ASSESSMENT OF THE INTEGRATED CHITINASE GENE STABILITY IN WHEAT LINES AFTER BIOBALLISTIC TRANSFORMATION

Аннотация

The synthesis of PR-proteins (pathogenesis related proteins), the most studied of which are chitinases
and β-1,3-glucanases, occurs in response to infection with pathogens in plants. Information about the exact role of
individual PR proteins within plant immunity makes it possible to use certain specific antifungal proteins for the
development of transgenic plants with increased resistance to fungal diseases. At the same time, it is important not
only to obtain a plant with the desired trait, but also to fix in it the stable expression of the transferred gene and the
inheritance of the acquired trait in generations.
Herein we have studied the stability of the chitinase gene insertion in T1, T2 and T3 generations of transformed
wheat lines obtained by the method of cis-gene transfer. Primary transformed regenerant plants were obtained as a
result of bioballistic transformation of the chitinase gene into immature wheat germ of Saratovskaya 29 and
Kazakhstanskaya 19 varieties. Following screening of regenerant plants by PCR for the presence of the target gene
made it possible to select 6 lines presumably carrying the insert based on variety Saratovskaya 29 and 2 lines based
on variety Kazakhstanskaya 19.
The seed material of the selected regenerant plants was cultivated in soil conditions and the seeds of the
T1 generation were obtained. DNA amplification of 8 selected lines St-29№25, St -29№43, St-29№44, St-29№33,
St-29№26, St-29№35, Кz-19№1, Кz-19№2 with specific primers revealed insert-carrying lines, partially cleaved
lines and lines with a high degree of insert instability. According to the results of the T2 and T3 generations PCR
analysis, a complete absence of insertion in the St-29№35 and St-29№33 lines was revealed, a partial cleavage of the
trait in the St-29№43 and Kz-19№2 lines was revealed, and the stable inheritance of the chitinase gene in four lines
St-29№25, St-29№44, St-29№26 and Kz-19№1.was confirmed. These lines were selected as promising for in-depth
study of their resistance to fungal diseases and further replication.