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Hulless barley (Hordeum vulgare L. var. nudum), also called naked barley, is a unique variety of cultivated barley. The genome-wide specific length amplified fragment sequencing (SLAF-seq) method is a rapid deep sequencing technology that is used for the selection and identification of genetic loci or markers. In this study, we collected 300 hulless barley accessions and used the SLAF-seq method to identify candidate genes involved in plant height (PH) and tiller number (TN). We obtained a total of 1407 M paired-end reads, and 228,227 SLAF tags were developed. After filtering using an integrity threshold of >0.8 and a minor allele frequency of >0.05, 14,504,892 single-nucleotide polymorphisms (SNP) loci were screened out. The remaining SNPs were used for the construction of a neighbour-joining phylogenetic tree, and the three subcluster members showed no obvious differentiation among regional varieties. We used a genome wide association study approach to identify 1006 and 113 SNPs associated with TN and PH, respectively. Based on best linear unbiased predictors (BLUP), 41 and 29 SNPs associated with TN and PH, respectively. Thus, several of genes, including Hd3a and CKX5, may be useful candidates for the future genetic breeding of hulless barley. Taken together, our results provide insight into the molecular mechanisms controlling barley architecture, which is important for breeding and yield.

Infection of seeds of cereal plants with fusarium affects their optical luminescent properties. The spectral characteristics of excitation (absorption) in the range of 180–700 nm of healthy and infected seeds of wheat, barley and oats were measured. The greatest difference in the excitation spectra of healthy and infected seeds was observed in the short-wave range of 220–450 nm. At the same time, the excitation characteristics of infected seeds were higher than those of healthy ones, and the integral parameter Η in the entire range was 10–56% higher. A new maximum appeared at the wavelength of 232 nm and the maximum value increased by 362 nm. The spectral characteristics were measured when excited by radiation at wavelengths of 232, 362, 424, 485, 528 nm and the luminescence fluxes were calculated. It is established that the photoluminescence fluxes Φ in the short-wave ranges of 290–380 nm increase by 1.58–3.14 times and 390–550 nm-by 1.44–2.54 times. The fluxes in longer wavelength ranges do not change systematically and less significantly: for wheat, they decrease by 12% and increase by 19%, for barley, they decrease by 10% and increase by 33%. The flux decreases by 43–71% for oats. Based on the results obtained for cereal seeds, it is possible to further develop a method for detecting fusarium infection with absolute measurements of photoluminescence fluxes in the range of 290–380 nm, or when measuring photoluminescence ratios: for wheat seeds when excited with wavelengths of 424 nm and 232 nm (Φ424/Φ232); for barley seeds–when excited with wavelengths of 485 nm and 232 nm (Φ485/Φ232) and for oat seeds–when excited with wavelengths of 424 nm and 362 nm (Φ424/Φ362).

Ageratina adenophora originating from central America has flooded forests, pastures, and farmland in more than 40 tropical and subtropical countries, causing huge ecological disasters and economic losses. In this paper, we intended to use a complex inoculum composed of Pseudomonas putita and Clostridium thermocellum to in-situ compost A. adenophora debris and then to compare the phytotoxicity of extracts from uncomposted and composted A. adenophora (UCA and CA respectively) to barley seed germination and young seedling growth. A field experiment was finally conducted to reveal the effects of UCA and CA on barley nutrients uptake, yield, grain quality, soil enzyme activities, microbial biomass and biodiversity. In-situ composting sharply decreased 4,7-dimethyl-1-(propan-2-ylidene)-1,4,4a,8a-tetrahydronaphthalene- 2,6(1H,7H)-dione(DTD) and 6-hydroxy-5-isopropyl-3,8-dimethyl-4a,5,6,7,8,8a-hexahydronaphthal en-2(1 H)-one(HHO) from 2096.3 and 743.7 mg kg-1 in uncomposted A. adenophora to 194.4 and 68.19 mg kg-1 in composted A. adenophora. UCAE showed negative influences on seed germination performances (except lower rates on germination percentage). The mechanism may be the inhibition of bio-macromolecules hydrolysis (including proteins, starch, and phytin) in endosperms and their hydrolysates for forming new plants. CAE promoted seed germination and seedling growth, increased chlorophyll levels in leaves, and stimulated dehydrogenase and nitrate reductase activities in plants, while UCAE got opposite performance. Compared with chemical fertilizers, application of CA in combination with chemical fertilizers significantly improved plant nutrient uptake (nitrogen, phosphorus, and potassium), yield, grain quality, quantity of 16S rDNA sequences, richness and diversity of bacterial communities in contrast to UCA which behaved otherwise. Taken together, the use of the microbial agent to in-situ compost A. adenophora may be an effective approach for agricultural use of A. adenophora debris as a plant-friendly organic fertilizer, being undoubtedly worth advocating.

The vast majority of plant viruses are unenveloped, i.e., they lack a lipid bilayer that is characteristic of most animal viruses. The interactions between plant viruses, and between viruses and surfaces, properties that are essential for understanding their infectivity and to their use as bionanomaterials, are largely controlled by their surface charge, which depends on pH and ionic strength. They may also depend on the charge of their contents, i.e., of their genes or–in the instance of virus-like particles–encapsidated cargo such as nucleic acid molecules, nanoparticles or drugs. In the case of enveloped viruses, the surface charge of the capsid is equally important for controlling its interaction with the lipid bilayer that it acquires and loses upon leaving and entering host cells. We have previously investigated the charge on the unenveloped plant virus Cowpea Chlorotic Mottle Virus (CCMV) by measurements of its electrophoretic mobility. Here we examine the electrophoretic properties of a structurally and genetically closely related bromovirus, Brome Mosaic Virus (BMV), of its capsid protein, and of its empty viral shells, as functions of pH and ionic strength, and compare them with those of CCMV. From measurements of both solution and gel electrophoretic mobilities (EMs) we find that the isoelectric point (pI) of BMV (5.2) is significantly higher than that of CCMV (3.7), that virion EMs are essentially the same as those of the corresponding empty capsids, and that the same is true for the pIs of the virions and of their cleaved protein subunits. We discuss these results in terms of current theories of charged colloidal particles and relate them to biological processes and the role of surface charge in the design of new classes of drug and gene delivery systems.

The structure of barley varieties were studied, using structured and semi-structured queries, at Legambo, Tenta and Worailu districts of South Wollo, Ethiopia. Eight local barley varieties (Belg, Ginbot, Sene/Nech, Tikur, Holker, Traveler Tegadime and Temezhi) were identified, and got their names found on seed color and planting season. According to farmers, Tegadime is the production leader among all, but the source of seeds and the market chain are the limiting factors for its popularity and this is why it’s not famous is because of the low price of the seed. Thus, Sene/Nech found to be popular and shared 46.91% at Tenta, 48.47% at Legambo and 51.55% at Wereilu followed by Tikur and Ginbote. High barley diversity was noted at Tenta (E = 0.773) followed by Wereilu (E = 0.678) and Legambo (E = 0.606). Sene/Nech (0.67), Belg (0.62), Tegadime (0.59), Tikur (0.55) and Ginbote (0.54) were found to be shared, but Traveler, Holker and Temezhi were rarely found. At farm, most farmers were plowing twice before sowing using horse. Biological fertilizer usages were well practice at Tenta, Legambo and Were’ilu, respectively. While, inorganic fertilizer usage was better at Wereilu, but none at Legambo. Pest management was better at Wereilu and hand weeding is a common system, but low at Legambo, and mowing by sickle, threshing by horse and store in Gotera were a shared practice. Farmers use outdated tools for agricultural practice and the yield is losing due to unavailable of update machinery. So, different managing approaches and new harvesting technologies should address.

In filamentous fungi, gene silencing by RNA interference (RNAi) shapes many biological processes, including pathogenicity. Recently, fungal small RNAs (sRNAs) have been shown to act as effectors that disrupt gene activity in interacting plant hosts, thereby undermining their defence responses. We show here that the devastating mycotoxin-producing ascomycete Fusarium graminearum (Fg) utilizes DICER-like (DCL)-dependent sRNAs to target defence genes in two Poaceae hosts, barley (Hordeum vulgare, Hv) and Brachypodium distachyon (Bd). We identified 104 Fg-sRNAs with sequence homology to host genes that were repressed during interactions of Fg and Hv, while they accumulated in plants infected by the DCL double knock-out (dKO) mutant PH1-dcl1/2. The strength of target gene expression correlated with the abundance of the corresponding Fg-sRNA. Specifically, the abundance of three tRNA-derived fragments (tRFs) targeting immunity-related Ethylene overproducer 1-like 1 (HvEOL1) and three Poaceae orthologues of Arabidopsis thaliana BRI1-associated receptor kinase 1 (HvBAK1, HvSERK2 and BdSERK2) was dependent on fungal DCL. Additionally, RNA-ligase-mediated Rapid Amplification of cDNA Ends (RLM-RACE) identified infection-specific degradation products for the three barley gene transcripts, consistent with the possibility that tRFs contribute to fungal virulence via targeted gene silencing.

Background Hordeum brevisubulatum, known as fine perennial forage, is used for soil salinity improvement in northern China. Chloroplast (cp) genome is an ideal model for assessing its genome evolution and the phylogenetic relationships. We de novo sequenced and analyzed the cp genome of H. brevisubulatum, providing a fundamental reference for further studies in genetics and molecular breeding. Results The cp genome of H. brevisubulatum was 137,155 bp in length with a typical quadripartite structure. A total of 130 functional genes were annotated and the gene of accD was lost in the process of evolution. Among all the annotated genes, 16 different genes harbored introns and the genes of ycf3 and rps12 contained two introns. Parity rule 2 (PR2) plot analysis showed that majority of genes had a bias toward T over A in the coding strand in all five Hordeum species, and a slight G over C in the other four Hordeum species except for H. bogdanil. Additionally, 52 dispersed repeat sequences and 182 simple sequence repeats were identified. Moreover, some unique SSRs of each species could be used as molecular markers for further study. Compared to the other four Hordeum species, H. brevisubulatum was most closely related to H. bogdanii and its cp genome was relatively conserved. Moreover, inverted repeat regions (IRa and IRb) were less divergent than other parts and coding regions were relatively conserved compared to non-coding regions. Main divergence was presented at the SSC/IR border. Conclusions This research comprehensively describes the architecture of the H. brevisubulatum cp genome and improves our understanding of its cp biology and genetic diversity, which will facilitate biological discoveries and cp genome engineering.

Metabolites play a key role in plants as they are routing plant developmental processes and are involved in biotic and abiotic stress responses. Their analysis can offer important information on the underlying processes. Regarding plant breeding, metabolite concentrations can be used as biomarkers instead of or in addition to genetic markers to predict important phenotypic traits (metabolic prediction). In this study, we applied a genome-wide association study (GWAS) in a wild barley nested association mapping (NAM) population to identify metabolic quantitative trait loci (mQTL). A set of approximately 130 metabolites, measured at early and late sampling dates, was analysed. For four metabolites from the early and six metabolites from the late sampling date significant mQTL (grouped as 19 mQTL for the early and 25 mQTL for the late sampling date) were found. Interestingly, all of those metabolites could be classified as sugars. Sugars are known to be involved in signalling, plant growth and plant development. Sugar-related genes, encoding mainly sugar transporters, have been identified as candidate genes for most of the mQTL. Moreover, several of them co-localized with known flowering time genes like Ppd-H1, HvELF3, Vrn-H1, Vrn-H2 and Vrn-H3, hinting on the known role of sugars in flowering. Furthermore, numerous disease resistance-related genes were detected, pointing to the signalling function of sugars in plant resistance. An mQTL on chromosome 1H in the region of 13 Mbp to 20 Mbp stood out, that alone explained up to 65% of the phenotypic variation of a single metabolite. Analysis of family-specific effects within the diverse NAM population showed the available natural genetic variation regarding sugar metabolites due to different wild alleles. The study represents a step towards a better understanding of the genetic components of metabolite accumulation, especially sugars, thereby linking them to biological functions in barley.

Celiac disease (CD) is triggered by ingestion of gluten-containing cereals such as wheat, barley, rye and in some cases oat. The only way for affected individuals to avoid symptoms of this condition is to adopt a gluten-free diet. Thus, gluten-free foodstuffs need to be monitored in order to ensure their innocuity. For this purpose, commercial immunoassays based on recognition of defined linear gluten sequences are currently used. These immunoassays are designed to detect or quantify total gluten regardless of the cereal, and often result in over or underestimation of the exact gluten content. In addition, Canadian regulations require a declaration of the source of gluten on the label of prepackaged foods, which cannot be done due to the limitations of existing methods. In this study, the development of new antibodies targeting discrimination of gluten sources was conducted using synthetic peptides as immunization strategy. Fourteen synthetic peptides selected from unique linear amino acid sequences of gluten were bioconjugated to Concholepas concholepas hemocyanin (CCH) as protein carrier, to elicit antibodies in rabbit. The resulting polyclonal antibodies (pAbs) successfully discriminated wheat, barley and oat prolamins during indirect ELISA assessments. pAbs raised against rye synthetic peptides cross-reacted evenly with wheat and rye prolamins but could still be useful to successfully discriminate gluten sources in combination with the other pAbs. Discrimination of gluten sources can be further refined and enhanced by raising monoclonal antibodies using a similar immunization strategy. A methodology capable of discriminating gluten sources, such as the one proposed in this study, could facilitate compliance with Canadian regulations on this matter. This type of discrimination could also complement current immunoassays by settling the issue of over and underestimation of gluten content, thus improving the safety of food intended to CD and wheat-allergic patients.

Ethiopia is recognized as a center of diversity for barley, and its landraces are known for the distinct genetic features compared to other barley collections. The genetic diversity of Ethiopian barley likely results from the highly diverse topography, altitude, climate conditions, soil types, and farming systems. To get detailed information on the genetic diversity a panel of 260 accessions, comprising 239 landraces and 21 barley breeding lines, obtained from the Ethiopian biodiversity institute (EBI) and the national barley improvement program, respectively were studied for their genetic diversity using the 50k iSelect single nucleotide polymorphism (SNP) array. A total of 983 highly informative SNP markers were used for structure and diversity analysis. Three genetically distinct clusters were obtained from the structure analysis comprising 80, 71, and 109 accessions, respectively. Analysis of molecular variance (AMOVA) revealed the presence of higher genetic variation (89%) within the clusters than between the clusters (11%), with moderate genetic differentiation (PhiPT = 0.11) and five accessions were detected as first-generation migrants using Monte Carlo resampling methods. The Mantel test revealed that the genetic distance between accessions is poorly associated with their geographical distance. Despite the observed weak correlation between geographic distance and genetic differentiation, for some regions like Gonder, Jimma, Gamo-Gofa, Shewa, and Welo, more than 50% of the landraces derived from these regions are assigned to one of the three clusters.

Small RHO-type G-proteins act as signaling hubs and master regulators of polarity in eukaryotic cells. Their activity is tightly controlled, as defective RHO signaling leads to aberrant growth and developmental defects. Two major processes regulate G-protein activity: canonical shuttling between different nucleotide bound states and posttranslational modification (PTM), of which the latter can support or suppress RHO signaling, depending on the individual PTM. In plants, regulation of Rho of plants (ROPs) signaling activity has been shown to act through nucleotide exchange and GTP hydrolysis, as well as through lipid modification, but there is little data available on phosphorylation or ubiquitination of ROPs. Hence, we applied proteomic analyses to identify PTMs of the barley ROP RACB. We observed in vitro phosphorylation by barley ROP binding kinase 1 and in vivo ubiquitination of RACB. Comparative analyses of the newly identified RACB phosphosites and human RHO protein phosphosites revealed conservation of modified amino acid residues, but no overlap of actual phosphorylation patterns. However, the identified RACB ubiquitination site is conserved in all ROPs from Hordeum vulgare, Arabidopsis thaliana and Oryza sativa and in mammalian Rac1 and Rac3. Point mutation of this ubiquitination site leads to stabilization of RACB. Hence, this highly conserved lysine residue may regulate protein stability across different kingdoms.

Concerns about the negative effects of declining agricultural biodiversity due to modern agricultural practices and climatic constraints in various parts of the world, including Iran, on the sustainability of agricultural ecosystems are increasingly growing. However, the historical knowledge of temporal and spatial biodiversity is lacking. To determine the value and trend of crop diversity in Iran, we used biodiversity indices based on the area under rainfed and irrigated crops and total cropland area from 1991 to 2018. There were large fluctuations in the amount of cultivated area in the past 30 years, peaking around 2005 to 2007 with about 13.1 million cultivated hectares. However, no general trend in increase or decrease of total cultivated land was shown. The crop species diversity of irrigated cropland was higher than the rainfed and total cropland. The Shannon diversity index showed a constant trend with a negligible slope, but species richness was increased, which was related to the rise in the area of some crop species in recent years. The area of wheat and barley had a significant impact on crop diversity, so Shannon diversity index reduced with their dominance. Overall, this study revealed that the Iranian agricultural system relies on wheat and barley. We warn that by increasing the area of these crops and the prevalence of monoculture, the probability of damage from external factors such as sudden weather changes or the spread of diseases will increase, leading to instability and production risks in the future.

Water deficit is one of the major limitations to food production worldwide and most climate change scenarios predict an aggravation of the situation. To face the expected increase in drought stress in the coming years, breeders are working to elucidate the genetic control of barley growth and productivity traits under water deficit. Barley is known as a relatively drought tolerant crop and genetic variability was observed for drought tolerance traits. The objectives of the present study were the quantification of morphological and physiological responses in a collection of 209 spring barley genotypes to drought stress, and the genetic analysis by genome-wide association study to find quantitative trait loci (QTL) and the allele contributions for each of the investigated traits. In six pot experiments, 209 spring barley genotypes were grown under a well-watered and water-limited regime. Stress phases were initiated individually for each genotype at the beginning of tillering and spiking for the vegetative- and the generative stage experiments, respectively, and terminated when the transpiration rates of stress treatments reached 10% of the well-watered control. After the stress phase, a total of 42 productivity related traits such as the dry matter of plant organs, tiller number, leaf length, leaf area, amount of water soluble carbohydrates in the stems, proline content in leaves and osmotic adjustment of corresponding well-watered and stressed plants were analysed, and QTL analyses were performed to find marker-trait associations. Significant water deficit effects were observed for almost all traits and significant genotype x treatment interactions (GxT) were observed for 37 phenotypic traits. Genome-wide association studies (GWAS) revealed 77 significant loci associated with 16 phenotypic traits during the vegetative stage experiment and a total of 85 significant loci associated with 13 phenotypic traits during the generative stage experiment for traits such as leaf area, number of green leaves, grain yield, harvest index and stem length. For traits with significant GxT interactions, genotypic differences for relative values were analysed using one way ANOVA. More than 110 loci for GxT interaction were found for 17 phenotypic traits explaining in many cases more than 50% of the genetic variance.

Soil salinity imposes an agricultural and economic burden that may be alleviated by identifying the components of salinity tolerance in barley, a major crop and the most salt tolerant cereal. To improve our understanding of these components, we evaluated a diversity panel of 377 two-row spring barley cultivars during both the vegetative, in a controlled environment, and the reproductive stages, in the field. In the controlled environment, a high-throughput phenotyping platform was used to assess the growth-related traits under both control and saline conditions. In the field, the agronomic traits were measured from plots irrigated with either fresh or saline water. Association mapping for the different components of salinity tolerance enabled us to detect previously known associations, such as HvHKT1;5. Using an "interaction model", which took into account the interaction between treatment (control and salt) and genetic markers, we identified several loci associated with yield components related to salinity tolerance. We also observed that the two developmental stages did not share genetic regions associated with the components of salinity tolerance, suggesting that different mechanisms play distinct roles throughout the barley life cycle. Our association analysis revealed that genetically defined regions containing known flowering genes (Vrn-H3, Vrn-H1, and HvNAM-1) were responsive to salt stress. We identified a salt-responsive locus (7H, 128.35 cM) that was associated with grain number per ear, and suggest a gene encoding a vacuolar H+-translocating pyrophosphatase, HVP1, as a candidate. We also found a new QTL on chromosome 3H (139.22 cM), which was significant for ear number per plant, and a locus on chromosome 2H (141.87 cM), previously identified using a nested association mapping population, which associated with a yield component and interacted with salinity stress. Our study is the first to evaluate a barley diversity panel for salinity stress under both controlled and field conditions, allowing us to identify contributions from new components of salinity tolerance which could be used for marker-assisted selection when breeding for marginal and saline regions. Refereed/Peer-reviewed

We report the earliest and the most abundant archaeobotanical assemblage of southwest Asian grain crops from Early Bronze Age Central Asia, recovered from the Chap II site in Kyrgyzstan. The archaeobotanical remains consist of thousands of cultivated grains dating to the mid-late third millennium BCE. The recovery of cereal chaff and weeds suggest local cultivation at 2000 m.a.s.l., as crops first spread to the mountains of Central Asia. The site's inhabitants possibly cultivated two types of free-threshing wheats, glume wheats, and hulled and naked barleys. Highly compact caryopses of wheat and barley grains represent distinct morphotypes of cereals adapted to highland environments. While additional macrobotanical evidence is needed to confirm the presence of glume wheats at Chap II, the possible identification of glume wheats at Chap II may represent their most eastern distribution in Central Asia. Based on the presence of weed species, we argue that the past environment of Chap II was characterized by an open mountain landscape, where animal grazing likely took place, which may have been further modified by people irrigating agricultural fields. This research suggests that early farmers in the mountains of Central Asia cultivated compact morphotypes of southwest Asian crops during the initial eastward dispersal of agricultural technologies, which likely played a critical role in shaping montane adaptations and dynamic interaction networks between farming societies across highland and lowland cultivation zones.

The quality of sourdough bread mainly depends on metabolic activities of lactic acid bacteria (LAB). The exopolysaccharides (EPS) produced by LAB affect positively the technological and nutritional properties of the bread, while phytases improve the bioavailability of the minerals by reducing its phytate content. In the present study, a pool of 152 cereal-sourced LAB were screened for production of phytases and EPS for potential use as sourdough starter cultures for the baking industry. There was large heterogeneity in the phytase activity observed among the screened isolates, with 95% showing the ability to degrade sodium phytate on plates containing Sourdough Simulation Medium (SSM). The isolates Lactobacillus brevis LD65 and Lactobacillus plantarum PB241 showed the highest enzymatic activity, while the isolates ascribed to Weissella confusa were characterized by low or no phytase activity. Only 18% of the screened LAB produced EPS, which were distinguished as ropy or mucoid phenotypes on SSM supplemented with sucrose. Almost all the EPS producers carried one or more genes (epsD/E and/or epsA) involved in the production of heteropolysaccharides (HePS), whereas the isolates ascribed to Leuconostoc citreum and W. confusa carried genes involved in the production of both HePS and homopolysaccharides (HoPS). Monosaccharide composition analysis of the EPS produced by a selected subset of isolates revealed that all the HePS included glucose, mannose, and galactose, though at different ratios. Furthermore, a few isolates ascribed to L. citreum and W. confusa and carrying the gtf gene produced β-glucans after fermentation in an ad hoc formulated barley flour medium. Based on the overall results collected, a subset of candidate sourdough starter cultures for the baking industry was selected, including Lb. brevis LD66 and L. citreum PB220, which showed high phytase activity and positive EPS production.

Whereas the role of arbuscular mycorrhizal fungi (AMF) in plant growth improvement has been well described in agroecosystems, little is known about the effect of environmental factors on AMF root colonization status of barley, the fourth most important cereal crop all over the world. In order to understand the influence of environmental factors, such as climatic and soil physico-chemical properties, on the spontaneous mycorrhizal ability of barley (Hordeum vulgare L.), a field investigation was conducted in 31 different sites in sub-humid, upper and middle semi-arid areas of Northern Tunisia. Mycorrhizal root colonization of H. vulgare varied considerably among sites. Principal component analysis showed that barley mycorrhization is influenced by both climatic and edaphic factors. A partial least square structural equation modelling (PLS-SEM) revealed that 39% (R²) of the total variation in AMF mycorrhizal rate of barley roots was mainly explained by chemical soil properties and climatic characteristics. Whereas barley root mycorrhizal rates were inversely correlated with soil organic nitrogen (ON), available phosphorus amounts (P), altitude (Z), average annual rainfall (AAR), they were directly correlated with soil pH and temperature. Our results indicated that AMF root colonization of barley was strongly related to climatic characteristics than chemical soil properties. The current study highlights the importance of the PLS-SEM to understand the interactions between climate, soil properties and AMF symbiosis of barley in field conditions.

We estimated the risk of selecting for herbicide resistance in 101 weed species known to occur in wheat and barley crops on farms in New Zealand. A protocol was used that accounts for both the risk that different herbicides will select for resistance and each weed's propensity to develop herbicide resistance based on the number of cases worldwide. To provide context we documented current herbicide use patterns. Most weeds (55) were low-risk, 30 were medium-risk and 16 high-risk. The top ten scored weeds were Echinochloa crus-galli, Poa annua, Lolium multiflorum, Erigeron sumatrensis, Raphanus raphanistrum, Lolium perenne, Erigeron bonariensis, Avena fatua, Avena sterilis and Digitaria sanguinalis. Seven out of ten high-risk weeds were grasses. The most used herbicides were synthetic auxins, an enolpyruvylshikimate-phosphate synthase inhibitor, acetolactate synthase (ALS) inhibitors, carotenoid biosynthesis inhibitors, and long-chain fatty acid inhibitors. ALS-inhibitors were assessed as posing the greatest risk for more species than other modes-of-action. Despite pre-emergence herbicides being known to delay resistance, New Zealand farmers only applied flufenacet and terbuthlazine with high frequency. Based on our analysis, surveys for herbicide-resistant species should focus on the high-risk species we identified. Farmer extension efforts in New Zealand should address resistance evolution in cropping weeds.

Considering the complex nature of salinity tolerance mechanisms, the use of isogenic lines or mutants possessing the same genetic background albeit different tolerance to salinity is a suitable method for reduction of analytical complexity to study these mechanisms. In the present study, whole transcriptome analysis was evaluated using RNA-seq method between a salt-tolerant mutant line "M4-73-30" and its wild-type "Zarjou" cultivar at seedling stage after six hours of exposure to salt stress (300 mM NaCl). Transcriptome sequencing yielded 20 million reads for each genotype. A total number of 7116 transcripts with differential expression were identified, 1586 and 1479 of which were obtained with significantly increased expression in the mutant and the wild-type, respectively. In addition, the families of WRKY, ERF, AP2/EREBP, NAC, CTR/DRE, AP2/ERF, MAD, MIKC, HSF, and bZIP were identified as the important transcription factors with specific expression in the mutant genotype. The RNA-seq results were confirmed at several time points using qRT-PCR for some important salt-responsive genes. In general, the results revealed that the mutant accumulated higher levels of sodium ion in the root and decreased its transfer to the shoot. Also, the mutant increased the amount of potassium ion leading to the maintenance a high ratio [K+]/[Na+] in the shoot compared to its wild-type via fast stomata closure and consequently transpiration reduction under the salt stress. Moreover, a reduction in photosynthesis and respiration was observed in the mutant, resulting in utilization of the stored energy and the carbon for maintaining the plant tissues, which is considered as a mechanism of salt tolerance in plants. Up-regulation of catalase, peroxidase, and ascorbate peroxidase genes has resulted in higher accumulation of H2O2 in the wild-type compared to the mutant. Therefore, the wild-type initiated rapid ROS signals which led to less oxidative scavenging in comparison with the mutant. The mutant increased expression in the ion transporters and the channels related to the salinity to maintain the ion homeostasis. In overall, the results demonstrated that the mutant responded better to the salt stress under both osmotic and ionic stress phases and lower damage was observed in the mutant compared to its wild-type under the salt stress.

Accurately estimating cause-specific mortality for immature insect herbivores is usually difficult. The insects are exposed to abiotic and biotic mortality factors, causing cadavers to simply disappear before cause of mortality can be recorded. Also, insect herbivores are often highly mobile on hosts, making it difficult to follow patterns for individuals through time. In contrast, the wheat stem sawfly, Cephus cinctus Norton, spends its entire egg, larval, and pupal period inside a host stem. Therefore, with periodic sampling stage-specific causes of mortality can be ascertained. Consequently, we examined C. cinctus mortality in eight barley, Hordeum vulgare L., cultivars in two locations in Montana from 2016 to 2018 by collecting stem samples from stem elongation to crop maturity at weekly intervals, and collecting overwintered barley stubs the following spring and summer from the same plots. If larvae were present, we examined larval status—dead or alive—and categorized dead individuals into one of 5 mortality categories: plant defense, cannibalism, parasitism, pathogens, and unknown factors. We used multiple decrement life tables to estimate cause-specific mortality and irreplaceable mortality (the proportion of mortality from a given cause that cannot be replaced by other causes of mortality). Plant defense (antibiosis) caused 85.7 ± 3.6%, cannibalism (governed by antixenosis) caused 70.1 ± 7.6%, parasitism caused 13.8 ± 5.9%, unknown factors caused 38.5 ± 7.6%, and pathogens caused 14.7 ± 8.5% mortality in the presence of all causes of mortality. Similarly, irreplaceable mortality due to plant defense was 22.3 ± 6.4%, cannibalism was 29.1± 4.2%, unknown factors was 6.2 ± 1.8%, pathogens was 0.9 ± 0.5%, and parasitism was 1. 5 ± 0. 6%. Antibiosis traits primarily killed newly emerged larvae, while other traits supported more favorable oviposition decisions by females, increasing mortality by obligate cannibalism. Our results suggest that breeding barley for resistance to C. cinctus targeting both categories of traits (antibiosis and antixenosis) is a highly valuable tactic for management of this important pest.

The high toxicity of cadmium (Cd) and its ready uptake by plants has become a major agricultural problem. To investigate the genetic architecture and genetic regulation of Cd tolerance in barley, we conducted quantitative trait loci (QTL) analysis in the phenotypically polymorphic Oregon Wolfe Barley (OWB) mapping population, derived from a cross between Rec and Dom parental genotypes. Through evaluating the Cd tolerance of 87 available doubled haploid lines of the OWB mapping population at the seedling stage, one minor and one major QTL were detected on chromosomes 2H and 6H, respectively. For chlorosis and necrosis traits, the major QTL explained 47.24% and 38.59% of the phenotypic variance, respectively. RNA-Seq analysis of the parental seedlings under Cd treatment revealed 542 differentially expressed genes between Cd-tolerant Rec and Cd-susceptible Dom genotypes. By analyzing sequence variations in transcribed sequences of the parental genotypes, 155,654 SNPs and 1,525 InDels were identified between the two contrasting genotypes and may contribute to Cd tolerance. Finally, by integrating the data from the identified QTLs and RNA-Seq analysis, 16 Cd tolerance-related candidate genes were detected, nine of which were metal ion transporters. These results provide promising candidate genes for further gene cloning and improving Cd tolerance in barley.

Biophoton emission is a well-known phenomenon in living organisms, including plant species; however, the underlying mechanisms are not yet well elucidated. Nevertheless, non-invasive stress detection is of high importance when in plant production and plant research. Therefore, the aim of our work was to investigate, whether biophoton emission is suitable for the detection of cadmium stress in the early phase of stress evolution and to identify certain stress-related events that occur rapidly upon cadmium exposure of barley seedlings parallel to biophoton emission measurements. Changes of biophoton emission, chlorophyll content estimation index, ascorbate level, the activity of ascorbate- and guaiacol peroxidase enzymes and lipid oxidation were measured during seven days of cadmium treatment in barley (Hordeum vulgare L.) seedlings. The results indicate that the antioxidant enzyme system responded the most rapidly to the stress caused by cadmium and the lipid oxidation-related emission of photons was detected in cadmium-treated samples as early as one day after cadmium exposure. Furthermore, a concentration-dependent increase in biophoton emission signals indicating an increased rate of antioxidative enzymes and lipid oxidation was also possible to determine. Our work shows evidence that biophoton emission is suitable to identify the initial phase of cadmium stress effectively and non-invasively.

Different techniques have been applied in feed processing to improve ruminal degradation and nutrient utilization in ruminant. There are limited studies investigating how moist heating process impacts barley protein utilization and internal molecular structures. The objectives of this study were to investigate: 1) how moist heating affects barley protein chemical profiles and Cornell Net Carbohydrate and Protein System (CNCPS) subfractions, in situ rumen degradation parameters, and predicted intestinal protein supply and feed milk value; 2) how moist heating affects protein molecular spectral features; and 3) the relationship between protein molecular structure spectral features and protein chemical profiles and metabolic characteristics. The barley variety CDC cowboy samples collected from the research farm during two consecutive years were used. Half of each sample was kept as raw and the other half underwent moist heating. The advanced molecular spectroscopy (attenuated total reflectance-fourier transform infrared, ATR-FTIR) was used to detect the barley protein molecular structure spectral features. It was found that moist heating decreased the fractions of soluble protein and increased the moderately degradable protein and ingestible protein fractions. This further resulted in the changes of in situ rumen degradation parameters and intestinal protein digestion characteristics. The protein molecular structure spectral features detected by using ATR-FTIR spectroscopy can be used as potential predictors for protein related chemical and metabolic parameters.

Soil quality is the basis for the development of sustainable agriculture and may be used for evaluating the sustainability of soil management practices. Soil quality status and integrated soil quality index (SQI) in sampled 97 farmlands distributed in 7 barley agro-ecological areas of China were analyzed by using 13 soil chemical parameters. The results showed six principal components totally explained 72% variability for the 13 parameters and identified 9 parameters (includes pH, NH4+-N, NO3--N, available P, available K, exchangeable Mg, DTPA-Fe, DTPA-Cu and Cl-) with high factor loading values as the minimum data set (MDS) for assessing soil quality. Average soil quality of all farmlands is moderate (SQI = 0.62). The SQI of barley farmlands in 7 agro-ecological areas showed the following order: Inner Mongolia Plateau (0.75 ± 0.02) > Yunnan-Kweichow Plateau (0.72 ± 0.06) > Qinghai-Tibet Plateau (0.63 ± 0.08) > Yangtze Plain (0.62 ± 0.10) > Huanghuai Region (0.58 ± 0.09) > Northeast China Plain (0.56 ± 0.07) > Xinjiang Province (0.54 ± 0.07). Total 29 out of 97 farmlands in 7 areas have low SQI level (< 0.55). Hence, these farmlands require urgent attention for soil quality improvement through modification of the soil parameters in the MDS.

Lignocellulosic biomass such as barley straw is a renewable and sustainable alternative to traditional feeds and could be used as bioenergy sources; however, low hydrolysis rate reduces the fermentation efficiency. Understanding the degradation and colonization of barley straw by rumen bacteria is the key step to improve the utilization of barley straw in animal feeding or biofuel production. This study evaluated the hydrolysis of barley straw as a result of the inoculation by rumen fluid of camel and sheep. Ground barley straw was incubated anaerobically with rumen inocula from three fistulated camels (FC) and three fistulated sheep (FR) for a period of 72 h. The source of rumen inoculum did not affect the disappearance of dry matter (DMD), neutral detergent fiber (NDFD). Group FR showed higher production of glucose, xylose, and gas; while higher ethanol production was associated with cellulosic hydrolysates obtained from FC group. The diversity and structure of bacterial communities attached to barley straw was investigated by Illumina Mi-Seq sequencing of V4-V5 region of 16S rRNA genes. The bacterial community was dominated by phylum Firmicutes and Bacteroidetes. The dominant genera were RC9_gut_group,Ruminococcus,Saccharofermentans,Butyrivibrio,Succiniclasticum,Selenomonas, andStreptococcus, indicating the important role of these genera in lignocellulose fermentation in the rumen. Group FR showed higher RC9_gut_group and group FC revealed higherRuminococcus,Saccharofermentans, andButyrivibrio. Higher enzymes activities (cellulase and xylanase) were associated with group FC. Thus, bacterial communities in camel and sheep have a great potential to improve the utilization lignocellulosic material in animal feeding and the production of biofuel and enzymes.

The two alkaloids gramine and hordenine have been known for playing a role in the allelopathic ability in barley (Hordeum vulgare L.). These allelochemicals can be both found in leaves and roots in some barley cultivars whereas in others one seems to exclude the other. In this study eighteen accessions of barley from the Middle-East area, one accession from Tibet and the modern spring cultivar Barke, already used as parental donor in a nested associated mapping (NAM) population, were screened for their gramine, hordenine and N-methyltyramine (the direct precursor of hordenine) content in leaves, roots and exudates. Moreover, the toxicity of the three allelochemicals on root growth inhibition on lettuce (Lactuca sativa L.) was evaluated. Results of this study showed the preferential production of gramine and hordenine in leaves and roots, respectively, in the nineteen barley accessions. On the other hand, in the modern barley cultivar Barke, the highest content of hordenine in roots and the general lack of gramine suggests a favored biosynthesis of the former. Gramine was not detected in the root exudates. In additions, different metabolomic profiles were observed in wild relatives compared to modern barley genotypes. The results also showed the phytotoxic effects of the three compounds on root growth of lettuce seedlings, with a reduction in root length and an increase of root surface area and diameter. In conclusion, this study highlighted the impact of the domestication effects on the production and distribution of the two allelopathic alkaloids gramine and hordenine in barley.

Optimizing barley (hordeum vulgare L.) production in Idaho and other parts of the Pacific Northwest (PNW) should focus on farm resource management. The effect of post-harvest residue management on barley residue decomposition has not been adequately studied. Thus, the objective of this study was to determine the effect of residue placement (surface vs. incorporated), residue size (chopped vs. ground-sieved) and soil type (sand and sandy loam) on barley residue decomposition. A 50-day(d) laboratory incubation experiment was conducted at a temperature of 25°C at the Aberdeen Research and Extension Center, Aberdeen, Idaho, USA. Following the study, a Markov-Chain Monte Carlo (MCMC) modeling approach was applied to investigate the first-order decay kinetics of barley residue. An accelerated initial flush of residue carbon(C)-mineralization was measured for the sieved (Day 1) compared to chopped (Day 3 to 5) residues for both surface incorporated applications. The highest evolution of carbon dioxide (CO2)-C of 8.3 g kg-1 dry residue was observed on Day 1 from the incorporated-sieved application for both soils. The highest and lowest amount of cumulative CO2-C released and percentage residue decomposed over 50-d was observed for surface-chopped (107 g kg-1 dry residue and 27%, respectively) and incorporated-sieved (69 g kg-1 dry residue and 18%, respectively) residues, respectively. There were no significant differences in C-mineralization from barley residue based on soil type or its interactions with residue placement and size (p >0.05). The largest decay constant k of 0.0083 d-1 was calculated for surface-chopped residue where the predicted half-life was 80 d, which did not differ from surface sieved or incorporated chopped. In contrast, incorporated-sieved treatments only resulted in a k of 0.0054 d-1 and would need an additional 48 d to decompose 50% of the residue. Future residue decomposition studies under field conditions are warranted to verify the residue C-mineralization and its impact on residue management.

The cytokinin dehydrogenase gene HvCKX2.1 is the regulatory target for the most abundant heterochromatic small RNAs in drought-stressed barley caryopses. We investigated the diversity of HvCKX2.1 in 228 barley landraces and 216 wild accessions and identified 14 haplotypes, five of these with ten or more members, coding for four different protein variants. The third largest haplotype was abundant in wild accessions (51 members), but absent from the landrace collection. Protein structure predictions indicated that the amino acid substitution specific to haplotype 3 could result in a change in the functional properties of the HvCKX2.1 protein. Haplotypes 1-3 have overlapping geographical distributions in the wild population, but the average rainfall amounts at the collection sites for haplotype 3 plants are significantly higher during November to February compared to the equivalent data for plants of haplotypes 1 and 2. We argue that the likelihood that haplotype 3 plants were excluded from landraces by sampling bias that occurred when the first wild barley plants were taken into cultivation is low, and that it is reasonable to suggest that plants with haplotype 3 are absent from the crop because these plants were less suited to the artificial conditions associated with cultivation. Although the cytokinin signalling pathway influences many aspects of plant development, the identified role of HvCKX2.1 in the drought response raises the possibility that the particular aspect of cultivation that mitigated against haplotype 3 relates in some way to water utilization. Our results therefore highlight the possibility that water utilization properties should be looked on as a possible component of the suite of physiological adaptations accompanying the domestication and subsequent evolution of cultivated barley.

To estimate the prevalence of herbicide-resistant weeds, 87 wheat and barley farms were randomly surveyed in the Canterbury region of New Zealand. Over 600 weed seed samples from up to 10 mother plants per taxon depending on abundance, were collected immediately prior to harvest (two fields per farm). Some samples provided by agronomists were tested on an ad-hoc basis. Over 40,000 seedlings were grown to the 2–4 leaf stage in glasshouse conditions and sprayed with high priority herbicides for grasses from the three modes-of-action acetyl-CoA carboxylase (ACCase)-inhibitors haloxyfop, fenoxaprop, clodinafop, pinoxaden, clethodim, acetolactate synthase (ALS)-inhibitors iodosulfuron, pyroxsulam, nicosulfuron, and the 5-enolpyruvyl shikimate 3-phosphate synthase (EPSPS)-inhibitor glyphosate. The highest manufacturer recommended label rates were applied for the products registered for use in New Zealand, often higher than the discriminatory rates used in studies elsewhere. Published studies of resistance were rare in New Zealand but we found weeds survived herbicide applications on 42 of the 87 (48%) randomly surveyed farms, while susceptible reference populations died. Resistance was found for ALS-inhibitors on 35 farms (40%) and to ACCase-inhibitors on 20 (23%) farms. The number of farms with resistant weeds (denominator is 87 farms) are reported for ACCase-inhibitors, ALS-inhibitors, and glyphosate respectively as: Avena fatua (9%, 1%, 0% of farms), Bromus catharticus (0%, 2%, 0%), Lolium spp. (17%, 28%, 0%), Phalaris minor (1%, 6%, 0%), and Vulpia bromoides (0%, not tested, 0%). Not all farms had the weeds present, five had no obvious weeds prior to harvest. This survey revealed New Zealand’s first documented cases of resistance in P. minor (fenoxaprop, clodinafop, iodosulfuron) and B. catharticus (pyroxsulam). Twelve of the 87 randomly sampled farms (14%) had ALS-inhibitor chlorsulfuron-resistant sow thistles, mostly Sonchus asper but also S. oleraceus. Resistance was confirmed in industry-supplied samples of the grasses Digitaria sanguinalis (nicosulfuron, two maize farms), P. minor (iodosulfuron, one farm), and Lolium spp. (cases included glyphosate, haloxyfop, pinoxaden, iodosulfuron, and pyroxsulam, 9 farms). Industry also supplied Stellaria media samples that were resistant to chlorsulfuron and flumetsulam (ALS-inhibitors) sourced from clover and ryegrass fields from the North and South Island.