Wild Barley Accessions Research Articles

Mapping malting quality and yield characteristics in a north American two-rowed malting barley × wild barley advanced backcross population

Due to the stringent quality requirements imposed by the US malting barley industry, barley breeders have been reluctant to introduce exotic germplasm into cultivar development programs. To determine whether wild barley (Hordeum vulgare ssp. spontaneum) contains favorable alleles for yield and malting quality characteristics, we mapped quantitative trait loci (QTL) for heading date, height, lodging, yield, and nine malting parameters important to the malting and brewing industry. Traits were mapped in a wild x cultivated barley BC2-derived advanced backcross mapping population. Harrington, the recurrent parent, is a North American two-rowed malting barley cultivar, and OUH602, the donor parent, is a wild barley accession that exhibits resistance to multiple barley diseases. The 98 derived lines were grown in replicated field trials at St. Paul and Crookston, MN in 2009–2011. One to four QTL were identified for each trait, for a total of 36 QTL. Trangressive segregants for increased yield were identified and four lines had higher yield than Harrington across all environments; however, for yield QTL the OUH602 alleles decreased the trait value. Wild barley alleles had both positive and negative effects on the malting traits of diastatic power, free amino nitrogen, and soluble protein. Combined with the previously identified QTL associated with resistance to fungal diseases, this population represents a rich resource for barley breeding. To facilitate future breeding and genetics studies with this population, a set of pre-introgression lines composed of 28 BC2-derived and 6 BC3-derived lines were identified that collectively contain introgressions across the entire OUH602 genome.

Diversity of a wall-associated kinase gene in wild and cultivated barley.

Domestication of barley and other cereals was accompanied by an increase in seed size which has been ascribed to human selection, large seeds being preferred by early farmers or favoured by cultivation practices such as deep sowing. An alternative suggestion is that the increase in seed size was an indirect consequence of selection for plants with more vigorous growth. To begin to address the latter hypothesis we studied the diversity of HvWAK1, a wall-associated kinase gene involved in root proliferation, in 220 wild barley accessions and 200 domesticated landraces. A 3655-bp sequence comprising the gene and upstream region contained 69 single nucleotide polymorphisms (SNPs), one indel and four short tandem repeats. A network of 50 haplotypes revealed a complex evolutionary relationship, but with landraces largely restricted to two parts of the topology. SNPs in the HvWAK1 coding region resulted in nonsynonymous substitutions at nine positions in the translation product, but none of these changes were predicted to have a significant effect on the protein structure. In contrast, the region upstream of the coding sequence contained five SNPs that were invariant in the domesticated population, fixation of these SNPs decreasing the likelihood that the upstream of a pair of TATA boxes and transcription start sites would be used to promote transcription of HvWAK1. The sequence diversity therefore suggests that the cis-regulatory region of HvWAK1 might have been subject to selection during barley domestication. The extent of root proliferation has been linked with traits such as above-ground biomass, so selection for particular cis-regulatory variants of HvWAK1 would be consistent with the hypothesis that seed size increases during domestication were the indirect consequence of selection for plants with increased growth vigour.

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.

Stranger than fiction: the highly divergent alleles of Jekyll

Stranger than fiction: the highly divergent alleles of <i>Jekyll</i>

The highly divergent Jekyll genes, required for sexual reproduction, are lineage specific for the related grass tribes Triticeae and Bromeae.

SummaryPhylogenetically related groups of species contain lineage‐specific genes that exhibit no sequence similarity to any genes outside the lineage. We describe here that the Jekyll gene, required for sexual reproduction, exists in two much diverged allelic variants, Jek1 and Jek3. Despite low similarity, the Jek1 and Jek3 proteins share identical signal peptides, conserved cysteine positions and direct repeats. The Jek1/Jek3 sequences are located at the same chromosomal locus and inherited in a monogenic Mendelian fashion. Jek3 has a similar expression as Jek1 and complements the Jek1 function in Jek1‐deficient plants. Jek1 and Jek3 allelic variants were almost equally distributed in a collection of 485 wild and domesticated barley accessions. All domesticated barleys harboring the Jek1 allele belong to single haplotype J1‐H1 indicating a genetic bottleneck during domestication. Domesticated barleys harboring the Jek3 allele consisted of three haplotypes. Jekyll‐like sequences were found only in species of the closely related tribes Bromeae and Triticeae but not in other Poaceae. Non‐invasive magnetic resonance imaging revealed intrinsic grain structure in Triticeae and Bromeae, associated with the Jekyll function. The emergence of Jekyll suggests its role in the separation of the Bromeae and Triticeae lineages within the Poaceae and identifies the Jekyll genes as lineage‐specific.

Genetic dissection of grain elements predicted by hyperspectral imaging associated with yield-related traits in a wild barley NAM population

Enhancing the accumulation of essential mineral elements in cereal grains is of prime importance for combating human malnutrition. Biofortification by breeding holds great potential for improving nutrient accumulation in grains. However, conventional breeding approaches require element analysis of many grain samples, which causes high costs. Here we applied hyperspectral imaging to estimate the concentration of 15 grain elements (C, B, Ca, Cd, Cu, Fe, K, Mg, Mn, Mo, N, Na, P, S, Zn) in high-throughput in the wild barley nested association mapping (NAM) population HEB-25, comprising 1,420 BC1S3 lines derived from crossing 25 wild barley accessions with the cultivar ‘Barke’. Nutrient concentrations varied largely with a multitude of lines having higher micronutrient concentration than ‘Barke’. In a genome-wide association study (GWAS), we located 75 quantitative trait locus (QTL) hotspots, whereof many could be explained by major genes such as NO APICAL MERISTEM-1 (NAM-1) and PHOTOPERIOD 1 (Ppd-H1). The GWAS approach revealed exotic alleles that were able to increase grain element concentrations. Remarkably, a QTL linked to GIBBERELLIN 20 OXIDASE 2 (HvGA20ox2) significantly increased several grain elements without yield loss. We conclude that introgressing promising exotic alleles into elite breeding material can assist in improving the nutritional value of barley grains.

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.

Wild barley shows a wider diversity in genes regulating heading date compared with cultivated barley

Heading date (HD) is an important agronomic trait that influences plant adaptability to varying environment and, ultimately, grain yield. In this study, two doubled haploid (DH) populations were used to identify new QTL for HD. One of the DH populations is originated from a cross between an Australian malting barley cv. Franklin and a wild barley accession TAM407227 and the other one is from the cross between a Syrian wild barley SYR01 and an Australian malting barley cv. Gairdner. Using three times of sowing (TOS) differing in daylength and temperature, we investigated quantitative trait loci (QTL) controlling HD from both populations. Fourteen QTL were identified for HD from different populations and sowing dates. The expression of HD related genes varied with the TOS, suggesting a significant QTL × environment interaction. By comparing the positions of previously mapped HD genes and those of QTL detected in this population, we found that eleven of the fourteen QTL identified in this study were located at similar positions to those reported genes for HD. Among the three new potential QTL, one was located at 73.5 cM on chromosome 2H, explaining 19.2% and 4.6% HD of DH lines in spring and summer growing, respectively. The wild barley parent TAM407227 contributed the early maturity allele. HORVU2Hr1G088460 within the interval of QTL could be the candidate gene. The second new QTL was identified on chromosome 3H from a summer sowing trial and the third one on chromosome 4H affected HD of DH lines only under spring sowing condition. These new QTL identified will provide alternative genetic resources for plant breeders developing barley varieties with improved HD adaptability to varying environments.

Response of Tibetan Wild Barley Genotypes to Drought Stress and Identification of Quantitative Trait Loci by Genome-Wide Association Analysis.

Tibetan wild barley has been identified to show large genetic variation and stress tolerance. A genome-wide association (GWA) analysis was performed to detect quantitative trait loci (QTLs) for drought tolerance using 777 Diversity Array Technology (DArT) markers and morphological and physiological traits of 166 Tibetan wild barley accessions in both hydroponic and pot experiments. Large genotypic variation for these traits was found; and population structure and kinship analysis identified three subpopulations among these barley genotypes. The average LD (linkage disequilibrium) decay distance was 5.16 cM, with the minimum on 6H (0.03 cM) and the maximum on 4H (23.48 cM). A total of 91 DArT markers were identified to be associated with drought tolerance-related traits, with 33, 26, 16, 1, 3, and 12 associations for morphological traits, H+K+-ATPase activity, antioxidant enzyme activities, malondialdehyde (MDA) content, soluble protein content, and potassium concentration, respectively. Furthermore, 7 and 24 putative candidate genes were identified based on the reference Meta-QTL map and by searching the Barleymap. The present study implicated that Tibetan annual wild barley from Qinghai–Tibet Plateau is rich in genetic variation for drought stress. The QTLs detected by genome-wide association analysis could be used in marker-assisting breeding for drought-tolerant barley genotypes and provide useful information for discovery and functional analysis of key genes in the future.

Genotypic difference of cadmium tolerance and the associated microRNAs in wild and cultivated barley

Little study was performed to know how microRNAs (miRNAs) are responsive to cadmium (Cd) stress in barley (Hordeum vulgare). In this study, 16 small RNA libraries of shoot and root tissues from a wild barley accession (WB-1) and cultivated barley (Golden Promise) with contrasting Cd tolerance were constructed and sequenced. Moreover, a degradome library was constructed and analyzed to identify target genes of the miRNAs. Based on high-throughput sequencing, 216 conserved miRNAs (in 59 miRNA families) and 87 novel miRNAs were identified. A total of 238 target genes for 149 miRNAs (113 conserved and 36 novel miRNAs) were detected by the degradome analysis. Among these miRNAs, 45 miRNAs (40 conserved and 5 novel miRNAs) and 43 miRNAs (40 conserved and 3 novel miRNAs) showed differential expression in roots and shoots of two genotypes under Cd conditions. Compared with cultivar Golden Promise, the wild genotype WB-1 had genotype-dependent responses of miR156, miR159, miR166, miR167, miR171 and miR393, which regulate target genes including SPL, MYB, HD-Zip, ARF, GRAS and TIR. Correspondingly, WB-1 had lower Cd concentration and stronger Cd tolerance than Golden Promise. It indicates that miRNAs may play critical roles underlying genotypic difference of Cd tolerance in barley.

Identification of microRNAs responding to salt stress in barley by high-throughput sequencing and degradome analysis

A deep understanding of the mechanisms underlying salt tolerance should be helpful for breeding of salt-tolerant crop cultivars. Our previous study identified some wild barley accessions with high salt tolerance. In this study, small RNA and degradome sequencing was performed to identify salt-responsive microRNAs (miRNAs) and their target genes in a Tibetan wild barley accession XZ16 and a cultivar Golden Promise (GP), differing in salt tolerance. Under salt stress, XZ16 showed higher salt tolerance than GP in terms of biomass and shoot Na+ accumulation. A total of 278 and 320 miRNAs were found in roots and shoots of both genotypes, respectively. Among them, 40 and 51 miRNAs were salt-responsive in roots and shoots, respectively. In roots, miR156d, miR164a, miR393a, miR319a and miR172b targeting UGTs, NAC079, HvAFB2/HvTIR1, TCP4 and HvAP2, respectively, are probably responsible for salt tolerance. In shoots, miR159a, miR169i, miR319a/miR396e module and miR172b regulating MYB33, NHX1/LEA7, TCP4, GRFs and HvAP2, respectively, might contribute to salt tolerance. Compared with GP, XZ16 showed differential expression of miR156d, miR164a, miR169i, miR172b, miR319a, miR393a and a novel miRNA PC-miR124. We proposed that these differentially expressed miRNAs could account for the difference in salt tolerance of the two genotypes and drawn up a putative regulatory network of miRNAs to reveal molecular mechanisms of salt tolerance in barley.

Population Genetics Revealed a New Locus That Underwent Positive Selection in Barley.

Trait variation among natural populations and their cultivated relatives occurs due to evolutionary forces, including selection and drift. In the present study, we analyzed these forces at the locus level in a global barley diversity set using population genetics analysis. Genome-wide outlier loci detection found a locus on chromosome 2H at which a common single nucleotide polymorphism (SNP) marker SCRI_RS_170235 accounted for the highest diversity index (Fst) values between cultivars and landraces and between cultivars and wild accessions. For a population wide genetic analysis, we developed a Polymerase Chain Reaction (PCR)-based cleaved amplified polymorphic marker at the identified locus. Marker genotyping of 115 genotypes identified a characteristic distribution of polymorphisms among the cultivated, landraces, and wild barley accessions. Using this marker, we screened a library of wild barley introgression lines (IL) and selected IL S42IL-109 that carried the wild introgression of the outlier locus in cultivar ‘Scarlett’ background. A plethora of phenotypic evaluation was performed between the S42IL109 and ‘Scarlett’ to dissect the putative effect of the identified outlier locus. Comparison of S42IL109 and ‘Scarlett’ revealed significant difference in the development of phyllochron two (Phyl-2), phyllochron three (Phyl-3), and phyllochron four (Phyl-4). Across the three phyllochrons, it was consistently observed that S42IL109 developed successive leaves in a shorter time span, by one to two days, compared to ‘Scarlett’. These data suggest that outlier locus may influence phyllochron variation which underwent positive selection in barley.

Genome-wide Scan Using DArT Markers for Selection Footprints in Six-rowed Naked Barley from the Tibetan Plateau

As one of the world’s earliest domesticated crops, barley is a model species for the study of evolution and domestication. Domestication is an evolutionary process whereby a population adapts, through selection; to new environments created by human cultivation. We describe the genome-scanning of molecular diversity to assess the evolution of barley in the Tibetan Plateau. We used 667 Diversity Arrays Technology (DArT) markers to genotype 185 barley landraces and wild barley accessions from the Tibetan Plateau. Genetic diversity in wild barley was greater than in landraces at both genome and chromosome levels, except for chromosome 3H. Landraces and wild barley accessions were clearly differentiated genetically, but a limited degree of introgression was still evident. Significant differences in diversity between barley subspecies at the chromosome level were observed for genes known to be related to physiological and phenotypical traits, disease resistance, abiotic stress tolerance, malting quality and agronomic traits. Selection on the genome of six-rowed naked barley has shown clear multiple targets related to both its specific end-use and the extreme environment in Tibet. Our data provide a platform to identify the genes and genetic mechanisms that underlie phenotypic changes, and provide lists of candidate domestication genes for modified breeding strategies.

Genetic dissection of quantitative and qualitative traits using a minimum set of barley Recombinant Chromosome Substitution Lines

BackgroundExploring the natural occurring genetic variation of the wild barley genepool has become a major target of barley crop breeding programmes aiming to increase crop productivity and sustainability in global climate change scenarios. However this diversity remains unexploited and effective approaches are required to investigate the benefits that unadapted genomes could bring to crop improved resilience. In the present study, a set of Recombinant Chromosome Substitution Lines (RCSLs) derived from an elite barley cultivar ‘Harrington’ as the recurrent parent, and a wild barley accession from the Fertile Crescent ‘Caesarea 26–24’, as the donor parent (Matus et al. Genome 46:1010–23, 2003) have been utilised in field and controlled conditions to examine the contribution of wild barley genome as a source of novel allelic variation for the cultivated barley genepool.MethodsTwenty-eight RCSLs which were selected to represent the entire genome of the wild barley accession, were genotyped using the 9 K iSelect SNP markers (Comadran et al. Nat Genet 44:1388–92, 2012) and phenotyped for a range of morphological, developmental and agronomic traits in 2 years using a rain-out shelter with four replicates and three water treatments. Data were analysed for marker traits associations using a mixed model approach.ResultsWe identified lines that differ significantly from the elite parent for both qualitative and quantitative traits across growing seasons and water regimes. The detailed genotypic characterisation of the lines for over 1800 polymorphic SNP markers and the design of a mixed model analysis identified chromosomal regions associated with yield related traits where the wild barley allele had a positive response increasing grain weight and size. In addition, variation for qualitative characters, such as the presence of cuticle waxes on the developing spikes, was associated with the wild barley introgressions. Despite the coarse location of the QTLs, interesting candidate genes for the major marker-trait associations were identified using the recently released barley genome assembly.ConclusionThis study has highlighted the role of exotic germplasm to contribute novel allelic variation by using an optimised experimental approach focused on an exotic genetic library. The results obtained constitute a step forward to the development of more tolerant and resilient varieties.

Risk-management strategies and transpiration rates of wild barley in uncertain environments.

Regulation of the rate of transpiration is an important part of plants' adaptation to uncertain environments. Stomatal closure is the most common response to severe drought. By closing their stomata, plants reduce transpiration to better their odds of survival under dry conditions. Under mild to moderate drought conditions, there are several possible transpiration patterns that balance the risk of lost productivity with the risk of water loss. Here, we hypothesize that plant ecotypes that have evolved in environments characterized by unstable patterns of precipitation will display a wider range of patterns of transpiration regulation along with other quantitative physiological traits (QPTs), compared to ecotypes from less variable environments. We examined five accessions of wild barley (Hordeum vulgare ssp. spontaneum) from different locations in Israel (the B1K collection) with annual rainfall levels ranging from 100 to 900 mm, along with one domesticated line (cv. Morex). We measured several QPTs and morphological traits of these accessions under well-irrigated conditions, under drought stress and during recovery from drought. Our results revealed a correlation between precipitation-certainty conditions and QPT plasticity. Specifically, accessions from stable environments (very wet or very dry locations) were found to take greater risks in their water-balance regulation than accessions from areas in which rainfall is less predictable. Notably, less risk-taking genotypes recovered more quickly than more risk-taking ones once irrigation was resumed. We discuss the relationships between environment, polymorphism, physiological plasticity and fitness, and suggest a general risk-taking model in which transpiration-rate plasticity is negatively correlated with population polymorphism.

Root and leaf metabolite profiles analysis reveals the adaptive strategies to low potassium stress in barley

BackgroundPotassium (K) deficiency in arable land is one of the most important factors affecting crop productivity. Development of low K (LK) tolerant crop cultivars is regarded as a best economic and effective approach for solving the issue of LK. In previous studies, we found a wider variation of LK tolerance in the Tibetan wild barley accessions than cultivated barley. However, the mechanism of LK tolerance in wild barley is still elusive.ResultsIn this study, two wild barley genotypes (XZ153, LK tolerant and XZ141, LK sensitive) and one cultivar (LuDaoMai, LK tolerant) was used to investigate metabolome changes in response to LK stress. Totally 57 kinds of metabolites were identified in roots and leaves of three genotypes at 16 d after LK treatment. In general, accumulation of amino acids and sugars was enhanced in both roots and leaves, while organic acids were reduced under LK stress compared to the control. Meanwhile, the concentrations of the negatively charged amino acids (Asp and Glu) and most organic acids was reduced in both roots and leaves, but more positively charged amino acids (Lys and Gln) were increased in three genotypes under LK. XZ153 had less reduction than other two genotypes in biomass and chlorophyll content under LK stress and showed greater antioxidant capacity as reflected by more synthesis of active oxygen scavengers. Higher LK tolerance of XZ153 may also be attributed to its less carbohydrate consumption and more storage of glucose and other sugars, thus providing more energy for plant growth under LK stress. Moreover, phenylpropanoid metabolic pathway mediated by PAL differed among three genotypes, which is closely associated with the genotypic difference in LK tolerance.ConclusionsLK tolerance in the wild barley is attributed to more active phenylpropanoid metabolic pathway mediated by PAL, energy use economy by reducing carbohydrate consumption and storage of glucose and other sugars, and higher antioxidant defense ability under LK stress.

An Ancestral Allele of Pyrroline-5-carboxylate synthase1 Promotes Proline Accumulation and Drought Adaptation in Cultivated Barley.

Water scarcity is a critical threat to global crop production. Here, we used the natural diversity of barley (Hordeum vulgare) to dissect the genetic control of proline (Pro) mediated drought stress adaptation. Genetic mapping and positional cloning of a major drought-inducible quantitative trait locus (QPro.S42-1H) revealed unique allelic variation in pyrroline-5-carboxylate synthase (P5cs1) between the cultivated cultivar Scarlett (ssp. vulgare) and the wild barley accession ISR42-8 (ssp. spontaneum). The putative causative mutations were located in the promoter of P5cs1 across the DNA binding motifs for abscisic acid-responsive element binding transcription factors. Introgression line (IL) S42IL-143 carrying the wild allele of P5cs1 showed significant up-regulation of P5cs1 expression compared to Scarlett, which was consistent with variation in Pro accumulation under drought. Next, we transiently expressed promoter::reporter constructs of ISR42-8 and Scarlett alleles in Arabidopsis (Arabidopsis thaliana) mesophyll protoplasts. GUS expression analysis showed a significantly higher activation of the ISR42-8 promoter compared to Scarlett upon abscisic acid treatment. Notably, the ISR42-8 promoter activity was impaired in protoplasts isolated from the loss-of-function abf1abf2abf3abf4 quadruple mutant. A series of phenotypic evaluations demonstrated that S42IL-143 maintained leaf water content and photosynthetic activity longer than Scarlett under drought. These findings suggest that the ancestral variant of P5cs1 has the potential for drought tolerance and understanding drought physiology of barley and related crops.

Retrotransposon Insertion and DNA Methylation Regulate Aluminum Tolerance in European Barley Accessions.

Aluminum (Al) toxicity is a major stress factor limiting crop productivity in acid soil. Although there is great genotypic variation in tolerance to Al toxicity, the underlying molecular mechanisms are poorly understood. Here, we report that, in barley (Hordeum vulgare), the fourth largest cereal crop produced in the world, both retrotransposon insertion and DNA methylation are involved in regulating differential Al tolerance. HvAACT1 is a major gene responsible for citrate secretion from the roots for external detoxification of Al. A multiretrotransposon-like (MRL) sequence insertion at least 15.3 kb in length was detected in the upstream genomic region of HvAACT1 that displayed promoter activity and significantly enhanced HvAACT1 expression, especially in the root tips of Al-tolerant accessions. Furthermore, in a number of accessions with low levels of HvAACT1 expression, this MRL insertion was present but highly methylated. Geographical analysis showed that accessions with this MRL insertion are distributed mainly in European areas with acid soils. Two wild barley accessions were found to possess this MRL insertion, but with a high degree of methylation. These results indicate that the MRL insertion and its degree of DNA methylation influence HvAACT1 expression and that demethylation of this MRL insertion, which facilitates adaptation to acid soils, occurred following barley domestication. Moreover, our results indicate that barley accessions in East Asia and Europe have developed independent but equivalent strategies to withstand Al toxicity in acid soils.

Identification of microRNAs in response to aluminum stress in the roots of Tibetan wild barley and cultivated barley

BackgroundBarley is relatively sensitive to Aluminum (Al) toxicity among cereal crops, but shows a wide genotypic difference in Al tolerance. The well-known Al-tolerant mechanism in barley is related to Al exclusion mediated by a citrate transporter HvAACT1 (Al-activated citrate transporter 1). A 1-kb insertion in the promoter region of HvAACT1 gene results in a dramatic increase of its expression level, which only occurs in some Al-tolerant cultivars. However, Al-tolerant Tibetan wild barley accession XZ29 did not have the 1-kb insertion.ResultsWe confirmed that the expression of HvAACT1 and secretion of citrate and other organic acids did not explain the difference in Al-tolerant wild barley XZ29 and Al-sensitive cultivated barley Golden Promise. To identify microRNAs (miRNAs) and their target genes responsive to Al stress in barley roots, eight small RNA libraries with two biological replicates from these two genotypes exposed to control and Al-treated conditions were constructed and submitted to deep sequencing. A total of 342 miRNAs were identified in Golden Promise and XZ29, with 296 miRNAs being commonly shared in the two genotypes. Target genes of these miRNAs were obtained through bioinformatics prediction or degradome identification. Comparative analysis detected 50 miRNAs responsive to Al stress, and some of them were found to be exclusively expressed in XZ29 and associated with Al tolerance.ConclusionsmiRNAs exclusively expressing in the wild barley were identified and found to be associated with Al stress tolerance. The current results provide a model of describing the roles of some special miRNAs associated with Al tolerance in the Tibetan wild barley.

A Sodium Transporter HvHKT1;1 Confers Salt Tolerance in Barley via Regulating Tissue and Cell Ion Homeostasis.

Our previous studies showed that high salt tolerance in Tibetan wild barley accessions was associated with HvHKT1;1, a member of the high-affinity potassium transporter family. However, molecular mechanisms of HvHKT1;1 for salt tolerance and its roles in K+/Na+ homeostasis remain to be elucidated. Functional characterization of HvHKT1;1 was conducted in the present study. NaCl-induced transcripts of HvHKT1;1 were significantly higher in the roots of Tibetan wild barley XZ16 relative to other genotypes, being closely associated with its higher biomass and lower tissue Na+ content under salt stress. Heterologous expression of HvHKT1;1 in Saccharomyces cerevisiae (yeast) and Xenopus laevis oocytes showed that HvHKT1;1 had higher selectivity for Na+ over K+ and other monovalent cations. HvHKT1;1 was found to be localized at the cell plasma membrane of root stele and epidermis. Knock-down of HvHKT1;1 in barley led to higher Na+ accumulation in both roots and leaves, while overexpression of HvHKT1;1 in salt-sensitive Arabidopsis hkt1-4 and sos1-12 loss-of-function lines resulted in significantly less shoot and root Na+ accumulation. Additionally, microelectrode ion flux measurements and root elongation assay revealed that the transgenic Arabidopsis plants exhibited a remarkable capacity for regulation of Na+, K+, Ca2+ and H+ homeostasis under salt stress. These results indicate that HvHKT1;1 is critical in radial root Na+ transport, which eventually reduces shoot Na+ accumulation. Additionally, HvHKT1;1 may be indirectly involved in retention of K+ and Ca2+ in root cells, which also improves plant salt tolerance.