Most modern breeding programs aim to develop wheat (T. aestivum L.) varieties with a high grain protein content (GPC) due to its greater milling and cooking quality, and improved grain price. Here, we used a genome-wide association study (GWAS) to map single nucleotide polymorphisms (SNPs) associated with GPC in 93 spring bread wheat varieties developed by eight Russian Breeding Centers. The varieties were evaluated for GPC, grain weight per spike (GWS), and thousand-kernel weight (TKW) at six environments, and genotyped with 9351 polymorphic SNPs and two SNPs associated with the NAM-A1 gene. GPC varied from 9.8 to 20.0%, depending on the genotype and environment. Nearly 52% of the genotypes had a GPC > 14.5%, which is the threshold value for entry into high-class wheat varieties. Broad-sense heritability for GPC was moderate (0.42), which is due to the significant effect of environment and genotype × environment interactions. GWAS performed on mean GPC evaluated across six environments identified eleven significant marker-trait associations, of which nine were physically mapped on chromosome 6A. Screening of wheat varieties for allelic variants of the NAM-A1 gene indicated that 60% of the varieties contained the NAM-A1c allele, followed by 33% for NAM-A1d, and 5% for NAM-A1a alleles. Varieties with the NAM-A1d allele showed significantly (p < 0.01) smaller GPC than those with NAM-A1c and NAM-A1a. However, no significant differences between NAM-A1 alleles were observed for both GWS and TKW.

Heading time is an important agronomic trait affecting the adaptability and productivity of common wheat. In this study, 95 common wheat varieties from Russia and the late-maturing breeding line ‘Velut’ were tested for allelic diversity of genes having the strongest effect on heading. In this research, allelic variation at the Ppd-D1, Vrn-A1, Vrn-B1, Vrn-D1, and Vrn-B3 loci was tested. The Vrn-B1 and Vrn-B3 loci provided the largest contribution to genetic diversity. We found two novel allelic variants of the Vrn-B3 gene in the studied varieties. Ten varieties carried a 160 bp insertion in the promoter region, and the breeding line ‘Velut’ carried a 1617 bp insertion. These alleles were designated Vrn-B3e and Vrn-B3d, respectively. The analysis of the sequences showed the recent insertion of a retrotransposon homologous to the LTR retrotransposon (RLX_Hvul_Dacia_ RND-1) in the Vrn-B3d allele. Plants with the Vrn-B3e and the ‘Velut’ line with the Vrn-B3d allele headed later than the plants with the wild-type allele; among these plants, ‘Velut’ is the latest maturing wheat variety. Analysis of the gene expression of two groups of lines differing by the Vrn-B3 alleles (Vrn-B3d or vrn-B3) from the F2 population with ‘Velut’ as a parental line did not reveal a significant difference in the expression level between the groups. Additional research is required to study the reasons for the late maturation of the ‘Velut’ line. However, the studied wheat varieties could be used as a potential source of natural variation in genes controlling heading times.