SSR Analysis of Maternal and Paternal Lines Selected in the Don Region (Russia)

Abstract

Evaluation of DNA polymorphisms of breeding material of sunflower from L.A. Zhdanov Don Experimental Station of oil Crops of V.S. Pustovoit All-Russian Research Institute of Oil Crops represented by 17 maternal (CMS) lines and 12 paternal (Rf) lines was conducted. There were identified 35 allelic variants of CMS lines and 42 allelic variants of Rf lines with use of 11 SSR markers. It is shown that the level of genetic diversity of microsatellite loci of CMS lines is 1.2 times lower than that of Rf lines. The average number of alleles per SSR locus for maternal lines also lowers than for paternal lines. In addition, CMS lines and Rf lines differ in the frequency of alleles of SSR loci. Thus, the discriminatory capacity of the studied marker system for identification of cultivated sunflower lines was quite high and is suitable for certification of the lines.

SSR Analysis of Nuclear DNA of Annual and Perennial Sunflower Species (Helianthus L.)

“…Using sunflower map from NCBI database (https://www.ncbi.nlm.nih.gov/projects/mapview/map_sear ch.cgi?taxid=4232&query=Helianthus), we selected 52 SSR markers, with localization on all the 17 linkage groups (chromosomes) of the sunflower genome (Table 2). Genomic DNA was isolated from sunflower leaf tissue, with our modifications (Markin et al, 2016). PCR The PCR was carried out in 25 μL reaction mixture of the following composition: 67 mm Tris-HCl buffer, pH 8.8, 16 mM (NH4)2SO4, 2.5 mM MgSO4, 0.1 mM mercaptoethanol, 0.25 mM of each dNTP (dATP, dCTP, dTTP and dGTP), 400 nM primers, 2.5 units of Taq polymerase and 15 ng of DNA template.…”

“…The most effective and simplest molecular methods for assessing genetic polymorphism are PCR-based techniques. Among such techniques, the Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeats (SSR) markers are widely used, allowing rapid detection of the variability of a large number of genome loci (Sivolap and Solodenko, 1998;Sossey-Alaoui et al, 1998;1999;Markin et al, 2016;Suresha et al, 2017;Yang et al, 2018;Uma et al, 2018). The spectra of DNA fragments, obtained as a result of their amplification, can be used as genetic markers for species identification and barcoding, as well as for determining taxonomic differences between species.…”

Study of informative DNA markers of the Rf1 gene in sunflower for breeding practice

“…Total DNA isolation and PCR were carried out as it was described earlier (Markin et al 2016). Primers designed by Horn (Horn et al 2003) were used for the amplification of SCAR markers (HRG01 and HRG02)…”