Pirko Y. Intron length polymorphism of cytoskeleton protein genes as an effective tool for plant genotyping

The dissertation is devoted to the development and implementation of molecular genetic markers based on the use of intron length polymorphism of genes encoding cytoskeleton proteins (α-, β-, γ-tubulin and actin) for differentiation and genotyping of plants. In particular, the analysis of existing genomic databases of plants for the presence of the gene sequences encoding α-, β-, γ-tubulin and actin was done. The exon-intron structure of the gene sequences encoding α-, β-, γ-tubulin and actin was studied, sequences alignments was performed in order to obtain consensus sequences and to do designe of the primers for conserved regions of genomes surrounding gene introns. Both species-specific and universal (which can be used in the study of many plant species) degenerate primers were selected. Polymorphism of introns of α-, β-, γ-tubulin and actin genes at species, population, varietal and intra-varietal levels in different plant species was evaluated. Based on the results of the polymorphism analysis of the introns of the α-, β-, γ-tubulin and actin genes, genetic profiling of such economically valuable plant species as flax, rice, wheat, barley, tomato and potato was performed. Also, due to the evaluation of the intron length polymorphism of the β-tubulin and actin genes, genetic variability was studied and natural populations of Aegilops biuncialis from the Crimean Peninsula were differentiated. Using the intron length polymorphism analysis of actin and α-tubulin genes, tomato (Solanum lycopersicum) and potato (Solanum tuberosum) varieties were genotyped, and specific DNA profiles of the studied varieties were obtained. Interspecific and intravarietal of introns length polymorphism of β-tubulin and actin genes in different cultivars of flax was evaluated. The high efficiency of using ILP markers based on the assessment of intron length polymorphism of cytoskeletal protein genes in comparison with SSR markers analysis on genera Linum L., Quercus L., Ulmus L. was shown. The convenience and reliability of the TBP analysis for molecular genetic labeling of herbaceous and woody plants, as well as for studying certain aspects of intraspecific polymorphism of economically valuable, horticultural and forest-forming species have been demonstrated. Using the TBP method, unique patterns for 20 woody plant species were identified and molecular profiles of each of these species were created. For the first time, specific TBP profiles of microalgae were obtained, which allowed to clearly differentiate genotypes at different taxonomic levels and to confirm the homogeneity of individual samples belonging to one strain.