all vector elements was confirmed by polymerase chain reaction; the construct was transferred to Escherichia coli and Agrobacterium tumefaciens cells; β-glucuronidase gene expression was verified by histochemical analysis after Nicotiana rustica L transient genetic transformation. Conclusions. The created genetic construct can be used to edit the plant genome for both stable and transient genetic transformation to accumulate recombinant Cas9 protein. The guide RNA sequences may be subsequently transferred into such plants using either stable or transient genetic transformation or traditional crossing methods. Keywords: cloning, genetic construction, gus and bar genes, Cas9 endonuclease protein, transient expression.

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