Chromosome 2H Research Articles

Yellow Dwarf Virus Resistance in Barley: Phenotyping, Remote Imagery, and Virus-Vector Characterization.

Yellow dwarf viruses (YDVs) spread by aphids are some of the most economically important barley (Hordeum vulgare) virus-vector complexes worldwide. Detection and control of these viruses are critical components in the production of barley, wheat, and numerous other grasses of agricultural importance. Genetic control of plant diseases is often preferable to chemical control to reduce the environmental and economic cost of foliar insecticides. Accordingly, the objectives of this work were to (i) screen a barley population for resistance to YDVs under natural infection using phenotypic assessment of disease symptoms, (ii) implement drone imagery to further assess resistance and test its utility as a disease screening tool, (iii) identify the prevailing virus and vector types in the experimental environment, and (iv) perform a genome-wide association study to identify genomic regions associated with measured traits. Significant genetic differences were found in a population of 192 barley inbred lines regarding their YDV symptom severity, and symptoms were moderately to highly correlated with grain yield. The YDV severity measured with aerial imaging was highly correlated with on-the-ground estimates (r = 0.65). Three aphid species vectoring three YDV species were identified with no apparent genotypic influence on their distribution. A quantitative trait locus impacting YDV resistance was detected on chromosome 2H, albeit undetected using aerial imaging. However, quantitative trait loci for canopy cover and mean normalized difference vegetation index were successfully mapped using the drone. This work provides a framework for utilizing drone imagery in future resistance breeding efforts for YDVs in cereals and grasses, as well as in other virus-vector disease complexes.

Genome-wide association scan reveals the reinforcing effect of nano-potassium in improving the yield and quality of salt-stressed barley via enhancing the antioxidant defense system.

Salinity is one of themajor environmental factor that can greatly impact the growth, development, and productivity of barley. Our study aims to detect the natural phenotypic variation of morphological and physiological traits under both salinity and potassium nanoparticles (n-K) treatment. In addition tounderstanding the genetic basis of salt tolerance in barley is a critical aspect of plant breeding for stress resilience.Therefore, a foliar application of n-K was applied at the vegetative stage for 138 barley accessions to enhance salt stress resilience. Interestingly, barley accessions showed high significant increment under n-K treatment compared to saline soil. Based on genome-wide association studies (GWAS) analysis, causative alleles /reliable genomic regions were discovered underlying improved salt resilience through the application of potassium nanoparticles. On chromosome 2H, a highly significant QTN marker (A:C) was located at position 36,665,559 bp which is associated with APX, AsA, GSH, GS, WGS, and TKW under n-K treatment. Inside this region, our candidate gene is HORVU.MOREX.r3.2HG0111480 that annotated as NAC domain protein. Allelic variation detected that the accessions carrying C allele showed higher antioxidants (APX, AsA, and GSH) and barley yield traits (GS, WGS, and TKW) than the accessions carrying A allele, suggesting a positive selection of the accessions carrying C allele that could be used to develop barley varieties with improved salt stress resilience.

Exotic alleles from the wild Hordeum spontaneum contribute to drought tolerance and grain quality in modern Hordeum vulgare

Conventional breeding relies on genetic variability in crop germplasm and wild species. This study evaluated 137 barley Recombinant Chromosome Substitution Lines (RCSLs), originating from crossing a drought and salinity-tolerant wild Hordeum spontaneum with the high-quality malting H. vulgare cv. Harrington. The objectives of this work were to i) improve understanding of the genetic diversity and introgression levels in RCSLs using a comprehensive set of high-density single nucleotide polymorphism (SNP) markers; ii) investigate the phenotypic variability of physiological, agronomic, and malting quality traits in RCSLs under rainfed and fully irrigated conditions; iii) unravel the relationship between these traits and the resilience of crops in drought-prone regions; and iv) identify specific SNPs associated with both drought tolerance indices and malting quality traits. Variability in physiological, agronomic, and malting quality traits was examined under rainfed and irrigated conditions, aiming to identify specific SNPs associated with both drought tolerance indices and malting quality traits. Additionally, a subset of three recombinant lines and the parent cv. Harrington was tested in growth chambers under well-watered and water-limited conditions. Agronomic traits were influenced by genotype-by-environment interaction, with notable drought tolerance lines. The grain yield was significantly correlated with yield-related traits and carbon isotope discrimination in grains. Drought stress impacted agronomic and physiological traits, with a strong reduction in leaf gas exchange and chlorophyll and protein content. The H. spontaneum genome introgression into H. vulgare cv. Harrington varied across 137 RCSLs, averaging 13.0 %. Associations between traits and genetic markers were found, particularly for the stress tolerance index in chromosome 1H and for grain quality traits (alfa-amylase, wort color, and protein) in chromosomes 1H, 4H, and 5H. These findings uncovered valuable barley RCSLs that demonstrate drought tolerance, offering promising prospects for breeding programs targeted at improving stress resilience.

Genetic mapping of stripe rust resistance in a geographically diverse barley collection and selected biparental populations.

Barley stripe or yellow rust (BYR) caused by Puccinia striiformis f. sp. hordei (Psh) is a significant constraint to barley production. The disease is best controlled by genetic resistance, which is considered the most economical and sustainable component of integrated disease management. In this study, we assessed the diversity of resistance to Psh in a panel of international barley genotypes (n = 266) under multiple disease environments (Ecuador, India, and Mexico) using genome-wide association studies (GWASs). Four quantitative trait loci (QTLs) (three on chromosome 1H and one on 7H) associated with resistance to Psh were identified. The QTLs were validated by mapping resistance to Psh in five biparental populations, which detected key genomic regions on chromosomes 1H (populations Pompadour/Zhoungdamei, Pompadour/Zug161, and CI9214/Baudin), 3H (Ricardo/Gus), and 7H (Fumai8/Baronesse). The QTL RpshQ.GWA.1H.1 detected by GWAS and RpshQ.Bau.1H detected using biparental mapping populations co-located were the most consistent and stable across environments and are likely the same resistance region. RpshQ.Bau.1H was saturated using population CI9214/Baudin by enriching the target region, which placed the resistance locus between 7.9 and 8.1 Mbp (flanked by markers sun_B1H_03, 0.7 cM proximal to Rpsh_1H and sun_B1H_KASP_02, 3.2 cM distal on 1HS) in the Morex reference genome v.2. A Kompetitive Allele Specific PCR (KASP) marker sun_B1H_KASP_01 that co-segregated for RpshQ.Bau.1H was developed. The marker was validated on 50 Australian barley cultivars, showing well-defined allelic discrimination and presence in six genotypes (Baudin, Fathom, Flagship, Grout, Sakurastar, and Shepherd). This marker can be used for reliable marker-assisted selection and pyramiding of resistance to Psh and in diversifying the genetic base of resistance to stripe rust.

Novel QTL Hotspots for Barley Flowering Time, Plant Architecture, and Grain Yield

Barley (Hordeum vulgare L.) is one of the oldest cultivated grains and remains a significant crop globally. Barley breeders focus on developing high-yield cultivars resistant to biotic and abiotic stresses. Barley’s flowering time, regulated genetically and by environmental stimuli, significantly impacts all of its agronomic traits, including the grain yield and plant architecture. This study aimed to detect the quantitative trait loci (QTLs) affecting these traits in 273 two-row spring barley accessions from the USA, Kazakhstan, Europe, and the Middle East across two regions of Kazakhstan, evaluating their impact on grain yield. Genotypic data were obtained from 26,529 segregating single-nucleotide polymorphisms (SNPs), and field trial data for 273 accessions, which were obtained for six traits (heading time, maturity time, vegetation period, plant height, peduncle length, and grain yield) in two regions of Kazakhstan over three growth years. As a result of a genome-wide association study (GWAS), 95 QTLs were identified for 6 agronomic traits, including 58 QTLs linked with candidate genes and/or QTLs. The remaining 37 QTLs were putatively novel, with 13 of them forming 3 QTL hotspots on chromosomes 1H (5 QTLs in the interval of 13.4–41.4 Mbp), 3H (4 QTLs in 608.6–624.9 Mbp), and 6H (4 QTLs in 553.8–572.8 Mbp). These hotspots were pleiotropic, and targeting these regions would allow breeders to enhance multiple yield-associated traits.

Genome wide association study of seedling and adult plant leaf rust resistance in two subsets of barley genetic resources

Leaf rust (LR) caused by Puccinia hordei is a serious disease of barley worldwide, causing significant yield losses and reduced grain quality. Discovery and incorporation of new sources of resistance from gene bank accessions into barley breeding programs is essential for the development of leaf rust resistant varieties. To identify Quantitative Trait Loci (QTL) conferring LR resistance in the two barley subsets, the Generation Challenge Program (GCP) reference set of 142 accessions and the leaf rust subset constructed using the Focused Identification of Germplasm Strategy (FIGS) of 76 barley accessions, were genotyped to conduct a genome-wide association study (GWAS). The results revealed a total of 59 QTL in the 218 accessions phenotyped against barley leaf rust at the seedling stage using two P. hordei isolates (ISO-SAT and ISO-MRC), and at the adult plant stage in four environments in Morocco. Out of these 59 QTL, 10 QTL were associated with the seedling resistance (SR) and 49 QTL were associated with the adult plant resistance (APR). Four QTL showed stable effects in at least two environments for APR, whereas two common QTL associated with SR and APR were detected on chromosomes 2H and 7H. Furthermore, 39 QTL identified in this study were potentially novel. Interestingly, the sequences of 27 SNP markers encoded the candidate genes (CGs) with predicted protein functions in plant disease resistance. These results will provide new perspectives on the diversity of leaf rust resistance loci for fine mapping, isolation of resistance genes, and for marker-assisted selection for the LR resistance in barley breeding programs worldwide.

High resolution mapping of a novel non-transgressive hybrid susceptibility locus in barley exploited by P. teres f. maculata

Hybrid genotypes can provide significant yield gains over conventional inbred varieties due to heterosis or hybrid vigor. However, hybrids can also display unintended negative attributes or phenotypes such as extreme pathogen susceptibility. The necrotrophic pathogen Pyrenophora teres f. maculata (Ptm) causes spot form net blotch, which has caused significant yield losses to barley worldwide. Here, we report on a non-transgressive hybrid susceptibility locus in barley identified between the three parental lines CI5791, Tifang and Golden Promise that are resistant to Ptm isolate 13IM.3. However, F2 progeny from CI5791 × Tifang and CI5791 × Golden Promise crosses exhibited extreme susceptibility. The susceptible phenotype segregated in a ratio of 1 resistant:1 susceptible representing a genetic segregation ratio of 1 parental (res):2 heterozygous (sus):1 parental (res) suggesting a single hybrid susceptibility locus. Genetic mapping using a total of 715 CI5791 × Tifang F2 individuals (1430 recombinant gametes) and 149 targeted SNPs delimited the hybrid susceptibility locus designated Susceptibility to Pyrenophora teres 2 (Spt2) to an ~ 198 kb region on chromosome 5H of the Morex V3 reference assembly. This single locus was independently mapped with 83 CI5791 × Golden Promise F2 individuals (166 recombinant gametes) and 180 genome wide SNPs that colocalized to the same Spt2 locus. The CI5791 genome was sequenced using PacBio Continuous Long Read technology and comparative analysis between CI5791 and the publicly available Golden Promise genome assembly determined that the delimited region contained a single high confidence Spt2 candidate gene predicted to encode a pentatricopeptide repeat-containing protein.

Wheat bread making (WBM)-like seed proteins (WSPN): A new family of small prolamins in barley

Expression level of the gene Wheat Breadmaking (TaWBM) has been shown to be correlated with bread quality, and influences the rising of the bread loaf. We found that a family of the wbm homologous genes are present not only in wheat, but also in oat, rye and barley. However, a WBM protein has never been demonstrated experimentally. We have identified the existence of a family of WBM-like Seed Proteins (WSPN) from the prolamin fraction of barley flour, encoded by three closely linked genes on the long arm of chromosome 7H, analysing the gene expression by qPCR and relative protein levels by mass spectroscopy. All three genes are expressed specifically in seeds with no detectable expression in the leaves. The barley WSPNs share a common gene structure with a single exon containing an open reading frame in the size 253–313 bp, with the first 80 bp predicted by TargetP 2.0 to encode a sorting peptide. Furthermore, they all contain the conserved motif C–P-X-G-X4-C-X(4–8)-C-X-C. Structure prediction with Alphafold 2.0 suggest that this motif is a structurally conserved microdomain consisting of two antiparallel strands where the cysteines align.

Variability in Chromosome 1 of Select Moroccan Pyrenophora teres f. teres Isolates Overcomes a Highly Effective Barley Chromosome 6H Source of Resistance.

Barley net form net blotch (NFNB) is a destructive foliar disease caused by Pyrenophora teres f. teres. Barley line CIho5791, which harbors the broadly effective chromosome 6H resistance gene Rpt5, displays dominant resistance to P. teres f. teres. To genetically characterize P. teres f. teres avirulence/virulence on the barley line CIho5791, we generated a P. teres f. teres mapping population using a cross between the Moroccan CIho5791-virulent isolate MorSM40-3 and the avirulent reference isolate 0-1. Full genome sequences were generated for 103 progenies. Saturated chromosome-level genetic maps were generated, and quantitative trait locus (QTL) mapping identified two major QTL associated with P. teres f. teres avirulence/virulence on CIho5791. The most significant QTL mapped to chromosome (Ch) 1, where the virulent allele was contributed by MorSM40-3. A second QTL mapped to Ch8; however, this virulent allele was contributed by the avirulent parent 0-1. The Ch1 and Ch8 loci accounted for 27 and 15% of the disease variation, respectively, and the avirulent allele at the Ch1 locus was epistatic over the virulent allele at the Ch8 locus. As a validation, we used a natural P. teres f. teres population in a genome-wide association study that identified the same Ch1 and Ch8 loci. We then generated a new reference quality genome assembly of parental isolate MorSM40-3 with annotation supported by deep transcriptome sequencing of infection time points. The annotation identified candidate genes predicted to encode small, secreted proteins, one or more of which are likely responsible for overcoming the CIho5791 resistance. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 "No Rights Reserved" license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2024.

A Quantitative Trait Locus with a Major Effect on Root-Lesion Nematode Resistance in Barley.

Although the root-lesion nematode Pratylenchus thornei is known to affect barley (Hordeum vulgare L.), there have been no reports on the genetic control of P. thornei resistance in barley. In this research, P. thornei resistance was assessed for a panel of 46 barley mapping parents and for two mapping populations (Arapiles/Franklin and Denar/Baudin). With both populations, a highly significant quantitative trait locus (QTL) was mapped at the same position on the long arm of chromosome 7H. Single-nucleotide polymorphisms (SNPs) in this region were anchored to an RGT Planet pan-genome assembly and assayed on the mapping parents and other barley varieties. The results indicate that Arapiles, Denar, RGT Planet and several other varieties likely have the same resistance gene on chromosome 7H. Marker assays reported here could be used to select for P. thornei resistance in barley breeding. Analysis of existing barley pan-genomic and pan-transcriptomic data provided a list of candidate genes along with information on the expression and differential expression of some of those genes in barley root tissue. Further research is required to identify a specific barley gene that affects root-lesion nematode resistance.

QTL Analysis of β-Glucan Content and Other Grain Traits in a Recombinant Population of Spring Barley.

Barley with high grain β-glucan content is valuable for functional foods. The identification of loci for high β-glucan content is, thus, of great importance for barley breeding. Segregation mapping for the content in β-glucan and other barley grain components (starch, protein, lipid, ash, phosphorous, calcium, sodium) was performed using the progeny of the cross between Glacier AC38, a mutant with high amylose, and CDC Fibar, a high β-glucan waxy cultivar. The offspring of this cross showed transgressive segregation for β-glucan content. Linkage analysis based on single-nucleotide polymorphism (SNP) molecular markers was used for the genotyping of the parents and recombinant inbred lines (RILs). Two Quantitative Trait Loci (QTL) for β-glucan content and several QTL for other grain components were found. The former ones, located on chromosomes 1H and 7H, explained 27.9% and 27.4% of the phenotypic variance, respectively. Glacier AC38 provided the allele for high β-glucan content at the QTL on chromosome 1H, whereas CDC Fibar contributed the allele at the QTL on chromosome 7H. Their recombination resulted in a novel haplotype with higher β-glucan content, up to 18.4%. Candidate genes are proposed for these two QTL: HvCslF9, involved in β-glucan biosynthesis, for the QTL on chromosome 1H; Horvu_PLANET_7H01G069300, a gene encoding an ATP-Binding Cassette (ABC) transporter, for the QTL on chromosome 7H.

Mapping of quantitative trait loci for agronomic and morpho‐physiological traits under drought environments in spring barley (Hordeum vulgare L.)

AbstractBarley production is severely affected by drought caused by the unpredictable Mediterranean weather patterns, which include uneven rainfall and extreme temperatures. This leads to a decrease in crop yield. However, to tackle this issue, landraces and wild species are crucial sources of variation for stress adaptive traits. By incorporating these traits into improved varieties, we may see an increase in yield and stability under drought conditions. Seventy‐six quantitative traits loci (QTLs) identified traits were mapped using recombinant inbred lines (RIL) population Arta × Harmal‐2//Esp/1808‐4L, evaluated at six dry and semi‐dry areas over 3 years. The study investigated traits such as grain yield, biological yield, harvest index, kernel weight, seed per head, days to heading, kernel filling duration, growth vigour, growth habit, lodging and plant height. Numerous QTLs were discovered that are associated with various phenotypic traits related to grain yield, kernel yield, duration of filling period and days to heading. For areas with less than 250 mm/annum of rainfall, QTLs were identified on chromosome 2H for biological yield, days to heading, and kernel weight, on 1H for harvest index, and on 2H, 4H, and 5H for kernel weight. For semi‐dry areas with rainfall less than 450 mm, QTLs were found on chromosome 6H for grain yield, 2H and 5H for kernel weight, 1H and 6H for seed per head, and 2H for days to heading. Notably, these QTLs significantly explain more than 10% of phenotypic variation. The 2H chromosome was found to have the most important QTL and pleiotropic effect for yield and its components, such as kernel weight, days to heading, and biological yield. The cross Arta/Harmal was adapted, and mechanisms were developed to cope with drought stress, reflected by the significant and positive correlation of biological yield and harvest index with grain yield. Chromosomes 1H, 2H, 4H and 5H harbour more than 60% of mapped QTLs for dry areas. It is worth noting that the QTLs mentioned earlier, along with the kernel weight QTLs (QKW 1.5, QKW2.7b, QKW4.1, QKW6.7, QKW6.9), have consistently exhibited positive effects on crop yield in semi‐dry and dry areas, making them potential candidates for breeding drought‐tolerant crops. Genomic co‐localisation of the QTL for Arta/Harmal population suggested that selection for drought through linked markers can be an option for drought tolerance selection for barley in dry areas.

Registration of the “Woodies” multi–rust‐resistant barley germplasm

AbstractSelection for resistance to plant diseases is a continuous effort on the part of plant breeders. Sources of genetic resistance are often limited, despite considerable discovery efforts. Stem rust and stripe rust are two diseases of particular importance in barley (Hordeum vulgare L.) production. The present work aims to develop and deploy genotypes with resistance to these diseases that can be used in future breeding efforts. The Woodies, Woody‐1 (DH160733; Reg. no. GP‐218, PI 704479) and Woody‐2 (DH160754; Reg. no. GP‐219, PI 704480), are two doubled‐haploid genotypes produced via F1 anther culture named in honor of the late Lynn “Woody” Gallagher. These two‐row spring habit barley germplasm accessions were released by the Oregon Agricultural Experiment Station in 2023. These genotypes have demonstrated resistance to both stem and stripe rust at the seedling and adult plant stage in trials conducted between 2018 and 2023. The genetic basis of this resistance appears to be a novel quantitative trait locus conferring resistance to both diseases on chromosome 5H that is different from the known rpg4/Rpg5 complex for stem rust resistance found on the same chromosome. Seed can be requested from the Oregon State University Barley Breeding Program or from the NLGRP Germplasm repository.

Genome-wide association study on seed dormancy in barley

Abstract Seed dormancy is an important trait associated with pre-sprouting and malting quality in barley. Genome-wide association studies (GWASs) have been used to detect quantitative trait loci (QTLs) underlying complex traits in major crops. In the present study, we collected 295 barley (Hordeum vulgare L.) accessions from Australia, Europe, Canada and China. A total of 25,179 single nucleotide polymorphism (SNP)/diversity arrays technology sequence markers were used for population structure, linkage disequilibrium and GWAS analysis. Candidate genes within QTL regions were investigated, and their expression levels were analysed using RNAseq data. Five QTLs for seed dormancy were identified. One QTL was mapped on chromosome 1H, and one QTL was mapped on chromosome 4H, while three QTLs were located on chromosome 5H. This is the first report of a QTL on the short arm of chromosome 5H in barley. Molecular markers linked to the QTL can be used for marker-assisted selection in barley breeding programmes.

Genome‐wide association study of adult plant resistance to spot blotch in an elite Canadian two‐row barley germplasm collection

AbstractSpot blotch, caused by Bipolaris sorokiniana, is a growing concern for barley (Hordeum vulgare) production in most humid, temperate growing regions of the world. Under epidemic conditions, this disease can reduce yields significantly. Reduction of kernel plumpness is associated with lower malt extract and consequently grain quality. Enhanced resistance to this disease is a major goal of western Canadian two‐row barley breeding programmes; however, two‐row barley is considered to be more susceptible than six‐row barley. Plant–pathogen interactions observed for Canadian germplasm are polygenic, where quantitative breeding methods could be beneficial for the development of resistant germplasm. A germplasm panel consisting of 200 two‐row, spring barley genotypes with differential reaction to spot blotch was evaluated at Brandon, Manitoba (MB), and Melfort, Saskatchewan (SK), over eight site years. Genome‐wide association study was conducted using a 50k single‐nucleotide polymorphism (SNP) Illumina iSelect genotyping array. The most‐resistant germplasm was found to carry Midwest Six‐rowed Durable Resistant Haplotype (MSDRH) alleles at the loci located on chromosomes 1H (Rcs‐qtl‐1H‐11_10764), 3H (Rcs‐qtl‐3H‐11_10565) and 7H (Rcs‐qtl‐7H‐11_20162, Rcs5 locus), where the 3H locus was found to be the most under‐represented in Canadian germplasm. Additional marker‐trait associations within nurseries were identified on chromosomes 1H, 2H and 4H. Several of the SNP markers identified were found to be polymorphic within the Canadian two‐row germplasm panel and thus could be useful for enhancing spot blotch resistance in two‐row barley for development of resistant cultivars.

Genome-Wide Association Study of Host Resistance to Hessian Fly in Barley.

The Hessian fly (HF), Mayetiola destructor (Diptera: Cecidomyiidae), is one of the most devastating insect pests of cereals including wheat, barley, and rye. Although wheat is the preferred host for HF, this continuously evolving pest has been emerging as a threat to barley production. However, characterization and identification of genetic resistance to HF has not been conducted in barley. In the present study, we used a genome-wide association study (GWAS) to identify barley resistance loci to HF using a geographically diverse set of 234 barley accessions. The results showed that around 90% of barley lines were highly susceptible, indicating a significant vulnerability to HF in barley, and a total of 29 accessions were resistant, serving as potential resistance resources. GWAS with a mixed linear model revealed two marker-trait associations, both on chromosome 4H. The resistance loci and associated markers will facilitate barley improvement and development for breeders. In addition, our results are fundamental for genetic studies to understand the HF resistance mechanism in barley.

The Hessian fly resistance gene HvRHF1 is localized in an NBS-LRR gene cluster in barley.

Hessian fly (Mayetiola destructor Say) is a significant pest in cereal crops, causing substantial yield losses worldwide. While host resistance is the most efficient method for pest control, research on genetic characterization of Hessian fly resistance in barley (Hordeum vulgare L.) has been limited, and the underlying resistance mechanism remains largely unknown. In this study, we conducted fine mapping of a crucial Hessian fly resistance locus, known as HvRHF1, using a biparental population. Assisted with genetic markers and robust phenotyping assay, we pinpointed the HvRHF1 gene to an ~ 82kb region on chromosome 4H. Gene prediction and annotation revealed that the HvRHF1 locus comprises three complete NBS-LRR genes, which are characteristic of disease resistance genes. As a result, our study not only provides valuable resources for resistance in barley and genetic tools for breeding, but also identifies candidate genes that lay the foundation for cloning HvRHF1. This endeavor will significantly contribute to our understanding of the molecular mechanisms underlying cereal resistance to Hessian fly.

High-resolution mapping of Ryd4Hb, a major resistance gene to Barley yellow dwarf virus from Hordeum bulbosum

Key messageWe mapped Ryd4Hb in a 66.5 kbp interval in barley and dissociated it from a sublethality factor. These results will enable a targeted selection of the resistance in barley breeding.Virus diseases are causing high yield losses in crops worldwide. The Barley yellow dwarf virus (BYDV) complex is responsible for one of the most widespread and economically important viral diseases of cereals. While no gene conferring complete resistance (immunity) has been uncovered in the primary gene pool of barley, sources of resistance were searched and identified in the wild relative Hordeum bulbosum, representing the secondary gene pool of barley. One such locus, Ryd4Hb, has been previously introgressed into barley, and was allocated to chromosome 3H, but is tightly linked to a sublethality factor that prevents the incorporation and utilization of Ryd4Hb in barley varieties. To solve this problem, we fine-mapped Ryd4Hb and separated it from this negative factor. We narrowed the Ryd4Hb locus to a corresponding 66.5 kbp physical interval in the barley ‘Morex’ reference genome. The region comprises a gene from the nucleotide-binding and leucine-rich repeat immune receptor family, typical of dominant virus resistance genes. The closest homolog to this Ryd4Hb candidate gene is the wheat Sr35 stem rust resistance gene. In addition to the fine mapping, we reduced the interval bearing the sublethality factor to 600 kbp in barley. Aphid feeding experiments demonstrated that Ryd4Hb provides a resistance to BYDV rather than to its vector. The presented results, including the high-throughput molecular markers, will permit a more targeted selection of the resistance in breeding, enabling the use of Ryd4Hb in barley varieties.

Identification of a novel and plant height-independent QTL for coleoptile length in barley and validation of its effect using near isogenic lines

Key messageThis study reported the identification and validation of novel QTL conferring coleoptile length in barley and predicted candidate genes underlying the largest effect QTL based on orthologous analysis and comparison of the whole genome assemblies for both parental genotypes of the mapping population.Coleoptile length (CL) is one of the most important agronomic traits in cereal crops due to its direct influence on the optimal depth for seed sowing which facilitates better seedling establishment. Varieties with longer coleoptiles are preferred in drought-prone areas where less moisture maintains at the top layer of the soil. Compared to wheat, genetic study on coleoptile length is limited in barley. Here, we reported a study on detecting the genomic regions associated with CL in barley by assessing a population consisting of 201 recombinant inbred lines. Four putative QTL conferring CL were consistently identified on chromosomes 1H, 5H, 6H, and 7H in each of the trials conducted. Of these QTL, the two located on chromosomes 5H and 6H (designated as Qcl.caf-5H and Qcl.caf-6H) are likely novel and Qcl.caf-5H showed the most significant effect explaining up to 30.9% of phenotypic variance with a LOD value of 15.1. To further validate the effect of this putative QTL, five pairs of near isogenic lines (NILs) were then developed and assessed. Analysis of the NILs showed an average difference of 21.0% in CL between the two isolines. Notably, none of the other assessed morphological characteristics showed consistent differences between the two isolines for each pair of the NILs. Candidate genes underlying the Qcl.caf-5H locus were also predicted by employing orthologous analysis and comparing the genome assemblies for both parental genotypes of the mapping population in the present study. Taken together, these findings expand our understanding on genetic basis of CL and will be indicative for further gene cloning and functional analysis underly this locus in barley.

A wild barley nested association mapping population shows a wide variation for yield-associated traits to be used for breeding in Australian environment

This study aimed to identify wild barley alleles controlling grain size and weight with the potential to improve barley yield in Australia and worldwide. The HEB-25 nested association mapping population was used, which samples 25 different wild barley accessions in a ‘Barke’ genetic background. The HEB-25 population was evaluated in field conditions at Strathalbyn in South Australia in 2015 and 2016. Seven yield component traits reflecting ear length, grain number per ear and grain dimension were measured. Among 114 quantitative trait loci (QTL) identified for the seven traits in both years, many co-localise with known genes controlling flowering and spike morphology. There were 18 QTL hotspots associated with four loci or more, of which one at the beginning of chromosome 5H had wild alleles that increased both grain number per ear and thousand-grain weight. A wide range of effects was found for wild alleles for each trait across all QTL identified, providing a rich source of genetic diversity that barley breeders can exploit to enhance barley yield.