Chromosome 2H Research Articles

The draft genome of a wild barley genotype reveals its enrichment in genes related to biotic and abiotic stresses compared to cultivated barley.

SummaryWild barley (Hordeum spontaneum) is the progenitor of cultivated barley (Hordeum vulgare) and provides a rich source of genetic variations for barley improvement. Currently, the genome sequences of wild barley and its differences with cultivated barley remain unclear. In this study, we report a high‐quality draft assembly of wild barley accession (AWCS276; henceforth named as WB1), which consists of 4.28 Gb genome and 36 395 high‐confidence protein‐coding genes. BUSCO analysis revealed that the assembly included full lengths of 95.3% of the 956 single‐copy plant genes, illustrating that the gene‐containing regions have been well assembled. By comparing with the genome of the cultivated genotype Morex, it is inferred that the WB1 genome contains more genes involved in resistance and tolerance to biotic and abiotic stresses. The presence of the numerous WB1‐specific genes indicates that, in addition to enhance allele diversity for genes already existing in the cultigen, exploiting the wild barley taxon in breeding should also allow the incorporation of novel genes. Furthermore, high levels of genetic variation in the pericentromeric regions were detected in chromosomes 3H and 5H between the wild and cultivated genotypes, which may be the results of domestication. This H. spontaneum draft genome assembly will help to accelerate wild barley research and be an invaluable resource for barley improvement and comparative genomics research.

Identifying a novel powdery mildew resistance gene in a barley landrace from Morocco

Powdery mildew is a barley foliar disease that causes great loss in yield. Because of the limited number of effective resistance genes, efforts to identify new sources of resistance are frequently focused on genetically diversified landraces. The goal of this study was to characterise the powdery mildew resistance gene in barley line 2553-3 selected from the Moroccan landrace. Phytopathological testing against a set of differential pathogen isolates revealed different pattern responses of this gene from those of other known resistance genes. F2 and F2:3 (2553-3 × Manchuria) mapping populations were employed to investigate resistance inheritance. Two approaches were applied for the linkage analysis: in the first approach, 22 resistant and 21 susceptible homozygous F2 plants genotyped by the DArTseq platform (Diversity Arrays Technology, Pty. Ltd.) were used; in the second, 94 F2 plants were genotyped by converted DArTseq markers and SSRs. Both analyses delineated a new resistance gene on the short arm of chromosome 2H. The authors propose MlMor as a gene symbol for newly characterized powdery mildew resistance genes in barley line 255-3-3. The results presented herein provide a good foundation for the development of closer linkage markers and MAS breeding.

Mapping of agronomic traits, disease resistance and malting quality in a wide cross of two-row barley cultivars.

Wide crosses between genetically diverged parents may reveal novel loci for crop improvement that are not apparent in crosses between elite cultivars. The landrace Chevallier was a noted malting barley first grown in 1820. To identify potentially novel alleles for agronomic traits, Chevallier was crossed with the modern malting cultivar NFC Tipple generating two genetically diverse recombinant inbred line populations. Genetic maps were produced using genotyping-by-sequencing and 384-SNP genotyping, and the populations were phenotyped for agronomic traits to allow the identification of quantitative trait loci (QTL). Within the semi-dwarf 1 (sdw1) region on chromosome 3H Chevallier conferred increased plant height and reduced tiller number, with QTL for these traits explaining 79.4% and 35.2% of the phenotypic variance observed, respectively. Chevallier was also associated with powdery mildew susceptibility, with a QTL on 1H accounting for up to 19.1% of the variance and resistance at this locus most likely resulting from an Mla variant from Tipple. Two novel QTL for physiological leaf spotting were identified on 3H and 7H, explaining up to 17.1% of the variance and with the Chevallier allele reducing symptom severity on 7H. Preliminary micromalting analysis was also undertaken to compare the malting characteristics of Chevallier and Tipple. Chevallier malt contained significantly lower levels of both α-amylase and wort β-glucan than Tipple malt, however no significant differences were observed for the remaining malting parameters measured. This suggests that the most obvious improvements in barley since the introduction of Chevallier are for agronomic traits such as height, yield and lodging resistance rather than for malting characteristics. Overall, our results demonstrate that this wide cross between Chevallier and Tipple may provide a source of novel QTL for barley breeding.

The search for candidate genes associated with natural variation of grain Zn accumulation in barley.

Combating hidden hunger through molecular breeding of nutritionally enriched crops requires a better understanding of micronutrient accumulation. We studied natural variation in grain micronutrient accumulation in barley (Hordeum vulgare L.) and searched for candidate genes by assessing marker-trait associations (MTAs) and by analyzing transcriptional differences between low and high zinc (Zn) accumulating cultivars during grain filling. A collection of 180 barley lines was grown in three different environments. Our results show a pronounced variation in Zn accumulation, which was under strong genotype influence across different environments. Genome-wide association mapping revealed 13 shared MTAs. Across three environments, the most significantly associated marker was on chromosome 2H at 82.8 cM and in close vicinity to two yellow stripe like (YSL) genes. A subset of two pairs of lines with contrasting Zn accumulation was chosen for detailed analysis. Whole ears and flag leaves were analyzed 15 days after pollination to detect transcriptional differences associated with elevated Zn concentrations in the grain. A putative α-amylase/trypsin inhibitor CMb precursor was decidedly higher expressed in high Zn cultivars in whole ears in all comparisons. Additionally, a gene similar to barley metal tolerance protein 5 (MTP5) was found to be a potential candidate gene.

Deciphering the genetic basis for vitamin E accumulation in leaves and grains of different barley accessions

Tocopherols and tocotrienols, commonly referred to as vitamin E, are essential compounds in food and feed. Due to their lipophilic nature they protect biomembranes by preventing the propagation of lipid-peroxidation especially during oxidative stress. Since their synthesis is restricted to photosynthetic organisms, plant-derived products are the major source of natural vitamin E. In the present study the genetic basis for high vitamin E accumulation in leaves and grains of different barley (Hordeum vulgare L.) accessions was uncovered. A genome wide association study (GWAS) allowed the identification of two genes located on chromosome 7H, homogentisate phytyltransferase (HPT-7H) and homogentisate geranylgeranyltransferase (HGGT) that code for key enzymes controlling the accumulation of tocopherols in leaves and tocotrienols in grains, respectively. Transcript profiling showed a correlation between HPT-7H expression and vitamin E content in leaves. Allele sequencing allowed to decipher the allelic variation of HPT-7H and HGGT genes corresponding to high and low vitamin E contents in the respective tissues. Using the obtained sequence information molecular markers have been developed which can be used to assist smart breeding of high vitamin E barley varieties. This will facilitate the selection of genotypes more tolerant to oxidative stress and producing high-quality grains.

Genome‐Wide Association Mapping of Seedling Net Form Net Blotch Resistance in an Ethiopian and Eritrean Barley Collection

ABSTRACTThe net form of net blotch, caused by Pyrenophora teres f. teres (Ptt), is an economically important disease of barley (Hordeum vulgare L.) that is best managed by breeding for resistance. Previous linkage mapping and genome‐wide association studies (GWAS) have identified many resistance quantitative trait loci (QTL) in barley. Here we report eight novel resistance QTL identified in a unique collection of barley landraces from Ethiopia and Eritrea (EEBC) using a GWAS approach. Lines from the EEBC were genotyped with the Illumina barley iSelect chip and phenotyped with four Ptt isolates. A GWAS was conducted using the efficient mixed model association (EMMA) method with principal components and kinship to control false positives. Using a significance threshold of false discovery rate (FDR) of 0.05, two marker–trait associations (MTAs) were found on chromosomes 6H and 5H, respectively. Since this is a stringent method, markers in the lower 0.1 percentile of the p value distribution were also considered significant, identifying 20 unique MTAs. The 20 MTAs corresponded to 15 unique genomic loci, based on analysis of local linkage disequilibrium. Of the 15 QTL identified, one was identified in chromosome 1H, six in 2H, three each in 3H and 5H, and two in 6H. Eight of these QTL appear to be novel and thus could be of value to barley breeding programs to introgress novel net blotch resistance.

Coincidence of QTLs determining foliar phosphorus fractionation patterns and phosphorus utilization efficiency in barley under low phosphorus stress

Plants can optimize the allocation of phosphorus (P) among their foliar P fractions to increase the P utilization efficiency (PUE). Identifying the genetic relationships between foliar P fractionation and PUE could provide opportunities to improve the P efficiency of barley (Hordeum vulgare L.). The differences in the concentrations and proportions of inorganic P, ester P, nucleic acid P and insoluble P between a wild barley cultivar CN4027 and a commercial cultivar Baudin were studied, and their quantitative trait loci (QTLs) at normal P (NP) and low P (LP) fertilisations were mapped. The PUE that was determined in the previous study was used to analyze the relationship between PUE and foliar P fractionation in this research. Both cultivars of barley could increase their metabolic P fractions as an LP stress tolerance strategy to ensure their continued metabolic activities in response to LP stress. Cultivar CN4027 showed higher nucleic acid P concentration (NPC), nucleic acid P proportion (NPP) and insoluble P proportion (IPP) than cultivar Baudin under LP stress. This abundant organic P (Po) pool of CN4027 ensured the normal functioning of its metabolic pathways under LP stress. The close relationships between the foliar P fractionation and PUE could be explained by two QTL clusters, Cl-3H.02 and Cl-5H.01. The QTL cluster Cl-3H.02 is flanked by the markers bPb3256099-bPb3255630 on chromosome 3H and controls the ester P concentration (EPC), ester P proportion (EPP), insoluble P concentration (IPC) and IPP. The QTL cluster Cl-3H.02 might have great potential for the future genetic improvement of barley PUE and may offer clues for the genetic relationships between the foliar P fractionation and PUE.

Characterization of genes required for both Rpg1 and rpg4-mediated wheat stem rust resistance in barley

BackgroundPuccinia graminis f. sp. tritici (Pgt) race TTKSK and its lineage pose a threat to barley production world-wide justifying the extensive efforts to identify, clone, and characterize the rpg4-mediated resistance locus (RMRL), the only effective resistance to virulent Pgt races in the TTKSK lineage. The RMRL contains two nucleotide-binding domain and leucine-rich repeat (NLR) resistance genes, Rpg5 and HvRga1, which are required for resistance. The two NLRs have head-to-head genome architecture with one NLR, Rpg5, containing an integrated C-terminal protein kinase domain, characteristic of an “integrated sensory domain” resistance mechanism. Fast neutron mutagenesis of line Q21861 was utilized in a forward genetics approach to identify genetic components that function in the RMRL or Rpg1 resistance mechanisms, as Q21861 contains both genes. A mutant was identified that compromises both RMRL and Rpg1-mediated resistances and had stunted seedling roots, designated required for P. graminis resistance 9 (rpr9).ResultsThe rpr9 mutant generated in the Q21861 background was crossed with the Swiss landrace Hv584, which carries RMRL but contains polymorphism across the genome compared to Q21861. To map Rpr9, a Hv584 x rpr9 F6:7 recombinant inbred line (RIL) population was developed. The RIL population was phenotyped with Pgt race QCCJB. The Hv584 x rpr9 RIL population was genotyped with the 9 k Illumina Infinium iSelect marker panel, producing 2701 polymorphic markers. A robust genetic map consisting of 563 noncosegregating markers was generated and used to map Rpr9 to an ~ 3.4 cM region on barley chromosome 3H. The NimbleGen barley exome capture array was utilized to capture rpr9 and wild type Q21861 exons, followed by Illumina sequencing. Comparative analysis, resulting in the identification of a 1.05 Mbp deletion at the chromosome 3H rpr9 locus. The identified deletion contains ten high confidence annotated genes with the best rpr9 candidates encoding a SKP1-like 9 protein and a F-box family protein.ConclusionGenetic mapping and exome capture rapidly identified candidate gene/s that function in RMRL and Rpg1 mediated resistance pathway/s. One or more of the identified candidate rpr9 genes are essential in the only two known effective stem rust resistance mechanisms, present in domesticated barley.

Host/nonhost status and genetics of resistance in barley against three pathotypes of Magnaporthe blast fungi

Blast disease, caused by the Magnaporthe oryzae/grisea species complex, occurs in a wide range of wild and cultivated gramineous plant species including rice, wheat and barley. We inoculated a collection of cultivated (Hordeum vulgare ssp. vulgare L.) and wild (ssp. spontaneum) barley accessions with M. oryzae Oryza pathotype (MoO), Triticum pathotype (MoT) and Pennisetum pathotype (MsP) to quantify the host status of barley, and to identify sources of blast resistance. Unlike wheat, the barley gene pool is rich with sources of complete and partial resistance against Magnaporthe. Cultivated barley appeared a nonhost to MsP, whereas wild barley showed some degree of susceptibility. All 153 tested rice accessions were resistant to the MoT isolate, suggesting that rice is nonhost to this pathotype. Inoculation of L94/Vada and Vada/SusPtrit RIL mapping populations with MoO and/or MoT isolates of M. oryzae indicated one large effect QTL, designated as Rmoq1, on the short arm of chromosome 7H against the MoT isolate PY 67.1 in both populations. Resistance in L94 to the MoO isolate was due to a different QTL, located at 5H. An association mapping panel of West European barley cultivars also suggested that most QTLs were pathotype specific. Six blast resistance genes found in the biparental and association mapping studies coincided with map positions of powdery mildew resistance genes viz. Mlt, Mla6, Mlg, mlo, Mlj, and Mlhb genes. Our QTL and association mapping analyses do not support the association of the mlo resistance gene with enhanced susceptibility to M. oryzae as reported in literature.

Genome-Wide Identification, Characterization and Expression Analysis of Xyloglucan Endotransglucosylase/Hydrolase Genes Family in Barley (Hordeum vulgare).

Xyloglucan endotransglucosylase/hydrolases (XTHs)—a family of xyloglucan modifying enzymes—play an essential role in the construction and restructuring of xyloglucan cross-links. However, no comprehensive study has been performed on this gene family in barley. A total of 24 HvXTH genes (named HvXTH1-24) and an EG16 member were identified using the recently completed genomic database of barley (Hordeum vulgare). Phylogenetic analysis showed that 24 HvXTH genes could be classified into three phylogenetic groups: (I/II, III-A and III-B) and HvXTH15 was in the ancestral group. All HvXTH protein members—except HvXTH15—had a conserved N-glycosylation site. The genomic location of HvXTHs on barley chromosomes showed that the 24 genes are unevenly distributed on the 7 chromosomes, with 10 of them specifically located on chromosome 7H. A structure-based sequence alignment demonstrates that each XTH possesses a highly conserved domain (ExDxE) responsible for catalytic activity. Expression profiles based on the barley genome database showed that HvXTH family members display different expression patterns in different tissues and at different stages. This study is the first systematic genomic analysis of the barley HvXTH gene family. Our results provide valuable information that will help to elucidate the roles of HvXTH genes in the growth and development of barley.

ASSOCIATION MAPPING OF MORPHOLOGICAL AND PHYSIOLOGICAL TRAITS OF FLAG LEAF RELATED TO DROUGHT TOLERANCE IN BARLEY

Association mapping has proven to be a powerful approach for dissecting the genetic basis of complex traits. In this study, QTLs controlling flag leaf characteristics under drought stress were detected in a set of 148 modern spring barley cultivars using AM analysis. Flag leaf length (FLL), flag leaf width (FLW), relative water content (RWC), chlorophyll content, and maximum quantum efficiency of PSII (Fv/Fm) which are important in photosynthetic rate, were evaluated under normal irrigation and drought stress conditions at grain filling stage. Population structure was estimated using Structure 2.3 and linkage disequilibrium (LD) was estimated by the ‘Full Matrix LD’ using Tassel 5.0 . Significant marker/trait associations were investigated based on K-Q matrix using Tassel 3.0 . The analysis of population structure divided the cultivars into two sub-groups. Significant LD values (P < 0.01) between polymorphic sites with regions of high and low LD were observed. A total of 84 significant putative genomic regions were identified, which delineated into 37 QTLs under two water treatments. Two stable QTLs on 2H and 3H were detected for FLL in drought stress treatment. A QTL for FLL were detected on 2H in normal treatment, which alone explained around 11% of phenotypic variance of FLL. This QTL was also associated with the expression of FLW and explained around 7.5% of phenotypic variance. The results suggest that major loci are located on chromosomes 2H, 3H, 4H and 5H involved in the development of flag leaf characteristics and could be used as selection criteria in barley breeding for drought tolerance.

Desynaptic5 carries a spontaneous semi-dominant mutation affecting Disrupted Meiotic cDNA 1 in barley.

Despite conservation of the process of meiosis, recombination landscapes vary between species, with large genome grasses such as barley (Hordeum vulgare L.) exhibiting a pattern of recombination that is very heavily skewed to the ends of chromosomes. We have been using a collection of semi-sterile desynaptic meiotic mutant lines to help elucidate how recombination is controlled in barley and the role of the corresponding wild-type (WT) meiotic genes within this process. Here we applied a combination of genetic segregation analysis, cytogenetics, and immunocytology to genetically map and characterize the meiotic mutant desynaptic5 (des5). We identified an exonic insertion in the positional candidate ortholog of Disrupted Meiotic cDNA 1 (HvDMC1) on chromosome 5H of des5. des5 exhibits a severe meiotic phenotype with disturbed synapsis, reduced crossovers, and chromosome mis-segregation. The meiotic phenotype and reduced fertility of des5 is similarly observed in Hvdmc1RNAi transgenic plants and HvDMC1p:GusPlus reporter lines show DMC1 expression specifically in the developing inflorescence. The des5 mutation maintains the reading frame of the gene and exhibits semi-dominance with respect to recombination in the heterozygote indicating the value of non-knockout mutations for dissection of the control of recombination in the early stages of meiosis.

High-Density Mapping of Triple Rust Resistance in Barley Using DArT-Seq Markers.

The recent availability of an assembled and annotated genome reference sequence for the diploid crop barley (Hordeum vulgare L.) provides new opportunities to study the genetic basis of agronomically important traits such as resistance to stripe [Puccinia striiformis f. sp. hordei (Psh)], leaf [P. hordei (Ph)], and stem [P. graminis f. sp. tritici (Pgt)] rust diseases. The European barley cultivar Pompadour is known to possess high levels of resistance to leaf rust, predominantly due to adult plant resistance (APR) gene Rph20. We developed a barley recombinant inbred line (RIL) population from a cross between Pompadour and the leaf rust and stripe rust susceptible selection Biosaline-19 (B-19), and genotyped this population using DArT-Seq genotyping by sequencing (GBS) markers. In the current study, we produced a high-density linkage map comprising 8,610 (SNP and in silico) markers spanning 5957.6 cM, with the aim of mapping loci for resistance to leaf rust, stem rust, and stripe rust. The RIL population was phenotyped in the field with Psh (Mexico and Ecuador) and Ph (Australia) and in the greenhouse at the seedling stage with Australian Ph and Pgt races, and at Wageningen University with a European variant of Psh race 24 (PshWUR). For Psh, we identified a consistent field QTL on chromosome 2H across all South American field sites and years. Two complementary resistance genes were mapped to chromosomes 1H and 4H at the seedling stage in response to PshWUR, likely to be the loci rpsEm1 and rpsEm2 previously reported from the cultivar Emir from which Pompadour was bred. For leaf rust, we determined that Rph20 in addition to two minor-effect QTL on 1H and 3H were effective at the seedling stage, whilst seedling resistance to stem rust was due to QTL on chromosomes 3H and 7H conferred by Pompadour and B-19, respectively.

Dissecting the Genetic Basis of Grain Size and Weight in Barley (Hordeum vulgare L.) by QTL and Comparative Genetic Analyses.

Grain size and weight are crucial components of barley yield and quality and are the target characteristics of domestication and modern breeding. Despite this, little is known about the genetic and molecular mechanisms of grain size and weight in barley. Here, we evaluated nine traits determining grain size and weight, including thousand grain weight (Tgw), grain length (Gl), grain width (Gw), grain length-width ratio (Lwr), grain area (Ga), grain perimeter (Gp), grain diameter (Gd), grain roundness (Gr), and factor form density (Ffd), in a double haploid (DH) population for three consecutive years. Using five mapping methods, we successfully identified 60 reliable QTLs and 27 hotspot regions that distributed on all chromosomes except 6H which controls the nine traits of grain size and weight. Moreover, we also identified 164 barley orthologs of 112 grain size/weight genes from rice, maize, wheat and 38 barley genes that affect grain yield. A total of 45 barley genes or orthologs were identified as potential candidate genes for barley grain size and weight, including 12, 20, 9, and 4 genes or orthologs for barley, rice, maize, and wheat, respectively. Importantly, 20 of them were located in the 14 QTL hotspot regions on chromosome 1H, 2H, 3H, 5H, and 7H, which controls barley grain size and weight. These results indicated that grain size/weight genes of other cereal species might have the same or similar functions in barley. Our findings provide new insights into the understanding of the genetic basis of grain size and weight in barley, and new information to facilitate high-yield breeding in barley. The function of these potential candidate genes identified in this study are worth exploring and studying in detail.

Genome-wide association of barley plant growth under drought stress using a nested association mapping population

BackgroundBarley (Hordeum vulgare L.) is the fourth most important cereal crop worldwide. Barley production is compromised by many abiotic stresses including drought. Wild barley is a valuable source of alleles that can improve adaptation of cultivated barley to drought stress.ResultsIn the present study, a nested association mapping population named HEB-25, consisting of 1420 BC1S3 lines that were developed by crossing 25 different wild barley accessions to the elite barley cultivar ‘Barke’, was evaluated under both control and drought-stressed conditions in the Australian Plant Phenomics Facility, University of Adelaide. Overall, 14 traits reflecting the performance of individual plants in each treatment were calculated from non-destructive imaging over time and destructive end-of-experiment measurements. For each trait, best linear unbiased estimators (BLUEs) were calculated and used for genome-wide association study (GWAS) analysis. Among the quantitative trait loci (QTL) identified for the 14 traits, many co-localise with known inflorescence and developmental genes. We identified a QTL on chromosome 4H where, under drought and control conditions, wild barley alleles increased biomass by 10 and 17% respectively compared to the Barke allele.ConclusionsAcross all traits, QTL which increased phenotypic values were identified, providing a wider range of genetic diversity for the improvement of drought tolerance in barley.

Harnessing Novel Diversity From Landraces to Improve an Elite Barley Variety.

The Spanish Barley Core Collection (SBCC) is a source of genetic variability of potential interest for breeding, particularly for adaptation to Mediterranean environments. Two backcross populations (BC2F5) were developed using the elite cultivar Cierzo as the recurrent parent. The donor parents, namely SBCC042 and SBCC073, were selected from the SBCC lines due to their outstanding yield in drought environments. Flowering time, yield and drought-related traits were evaluated in two field trials in Zaragoza (Spain) during the 2014–15 and 2015–16 seasons and validated in the 2017–18 season. Two hundred sixty-four lines of each population were genotyped with the Barley Illumina iSelect 50k SNP chip. Genetic maps for each population were generated. The map for SBCC042 × Cierzo contains 12,893 SNPs distributed in 9 linkage groups. The map for SBCC073 × Cierzo includes 12,026 SNPs in 7 linkage groups. Both populations shared two QTL hotspots. There are QTLs for flowering time, thousand-kernel weight (TKW), and hectoliter weight on a segment of 23 Mb at ~515 Mb on chromosome 1H, which encompasses the HvFT3 gene. In both populations, flowering was accelerated by the landrace allele, which also increased the TKW. In the same region, better soil coverage was contributed by SBCC042 but coincident with a lower hectoliter weight. The second large hotspot was on chromosome 6H and contained QTLs with wide intervals for grain yield, plant height and TKW. Landrace alleles contributed to increased plant height and TKW and reduced grain yield. Only SBCC042 contributed favorable alleles for “green area,” with three significant QTLs that increased ground coverage after winter, which might be exploited as an adaptive trait of this landrace. Some genes of interest found in or very close to the peaks of the QTLs are highlighted. Strategies to deploy the QTLs found for breeding and pre-breeding are proposed.

HvPAA1 Encodes a P-Type ATPase, a Novel Gene for Cadmium Accumulation and Tolerance in Barley (Hordeum vulgare L.).

The identification of gene(s) that are involved in Cd accumulation/tolerance is vital in developing crop cultivars with low Cd accumulation. We developed a doubled haploid (DH) population that was derived from a cross of Suyinmai 2 (Cd-sensitive) × Weisuobuzhi (Cd-tolerant) to conduct quantitative trait loci (QTL) mapping studies. We assessed chlorophyll content, traits that are associated with development, metal concentration, and antioxidative enzyme activity in DH population lines and parents under control and Cd stress conditions. A single QTL, designated as qShCd7H, was identified on chromosome 7H that was linked to shoot Cd concentration; qShCd7H explained 17% of the phenotypic variation. Comparative genomics, map-based cloning, and gene silencing were used in isolation, cloning, and functional characterization of the candidate gene. A novel gene HvPAA1, being related to shoot Cd concentration, was identified from qShCd7H. Sequence comparison indicated that HvPAA1 carried seven domains with an N-glycosylation motif. HvPAA1 is predominantly expressed in shoots. Subcellular localization verified that HvPAA1 is located in plasma membrane. The silencing of HvPAA1 resulted in growth inhibition, greater Cd accumulation, and a significant decrease in Cd tolerance. We conclude HvPAA1 is a novel plasma membrane-localized ATPase that contributes to Cd tolerance and accumulation in barley. The results provide us with new insights that may aid in the screening and development of Cd-tolerant and low-Cd-accumulation crops.

Composition and random elimination of paternal chromosomes in a large population of wheat\u2009\xd7\u2009barley (Triticum aestivum L. \xd7 Hordeum vulgare L.) hybrids

Key messageStatistical analysis of the chromosomal composition in a population of 210 primary plants regenerated from two intergeneric wheat–barley cross combinations revealed the random nature of uniparental elimination for barley chromosomes.Uniparental chromosome elimination is a common process in interspecific and intergeneric cereal hybrids. To characterize the frequency of paternal chromosomes, a population of 218 independent green plants was generated from two wheat (♀) × barley (♂) cross combinations via embryo rescue. The chromosomal composition of 210 primary plants was analyzed with chromosome-specific DNA markers representing all seven barley chromosomes. The analysis revealed an equal proportion of haploid and full hybrids (20.5% and 19.5%, respectively), while the rest of the population contained hypoploids (partial hybrids) with no preference for any possible numbers (one to six) of barley chromosome additions. Contrary to the previous reports, there was no statistical bias or preferential elimination for any individual barley chromosome (1H–7H) in this population. The reasons for the apparent contradiction and the implications of the above findings for cereal breeding are discussed.

Physical organization of repetitive sequences and chromosome diversity of barley revealed by fluorescence in situ hybridization (FISH).

Fluorescence in situ hybridization (FISH) using oligonucleotides is a simple and convenient method for chromosome research. In this study, 34 of 46 previously developed oligonucleotides produced signals in barley. Together with two plasmid clones and one PCR-amplified cereal centromere repeat (CCS1) probe, 37 repetitive sequences were chromosomally located produced three types of signals covering different positions on the chromosomes. The centromeric and pericentric regions had a more complex genomic organization and sequence composition probably indicative of higher contents of heterochromatin. An efficient multi-plex probe containing eight oligonucleotides and a plasmid clone of 45S rDNA was developed. Thirty-three barley karyotypes were developed and compared. Among them, 11 irradiation-induced mutants of cultivar 08-49 showed no chromosomal variation, whereas 22 cultivar and landrace accessions contained 28 chromosomal polymorphisms. Chromosome 4H was the most variable and 6H was the least variable based on chromosome polymorphic information content (CPIC). Five polymorphic chromosomes (1H-2, 2H-1, 3H-3, 5H-2, and 6H-2) were dominant types, each occurring in more than 50% of accessions. The multi-plex probe should facilitate identification of further chromosomal polymorphisms in barley.

Mapping of barley susceptibility/resistance QTL against spot form net blotch caused by Pyrenophora teres f. maculata using RIL populations.

Spot form net blotch (SFNB) caused by the necrotrophic fungal pathogen Pyrenophora teres f. maculata (Ptm) is an important disease of barley worldwide including the major barley production regions of North America. To characterize SFNB resistance/susceptibility quantitative trait loci (QTL), three recombinant inbred line (RIL) populations were developed from crosses between the malting barley cultivars, Tradition (six row) and Pinnacle (two row), and the two world barley core collection lines, PI67381 and PI84314. Tradition and Pinnacle were susceptible to many North American Ptm isolates, while PI67381 and PI84314 carry resistances to diverse Ptm isolates from across the globe. The RIL populations, Tradition/PI67381, Pinnacle/PI67381, and Pinnacle/PI84314 were genotyped using polymerase chain reaction-mediated genotype-by-sequencing single nucleotide polymorphism marker panels and phenotyped at the seedling stage with six geographically distinct Ptm isolates: FGOB10Ptm-1 (North Dakota, USA), Pin-A14 (Montana, USA), Cel-A17 (Montana, USA), SG1 (Australia), NZKF2 (New Zealand) and DEN2.6 (Denmark). The goal was to determine if the susceptible elite lines contained common susceptibility genes/QTL or if the resistant lines had common resistant genes/QTL effective against diverse Ptm isolates. The QTL analyses identified a total of 12 resistance and/or susceptibility loci on chromosomes 2H, 3H, 4H, 6H, and 7H of which three had not been previously reported. Common major QTL were detected on chromosome 2H (R2 = 14-40%) and 7H (R2 = 24-80%) in all three RIL populations, suggesting underlying genes with broad resistance specificity. The major 7H QTL was shown to be a dominant susceptibility gene in both susceptible malting barley varieties.