Barley Spikes Research Articles

Restoration of the spike architectonics in ancient barley excavated at the twelfth-century settlement of Usvyaty

Background. The data are presented on the architectonics of ancient barley spikes from the 12th century, excavated in 2019 at Usvyaty Settlement. Modern molecular genetics approaches were used to study domestication genes (Btr1, Btr2, and Vrs) in ancient and contemporary barleys (germplasm accessions preserved at VIR).Materials and methods. The carbonized kernels found by archaeologists during the excavations at Usvyaty were analyzed. Primers for domestication genes were designed, and PCR was performed on contemporary and ancient barley grains. Ancient kernels were studied in accordance with the rules established for organizing a paleogenetics laboratory, which excluded any contamination with contemporary DNA. Fragments of domestication genes from contemporary and ancient barley grain samples underwent Sanger sequencing. Results. Ancient DNA was isolated and enriched. The analysis of domestication gene sequences made it possible to reconstruct the ancient barley spike’s features. Conclusion. The ancient cereal crop architectonics was restored to ascertain a brittle two-row spike of ancient barley

Development and Validation of a TaqMan Multiplex Real-Time PCR Assay for High-Throughput Quantification of Fusarium graminearum Biomass in Barley Spikes and Grains

Fusarium head blight (FHB) is a devastating disease primarily caused by Fusarium graminearum that commonly affects North American small grains. Although the disease causes visible symptoms, the more pressing concern is the contamination of grain with trichothecene toxins, especially deoxynivalenol (DON), which can cause toxicity in humans and animals by inhibiting protein biosynthesis. The DON levels are more critical when breeding for resistance, but this does not account for pathogen infection and disease severity. To link DON and Fusarium infection, we optimized and validated a real-time multiplex quantitative PCR protocol for high-throughput quantification of Fusarium biomass based on the Tri5 gene, which encodes trichodiene synthase, the first enzyme involved in trichothecene toxin production. The primers and probes were designed to measure the abundance of Tri5 and normalized to the barley actin gene. The assay is repeatable, robust, and sensitive, up to 0.003 ng/µl of F. graminearum and 0.3 ng/µl of barley DNA. The method is cost-effective and has the potential to quantify Fusarium biomass, monitor disease progression, and determine DON levels in barley spikes and harvested grains. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

A transcriptional atlas identifies key regulators and networks for the development of spike tissues in barley

Grain number and size determine grain yield in crops and are closely associated with spikelet fertility and grain filling in barley (Hordeum vulgare). Abortion of spikelet primordia within individual barley spikes causes a 30%-50% loss in the potential number of grains during development from the awn primordium stage to the tipping stage, after that grain filling is the primary factor regulating grain size. To identify transcriptional signatures associated with spike development, we use a six-rowed barley cultivar (Morex) to develop a spatiotemporal transcriptome atlas containing 255 samples covering 17 stages and 5 positions along the spike. We identify several fundamental regulatory networks, in addition to key regulators of spike development and morphology. Specifically, we show HvGELP96, encoding a GDSL domain-containing protein, as a regulator of spikelet fertility and grain number. Our transcriptional atlas offers a powerful resource to answer fundamental questions in spikelet development and degeneration in barley.

Detection and characterization of spike architecture based on deep learning and X-ray computed tomography in barley

BackgroundSpike is the grain-bearing organ in cereal crops, which is a key proxy indicator determining the grain yield and quality. Machine learning methods for image analysis of spike-related phenotypic traits not only hold the promise for high-throughput estimating grain production and quality, but also lay the foundation for better dissection of the genetic basis for spike development. Barley (Hordeum vulgare L.) is one of the most important crops globally, ranking as the fourth largest cereal crop in terms of cultivated area and total yield. However, image analysis of spike-related traits in barley, especially based on CT-scanning, remains elusive at present.ResultsIn this study, we developed a non-invasive, high-throughput approach to quantitatively measuring the multitude of spike architectural traits in barley through combining X-ray computed tomography (CT) and a deep learning model (UNet). Firstly, the spikes of 11 barley accessions, including 2 wild barley, 3 landraces and 6 cultivars were used for X-ray CT scanning to obtain the tomographic images. And then, an optimized 3D image processing method was used to point cloud data to generate the 3D point cloud images of spike, namely ‘virtual’ spike, which is then used to investigate internal structures and morphological traits of barley spikes. Furthermore, the virtual spike-related traits, such as spike length, grain number per spike, grain volume, grain surface area, grain length and grain width as well as grain thickness were efficiently and non-destructively quantified. The virtual values of these traits were highly consistent with the actual value using manual measurement, demonstrating the accuracy and reliability of the developed model. The reconstruction process took 15 min approximately, 10 min for CT scanning and 5 min for imaging and features extraction, respectively.ConclusionsThis study provides an efficient, non-invasive and useful tool for dissecting barley spike architecture, which will contribute to high-throughput phenotyping and breeding for high yield in barley and other crops.

Positional cloning identified HvTUBULIN8 as the candidate gene for round lateral spikelet (RLS) in barley (Hordeum vulgare L.)

Key messageMap-based cloning, subcellular localization, virus-induced-gene-silencing and transcriptomic analysis reveal HvTUB8 as a candidate gene with pleiotropic effects on barley spike and leaf development via ethylene and chlorophyll metabolism.Barley lateral spikelet morphology and grain shape play key roles in grain physical quality and yield. Several genes and QTLs for these traits have been cloned or fine mapped previously. Here, we report the phenotypic and genotypic analysis of a barley mutant with round lateral spikelet (rls) from cv. Edamai 934. rls had round lateral spikelet, short but round grain, shortened awn, thick glume and dark green leaves. Histocytologic and ultrastructural analysis revealed that the difference of grain shape of rls was caused by change of cell arrangement in glume, and the dark leaf color resulted from enlarged chloroplast. HvTUBULIN8 (HvTUB8) was identified as the candidate gene for rls by combination of RNA-Seq, map-based-cloning, virus-induced-gene-silencing (VIGS) and protein subcellular location. A single G-A substitution at the third exon of HvTUB8 resulted in change of Cysteine 354 to tyrosine. Furthermore, the mutant isoform Hvtub8 could be detected in both nucleus and cytoplasm, whereas the wild-type protein was only in cytoplasm and granular organelles of wheat protoplasts. Being consistent with the rare phenotype, the “A” allele of HvTUB8 was only detected in rls, but not in a worldwide barley germplasm panel with 400 accessions. VIGS confirmed that HvTUB8 was essential to maintain spike integrity. RNA-Seq results suggested that HvTUB8 may control spike morphogenesis via ethylene homeostasis and signaling, and control leaf color through chlorophyll metabolism. Collectively, our results support HvTUB8 as a candidate gene for barley spike and leaf morphology and provide insight of a novel mechanism of it in barley development.

Genome-Wide Analysis of Snf2 Gene Family Reveals Potential Role in Regulation of Spike Development in Barley

Sucrose nonfermenting 2 (Snf2) family proteins, as the catalytic core of ATP-dependent chromatin remodeling complexes, play important roles in nuclear processes as diverse as DNA replication, transcriptional regulation, and DNA repair and recombination. The Snf2 gene family has been characterized in several plant species; some of its members regulate flower development in Arabidopsis. However, little is known about the members of the family in barley (Hordeum vulgare). Here, 38 Snf2 genes unevenly distributed among seven chromosomes were identified from the barley (cv. Morex) genome. Phylogenetic analysis categorized them into 18 subfamilies. They contained combinations of 21 domains and consisted of 3 to 34 exons. Evolution analysis revealed that segmental duplication contributed predominantly to the expansion of the family in barley, and the duplicated gene pairs have undergone purifying selection. About eight hundred Snf2 family genes were identified from 20 barley accessions, ranging from 38 to 41 genes in each. Most of these genes were subjected to purification selection during barley domestication. Most were expressed abundantly during spike development. This study provides a comprehensive characterization of barley Snf2 family members, which should help to improve our understanding of their potential regulatory roles in barley spike development.

Spikelet abortion in six-rowed barley is mainly influenced by final spikelet number, with potential spikelet number acting as a suppressor trait.

The potential to increase barley grain yield lies in the indeterminate nature of its inflorescence meristem, which produces spikelets, the basic reproductive unit in grasses that are linked to reproductive success. During early reproductive growth, barley spikes pass through the maximum yield potential-a stage after which no new spikelet ridges are produced. Subsequently, spikelet abortion (SA), a phenomenon in which spikelets abort during spike growth, imposes a bottleneck for increasing the grain yield potential. Here, we studied the potential of main culm spikes by counting potential spikelet number (PSN) and final spikelet number (FSN), and computed the corresponding SA (%) in a panel of 417 six-rowed spring barleys. Our phenotypic data analyses showed a significantly large within- and across-years genotypic variation with high broad-sense heritability estimates for all the investigated traits, including SA. Asian accessions displayed the lowest SA, indicating the presence of favourable alleles that may be exploited in breeding programs. A significantly negative Pearson's product-moment correlation was observed between FSN and SA. Our path analysis revealed that PSN and FSN explain 93% of the observed phenotypic variability for SA, with PSN behaving as a suppressor trait that magnifies the effect of FSN. Based on a large set of diverse barley accessions, our results provide a deeper phenotypic understanding of the quantitative genetic nature of SA, its association with traits of high agronomic importance, and a resource for further genetic analyses.

Analysis of barley mutants ert-c.1 and ert-d.7 reveals two loci with additive effect on plant architecture

Main conclusionBoth mutant ert-c.1 and ert-d.7 carry T2-T3 translocations in the Ert-c gene. Principal coordinate analyses revealed the translocation types and translocation breakpoints. Mutant ert-d.7 is an Ert-cErt-d double mutant.Mutations in the Ert-c and Ert-d loci are among the most common barley mutations affecting plant architecture. The mutants have various degrees of erect and compact spikes, often accompanied with short and stiff culms. In the current study, complementation tests, linkage mapping, principal coordinate analyses and fine mapping were conducted. We conclude that the original ert-d.7 mutant does not only carry an ert-d mutation but also an ert-c mutation. Combined, mutations in Ert-c and Ert-d cause a pyramid-dense spike phenotype, whereas mutations in only Ert-c or Ert-d give a pyramid and dense phenotype, respectively. Associations between the Ert-c gene and T2-T3 translocations were detected in both mutant ert-c.1 and ert-d.7. Different genetic association patterns indicate different translocation breakpoints in these two mutants. Principal coordinate analysis based on genetic distance and screening of recombinants from all four ends of polymorphic regions was an efficient way to narrow down the region of interest in translocation-involved populations. The Ert-c gene was mapped to the marker interval of 2_0801to1_0224 on 3HL near the centromere. The results illuminate a complex connection between two single genes having additive effects on barley spike architecture and will facilitate the identification of the Ert-c and Ert-d genes.

Fusarium tricinctum associated with Head Blight on barley in Argentina: pathogenicity and potential degradation of the different hordein fractions

The objective of the present work was to evaluate the pathogenicity of Fusarium tricinctum in barley and to determine the potential effects on grain quality, more specifically regarding the hordein fractions. For this purpose, Koch´s postulates were carried out to evaluate the pathogenicity of F. tricinctum in barley, under greenhouse conditions. Furthermore, the ability of F. tricinctum to degrade hordeins was tested by a sequential extraction method, and separated by polyacrilamide gel electrophoresis (SDS-PAGE). Finally, the relative abundance of each hordein fractions (D, C, and B-hordeins) was measured. The results demonstrated that F. tricinctum was pathogenic when barley spikes were inoculated, also showing that F. tricinctum could degrade barley C-hordeins by 21.80% of the total C-hordein content.

Use of spike moisture content to define physiological maturity and quantify progress through grain development in wheat and barley

A single measurement is useful for determining how far a crop has progressed through grain development, and whether it has reached physiological maturity. Grain development is commonly assessed by using subjective, qualitative methods that describe the look and feel of the kernel or the colour of the straw. Physiological maturity in cereal crops can be determined more accurately by the grain moisture content; however, the moisture content of whole spikes is potentially quicker and easier to assess than that of individual kernels, and with a greater degree of accuracy. This experiment aimed to characterise the moisture dynamics of whole intact wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) spikes during the grain development phase and identify the spike moisture content corresponding to physiological maturity for both species. The dry matter and water contents of whole spikes of five wheat and five barley cultivars sown over six dates were determined at weekly intervals throughout the period of grain development from anthesis to harvest ripeness. Use of regression analysis determined that the spike moisture content at physiological maturity was 43% (95% confidence interval 41–45%) for wheat and 50% (95% confidence interval 49–51%) for barley, irrespective of differences in cultivar morphology, phenology and growing conditions. We demonstrate that progression through kernel development in wheat and barley can be assessed objectively and quantitatively by using spike moisture content, and we provide guidelines for accurate determination of the grain development stage using spike moisture.

Assessing frost damage in barley using terahertz imaging.

Frost is estimated to cost Australian grain growers $ 360 million in direct and indirect losses every year. Assessing frost damage manually in barley is labor intensive and involves destructive sampling. To mitigate against significant economic loss, it is crucial that assessment decisions on whether to cut for hay or continue to harvest are made soon after frost damage has occurred. In this paper, we propose a non-destructive technique by using raster-scan terahertz imaging. Terahertz waves can penetrate the spike to determine differences between frosted and unfrosted grains. With terahertz raster-scan imaging, conducted in both transmission and reflection at 275 GHz, frosted and unfrosted barley spikes show significant differences. In addition, terahertz imaging allows to determine individual grain positions. The emergence of compact terahertz sources and cameras would enable field deployment of terahertz non-destructive inspection for early frost damage.

Genetic analysis reveals four interacting loci underlying awn trait diversity in barley (Hordeum vulgare)

Barley (Hordeum vulgare) awns contribute to grain yield, but the genetic basis of awn development remains largely unclear. Five barley lines differing in awn traits and row types were used to create four F2 populations. Genetic analyses revealed that four pairs of genes were involved in awn development: A/a (awnless/awned), B/b (awnless/awned), H/h (hooded/straight), and L/l (long/short). Of these four loci, A, H and L functioned on both central rows (CR) and lateral rows (LR) of the barley spikes, while B exhibited effect only on LR. A and B had duplicate effects on LR, and both showed dominant epistasis to loci H and L, whereas H was epistatic to L. Meanwhile, A and B were found to be genetically linked, with a row-type locus V located between them. The genetic distances of A-V and B-V were estimated to be 9.6 and 7.7 cM, respectively. Literature search suggested that A, H and V may correspond to the reported Lks1, Kap1 and Vrs1, respectively, whereas B is a novel gene specifically controlling awn development on LR, designated as Lsa1 for lateral spikelet awnless 1. The only barley homolog of wheat awn inhibitor gene B1, HORVU2Hr1G077570, is a potential candidate of Lsa1.

Genome-wide characterization and expression analyses of the auxin/indole-3-acetic acid (Aux/IAA) gene family in barley (Hordeum vulgare L.)

Aux/IAA genes are early auxin-responsive genes and essential for auxin signaling transduction. There is little information about Aux/IAAs in the agriculturally important cereal, barley. Using in silico method, we identified and subsequently characterized 36 Aux/IAAs from the barley genome. Based on their genomic sequences and the phylogenic relationship with Arabidopsis and rice Aux/IAA, the 36 HvIAAs were categorized into two major groups and 14 subgroups. The indication of the presence or absence of these domains for the biological functions and acting mechanisms was discussed. The cis-element distributions in HvIAA promoters suggests that the HvIAAs expressions may not only regulated by auxin (the presence of AuxREs and TGA-element) but also by other hormones and developmental and environmental cues. We then studied the HvIAAs expression in response to NAA (1-Naphthaleneacetic acid) using quantitative real-time PCR (qRT-PCR). Like the promoter analysis, only 14 HvIAAs were upregulated by NAA over two-fold at 4 h. HvIAAs were clustered into three groups based on the spatiotemporal expression data. We confirmed by qRT-PCR that most HvIAAs, especially HvIAA3, HvIAA7, HvIAA8, HvIAA18, HvIAA24 and HvIAA34, are expressed in the developing barley spike compared within seedling, suggesting their roles in regulating spike development. Taken together, our data provide a foundation for further revealing the biological function of these HvIAAs.

Construction of a high-density genetic map and fine mapping of a candidate gene locus for a novel branched-spike mutant in barley

A Yunnan branched-spike (Ynbs) barley mutant is useful for study of the genetic mechanisms underlying variation in barley spike architecture. In the current study, a mutant (Ynbs-1), a recombinant inbred line (RIL-1), and a cultivar (BDM-8) were used as parents to develop populations. Ynbs-1 exhibits typical branched spike, whereas the others exhibit six-row spike. Genetic analysis on their F1, F2 and F3 populations showed that one recessive gene is responsible for the branched spike trait. SLAF marker generated from specific locus amplified fragment sequencing (SLAF-seq) was used to genotype the populations. A high-density genetic map of barley was constructed using 14,348 SLAF markers, which covered all 7 chromosomes at 1,347.44 cM in length with an average marker density of 0.09 cM between adjacent markers. Linkage analysis of the branched-spike trait using the genetic map indicated that branched spike trait in the Ynbs-1 is controlled by single locus on chromosome 2H at the interval between 65.00 and 65.47 cM that is flanked by Marker310119 and Marker2679451. Several candidate genes that may be responsible for barley multiple-spikelet degeneration, single-floret spikelet increase and seed set rate decrease were identified in the region. The high-density genetic map and the gene locus revealed in this study provide valuable information for elucidating the genetic mechanism of spike branching in barley.

First Report of Aspergillus parasiticus on Barley Grain in Serbia

Barley (Hordeum vulgare L.) is a secondary grain crop in Serbia used commercially for animal feed, seed, and human food applications. The production of barley in the 2016 to 2017 growing season reached a record yield of almost 400,000 metric tons (USDA 2017). Aspergillus contamination has been rare in the agroecological conditions of cereal-growing areas in Serbia. Changes in climatic factors, such as occurrence of high temperatures and prolonged droughts, increased frequency of Aspergillus spp. Species Aspergillus parasiticus was isolated from maize grain for the first time in Serbia in 2012 and from wheat grains in 2017 (Nikolic et al. 2018). We hypothesized that these pathogens can also be present in barley fields in Serbia. Barley spikes exhibiting bleaching were sampled at the beginning of June 2017 grown in northern Serbia. In severe infections, barley spikes get a dry look with awns that stand upright and firm. The incidence of the disease of the bleached spiked in the field was 15 to 20%. From each representative sample, 100 shriveled grains were collected. After surface sterilization with bleach/distilled water 1:3, 100 grains per sample (10 per Petri dish) were placed on potato dextrose agar and incubated at 25°C for 7 days. After isolation, 50% of isolates were identified as Alternaria spp., 20% as Fusarium spp., approximately 15% as Aspergillus spp. In order to reliably identify individual species of fungi, the fragments of colonies were transferred to malt extract agar (MEA) and Czapek yeast agar (CYA) and incubated in the dark for 7 days. The fungal colonies were dark green. The reverse side was pale yellow. The average colony diameter was 65 mm. Conidia were spherical and rough with thick walls. The average size of conidia was 5.1 µm. Colonies were floccose and thin on MEA and CYA. Based on growth and morphological characteristics, isolates were determined as A. parasiticus (Pitt and Hocking 2009). Molecular detection of Aspergillus species was done by using PCR-restriction fragment length polymorphism analysis of the aflR-aflJ (genes for aflatoxin biosynthesis) intergenic spacer. The restriction enzyme BglII was able to cut the PCR product of A. parasiticus at one restriction site, resulting in two fragments of 363 and 311 bp (El Khoury et al. 2011). A. parasiticus CBS 100926 was used as a reference isolate. The pathogenicity of 20 isolates was verified on a group of 20 randomly selected spikes in four replicates (Mesterhazy et al. 1999). A 7-day-old culture of each isolate was used for the preparation of the spore suspension (1 × 10⁶ spores/ml). Inoculation was carried out after 50% of plants reached the anthesis stage. Groups of 20 selected spikes were sprayed from all sides with 20 ml of fungal spore suspension. Control spikes were inoculated by applying an equal amount of sterile distilled water. The infection rate was estimated after 3 weeks on a 1 to 7 scale, with 1 = 0 to 5%, 2 = 5 to 15%, 3 = 15 to 30%, 4 = 30 to 50%, 5 = 50 to 75%, 6 = 75 to 90%, and 7 = 90 to 100% infected spike area. The average infection rate was 3.2. The pathogen was reisolated from the inoculated spikes and identified as A. parasiticus, with the aim to confirm Koch’s postulates. Developed symptoms were similar to those observed on spikes collected from the field. Control spikes did not show any symptoms of the disease. These results confirmed the pathogenicity of A. parasiticus on H. vulgare. To the best of our knowledge, this is the first report of the occurrence of A. parasiticus on barley grain in Serbia. Because A. parasiticus is known to be a severe aflatoxin producer and climatic changes can increase the frequency of this fungus, further studies are necessary to improve strategies for food safety and quality.

Spike developmental stages and ABA role in spikelet primordia abortion contribute to the final yield in barley (Hordeum vulgare L.)

BackgroundSalinity is a significant environmental stress factor limiting crops productivity. Barley (Hordeum vulgare L.) has a natural tolerance to salinity stress, making it an interesting study object in stress biology research. In the present study, for the first time the effect of salinity stress on barley inflorescence developmental stages was investigated. Five spring barley genotypes irrigated with saline water (12.5 ds/m NaCl) were compared to controls treated with normal tap water. We measured abscisic acid (ABA) concentrations in the apical, central and basal sections of the immature inflorescence at green anther (GA) stage. The role of ABA in spikelet primordia development, atrophy and abortion and final yield was evaluated.ResultsA time course experiment starting from double ridge until green anther (GA) stages revealed that salinity reduced the length of spike developmental stages in all genotypes causing shortened of the plant life cycle. The shortened plant life cycle negatively affected plant height and number of tillers/plant. Salinity also affected spikelet primordia development. In both control and salinity treated plants apical spikelet abortion started in late awn primordium (AP) stage. However, under salinity treatment, significantly more spikelets were aborted, thus directly affecting plant yield potential. ABA, which plays a role in the spikelet/floret abortion process, was markedly elevated in the base and apex of salt treated spikes correlating with an increased spikelet abortion in these regions.ConclusionsOverall, salinity treatment reduced all plant and yield-related parameters investigated and turned some of the correlations among them from positive to negative or vice versa. Investigations of ABA role in floral development and phase duration of barley spike showed that, ABA regulates the spikelet/floret abortion process affecting the yield potential under salinity and control conditions.

Crosstalk among hormones in barley spike contributes to the yield

Key messageThe hormonal ratios along the barley spike regulate the development, atrophy and abortion of the spikelets and could be the mechanism by which the barley spike adapts its yield potential.Barley (Hordeum vulgare L.) is one of the oldest cereal crops known to be cultivated since about 10,000 years. The inflorescence of cultivated barley is an indeterminate spike that produces three single-flowered spikelets at each rachis node which make it unique among the grasses. The yield production in barley is predominantly controlled by very important parameters such as number of tillers and number of spikelets per spike. These two parameters are negatively correlated. Therefore, studying the biological and genetics of the spikelet development during the spike developmental stages is essential for breeding programs. Here we summarize our current understanding of the crosstalk between hormones such as auxin, cytokinin, gibberellin and abscisic acid along the spike and what is their role in regulating spike and spikelet development in barley. We conclude that the hormonal ratios at the apical, central, and basal sections of the spike not only regulate the spike developmental stages, but also the development, atrophy, and abortion of the spikelets. This hormonal dependent modification of the grain number along the spike could be the mechanism by which the barley spike adapts its yield potential.

The dependence of barley spike length on the effects of foliar fertilization by micro fertilizers

The dependence of barley spike length on the effects of foliar fertilization by micro fertilizers

The dependence of the grains number in spring barley spikes on the effect of mineral fertilization

The dependence of the grains number in spring barley spikes on the effect of mineral fertilization

Marker-trait associations in two-rowed spring barley accessions from Kazakhstan and the USA

In this study, phenotyping and single nucleotide polymorphism (SNP) genotyping data of 272 accessions of two-rowed spring barley from the USA along with 94 accessions from Kazakhstan were assessed in field trials at six breeding organizations in Kazakhstan to evaluate the performance of the USA samples over three years (2009–2011). The average grain yield over the six locations was not significantly higher in Kazakh accessions in comparison to the USA samples. Twenty four samples from Montana, Washington, the USDA station in Aberdeen Idaho, and the Anheuser-Busch breeding programs showed heavier average yield than the local standard cultivar “Ubagan”. Principal Coordinate analysis based on two sets of SNP data suggested that Kazakh accessions were closest to the USA accessions among eight groups of samples from different parts of the World, and within five US barley origin groups the samples from Montana and Washington perfectly matched six groups of Kazakh breeding origins. A genome-wide association study (GWAS) using data from eighteen field trials allowed the identification of ninety one marker-trait associations (MTA) in two or more environments for nine traits, including key characters such as heading time (HT), number of kernels per spike (NKS), and thousand grain weight (TGW). Our GWAS allowed the identification of eight MTA for HT and NKS, and sixteen MTA for TGW, when those MTA were linked to mapped SNPs. Based on comparisons of chromosomal positions of MTA identified in this study, and positions of known genes and quantitative trait loci for HT, NKS and TGW, it was suggested that MTA for HT on chromosome 2H (at 158.2 cM, 11_21414), MTA for NKS on 5H (at 118.6 cM, 11_20298), and two MTA for TGW on chromosome 4H (at 94.7 cM, 12_30718, and at 129.3 cM, 11_20013) were potentially new associations in barley. GWAS suggested that six MTA for HT, including two on chromosome 1H, two on chromosome 3H, and one each on chromosomes 4H and 6H, had useful pleiotropic effects for improving barley spike traits.