Variation of morphogenetic features of Arabidopsis thaliana (L.) Heynh. leaves after Agrobacterium-mediated transformation

?Lugansk National Agrarian University, This e-mail address is being protected from spambots. You need JavaScript enabled to view it.
?Institute of Cell Biology and Genetic Engineering, NAS of Ukraine
?National University of Bioresources and Nature

Formation of plants leaf is one of the important processes, which is fundamental for the vital functions of a plant and in determination of its productivity. Understanding of the leaf plates genetic control is of great significance for programmed modeling of plants with desired characteristics and will help avoiding undesirable consequences in the creation of transgenic plants.

The aim of our work was obtaining Arabidopsis plants resistant to phosphinothricin, studying the functioning of the transferred genes in the new genetic environment of plant genome and determining the genetic construction on a form of leaf plate. The pICBV19 plasmid and Agrobacterium tumefaciens (GV3101 strain) were used for transformation. T-DNA region of plasmid contained the coding sequence of the gene of resistance to phosphinothricin (bar gene), 35S promoter, the promoter of nopalinsynthase (NOSpro), the terminator of nopalinsynthase (NOSter), the terminator of oktapinsynthase (OCSter), the ?-glucuronidase gene (GUS). Transformation was performed by the modified method of Clough J. S. and Viktorek-Smahur A. The Arabidopsis of Landsberg (N1298) ecotype and a Landsberg erecta (NW20), (NASC, UK) line were used as a plant material. The plants were grown by the standard technique, treated with herbicide BASTA in concentration of 1.25 mg/l at the stage of four rosette leaves; re-treatment was performed 10 days after. The experiments resulted in obtaining three lines of Landsberg ecotype (lines 1, 2, 3) and one line of Landsberg erecta (line 4) resistant to phosphinothricin. The splitting, observed in the T1 generation for expression of introduced gene bar, was 3:1 in one of the Landsberg lines, so these plants contained one copy of the transferred gene. In two other lines the hypothesis of monogenic cleavage was not confirmed, probably indicating the presence of multiple copies of the bar gene in plants. The 3:1 splitting in the Landsberg erecta line confirms the presence of one copy of the gene of resistance to phosphinothricin. Nineteen quantitative traits (leaf length, leaf plate length, petiole length, plate width, the ratio of the plate length to its width, flowering term) in the first, third, the last rosette leaf and stem leaf were considered in order to determine the effect of the insertions of -DNA on the leaf form. Transgenic lines with one copy of the transferred gene did not differ from the control sample in the form of the rosette leaf and stem. At the same time, genetic construction changed the quantitative values of leaf characteristics in two Landsberg lines, which are likely to contain multiple copies of this gene. The "leaf length" trait was significantly increased, by 14.6% (line 2) and 17.0% (line 3) in the stage of the first rosette leaf.