BackgroundUnderstanding the genetic basis of frost tolerance (FT) in wheat (Triticum aestivum L.) is essential for preventing yield losses caused by frost due to cellular damage, dehydration and reduced metabolism. FT is a complex trait regulated by a number of genes and several gene families. Availability of the wheat genomic sequence opens new opportunities for exploring candidate genes diversity for FT. Therefore, the objectives of this study were to identity SNPs and insertion-deletion (indels) in genes known to be involved in frost tolerance and to perform association genetics analysis of respective SNPs and indels on FT.ResultsHere we report on the sequence analysis of 19 candidate genes for FT in wheat assembled using the Chinese Spring IWGSC RefSeq v1.0. Out of these, the tandem duplicated C-repeat binding factors (CBF), i.e. CBF-A3, CBF-A5, CBF-A10, CBF-A13, CBF-A14, CBF-A15, CBF-A18, the vernalisation response gene VRN-A1, VRN-B3, the photoperiod response genes PPD-B1 and PPD-D1 revealed association to FT in 235 wheat cultivars. Within six genes (CBF-A3, CBF-A15, VRN-A1, VRN-B3, PPD-B1 and PPD-D1) amino acid (AA) substitutions in important protein domains were identified. The amino acid substitution effect in VRN-A1 on FT was confirmed and new AA substitutions in CBF-A3, CBF-A15, VRN-B3, PPD-B1 and PPD-D1 located at highly conserved sites were detected. Since these results rely on phenotypic data obtained at five locations in 2 years, detection of significant associations of FT to AA changes in CBF-A3, CBF-A15, VRN-A1, VRN-B3, PPD-B1 and PPD-D1 may be exploited in marker assisted breeding for frost tolerance in winter wheat.ConclusionsA set of 65 primer pairs for the genes mentioned above from a previous study was BLASTed against the IWGSC RefSeq resulting in the identification of 39 primer combinations covering the full length of 19 genes. This work demonstrates the usefulness of the IWGSC RefSeq in specific primer development for highly conserved gene families in hexaploid wheat and, that a candidate gene association genetics approach based on the sequence data is an efficient tool to identify new alleles of genes important for the response to abiotic stress in wheat.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4795-6) contains supplementary material, which is available to authorized users.

Two major genes controlling leaf pubescence were mapped on chromosomes 4BL (Hl1) and 7BS (Hl2 Aesp ) in wheat (Saratovskaya 29) and a wheat/Aegilops introgression line (102/ 00 I ), respectively, together with quantitative trait loci (QTLs) determining hairiness of the leaf margin (QHl.ipk-4B, QHl.ipk-4D) and auricle (QPa.ipk-4B, QPa.ipk-4D) on the long arms of chromosomes 4B and 4D, respectively. The QTLs on chromosome 4D were contributed by a synthetic wheat and, therefore, originated from Aegilops tauschii. The homoeologous group 4 wheat/ A. tauschii genes/QTLs detected in the present study were aligned with the barley pubescence genes Hln/Hsh and Hs b and the hairy peduncle rye gene Hp1. The locus seems to be pleiotropically responsible for the pubescence of diVerent plant organs in diVerent species of the Triticeae. Another homoeologous series may be present on the short arms of the homoeologous group 7 chromosomes, based on the results of an allelic test cross between the Chinese local cultivar Hong-mang-mai carrying Hl2 and the wheat/ Aegilops speltoides introgression line 102/00 I .

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Microsatellite markers were used to map the major genes Bg (determining black glume colour), Rg1 and Rg3 (red glume), and a locus determining smokey-grey coloured glume to the distal ends of the short arms of the homoeologous group 1 chromosomes, proximally (or closely linked) to Xgwm1223 and distal to Xgwm0033. On this basis, we propose that these genes represent a set of homoeoloci, designated Rg-A1, Rg-B1, and Rg-D1. Rg3 and Bg appear to be variant alleles of Rg-A1. Both Rg3 and Bg are closely linked with the major glume pubescence gene Hg. Similarly, the hexaploid wheat smokey-grey glume gene and Rg2 represent alleles at Rg-D1. The microsatellite markers linked to the Rg genes were used to analyse a phenotypically and genotypically characterized set of Siberian spring wheats. A coincidence between the presence of the 264-bp allele of Xgwm0136 and Rg-A1b (Rg3) was observed; so Xgwm0136 can probably be used as a diagnostic marker for this gene.