Identification of genetic loci for early maturity in spring bread wheat using the association analysis and gene dissection.

Background: Early maturity in spring bread wheat is highly desirable in the regions where it enables the plants to evade high temperatures and plant pathogens at the end of the growing season. Methods: To reveal the genetic loci responsible for the maturity time association analysis was carried out based on phenotyping for an 11-year period and high-throughput SNP genotyping of a panel of the varieties contrasting for this trait. The expression of candidate genes was verified using qPCR. The association between the SNP markers and the trait was validated using the biparental F2:3 population. Results: Our data showed that under long-day conditions, the period from seedling to maturity is mostly influenced by the time from heading to maturity, rather than the heading time. The QTLs associated with the trait were located on 2A, 3B, 4A, 5B, 7A and 7B chromosomes with the 7BL locus being the most significant and promising for its SNPs accelerated the maturity time by about 9 days. Gene dissection in this locus detected a number of candidates, the best being TraesCS7B02G391800 (bZIP9) and TraesCS7B02G412200 (photosystem II reaction center). The two genes are predominantly expressed in the flag leaf while flowering. The effect of the SNPs was verified in F2:3 population and confirmed the association of the 4A, 5B and 7BL loci with the maturity time.

Novel Genetic Loci from Triticum timopheevii Associated with Gluten Content Revealed by GWAS in Wheat Breeding Lines.

The content and quality of gluten in wheat grain is a distinctive characteristic that determines the final properties of wheat flour. In this study, a genome-wide association study (GWAS) was performed on a wheat panel consisting of bread wheat varieties and the introgression lines (ILs) obtained via hybridization with tetraploid wheat relatives. A total of 17 stable quantitative trait nucleotides (QTNs) located on chromosomes 1D, 2A, 2B, 3D, 5A, 6A, 7B, and 7D that explained up to 21% of the phenotypic variation were identified. Among them, the QTLs on chromosomes 2A and 7B were found to contain three and six linked SNP markers, respectively. Comparative analysis of wheat genotypes according to the composition of haplotypes for the three closely linked SNPs of chromosome 2A indicated that haplotype TT/AA/GG was characteristic of ten ILs containing introgressions from T. timopheevii. The gluten content in the plants with TT/AA/GG haplotype was significantly higher than in the varieties with haplotype GG/GG/AA. Having compared the newly obtained data with the previously reported quantitative trait loci (QTLs) we inferred that the locus on chromosome 2A inherited from T. timopheevii is potentially novel. The introgression lines containing the new locus can be used as sources of genetic factors to improve the quality traits of bread wheat.

Multivariate Genome-Wide Association Study of Concentrations of Seven Elements in Seeds Reveals Four New Loci in Russian Wheat Lines.

Wheat is a cereal grain that plays an important role in the world's food industry. The identification of the loci that change the concentration of elements in wheat seeds is an important challenge nowadays especially for genomic selection and breeding of novel varieties. In this study, we performed a multivariate genome-wide association study (GWAS) of the seven traits-concentrations of Zn, Mg, Mn, Ca, Cu, Fe, and K in grain-of the Russian collection of common wheat Triticum aestivum (N = 149 measured in two years in two different fields). We replicated one known locus associated with the concentration of Zn (IAAV1375). We identified four novel loci-BS00022069_51 (associated with concentrations of Ca and K), RFL_Contig6053_3082 (associated with concentrations of Fe and Mn), Kukri_rep_c70864_329 (associated with concentrations of all elements), and IAAV8416 (associated with concentrations of Fe and Mn)-three of them were located near the genes TraesCS6A02G375400, TraesCS7A02G094800, and TraesCS5B02G325400. Our result adds novel information on the loci involved in wheat grain element contents and may be further used in genomic selection.

Identification of QTLs for Grain Protein Content in Russian Spring Wheat Varieties.

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.

Regions of Chromosome 2A of Bread Wheat (Triticum aestivum L.) Associated with Variation in Physiological and Agronomical Traits under Contrasting Water Regimes.

Understanding the genetic architecture of drought tolerance is of great importance for overcoming the negative impact of drought on wheat yield. Earlier, we discovered the critical role of chromosome 2A for the drought-tolerant status of wheat spring cultivar Saratovskaya 29. A set of 92 single-chromosome recombinant double haploid (SCRDH) lines were obtained in the genetic background of Saratovskaya 29. The lines carry fragments of chromosome 2A from the drought-sensitive cultivar Yanetzkis Probat. The SCRDH lines were used to identify regions on chromosome 2A associated with the manifestation of physiological and agronomical traits under distinct water supply, and to identify candidate genes that may be associated with adaptive gene networks in wheat. Genotyping was done with Illumina Infinium 15k wheat array using 590 SNP markers with 146 markers being polymorphic. In four identified regions of chromosome 2A, 53 out of 58 QTLs associated with physiological and agronomic traits under contrasting water supply were mapped. Thirty-nine candidate genes were identified, of which 18 were transcription factors. The region 73.8-78.1 cM included the largest number of QTLs and candidate genes. The variation in SNPs associated with agronomical and physiological traits revealed among the SCRDH lines may provide useful information for drought related marker-assisted breeding.

Genome-wide association study of leaf rust resistance in Russian spring wheat varieties.

BACKGROUND: Leaf rust (Puccinia triticina Eriks.) is one of the most dangerous diseases of common wheat worldwide. Three approaches: genome-wide association study (GWAS), marker-assisted selection (MAS) and phytopathological evaluation in field, were used for assessment of the genetic diversity of Russian spring wheat varieties on leaf rust resistance loci and for identification of associated molecular markers. RESULTS: The collection, consisting of 100 Russian varieties of spring wheat, was evaluated over three seasons for resistance to the native population of leaf rust specific to the West Siberian region of Russia. The results indicated that most cultivars showed high susceptibility to P. triticina, with severity ratings (SR) of 60S-90S, however some cultivars showed a high level of leaf rust resistance (SR < 20MR-R). Based on the results of genome-wide association studies (GWAS) performed using the wheat 15 K genotyping array, 20 SNPs located on chromosomes 6D, 6A, 6B, 5A, 1B, 2A, 2B and 7A were revealed to be associated with leaf rust resistance. Genotyping with markers developed for known leaf rust resistance genes showed that most of the varieties contain genes Lr1, Lr3a, Lr9, Lr10, Lr17a, Lr20, Lr26 and Lr34, which are not currently effective against the pathogen. In the genome of three wheat varieties, gene Lr6Ai = 2 inherited from Th. intermedium was detected, which provides complete protection against the rust pathogen. It has been suggested that the QTL mapped to the chromosome 5AS of wheat cultivar Tulaikovskaya-zolotistaya, Tulaikovskaya-10, Samsar, and Volgouralskaya may be a new, previously undescribed locus conferring resistance to leaf rust. Obtained results also indicate that chromosome 1BL of the varieties Sonata, Otrada-Sibiri, Tertsiya, Omskaya-23, Tulaikovskaya-1, Obskaya-14, and Sirena may contain an unknown locus that provides a resistance response to local population. CONCLUSIONS: This study provides new insights into the genetic basis of resistance to leaf rust in Russian spring wheat varieties. The SNPs significantly associated with leaf rust resistance can be used for the development and application of diagnostic markers in marker-assisted selection schemes.

Detection of Genomic Regions Associated with Resistance to Stem Rust in Russian Spring Wheat Varieties and Breeding Germplasm.

Stem rust caused by Puccinia graminis f. sp. tritici Eriks. is a dangerous disease of common wheat worldwide. Development and cultivation of the varieties with genetic resistance is one of the most effective and environmentally important ways for protection of wheat against fungal pathogens. Field phytopathological screening and genome-wide association study (GWAS) were used for assessment of the genetic diversity of a collection of spring wheat genotypes on stem rust resistance loci. The collection consisting of Russian varieties of spring wheat and introgression lines with alien genetic materials was evaluated over three seasons (2016, 2017 and 2018) for resistance to the native population of stem rust specific to the West Siberian region of Russia. The results indicate that most varieties displayed from moderate to high levels of susceptibility to P. graminis; 16% of genotypes had resistance or immune response. In total, 13,006 single-nucleotide polymorphism (SNP) markers obtained from the Infinium 15K array were used to perform genome-wide association analysis. GWAS detected 35 significant marker-trait associations (MTAs) with SNPs located on chromosomes 1A, 2A, 2B, 3B, 5A, 5B, 6A, 7A and 7B. The most significant associations were found on chromosomes 7A and 6A where known resistance genes Sr25 and Sr6Ai = 2 originated from Thinopyrum ssp. are located. Common wheat lines containing introgressed fragments from Triticum timopheevii and Triticum kiharae were found to carry Sr36 gene on 2B chromosome. It has been suggested that the quantitative trait loci (QTL) mapped to the chromosome 5BL may be new loci inherited from the T. timopheevii. It can be inferred that a number of Russian wheat varieties may contain the Sr17 gene, which does not currently provide effective protection against pathogen. This is the first report describing the results of analysis of the genetic factors conferring resistance of Russian spring wheat varieties to stem rust.

Identification of new heading date determinants in wheat 5B chromosome.

BACKGROUND: Variability of heading date may assist in wheat adaptation to local environments. Thereafter, discovery of new heading date determinants is important for cereal improvement. In this study we used common wheat cultivar Chinese Spring (CS) and the substitution line of CS with 5B chromosome from T. dicoccoides (CS-5Bdic), different in their heading date by two weeks, to detect determinants of heading date on 5B chromosome. RESULTS: The possible influence of the VRN-B1 gene, the most powerful regulator of flowering, located on 5B chromosome, to differences in heading time between CS and CS-5Bdic was studied. The sequencing of this gene from CS-5Bdic showed that an insertion of a nucleotide triplet produced an additional amino acid in the corresponding protein. No changes in the transcription levels of each homoeologous VRN-1 loci were found in CS-5Bdic by comparison with CS. To ascertain the loci determining heading date difference, a set of 116 recombinant inbred 5В chromosomal lines as a result of hybridization of CS with CS-5Bdic were developed and their heading dates were estimated. Using the Illumina Infinium 15 k Wheat platform, 379 5B-specific polymorphic markers were detected and a genetic map with 82 skeletal markers was constructed. Phenotype (heading date) - genotype association analysis revealed seventy eight markers in pericentromeric region of 5B chromosome significantly associated with heading date variation. Based on this estimation and synteny with model crop genomes we identified the three best candidate genes: WRKY, ERF/AP2 and FHY3/FAR1. CONCLUSIONS: We supposed that the difference in activity of WRKY, ERF/AP2 and/or FHY3/FAR1 transcription factors between CS and CS-5Bdic to be a probable reason for the observed difference in heading dates. Data obtained in this study provide a good basis for the subsequent investigation of heading time pathways in wheat.

Wheat genome structure: translocations during the course of polyploidization.

The genomic organization of Triticum timopheevii (2n=28, AtAtGG) was compared with hexaploid wheat T. aestivum (2n=42, AABBDD) by comparative mapping using microsatellites derived from bread wheat. Genetic maps for the two crosses T. timopheevii var. timopheevii x T. timopheevii var. typica and T. timopheevii K-38555xT. militinae were constructed. On the first population, 121 loci were mapped, and on the second population 103 loci. The transferability of the wheat markers to T. timopheevii was generally better for the A genome-specific markers (76-78% produced amplification products; 26 and 29% were polymorphic) than for B genome-specific markers (54% produced amplification products; 14 and 16% were polymorphic). Of the D genome-specific markers, one third produced amplification products in T. timopheevii, but only 5 and 2% were polymorphic in the corresponding mapping populations. The maps constructed confirmed the previously described translocation between chromosome arms 6AtS and 1GS and revealed at least two yet unknown rearrangements on chromosomes 4At and 6At09. The presence of other translocations and rearrangements between T. timopheevii and T. aestivum was demonstrated by a variety of markers mapping to nonhomoeologous positions.