Development of ILP-markers for Aegilops tauschii and their application in molecular genetic analysis

Abstract

To develop marker systems based on the study of intron length polymorphism of various genes (ILP) in Ae. tauschii, to test the possibility of their use for genetic differentiation of Ae. tauschii and Ae. biuncialis. Methods. The NCBI database was used to obtain EST sequences, online tools CD-HIT, BLAST and Primer3Plus (for the development of primers). The polymerase chain reaction (PCR) method with the developed primers was done. Amplified fragments were separated by non-denaturing polyacrylamide gel electrophoresis and stained with silver. Results. Molecular markers Aet_ILP1, Aet_ILP6 were developed and tested for different genotypes of Ae. tauschii and Ae. biuncialis. No intraspecific differentiation was observed in all studied samples, but amplicons of introns of different species differed significantly. It was also found that the studied species had one copy of the HO222074.1 gene and 1 to 3 copies of the CX244643.1 gene. Conclusions. The obtained results indicate a low level of intraspecific variability of the developed ILP markers and the possibility of their use for interspecific differentiation of Ae. tauschii and Ae. biuncialis.