B Chromosomes Research Articles

Chromosomal Location of the <i>b</i>-<i>Amy-A1</i> Gene and Distribution of Its Alleles in Winter Common Wheat Culture

The analysis of the chromosomal location of the b-Amy-A1 gene, which controls the synthesis of wheat beta-amylase isoenzymes, was carried out by assessing the linkage with the B1 (Awnless) marker gene located in the long arm of chromosome 5A. Results based on F2 and F→∞ Delta × Selection 2092 study. The b-Amy-A1 locus showed 6.55 ± 2.10% linkage to the B1 gene. In parallel, alleles of the Rht8 locus, which causes dwarfing of plants, were identified and its linkage to the C gene (Compact spike) was established in 22.59 ± 6.28% (F2) and 24.22 ± 1.18% (F→∞) of recombination in chromosome 2D. The Delta variety carries the Rht8a allele, and the Selection 2092 accession carries the Rht8c allele. A collection of winter common wheat varieties created and released in the Russia was studied. The selected cultivars carried identical b-Amy-B1a and b-Amy-D1a alleles of the 4B and 4D chromosome loci. At the same time, the most common b-Amy-A1a and b-Amy-A1b alleles of chromosome 5A differed. It was found that in the direction from south to north of European part, the frequency of the b-Amy-A1a allele increases from 24.2% (Northern Caucasus) to 72.50% (Moscow–Ulyanovsk regions). Accordingly, the frequency of the alternative allele b-Amy-A1b decreased from 75.8% (Northern Caucasus) to 25.0% (Moscow–Ulyanovsk regions). In the Rostov region, the ratio of the occurrence of alleles b-Amy-A1a to b-Amy-A1b was 42.5 to 57.5%. At the same time, already in the Central Chernozem region+, a significant dominance of the b-Amy-A1a allele over b-Amy-A1b was manifested. Using the collection of varieties as a population of F→∞ demonstrated the linkage of b-Amy-A1 with B1 value in 5.56 ± 1.90% recombination.

Genome wide association study for stripe rust resistance in elite spring bread wheat genotypes (Triticum aestivum L.) in Morocco

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases affecting wheat (Triticum aestivum L.) worldwide. Developing and deploying resistant cultivars is the most cost-effective and environmentally friendly method of combating this disease. However, because the pathogen continues to mutate, resistance genes are quickly overpowered by newly virulent Pst races, requiring the continuous identification and introduction of resistance genes to create resistant wheat varieties. To identify closely linked markers for possible use in marker-assisted selection a genome-wide association study was conducted using rust data at adult plant stages for 200 elite spring bread wheat genotypes and 13151 SNP markers. Stripe rust resistance was evaluated under rainfed conditions at the ICARDA Merchouch station in Morocco during the 2019–2020 and 2020–2021 crop seasons. The genotypes were classified into five distinct categories based on the coefficient of infection (CI), 18 genotypes were highly resistant, while 42 were moderately resistant (21%), additionally, 73 were intermediate (37%), while 26 were moderately susceptible (13%), and the remaining 41 genotypes highly susceptible. Thirteen markers were associated with stripe rust resistance through GWAS analysis using a general linear model. Four of these markers for adult plant resistance (Kukri c8390 1515, CAP7 c5481 96, Excalibur c35203 334, and BS00017187 51) on chromosomes 1B, 5B, 3B, and 7B, correspondingly, are of particular importance and could be used for marker-assisted selection to pyramid rust resistance genes in wheat breeding programs.

Identification of solvent retention capacity quantitative trait loci by combined linkage and association mapping in wheat (Triticum aestivum L.)

AbstractSolvent retention capacity (SRC) is an important tool for assessing wheat quality by estimating the biochemical characteristics of flour components. Although it has been widely used in wheat breeding programs, its genetic architecture is still poorly understood. Hence, the objective of this study was to find quantitative trait loci (QTLs) associated with four SRC traits, namely sucrose SRC, sodium carbonate SRC, water SRC and lactic acid SRC, in a recombinant inbred line population consisting of 173 lines descended from Shannong 01–35/Gaocheng 9411 and an association panel of 205 wheat varieties. The 90K single nucleotide polymorphism array was used to genotype both populations. SRC was primarily influenced by genotype, with broad‐sense heritability (H2) ranging from 59.72% to 80.76%. Linkage mapping revealed seven major QTLs on chromosomes 1B, 4B, 5B, and 6B. Among them, QSc1B.6‐27 was repeatedly identified in two different environments, with a 14.7% phenotypic variance explained value. Additionally, 24 significant QTLs, consisting of 57 marker‐trait associations, were identified using association mapping across four environments. Nineteen of them explained more than 10.0% of the phenotypic variance, with QSU6A.141 explaining the most (20.1%). Two common chromosomal regions were discovered when combining the two mapping populations. These regions were located between IACX5803 and BS00067000_51 on chromosome 1B and between IACX557 and CAP11_c3631_75 on chromosome 4B, respectively. In addition, four candidate genes associated with wheat SRC were identified. These findings shed light on the complex genetic mechanisms underlying wheat SRC and may help to genetically enhance wheat quality.

Fibrosis-the tale of H3K27 histone methyltransferases and demethylases.

Fibrosis, or excessive scarring, is characterized by the emergence of alpha-smooth muscle actin (αSMA)-expressing myofibroblasts and the excessive accumulation of fibrotic extracellular matrix (ECM). Currently, there is a lack of effective treatment options for fibrosis, highlighting an unmet need to identify new therapeutic targets. The acquisition of a fibrotic phenotype is associated with changes in chromatin structure, a key determinant of gene transcription activation and repression. The major repressive histone mark, H3K27me3, has been linked to dynamic changes in gene expression in fibrosis through alterations in chromatin structure. H3K27-specific homologous histone methylase (HMT) enzymes, Enhancer of zeste 1 and 2 (EZH1, EZH2), which are the alternative subunits of the Polycomb Repressive Complex 2 (PRC2) and demethylase (KDM) enzymes, Ubiquitously transcribed tetratricopeptide repeat, X chromosome (UTX), and Lysine demethylase 6B (KDM6B), are responsible for regulating methylation status of H3K27me3. In this review, we explore how these key enzymes regulate chromatin structure to alter gene expression in fibrosis, highlighting them as attractive targets for the treatment of fibrosis.

Consensus linkage map construction and QTL mapping for eight yield-related traits in wheat using the BAAFS Wheat 90K SNP array

Identifying stable quantitative trait loci (QTLs) for yield-related traits across populations and environments is crucial for wheat breeding and genetic studies. Consensus maps also play important roles in wheat genetic and genomic research. In the present study, a wheat consensus map was constructed using a doubled haploid (DH) population derived from Jinghua 1×Xiaobaidongmai (JX), an F2 population derived from L43×Shanxibaimai (LS) and the BAAFS Wheat 90K SNP array single nucleotide polymorphism (SNP) array. A total of 44,503 SNP markers were mapped on the constructed consensus map, and they covered 5,437.92 cM across 21 chromosomes. The consensus map showed high collinearity with the individual maps and the wheat reference genome IWGSC RefSeq v2.1. Phenotypic data on eight yield-related traits were collected in the JX population, as well as the F2:3 and F2:4 populations of LS, in six, two and two environments, respectively, and those data were used for QTL analysis. Inclusive composite interval mapping (ICIM) identified 32 environmentally stable QTLs for the eight yield-related traits. Among them, four QTLs (QPH.baafs-4B, QKNS.baafs-4B, QTGW.baafs-4B, and QSL.baafs-5A.3) were detected across mapping populations and environments, and nine stable QTLs (qKL.baafs-1D, QPH.baafs-2B, QKNS.baafs-3D, QSL.baafs-3D, QKW.baafs-4B, QPH.baafs-5D, QPH.baafs-6A.1, QSL.baafs-6A, and QSL.baafs-6D) are likely to be new. The physical region of 17.25–44.91 Mb on chromosome 4B was associated with six yield-related traits, so it is an important region for wheat yield. The physical region around the dwarfing gene Rht24 contained QTLs for kernel length (KL), kernel width (KW), spike length (SL), and thousand-grain weight (TGW), which are either from a pleiotropic effect of Rht24 or closely linked loci. For the stable QTLs, 254 promising candidate genes were identified. Among them, TraesCS5A03G1264300, TraesCS1B03G0624000 and TraesCS6A03G0697000 are particularly noteworthy since their homologous genes have similar functions for the corresponding traits. The constructed consensus map and the identified QTLs along with their candidate genes will facilitate the genetic dissection of wheat yield-related traits and accelerate the development of wheat cultivars with desirable plant morphology and high yield.

From Grain to Genome: Investigating Arsenic Levels in Triticum turgidum ssp durum Desf. Using GWAS

Producing safe and high-quality food is increasingly important, and developing durum wheat varieties with low toxicity is crucial to meeting this demand. Durum wheat breeders can achieve this goal by developing new varieties that are either more resistant to arsenic uptake or better adapted to grow in areas with high arsenic levels. High levels of arsenic can pose serious health hazards, which makes it critical to evaluate the arsenic levels. Therefore, this study evaluated the arsenic levels in diverse durum wheat genotypes, including Turkish-released cultivars and local landraces. The results showed that all genotypes had significantly low and non-toxic levels of arsenic, with an average concentration of 5.24 μg/kg. These concentrations were much lower than the minimum reported in numerous published research studies and well below the risky international standard limits for durum wheat grain (0.1 mg/kg). The study also identified two significant marker-trait associations linked to arsenic contents located on chromosomes 4A and 7B, which explained 11-17% of the phenotypic variation. These findings provide valuable insights into the arsenic levels in durum wheat genotypes and highlight the need for ongoing monitoring to ensure safe and healthy food for consumers. By conducting collaborative genome-wide association studies and employing marker-assisted selection, durum wheat breeders can accelerate the creation of new varieties that have reduced arsenic levels by identifying alleles linked to arsenic content. This study emphasizes the importance of developing low-toxicity durum wheat varieties to ensure the safety and quality of our food supply. The findings can inform breeding programs to develop such varieties and contribute to sustainable agriculture. While the study’s methodology was robust, further research is necessary to confirm and validate the genetic factors contributing to variation in arsenic content among different durum wheat genotypes.

Multi-locus genome-wide association study of fusarium head blight in relation to days to anthesis and plant height in a spring wheat association panel.

Fusarium head blight (FHB) is a highly destructive fungal disease of wheat to which host resistance is quantitatively inherited and largely influenced by the environment. Resistance to FHB has been associated with taller height and later maturity; however, a further understanding of these relationships is needed. An association mapping panel (AMP) of 192 predominantly Canadian spring wheat was genotyped with the wheat 90K single-nucleotide polymorphism (SNP) array. The AMP was assessed for FHB incidence (INC), severity (SEV) and index (IND), days to anthesis (DTA), and plant height (PLHT) between 2015 and 2017 at three Canadian FHB-inoculated nurseries. Seven multi-environment trial (MET) datasets were deployed in a genome-wide association study (GWAS) using a single-locus mixed linear model (MLM) and a multi-locus random SNP-effect mixed linear model (mrMLM). MLM detected four quantitative trait nucleotides (QTNs) for INC on chromosomes 2D and 3D and for SEV and IND on chromosome 3B. Further, mrMLM identified 291 QTNs: 50 (INC), 72 (SEV), 90 (IND), 41 (DTA), and 38 (PLHT). At two or more environments, 17 QTNs for FHB, DTA, and PLHT were detected. Of these 17, 12 QTNs were pleiotropic for FHB traits, DTA, and PLHT on chromosomes 1A, 1D, 2D, 3B, 5A, 6B, 7A, and 7B; two QTNs for DTA were detected on chromosomes 1B and 7A; and three PLHT QTNs were located on chromosomes 4B and 6B. The 1B DTA QTN and the three pleiotropic QTNs on chromosomes 1A, 3B, and 6B are potentially identical to corresponding quantitative trait loci (QTLs) in durum wheat. Further, the 3B pleiotropic QTN for FHB INC, SEV, and IND co-locates with TraesCS3B02G024900 within the Fhb1 region on chromosome 3B and is ~3 Mb from a cloned Fhb1 candidate gene TaHRC. While the PLHT QTN on chromosome 6B is putatively novel, the 1B DTA QTN co-locates with a disease resistance protein located ~10 Mb from a Flowering Locus T1-like gene TaFT3-B1, and the 7A DTA QTN is ~5 Mb away from a maturity QTL QMat.dms-7A.3 of another study. GWAS and QTN candidate genes enabled the characterization of FHB resistance in relation to DTA and PLHT. This approach should eventually generate additional and reliable trait-specific markers for breeding selection, in addition to providing useful information for FHB trait discovery.

Genome-Wide Association Study for Fusarium Head Blight Resistance in Common Wheat from China

As a devastating wheat disease, Fusarium head blight (FHB) causes large losses in yield and quality. The purpose of this study was to detect the genetic loci for FHB resistance based on a genome-wide association study (GWAS) using two panels genotyped by the Wheat 660 K SNP array and Wheat 90 K SNP array. Phenotypic investigation results showed that 43.6–51.6% of the test varieties showed an infection type of more than 3, indicating that substantial improvement in the level of resistance was urgently required. GWAS detected 1264 SNPs associated with FHB response distributed on all wheat chromosomes. Fourteen QTL were stably detected in two environments. gFHB-1B (582.9~607.9 Mb on chromosome 1B) and gFHB-5A (577.9~596.5 Mb on 5A) were two novel resistance loci that were detected in two wheat panels. Haplotype analysis of both gFHB-1B and gFHB-5A indicated that varieties with gFHB-1B_R and gFHB-5A_R had consistently less FHB than varieties carrying a one or no resistance QTL. The application frequency of the FHB-resistant alleles gFHB-1B and gFHB-5A showed a gradually decreasing trend in FHB resistance breeding. This study identified new genetic loci for the control of FHB and provides useful information for marker-assisted selection in wheat FHB resistance.

Identification of novel putative alleles related to important agronomic traits of wheat using robust strategies in GWAS

Principal component analysis (PCA) is widely used in various genetics studies. In this study, the role of classical PCA (cPCA) and robust PCA (rPCA) was evaluated explicitly in genome-wide association studies (GWAS). We evaluated 294 wheat genotypes under well-watered and rain-fed, focusing on spike traits. First, we showed that some phenotypic and genotypic observations could be outliers based on cPCA and different rPCA algorithms (Proj, Grid, Hubert, and Locantore). Hubert’s method provided a better approach to identifying outliers, which helped to understand the nature of these samples. These outliers led to the deviation of the heritability of traits from the actual value. Then, we performed GWAS with 36,000 single nucleotide polymorphisms (SNPs) based on the traditional approach and two robust strategies. In the conventional approach and using the first three components of cPCA as population structure, 184 and 139 marker-trait associations (MTAs) were identified for five traits in well-watered and rain-fed environments, respectively. In the first robust strategy and when rPCA was used as population structure in GWAS, we observed that the Hubert and Grid methods identified new MTAs, especially for yield and spike weight on chromosomes 7A and 6B. In the second strategy, we followed the classical and robust principal component-based GWAS, where the first two PCs obtained from phenotypic variables were used instead of traits. In the recent strategy, despite the similarity between the methods, some new MTAs were identified that can be considered pleiotropic. Hubert's method provided a better linear combination of traits because it had the most MTAs in common with the traditional approach. Newly identified SNPs, including rs19833 (5B) and rs48316 (2B), were annotated with important genes with vital biological processes and molecular functions. The approaches presented in this study can reduce the misleading GWAS results caused by the adverse effect of outlier observations.

Genetic dissection of protein and starch during wheat grain development using QTL mapping and GWAS.

Protein, starch, and their components are important for wheat grain yield and end-products, which are affected by wheat grain development. Therefore, QTL mapping and a genome-wide association study (GWAS) of grain protein content (GPC), glutenin macropolymer content (GMP), amylopectin content (GApC), and amylose content (GAsC) were performed on wheat grain development at 7, 14, 21, and 28 days after anthesis (DAA) in two environments using a recombinant inbred line (RIL) population of 256 stable lines and a panel of 205 wheat accessions. A total of 29 unconditional QTLs, 13 conditional QTLs, 99 unconditional marker-trait associations (MTAs), and 14 conditional MTAs significantly associated (p < 10-4) with four quality traits were found to be distributed on 15 chromosomes, with the phenotypic variation explained (PVE) ranging from 5.35% to 39.86%. Among these genomic variations, three major QTLs [QGPC3B, QGPC2A, and QGPC(S3|S2)3B] and SNP clusters on the 3A and 6B chromosomes were detected for GPC, and the SNP TA005876-0602 was stably expressed during the three periods in the natural population. The QGMP3B locus was detected five times in three developmental stages in two environments with 5.89%-33.62% PVE, and SNP clusters for GMP content were found on the 3A and 3B chromosomes. For GApC, the QGApC3B.1 locus had the highest PVE of 25.69%, and SNP clusters were found on chromosomes 4A, 4B, 5B, 6B, and 7B. Four major QTLs of GAsC were detected at 21 and 28 DAA. Most interestingly, both QTL mapping and GWAS analysis indicated that four chromosomes (3B, 4A, 6B, and 7A) were mainly involved in the development of protein, GMP, amylopectin, and amylose synthesis. Of these, the wPt-5870-wPt-3620 marker interval on chromosome 3B seemed to be most important because it played an important role in the synthesis of GMP and amylopectin before 7 DAA, in the synthesis of protein and GMP from 14 to 21 DAA, and in the development of GApC and GAsC from 21 to 28 DAA. Using the annotation information of IWGSC Chinese Spring RefSeq v1.1 genome assembly, we predicted 28 and 69 candidate genes for major loci from QTL mapping and GWAS, respectively. Most of them have multiple effects on protein and starch synthesis during grain development. These results provide new insights and information for the potential regulatory network between grain protein and starch synthesis.

Identification of Novel QTLs/Defense Genes in Spring Wheat Germplasm Panel for Seedling and Adult Plant Resistance to Stem Rust and Their Validation Through KASP Marker Assays.

Stem rust is one of the major diseases threatening wheat production globally. To identify novel resistance quantitative trait loci (QTLs), we performed 35K Axiom Array SNP genotyping assays on an association mapping panel of 400 germplasm accessions, including Indian landraces, in conjunction with phenotyping for stem rust at seedling and adult plant stages. Association analyses using three genome wide association study (GWAS) models (CMLM, MLMM, and FarmCPU) revealed 20 reliable QTLs for seedling and adult plant resistance. Among these 20 QTLs, five QTLs were found consistent with three models, i.e., four QTLs on chromosome 2AL, 2BL, 2DL, and 3BL for seedling resistance and one QTL on chromosome 7DS for adult plant resistance. Further, we identified a total of 21 potential candidate genes underlying QTLs using gene ontology analysis, including a leucine rich repeat receptor (LRR) and P-loop nucleoside triphosphate hydrolase, which have a role in pathogen recognition and disease resistance. Furthermore, four QTLs (Qsr.nbpgr-3B_11, QSr.nbpgr-6AS_11, QSr.nbpgr-2AL_117-6, and QSr.nbpgr-7BS_APR) were validated through KASP located on chromosomes 3B, 6A, 2A, and 7B. Out of these QTLs, QSr.nbpgr-7BS_APR was identified as a novel QTL for stem rust resistance which has been found effective in both seedling as well as the adult plant stages. Identified novel genomic regions and validated QTLs have the potential to be deployed in wheat improvement programs to develop disease resistant varieties for stem rust and can diversify the genetic basis of resistance.

Precisely mapping a major QTL for grain weight on chromosome 5B of the founder parent Chuanmai42 in the wheat-growing region of southwestern China.

QTgw.saas-5B was validated as a major thousand-grain weight-related QTL in a founder parent used for wheat breeding and then precisely mapped to a 0.6 cM interval. Increasing the thousand-grain weight (TGW) is considered to be one of the most important ways to improve yield, which is a core objective among wheat breeders. Chuanmai42, which is a wheat cultivar with high TGW and a high and stable yield, is a parent of more than 30 new varieties grown in southwestern China. In this study, a Chuanmai42-derived recombinant inbred line (RIL) population was used to dissect the genetic basis of TGW. A major QTL (QTgw.saas-5B) mapped to the Xgwm213-Xgwm540 interval on chromosome 5B of Chuanmai42 explained up to 20% of the phenotypic variation. Using 71 recombinants with a recombination in the QTgw.saas-5B interval identified from a secondary RIL population comprising 1818 lines constructed by crossing the QTgw.saas-5B near-isogenic line with the recurrent parent Chuannong16, QTgw.saas-5B was delimited to a 0.6cM interval, corresponding to a 21.83Mb physical interval in the Chinese Spring genome. These findings provide the foundation for QTgw.saas-5B cloning and its use in molecular marker-assisted breeding.

ZIP4 is required for normal progression of synapsis and for over 95% of crossovers in wheat meiosis.

Tetraploid (AABB) and hexaploid (AABBDD) wheat have multiple sets of similar chromosomes, with successful meiosis and preservation of fertility relying on synapsis and crossover (CO) formation only taking place between homologous chromosomes. In hexaploid wheat, the major meiotic gene TaZIP4-B2 (Ph1) on chromosome 5B, promotes CO formation between homologous chromosomes, whilst suppressing COs between homeologous (related) chromosomes. In other species, ZIP4 mutations eliminate approximately 85% of COs, consistent with loss of the class I CO pathway. Tetraploid wheat has three ZIP4 copies: TtZIP4-A1 on chromosome 3A, TtZIP4-B1 on 3B and TtZIP4-B2 on 5B. Here, we have developed single, double and triple zip4 TILLING mutants and a CRISPR Ttzip4-B2 mutant, to determine the effect of ZIP4 genes on synapsis and CO formation in the tetraploid wheat cultivar 'Kronos'. We show that disruption of two ZIP4 gene copies in Ttzip4-A1B1 double mutants, results in a 76-78% reduction in COs when compared to wild-type plants. Moreover, when all three copies are disrupted in Ttzip4-A1B1B2 triple mutants, COs are reduced by over 95%, suggesting that the TtZIP4-B2 copy may also affect class II COs. If this is the case, the class I and class II CO pathways may be interlinked in wheat. When ZIP4 duplicated and diverged from chromosome 3B on wheat polyploidization, the new 5B copy, TaZIP4-B2, could have acquired an additional function to stabilize both CO pathways. In tetraploid plants deficient in all three ZIP4 copies, synapsis is delayed and does not complete, consistent with our previous studies in hexaploid wheat, when a similar delay in synapsis was observed in a 59.3 Mb deletion mutant, ph1b, encompassing the TaZIP4-B2 gene on chromosome 5B. These findings confirm the requirement of ZIP4-B2 for efficient synapsis, and suggest that TtZIP4 genes have a stronger effect on synapsis than previously described in Arabidopsis and rice. Thus, ZIP4-B2 in wheat accounts for the two major phenotypes reported for Ph1, promotion of homologous synapsis and suppression of homeologous COs.

The putative vacuolar processing enzyme gene TaVPE3cB is a candidate gene for wheat stem pith-thickness

Key messageThe vacuolar processing enzyme gene TaVPE3cB is identified as a candidate gene for a QTL of wheat pith-thickness on chromosome 3B by BSR-seq and differential expression analyses.The high pith-thickness (PT) of the wheat stem could greatly enhance stem mechanical strength, especially the basal internodes which support the heavier upper part, such as upper stems, leaves and spikes. A QTL for PT in wheat was previously discovered on 3BL in a double haploid population of ‘Westonia’ × ‘Kauz’. Here, a bulked segregant RNA-seq analysis was applied to identify candidate genes and develop associated SNP markers for PT. In this study, we aimed at screening differentially expressed genes (DEGs) and SNPs in the 3BL QTL interval. Sixteen DEGs were obtained based on BSR-seq and differential expression analyses. Twenty-four high-probability SNPs in eight genes were identified by comparing the allelic polymorphism in mRNA sequences between the high PT and low PT samples. Among them, six genes were confirmed to be associated with PT by qRT-PCR and sequencing. A putative vacuolar processing enzyme gene TaVPE3cB was screened out as a potential PT candidate gene in Australian wheat ‘Westonia’. A robust SNP marker associated with TaVPE3cB was developed, which can assist in the introgression of TaVPE3cB.b in wheat breeding programs. In addition, we also discussed the function of other DEGs which may be related to pith development and programmed cell death (PCD). A five-level hierarchical regulation mechanism of stem pith PCD in wheat was proposed.

Natural variations of wheat EARLY FLOWERING 3 highlight their contributions to local adaptation through fine-tuning of heading time

We identified a large chromosomal deletion containing TaELF-B3 that confers early flowering in wheat. This allele has been preferred in recent wheat breeding in Japan to adapt to the environment. Heading at the appropriate time in each cultivation region can greatly contribute to stabilizing and maximizing yield. Vrn-1 and Ppd-1 are known as the major genes for vernalization requirement and photoperiod sensitivity in wheat. Genotype combinations of Vrn-1 and Ppd-1 can explain the variation in heading time. However, the genes that can explain the remaining variations in heading time are largely unknown. In this study, we aimed to identify the genes conferring early heading using doubled haploid lines derived from Japanese wheat varieties. Quantitative trait locus (QTL) analysis revealed a significant QTL on the long arm of chromosome 1B in multiple growing seasons. Genome sequencing using Illumina short reads and Pacbio HiFi reads revealed a large deletion of a ~ 500kb region containing TaELF-B3, an orthologue of Arabidopsis clock gene EARLY FLOWERING 3 (ELF3). Plants with the deleted allele of TaELF-B3 (ΔTaELF-B3 allele) headed earlier only under short-day vernalization conditions. Higher expression levels of clock- and clock-output genes, such as Ppd-1 and TaGI, were observed in plants with the ΔTaELF-B3 allele. These results suggest that the deletion of TaELF-B3 causes early heading. Of the TaELF-3 homoeoalleles conferring early heading, the ΔTaELF-B3 allele showed the greatest effect on the early heading phenotype in Japan. The higher allele frequency of the ΔTaELF-B3 allele in western Japan suggests that the ΔTaELF-B3 allele was preferred during recent breeding to adapt to the environment. TaELF-3 homoeologs will help to expand the cultivated area by fine-tuning the optimal timing of heading in each environment.

Genome-wide association study reveals the genetic variation and candidate gene for grain calcium content in bread wheat.

This study provides important information on the genetic basis of GCaC in wheat, thus contributing to breeding efforts to improve the nutrient quality of wheat. Calcium (Ca) plays important roles in the human body. Wheat grain provides the main diet for billions of people worldwide but is low in Ca content. Here, grain Ca content (GCaC) of 471 wheat accessions was determined in four field environments. A genome-wide association study (GWAS) was performed to reveal the genetic basis of GCaC using the phenotypic data form four environments and a wheat 660K single nucleotide polymorphism (SNP) array. Twelve quantitative trait locus (QTLs) for GCaC were identified on chromosomes 1A, 1D, 2A, 3B, 6A, 6D, 7A, and 7D, which was significant in at least two environments. Haplotype analysis revealed that the phenotypic difference between the haplotypes of TraesCS6D01G399100 was significant (P ≤ 0.05) across four environments, suggesting it as an important candidate gene for GCaC. This research enhances our understanding of the genetic architecture of GCaC for further improving the nutrient quality of wheat.

FHB resistance conferred by Fhb1 is under inhibitory regulation of two genetic loci in wheat (Triticum aestivum L.).

Two loci inhibiting Fhb1 resistance to Fusarium head blight were identified through genome-wide association mapping and validated in biparental populations. Fhb1 confers Fusarium head blight (FHB) resistance by limiting fungal spread within spikes in wheat (type II resistance). However, not all lines with Fhb1 display the expected resistance. To identify genetic factors regulating Fhb1 effect, a genome-wide association study for type II resistance was first performed with 72 Fhb1-carrying lines using the Illumina 90K iSelect SNP chip. Of 84 significant marker-trait associations detected, more than half were repeatedly detected in at least two environments, with the SNPs distributed in one region on chromosome 5B and one on chromosome 6A. This result was validated in a collection of 111 lines with Fhb1 and 301 lines without Fhb1. We found that these two loci caused significant resistance variations solely among lines with Fhb1 by compromising the resistance. In1, the inhibitory gene on chromosome 5B, was in close linkage with Xwgrb3860 in a recombinant inbred line population derived from Nanda2419 × Wangshuibai and a double haploid (DH) population derived from R-43 (Fhb1 near isogenic line) × Biansui7 (with Fhb1 and In1); and In2, the inhibitory gene on chromosome 6A, was mapped to the Xwgrb4113-Xwgrb4034 interval using a DH population derived from R-43 × PH8901 (with Fhb1 and In2). In1 and In2 are present in all wheat-growing areas worldwide. Their frequencies in China's modern cultivars are high but have significantly decreased in comparison with landraces. These findings are of great significance for FHB resistance breeding using Fhb1.

Genetic mapping of candidate loci for water‐deficit stress‐induced proline accumulation in bread wheat (Triticum aestivum)

AbstractProline is an important metabolite that plays an adaptive role in plants under water‐deficit stress. Understanding the genetic basis of water‐deficit‐induced proline accumulation remains elusive in crop plants, especially in wheat. Here, we investigated proline accumulation under control and water‐deficit conditions using a diversity panel which comprises of 150 bread wheat cultivars. Water‐deficit stress significantly increased proline accumulation than well‐water treated plants. Water‐deficit‐induced proline variability was dissected using genome‐wide association studies that identified significant marker‐trait associations, especially on 1A and 1B chromosomes. Population structure analysis revealed the cultivars originated from Europe were associated with higher proline content. Further, linkage disequilibrium analysis identified minor allele of haplotypes, and single markers were linked with higher proline accumulation under water‐deficit conditions. The identified candidate genes were mostly involved in ATP and ADP binding, protein and oligopeptide transporter activity and also found in defence response, carbohydrate and fatty acid biosynthetic processes. Next, an in silico transcript analysis found higher expression of candidate genes in shoot/leaves under water‐deficit stress conditions. Overall, the key genomic regions controlling water deficit‐induced proline accumulation can be utilized in improving plant adaptation to drought.

Proteomic analysis of near-isogenic lines reveals key biomarkers on wheat chromosome 4B conferring drought tolerance.

Drought is a major constraint for wheat production that is receiving increased attention due to global climate change. This study conducted isobaric tags for relative and absolute quantitation proteomic analysis on near-isogenic lines to shed light on the underlying mechanism of qDSI.4B.1 quantitative trait loci (QTL) on the short arm of chromosome 4B conferring drought tolerance in wheat. Comparing tolerant with susceptible isolines, 41 differentially expressed proteins were identified to be responsible for drought tolerance with a p-value of < 0.05 and fold change >1.3 or <0.7. These proteins were mainly enriched in hydrogen peroxide metabolic activity, reactive oxygen species metabolic activity, photosynthetic activity, intracellular protein transport, cellular macromolecule localization, and response to oxidative stress. Prediction of protein interactions and pathways analysis revealed the interaction between transcription, translation, protein export, photosynthesis, and carbohydrate metabolism as the most important pathways responsible for drought tolerance. The five proteins, including 30S ribosomal protein S15, SRP54 domain-containing protein, auxin-repressed protein, serine hydroxymethyltransferase, and an uncharacterized protein with encoding genes on 4BS, were suggested as candidate proteins responsible for drought tolerance in qDSI.4B.1 QTL. The gene coding SRP54 protein was also one of the differentially expressed genes in our previous transcriptomic study.

High-density genetic mapping of Fusarium head blight resistance and agronomic traits in spring wheat.

Fusarium head blight (FHB) has rapidly become a major challenge to successful wheat production and competitive end-use quality in western Canada. Continuous effort is required to develop germplasm with improved FHB resistance and understand how to incorporate the material into crossing schemes for marker-assisted selection and genomic selection. The aim of this study was to map quantitative trait loci (QTL) responsible for the expression of FHB resistance in two adapted cultivars and to evaluate their co-localization with plant height, days to maturity, days to heading, and awnedness. A large doubled haploid population of 775 lines developed from cultivars Carberry and AC Cadillac was assessed for FHB incidence and severity in nurseries near Portage la Prairie, Brandon, and Morden in different years, and for plant height, awnedness, days to heading, and days to maturity near Swift Current. An initial linkage map using a subset of 261 lines was constructed using 634 polymorphic DArT and SSR markers. QTL analysis revealed five resistance QTL on chromosomes 2A, 3B (two loci), 4B, and 5A. A second genetic map with increased marker density was constructed using the Infinium iSelect 90k SNP wheat array in addition to the previous DArT and SSR markers, which revealed two additional QTL on 6A and 6D. The complete population was genotyped, and a total of 6,806 Infinium iSelect 90k SNP polymorphic markers were used to identify 17 putative resistance QTL on 14 different chromosomes. As with the smaller population size and fewer markers, large-effect QTL were detected on 3B, 4B, and 5A that were consistently expressed across environments. FHB resistance QTL were co-localized with plant height QTL on chromosomes 4B, 6D, and 7D; days to heading on 2B, 3A, 4A, 4B, and 5A; and maturity on 3A, 4B, and 7D. A major QTL for awnedness was identified as being associated with FHB resistance on chromosome 5A. Nine small-effect QTL were not associated with any of the agronomic traits, whereas 13 QTL that were associated with agronomic traits did not co-localize with any of the FHB traits. There is an opportunity to select for improved FHB resistance within adapted cultivars by using markers associated with complementary QTL.