DEVELOPMENT OF ILP MARKERS FOR PFKFB4 AND RAS GENES OF AEGILOPS TAUSCHII

Abstract

The relevance of this research is determined by the prevalence of cereal crops, particularly common wheat, one of the ancestors of which is Aegilops tauschii. The development of new markers that can be used to study the genetic diversity of cereals is a current issue today. The aim of this study was to develop intron length polymorphism (ILP) markers to investigate the intron length polymorphism of PFKFB4 and RAS genes in Aegilops tauschii, and to assess the potential use of the developed markers for genetic differentiation of Ae. tauschii and Ae. biuncialis. EST sequences were obtained using the NCBI database, and online tools such as CD-HIT, BLAST, and Primer3Plus were employed for primer design. Molecular markers, Aet_ILP_1.1, Aet_ILP_1.2, and Aet_ILP_2, were developed and their efficiency was tested on samples of Ae. tauschii and Ae. biuncialis. Amplified fragments were separated using denaturing polyacrylamide gel electrophoresis and silver staining was performed. The Aet_ILP_1.1 marker differentiated fifteen Ae. biuncialis samples into two groups. The use of the Aet_ILP_1.2 marker revealed additional amplicons in certain Ae. tauschii and Ae. biuncialis samples on electropherograms. The Aet_ILP_2 marker visualized amplicons only in Ae. biuncialis samples. Overall, the results indicate a low level of intraspecific genetic variability in Ae. tauschii using the developed markers, however, they can be effectively utilized for assessing interspecific differentiation between Ae. tauschii and Ae. biuncialis.