Wild Barley Research Articles

Repetitive DNA is Functional and Encodes Parts of the Non-Coding RNA Repertoire.

This is a commentary on the article by Eviatar Nevo and Kexin Li entitled "Sympatric Speciation in Mole Rats and Wild Barley and Their Genome Repeatome Evolution: A Commentary", published recently in Advanced Genetics.

Epichloë bromicola from wild barley improves salt-tolerance of cultivated barley by altering physiological responses to salt stress.

Epichloë bromicola is a cultivable fungal endophyte that lives in symbiosis with wild barley (Hordeum brevisubulatum) to which it confers salt tolerance. This study tested the hypothesis that E. bromicola derived from wild barley has the potential to increase salt tolerance in cultivated barley under salt stress. To test this hypothesis, the growth response, physiological parameters, and metabolic profiles of barley plants inoculated with E. bromicola (E+) and those not inoculated with E. bromicola (E-) were compared under salt stress. Compared with E- barley plants, E+ barley plants had significantly increased plant height, shoot biomass, total biomass, chlorophyll content, osmotic synthesis, and accumulation of stress adaptation metabolites. E. bromicola increased the salt stress tolerance of cultivated barley, and the positive effects correlated with different salt stress conditions. These results suggest that E. bromicola has promising potential for enhancing the salt tolerance of barley. New insights into the mechanisms underlying this barley-fungal endophyte association are provided, and interesting questions regarding the role of E. bromicola in fungus-enhanced tolerance to salt stress in this symbiosis are raised.

The importance of the circadian system for adaptation to heat wave stress in wild barley (Hordeum spontaneum)

Acute heat episodes (heat waves) present some of the most significant challenges for plants growing in both natural and agricultural settings. Circadian (∼24 h) systems are molecular time-pacing mechanisms responsible for synchronizing a wide range of diverse physiological processes, including abiotic and biotic stress responses. Our aim in this paper was to determine whether circadian systems may also contribute to the capacity of plants to withstand acute heat episodes. We used chlorophyll fluorescence (F) to analyze circadian rhythms and plant vitality under non-stress and heat wave conditions in a panel of wild barley (Hordeum spontaneum) accessions from diverse environmental sites across the Southern Levant. Using average linear multi-models we identified correlations between rhythm attributes, plant vitality and eco-climate environment at the sites of origin of the wild barley accessions. Under non-stress conditions, the environment explained a significant portion of the variance in period time and amplitude. By contrast, in acute heat stress conditions while there was a shortening of period and decrease in amplitude, circadian traits did not directly correlate with eco-climate origins. However, plant vitality showed significant correlations with circadian plasticity in response to increasing temperatures. Unexpectedly, optimal circadian traits for heat stress resilience were not primarily associated with barley accessions from any particular climate, suggesting that thermotolerance capacity for consistently higher temperatures is not necessarily correlated with the capacity to withstand heat wave stress.

Genome architecture and diverged selection shaping pattern of genomic differentiation in wild barley.

Divergent selection of populations in contrasting environments leads to functional genomic divergence. However, the genomic architecture underlying heterogeneous genomic differentiation remains poorly understood. Here, we de novo assembled two high-quality wild barley (Hordeum spontaneum K. Koch) genomes and examined genomic differentiation and gene expression patterns under abiotic stress in two populations. These two populations had a shared ancestry and originated in close geographic proximity but experienced different selective pressures due to their contrasting micro-environments. We identified structural variants that may have played significant roles in affecting genes potentially associated with well-differentiated phenotypes such as flowering time and drought response between two wild barley genomes. Among them, a 29-bp insertion into the promoter region formed a cis-regulatory element in the HvWRKY45 gene, which may contribute to enhanced tolerance to drought. A single SNP mutation in the promoter region may influence HvCO5 expression and be putatively linked to local flowering time adaptation. We also revealed significant genomic differentiation between the two populations with ongoing gene flow. Our results indicate that SNPs and small SVs link to genetic differentiation at the gene level through local adaptation and are maintained through divergent selection. In contrast, large chromosome inversions may have shaped the heterogeneous pattern of genomic differentiation along the chromosomes by suppressing chromosome recombination and gene flow. Our research offers novel insights into the genomic basis underlying local adaptation and provides valuable resources for the genetic improvement of cultivated barley.

Identification of Genomic Loci Controlling Grain Macro and Micronutrient Variation in a Wild Barley (Hordeum vulgare spp. spontaneum) Diversity Panel

Nutrient deficiencies in humans are problematic on a global scale but are more prevalent in regions where high-quality and nutrient-dense foods are scarce. Developing nutrient-rich crops that thrive in these regions of the world would help alleviate the disparity. We leveraged the wild barley (Hordeum vulgare spp. spontaneum) Diversity Collection (WBDC) (N = 232) to characterize the variation in seed macronutrient (P, K, Ca, and Mg) and micronutrient (B, Cu, Fe, Mn, and Zn) contents found in this subspecies and to reveal chromosomal regions associated with these traits. Most micro- and macronutrients displayed variation in the WBDC and, except for boron and phosphorous, had a modest level of heritability (>0.5). Variation due to environment was significant (p < 0.001) for each element, except iron, and genotype was significant for all the tested nutrients, except boron. Thirty-seven marker–trait associations (MTAs) were detected for three (K, Ca, and Mg) of the four macro- and four (Cu, Fe, Mn, and Zn) of the five micronutrients. Several compelling candidate genes harbored within MTAs were also identified, including ABC transporters, NAC transcription factors, and bZIP transcription factors.

Compositional variances in cuticular lipids of wild and domesticated barley leaves and their impact on plant-environment interactions

One of the oldest cereal crops, barley is thought to have been domesticated ∼ 8000 years ago, in the Fertile Crescent. In this study, we explored the overlooked contribution of cuticular lipid metabolism to barley domestication by comparatively characterizing wild and domesticated barley variants. We revealed substantial phenotypic variances in plants’ overall morphology and in vegetative and reproductive tissues. Multiple microscopic approaches combined with gas chromatography-mass spectrometry (GC-MS) metabolite profiling indicated that wild barley leaves are more densely covered with epicuticular waxes compared to domesticated leaves with distinct compositions, but both variants contain a similar cuticle ultrastructure. Gene expression assays corroborated these observations showing higher transcript expression of key epicuticular wax biosynthetic genes in wild barley leaves, but similar expression patterns of cutin biosynthetic genes in leaves of both cultivars. Wild barley leaves also transpired water at higher rates apparently due to higher stomata density and conductance. Previous evidence claimed that barley leaf epicuticular waxes shape the interactions with Blumeria graminis f.sp. hordei (Bgh), the causal agent of powdery mildew in barely. However, in-vivo and in-vitro inoculation assays inferred that the disparate wax content and composition in wild and domesticated leaves had no apparent effect on Bgh pre-penetration processes. Altogether, our data provide novel insight into the compositional variances in cuticular lipids of wild and domesticated barley leaves and their impact on plant-environment interactions.

Genome-Wide Identification, Evolution and Expressional Analysis of OSCA Gene Family in Barley (Hordeum vulgare L.).

The hyperosmolality-gated calcium-permeable channel gene family (OSCA) is one kind of conserved osmosensors, playing a crucial role in maintaining ion and water homeostasis and protecting cellular stability from the damage of hypertonic stress. Although it has been systematically characterized in diverse plants, it is necessary to explore the role of the OSCA family in barley, especially its importance in regulating abiotic stress response. In this study, a total of 13 OSCA genes (HvOSCAs) were identified in barley through an in silico genome search method, which were clustered into 4 clades based on phylogenetic relationships with members in the same clade showing similar protein structures and conserved motif compositions. These HvOSCAs had many cis-regulatory elements related to various abiotic stress, such as MBS and ARE, indicating their potential roles in abiotic stress regulation. Furthermore, their expression patterns were systematically detected under diverse stresses using RNA-seq data and qRT-PCR methods. All of these 13 HvOSCAs were significantly induced by drought, cold, salt and ABA treatment, demonstrating their functions in osmotic regulation. Finally, the genetic variations of the HvOSCAs were investigated using the re-sequencing data, and their nucleotide diversity in wild barley and landrace populations were 0.4966 × 10-3 and 0.391 × 10-3, respectively, indicating that a genetic bottleneck has occurred in the OSCA family during the barley evolution process. This study evaluated the genomic organization, evolutionary relationship and genetic expression of the OSCA family in barley, which not only provides potential candidates for further functional genomic study, but also contributes to genetically improving stress tolerance in barley and other crops.

Natural diversity uncovers P5CS1 regulation and its role in drought stress tolerance and yield sustainability in barley.

Proline accumulation is one of the major responses of plants to many abiotic stresses. However, the significance of proline accumulation for drought stress tolerance remains enigmatic in crop plants. First, we examined the natural variation of the pyrolline-5-carboxylate synthase (P5CS1) among 49 barley genotypes. Allele mining identified a previously unknown allelic series that showed polymorphisms at 42 cis-elements across the P5CS1 promoter. Selected haplotypes had quantitative variation in P5CS1 gene expression and proline accumulation, putatively influenced by both abscisic acid-dependent and independent pathways under drought stress. Next, we introgressed the P5CS1 allele from a high proline accumulating wild barley accession ISR42-8 into cultivar Scarlett developing a near-isogenic line (NIL-143). NIL-143 accumulated higher proline concentrations than Scarlett under drought stress at seedling and reproductive stages. Under drought stress, NIL-143 showed less pigment damage, sustained photosynthetic health, and higher drought stress recovery compared to Scarlett. Further, the drought-induced damage to yield-related traits, mainly thousand-grain weight, was lower in NIL-143 compared with Scarlett in field conditions. Our data uncovered new variants of the P5CS1 promoter and the significance of the increased proline accumulation regulated by the P5CS1 allele of ISR42-8 in drought stress tolerance and yield stability in barley.

Genome resequencing and transcriptome profiling reveal molecular evidence of tolerance to water deficit in barley

IntroductionFrequent climate change-induced drought events are detrimental environmental stresses affecting global crop production and ecosystem health. Several efforts have facilitated crop breeding for resilient varieties to counteract stress. However, progress is hampered due to the complexity of drought tolerance; a greater variety of novel genes are required across varying environments. Tibetan annual wild barley is a unique and precious germplasm that is well adapted to abiotic stress and can provide elite genes for crop improvement in drought tolerance. ObjectivesTo identify the genetic basis and unique mechanisms for drought tolerance in Tibetan wild barley. MethodsWhole genome resequencing and comparative RNA-seq approaches were performed to identify candidate genes associated with drought tolerance via investigating the genetic diversity and transcriptional variation between cultivated and Tibetan wild barley. Bioinformatics, population genetics, and gene silencing were conducted to obtain insights into ecological adaptation in barley and functions of key genes. ResultsOver 20 million genetic variants and a total of 15,361 significantly affected genes were identified in our dataset. Combined genomic, transcriptomic, evolutionary, and experimental analyses revealed 26 water deficit resilience-associated genes in the drought-tolerant wild barley XZ5 with unique genetic variants and expression patterns. Functional prediction revealed Tibetan wild barley employs effective regulators to activate various responsive pathways with novel genes, such as Zinc-Induced Facilitator-Like 2 (HvZIFL2) and Peroxidase 11 (HvPOD11), to adapt to water deficit conditions. Gene silencing and drought tolerance evaluation in a natural barley population demonstrated that HvZIFL2 and HvPOD11 positively regulate drought tolerance in barley. ConclusionOur findings reveal functional genes that have been selected across barley’s complex history of domestication to thrive in water deficit environments. This will be useful for molecular breeding and provide new insights into drought-tolerance mechanisms in wild relatives of major cereal crops.

Sympatric Speciation in Mole Rats and Wild Barley and Their Genome Repeatome Evolution: A Commentary.

The theories of sympatric speciation (SS) and coding and noncoding (cd and ncd =repeatome) genome function are still contentious. Studies on SS in our two new models, "Evolution Canyon" and "Evolution Plateau", in Israel, divergent microclimatically and geologically-edaphically, respectively, indicated that in ecologically divergent microsites SS is a common speciation model across life from bacteria to mammals. Genomically, the intergenic ncd repeatome was and is still regarded by many biologists as "selfish," "junk," and non-functional. In contrast, it is considered by the encyclopedia of DNA elements discovery as biochemically functional and regulatory, and the transposable elements were considered earlier by Barbara McClintock as "controlling elements" of genes. Remarkably, it is found that repeated elements can statistically identify significantly, the five species of subterranean mole rats of Spalax ehrenbergi superspecies adapted to increasingly arid climatic trend southward in Israel. Moreover, it is first discovered in the SS studies in two distant taxa, subterranean mole rats and wild barley, and later also in spiny mice in Israel and subterranean zokors in China, that the noncoding repeatome is genomically mirroring the image of the protein-coding genome in divergent ecologies. It is shown that this mirroring image is statistically significant both within and between the ecologically divergent taxa supporting the hypothesis that much of the repeatome might be regulatory and selected as the protein-coding genome by the same ecological stresses.

The evolutionary patterns of barley pericentromeric chromosome regions, as shaped by linkage disequilibrium and domestication.

SUMMARYThe distribution of recombination events along large cereal chromosomes is uneven and is generally restricted to gene‐rich telomeric ends. To understand how the lack of recombination affects diversity in the large pericentromeric regions, we analysed deep exome capture data from a final panel of 815 Hordeum vulgare (barley) cultivars, landraces and wild barleys, sampled from across their eco‐geographical ranges. We defined and compared variant data across the pericentromeric and non‐pericentromeric regions, observing a clear partitioning of diversity both within and between chromosomes and germplasm groups. Dramatically reduced diversity was found in the pericentromeres of both cultivars and landraces when compared with wild barley. We observed a mixture of completely and partially differentiated single‐nucleotide polymorphisms (SNPs) between domesticated and wild gene pools, suggesting that domesticated gene pools were derived from multiple wild ancestors. Patterns of genome‐wide linkage disequilibrium, haplotype block size and number, and variant frequency within blocks showed clear contrasts among individual chromosomes and between cultivars and wild barleys. Although most cultivar chromosomes shared a single major pericentromeric haplotype, chromosome 7H clearly differentiated the two‐row and six‐row types associated with different geographical origins. Within the pericentromeric regions we identified 22 387 non‐synonymous SNPs, 92 of which were fixed for alternative alleles in cultivar versus wild accessions. Surprisingly, only 29 SNPs found exclusively in the cultivars were predicted to be ‘highly deleterious’. Overall, our data reveal an unconventional pericentromeric genetic landscape among distinct barley gene pools, with different evolutionary processes driving domestication and diversification.

Single-Nucleotide Polymorphisms in Bmy1 Intron III Alleles Conferring the Genotypic Variations in β-Amylase Activity under Drought Stress between Tibetan Wild and Cultivated Barley

β-amylase activity is related to the polymorphisms of Bmy1 intron III; however, no attention has been given to such relationships under environmental stresses such as drought. In this study, 73 cultivated barley genotypes and 52 Tibetan wild barley accessions were used to test the association between Bmy1 gene intron III polymorphisms and β-amylase activity under drought stress. Our results showed that three alleles, Bmy1.a, Bmy1.b, and Bmy1.c, existed in the examined barley genotypes. Tibetan wild barley had a higher proportion of Bmy1.b, whereas cultivated barley showed a higher proportion of Bmy1.a. Impressively, barley genotypes with Bmy1.b showed a significant increase in β-amylase activity under drought stress, compared with those with Bmy1.a or Bmy1.c, indicating that the Bmy1.b allele might provide more chances for developing barley cultivars with higher β-amylase activity under water stress than both the Bmy1.a and Bmy1.c alleles. Furthermore, the Tibetan wild barley XZ147, belonging to the Bmy1.b allele type, showed significantly higher β-amylase activity than Triumph under drought stress. This might be the result of the unique amino acid substitution M527 or the amino acid composition of R115, D165, A233, S347, and M527 of XZ147.

Genome architecture and diverged selection shaping pattern of genomic differentiation in wild barley

Genome architecture and diverged selection shaping pattern of genomic differentiation in wild barley

Cuticular transpiration is not affected by enhanced wax and cutin amounts in response to osmotic stress in barley.

The plant cuticle, which covers all aerial parts of plants in their primary developmental stage, is the major barrier against water loss from leaves. Accumulation of cutin and waxes has often been linked to drought tolerance. Here we investigated whether cutin and waxes play a role in the drought adaption of barley mimicked by osmotic stress acting on roots. We compared the cuticle properties of cultivated barley (Hordeum vulgare spp. vulgare) with wild barley (Hordeum vulgare spp. spontaneum), and tested whether wax and cutin composition or amount and cuticular transpiration could be future breeding targets for more drought-tolerant barley lines. In response to osmotic stress, accumulation of wax crystals was observed. This coincides with an increased wax and cutin gene expression and a total increase of wax and cutin amounts in leaves, which seems to be a general response triggered through root shoot signalling. Stomatal conductance decreased fast and significantly, whereas cuticular conductance remained unaffected in both wild and cultivated barley. The often-made conclusion that higher amounts of wax and cutin necessarily reduce cuticular transpiration and thus enhance drought tolerance is not always straightforward. To prevent water loss, stomatal regulation under water stress is much more important than regulation or adaptation of cuticular transpiration in response to drought.

Grain Disarticulation in Wild Wheat and Barley.

Our industrial-scale crop monocultures, which are necessary to provide grain for large-scale food and feed production, are highly vulnerable to biotic and abiotic stresses. Crop wild relatives have adapted to harsh environmental conditions over millennia; thus, they are an important source of genetic variation and crop diversification. Despite several examples where significant yield increases have been achieved through the introgression of genomic regions from wild relatives, more detailed understanding of the differences between wild and cultivated species for favorable and unfavorable traits is still required to harness these valuable resources. Recently, as an alternative to the introgression of beneficial alleles from the wild into domesticated species, a radical suggestion is to domesticate wild relatives to generate new crops. A first and critical step for the domestication of cereal wild relatives would be to prevent grain disarticulation from the inflorescence at maturity. Discovering the molecular mechanisms and understanding the network of interactions behind grain retention/disarticulation would enable the implementation of approaches to select for this character in targeted species. Brittle rachis 1 and Brittle rachis 2 are major genes responsible for grain disarticulation in the wild progenitors of wheat and barley that were the target of mutations during domestication. These two genes are only found in the Triticeae tribe and are hypothesized to have evolved by a duplication followed by neo-functionalization. Current knowledge gaps include the molecular mechanisms controlling grain retention in cereals and the genomic consequences of strong selection for this essential character.

Quantitative Proteomics Analysis Reveals Proteins Associated with High Melatonin Content in Barley Seeds under NaCl-Induced Salt Stress.

Soil salinization limits hull-less barley cultivation in the Qinghai-Tibet Plateau of China. However, some wild hull-less barley seeds accumulate high melatonin (MEL) during germination with improved salt tolerance; but the mechanism of melatonin-mediated salt tolerance in hull-less barley is not well understood at the protein level. This study investigated proteome changes resulting in high melatonin content in germinating hull-less barley seeds under high saline conditions. The proteome profiles of seed treatment with 240 mM-NaCl (N), water (H), and control (C) taken 7 days after germination were compared using the TMT-based quantitative proteomics. Our results indicate that salt stress-induced global changes in the proteomes of germinating hull-less barley seeds, altering the expression and abundance of proteins related to cell cycle and control, carbohydrate and energy metabolism, and amino acid transport and metabolism including proteins related to melatonin production. Furthermore, proteins associated with cellular redox homeostasis, osmotic stress response, and secondary metabolites derived primarily from amino acid metabolism, purine degradation, and shikimate pathways increased significantly in abundance and may contribute to the high melatonin content in seeds under salt stress. Consequently, triggering the robust response to oxidative stress occasioned by the NaCl-induced salt stress, improved seed germination and strong adaptation to salt stress.

A new major QTL for flag leaf thickness in barley (Hordeum vulgare L.)

BackgroundCarbohydrate accumulation of photosynthetic organs, mainly leaves, are the primary sources of grain yield in cereals. The flag leaf plays a vital role in seed development, which is probably the most neglected morphological characteristic during traditional selection processes.ResultsIn this experiment, four flag leaf morphological traits and seven yield-related traits were investigated in a DH population derived from a cross between a wild barley and an Australian malting barley cultivar. Flag leaf thickness (FLT) showed significantly positive correlations with grain size. Four QTL, located on chromosomes 1H, 2H, 3H, and 5H, respectively, were identified for FLT. Among them, a major QTL was located on chromosome 3H with a LOD value of 18.4 and determined 32% of the phenotypic variation. This QTL showed close links but not pleiotropism to the previously reported semi-dwarf gene sdw1 from the cultivated barley. This QTL was not reported before and the thick leaf allele from the wild barley could provide a useful source for improving grain yield through breeding.ConclusionsOur results also provided valuable evidence that source traits and sink traits in barley are tightly connected and suggest further improvement of barley yield potential with enhanced and balanced source and sink relationships by exploiting potentialities of the wild barley resources. Moreover, this study will provide a novel sight on understanding the evolution and development of leaf morphology in barley and improving barley production by rewilding for lost superior traits during plant evolution.

Yield Related Traits and Tolerance Indices to Screen Salinity Tolerant Genotypes in Cultivated and Wild Barley

Yield Related Traits and Tolerance Indices to Screen Salinity Tolerant Genotypes in Cultivated and Wild Barley

Six‐rowed wild‐growing barleys are hybrids of diverse origins

Crop-wild gene flow is common when domesticated plants and their wild relatives grow close to each other. The resultant hybrid forms appear as semi-domesticates and were sometimes considered as missing links between crops and their wild progenitors. Wild-growing barleys in Central and Eastern Asia, named Hordeum agriocrithon, show hallmark characters of both wild and domesticated forms. Their spikes disintegrate at maturity to disperse without human intervention, but bear lateral grains, which were favored by early farmers and are absent from other wild barleys. As an intermediate form, H. agriocrithon has been proposed several times as a progenitor of domesticated barley. Here, we used genome-wide marker data and whole-genome resequencing to show that all H. agriocrithon accessions of a major germplasm collection are hybrid forms that arose multiple times by admixture of diverse domesticated and wild populations. Although H. agriocrithon barleys have not played a special role in barley domestication, future analysis of the adaptative potential of bi-directional crop-wild gene flow in extant barleys may prove a fertile research field.

A Novel Mutator-Like Transposable Elements With Unusual Structure and Recent Transpositions in Barley (Hordeum vulgare).

Mutator-like transposable elements (MULEs) represent a unique superfamily of DNA transposons as they can capture host genes and cause higher frequency of mutations in some eukaryotes. Despite their essential roles in plant evolution and functional genomics, MULEs are not fully understood yet in many important crops including barley (Hordeum vulgare). In this study, we analyzed the barley genome and identified a new mutator transposon Hvu_Abermu. This transposon is present at extremely high copy number in barley and shows unusual structure as it contains three open reading frames (ORFs) including one ORF (ORF1) encoding mutator transposase protein and one ORF (ORFR) showing opposite transcriptional orientation. We identified homologous sequences of Hvu_Abermu in both monocots and dicots and grouped them into a large mutator family named Abermu. Abermu transposons from different species share significant sequence identity, but they exhibit distinct sequence structures. Unlike the transposase proteins which are highly conserved between Abermu transposons from different organisms, the ORFR-encoded proteins are quite different from distant species. Phylogenetic analysis indicated that Abermu transposons shared closer evolutionary relationships with the maize MuDR transposon than other reported MULEs. We also found phylogenetic incongruence for the Abermu transposons identified in rice and its wild species implying the possibility of horizontal transfer of transposon. Further comparison indicated that over 200 barley genes contain Abermu-related sequences. We analyzed the barley pan genomes and detected polymorphic Hvu_Abermu transposons between the sequenced 23 wild and cultivated barley genomes. Our efforts identified a novel mutator transposon and revealed its recent transposition activity, which may help to develop genetic tools for barley and other crops.