استفاده از کروماتوگرافی مایع فاز معکوس با کارایی بالا (RP-HPLC) در بررسی تنوع ژنتیکی گندم نان

This study was conducted to evaluate the capability of reversed-phase high performance liquid chromatography (RP-HPLC) of wheat (Triticum aestivum L.) storage proteins, and their genetic variability in different winter and spring isolines, cultivars and landraces. Gliadin proteins were extracted from the flour of five randomly selected seeds of each genotype. In this method, Nucleosil C18 300 A column (250×4.6 mm ID), equipped with a guard column and acetonitrile containing TFA as mobile phase was used. All selected conditions for RP-HPLC such as suitable velocity and resolution were sufficient to achieve the objectives of this study. Under these conditions, the number of gliadin components by RP-HPLC analysis was more than electrophoresis subunits. In addition, quantitative results of RP-HPLC facilitated the data analysis. Also gliadin analysis by RP-HPLC had a high potential in detecting rye (Secale cereale L.) genes, so that the presence of ω-secalins in the genome of Falat variety was easily detected. Therefore, it was concluded that RP-HPLC analysis of gliadin proteins is as efficient as electrophoresis, and could even replace it in some cases. The result of cluster analysis for gliadin polypeptides indicated the gradual increase of genetic variation from isolines to landraces. Generally speaking, among the landraces, Ali Abad, Aghda, Sefid Bafghi, Ghermez Bafghi, Shahdas and Sorkheh had greater genetic variations.