Designing of specific primers for different allelic forms of the DREB gene in bread wheat

Abstract

The DREB transcription factor gene, which is considered the main component of the signaling network that regulates stress tolerance, was downloaded from the D genome of wheat whose genome was read from the EnsemblPlants database within the framework of the International Consortium on Wheat Genome Sequencing (Triticum aestivum IWGSC), and the intron-exon composition, domain structure and allelic forms were determined. The AP2 domain, which is responsible for the binding of the DREB protein to the target site on DNA, was found to be located in exon 2, and 14 single nucleotide polymorphisms (SNPs) causing amino acid changes in this area of the gene were determined. For the C/T allele, which leads to the replacement of valine at the 31st amino acid in the DREB protein with isoleucine, 5 pairs of primers meeting the requirements of RT-PCR were designed and tested experimentally, and 2 pairs of primers were found to be effective for the determination of this allele. Determination of SNPs that cause overexpression of the DREB gene is important for the future development of tolerant wheat varieties that can produce high yields under stress conditions by using the site-specific editing capabilities of CRISPR-Cas genome technology.