Evaluation of the intracellular lipid-lowering effect of polyphenols extract from highland barley in HepG2 cells

Active ingredients from highland barley have received considerable attention as natural products for developing treatments and dietary supplements against obesity.In practical application,the research of food combinations is more significant than a specific food component.This study investigated the lipid-lowering effect of highland barley polyphenols via lipase assay in vitro and HepG2 cells induced by oleic acid(OA).Five indexes,triglyceride(TG),total cholesterol(T-CHO),low density lipoprotein-cholesterol(LDL-C),aspartate aminotransferase(AST),and alanine aminotransferase(ALT),were used to evaluate the lipidlowering effect of highland barley extract.We also preliminary studied the lipid-lowering mechanism by Realtime fluorescent quantitative polymerase chain reaction(q PCR).The results indicated that highland barley extract contains many components with lipid-lowering effects,such as hyperoside and scoparone.In vitro,the lipase assay showed an 18.4%lipase inhibition rate when the additive contents of highland barley extract were 100μg/m L.The intracellular lipid-lowering effect of highland barley extract was examined using 0.25 mmol/L OA-induced HepG2 cells.The results showed that intracellular TG,LDL-C,and T-CHO content decreased by 34.4%,51.2%,and 18.4%,respectively.ALT and AST decreased by 51.6%and 20.7%compared with the untreated hyperlipidemic HepG2 cells.q PCR results showed that highland barley polyphenols could up-regulation the expression of lipid metabolism-related genes such as PPARγand Fabp4.

Fecal microbiota transplantation:whole grain highland barley improves glucose metabolism by changing gut microbiota

Highland barley(HB)is a high-altitude cereal with rich nutritional components and potential health benefits.To clarify its hypoglycemic effect and mechanism,we investigated the effect of whole grain HB and fecal microbiota transplantation(FMT)on glucose metabolism and gut microbiota in high-fat diet and streptozotocin(HFD/STZ)-induced diabetic mice.The results showed that HB(40%)significantly decreased fasting blood glucose and the area under the glucose tolerance curve,significantly increased insulin secretion and improved insulin resistance in HFD/STZ-induced diabetic mice(P<0.05).Inflammatory factors and blood lipid indices were also significantly alleviated after 12 weeks of 40%HB intervention(P<0.05).Additionally,beneficial bacteria,such as Bifidobacterium and Akkermansia,were significantly enriched in the gut of diabetic mice after whole grain HB intervention.Meanwhile,the results of further FMT experiments verified that the fecal microbiota after the 40%HB intervention not only significantly increased the relative abundance of Bifidobacterium and Akkermansia but also effectively improved glucose metabolism and alleviated the inflammatory state in HFD/STZ-induced diabetic mice.Collectively,our study confirmed the bridge role of gut microbiota in improving glucose metabolism of whole grain HB,which could promote the development of precision nutrition.

Assessment of molecular markers and marker-assisted selection for drought tolerance in barley(Hordeum vulgare L.)

This review updates the present status of the field of molecular markers and marker-assisted selection(MAS),using the example of drought tolerance in barley.The accuracy of selected quantitative trait loci(QTLs),candidate genes and suggested markers was assessed in the barley genome cv.Morex.Six common strategies are described for molecular marker development,candidate gene identification and verification,and their possible applications in MAS to improve the grain yield and yield components in barley under drought stress.These strategies are based on the following five principles:(1)Molecular markers are designated as genomic‘tags’,and their‘prediction’is strongly dependent on their distance from a candidate gene on genetic or physical maps;(2)plants react differently under favourable and stressful conditions or depending on their stage of development;(3)each candidate gene must be verified by confirming its expression in the relevant conditions,e.g.,drought;(4)the molecular marker identified must be validated for MAS for tolerance to drought stress and improved grain yield;and(5)the small number of molecular markers realized for MAS in breeding,from among the many studies targeting candidate genes,can be explained by the complex nature of drought stress,and multiple stress-responsive genes in each barley genotype that are expressed differentially depending on many other factors.

Giving Food a Fiber Boost: Adding High Beta-Glucan Ingredient from Barley to Everyday Foods

Lack of dietary fiber contributes to many health issues, particularly chronic vascular diseases. Mixed linkage β-1.3 - 1.4 beta-glucan (beta-glucan, in this paper) is a confirmed beneficial ingredient for the human diet through reduction of cholesterol and the glycemic index. Barley contains the highest beta-glucan content of all the grains, and in this study, a percentage of flour from two high beta glucan lines was, each, added to an array of wheat-based food products to measure how it impacted total dietary fiber. Results showed that beta-glucan content was higher in all the products containing the added high beta-glucan flour, along with increased total dietary fiber content. Protein content in the food products is also increased with the higher protein in the barley flours added. Beta-glucan content in the barley-added products increased to 1.2% - 4.0% versus 0.2% - 0.5% in the pure wheat products, while the dietary fibers increased to 3.5% - 24.4% versus 2.1% - 9.1% in pure wheat product controls. This research provided experimental evidence that using a high beta-glucan barley ingredient in food can increase dietary fiber to benefit health.

Advances in studies on the physiological and molecular regulation of barley tillering

Tillering is a crucial trait closely associated with yield potential and environmental adaptation in cereal crops,regulated by the synergy of endogenous(genetic)and exogenous(environmental)factors.The physiological and molecular regulation of tillering has been intensively studied in rice and wheat.However,tillering research on barley is scarce.This review used the recent advances in bioinformatics to map all known and potential barley tiller development genes with their chromosomal genetic and physical positions.Many of them were mapped for the first time.We also discussed tillering regulation at genetic,physiological,and environmental levels.Moreover,we established a novel link between the genetic control of phytohormones and sugars with tillering.We provided evidence of how environmental cues and cropping systems help optimize the tiller number.This comprehensive review enhances the understanding of barley’s physiological and genetic mechanisms controlling tillering and other developmental traits.

A single nucleotide substitution in the MATE transporter gene regulates plastochron and the many noded dwarf phenotype in barley(Hordeum vulgare L.)

In higher plants,the shoot apical meristem produces lateral organs in a regular spacing(phyllotaxy)and timing(plastochron).The molecular analysis of mutants associated with phyllotaxy and plastochron would increase our understanding of the mechanism of shoot architecture formation.In this study,we identified mutant mnd8ynp5 that shows an increased rate of leaf emergence and a larger number of nodes in combination with a dwarfed growth habit from an EMS-treated population of the elite barley cultivar Yangnongpi 5.Using a map-based cloning strategy,the mnd8 gene was narrowed down to a 6.7-kb genomic interval on the long arm of chromosome 5H.Sequence analysis revealed that a C to T single-nucleotide mutation occurred at the first exon(position 953)of HORVU5Hr1G118820,leading to an alanine(Ala)to valine(Val)substitution at the 318th amino acid site.Next,HORVU5Hr1G118820 was defined as the candidate gene of MND8 encoding 514 amino acids and containing two multidrug and toxic compound extrusion(MATE)domains.It is highly homologous to maize Bige1and has a conserved function in the regulation of plant development by controlling the leaf initiation rate.Examination of modern barely varieties showed that Hap-1 was the dominant haplotype and was selected in barley breeding around the world.Collectively,our results indicated that mnd8ynp5 is a novel allele of the HORVU5Hr1G118820 gene that is possibly responsible for the shortened plastochron and many noded dwarf phenotype in barley.

Barley FASCIATED EAR genes determine inflorescence meristem size and yield traits

In flowering plants,the inflorescence meristem(IM)provides founder cells to form successive floral meristems,which are precursors of fruits and seeds.The activity and developmental progression of IM are thus critical for yield production in seed crops.In some cereals,such as rice(Oryza sativa)and maize(Zea mays),the size of undifferentiated IM,which is located at the inflorescence apex,is positively associated with yield traits such as spikelet number.However,the relationship between IM size and yieldrelated spike traits remains unknown in the Triticeae tribe.Here we report that IM size has a negative correlation with yield traits in barley(Hordeum vulgare).Three FASCIATED EAR(FEA)orthologs,HvFEA2,HvFEA3,and HvFEA4,regulate IM size and spike morphogenesis and ultimately affect yield traits.Three HvFEAs genes are highly expressed in developing spikes,and all three loss-of-function mutants exhibit enlarged IM size,shortened spikes,and reduced spikelet number,which may lead to reduced grain yield.Natural variations identified in HvFEAs indicate selection events during barley domestication.We further reveal that HvFEA4,as a transcription factor,potentially targets multiple pathways during reproductive development,including transcriptional control,phytohormone signaling,and redox status.The roles of barley FEA genes in limiting IM size and promoting spikelet formation suggest the potential of increasing yield by manipulating IM activity.

Salicylic Acid Application Mitigates Oxidative Damage and Improves the Growth Performance of Barley under Drought Stress

Drought is a severe environmental constraint,causing a significant reduction in crop productivity across the world.Salicylic acid(SA)is an important plant growth regulator that helps plants cope with the adverse effects induced by various abiotic stresses.The current study investigated the potential effects of SA on drought tolerance efficacy in two barley(Hordeum vulgare)genotypes,namely BARI barley 5 and BARI barley 7.Ten-day-old barley seedlings were exposed to drought stress by maintaining 7.5%soil moisture content in the absence or presence of 0.5,1.0 and 1.5 mM SA.Drought exposure led to severe damage to both genotypes,as indicated by phenotypic aberrations and reduction of dry biomass.On the other hand,the application of SA to drought-stressed plants protected both barley genotypes from the adverse effects of drought,which was reflected in the improvement of phenotypes and biomass production.SA supplementation improved relative water content and proline levels in drought-stressed barley genotypes,indicating the osmotic adjustment functions of SA under water-deficit conditions.Drought stress induced the accumulation of reactive oxygen species(ROS),such as hydrogen peroxide(H2O2)and superoxide(O_(2)•^(−)),and the lipid peroxidation product malondialdehyde(MDA)in the leaves of barley plants.Exogenous supply of SA reduced oxidative damage by restricting the accumulation of ROS through the stimulation of the activities of key antioxidant enzymes,including superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),ascorbate peroxidase(APX)and glutathione peroxidase(GPX).Among the three-applied concentrations of SA,0.5 mM SA exhibited better mitigating effects against drought stress considering the phenotypic performance and biochemical data.Furthermore,BARI barley 5 showed better performance under drought stress than BARI barley 7 in the presence of SA application.Collectively,our results suggest that SA played a crucial role in improving water status and antioxidant defense strategy to protect barley plants from the deleterious effects of water deficiency.

Effects of different thermal processing methods on bioactive components,phenolic compounds,and antioxidant activities of Qingke(highland hull-less barley)

Qingke(highland hull-less barley)is a grain replete with substantial nutrients and bioactive ingredients.In this study,we evaluated the effects of boiling(BO),steaming(ST),microwave baking(MB),far-infrared baking(FB),steam explosion(SE),and deep frying(DF)on bioactive components,phenolic compounds,and antioxidant activities of Qingke compared with the effects of traditional roast(TR).Results showed that the soluble dietary fiber,beta-glucan and water-extractable pentosans of Qingke in dry heat processes of TR,SE,MB and FB had a higher content compared with other thermal methods and had a better antioxidant activity of hydroxyl radical scavenging and a better reduction capacity,while those in wet heat processes of BO and ST had a better antioxidant activity of ABTS radical scavenging and a better Fe^(2+) chelating ability.DF-and SE-Qingke had a higher content of tocopherol,phenolic,and flavonoid.Overall,6 free phenolic compounds and 12 bound phenolic compounds of Qingke were identified,and free phenolic compounds suffered more damage during thermal processing.Principal component analysis showed that SE had more advantages in retaining and improving the main biological active ingredients of Qingke,and it may be the best method for treating Qingke.

Evaluation of Chlorophyll Content and Fluorescence Parameters as Indicators of Drought Tolerance in Barley

Drought is a major abiotic stress that severely affects food production worldwide. Agronomic and physiological traits associated with drought tolerance are suitable indicators for selection of drought tolerance genotypes to reduce the impact of water deficit on crop yield in breeding program. The objective of this study was to identify indicators related to drought tolerance through analysis of photosynthetic traits in barley （Hordeum vulgare L.）. These traits included chlorophyll content, initial fluorescence （Fo）, maximum primary yield of photochemistry of photosystem Ⅱ （Fv /Fo） and maximum quantum yield of photosystem Ⅱ （Fv/Fm）. Four genotypes （Tadmor, Arta, Morocco9-75 and WI2291） variable in drought tolerance were used to investigate the correlation between these traits and drought tolerance. The results reflected that all of these traits were affected negatively in the four genotypes at different levels of post-anthesis drought stress, but the decrease in drought tolerant genotypes was much less than that of drought sensitive genotypes. The results further revealed that the components of the photosynthetic apparatus could be damaged significantly in drought sensitive genotypes, while drought tolerant genotypes were relatively less affected. On the other hand, the values of chlorophyll content, Fo, Fv/Fo and Fv/Fm in drought tolerance genotypes were significantly higher than those in drought sensitive genotypes under drought stress. It was concluded that chlorophyll content, Fo, Fv/Fo and Fv/Fm could be considered as reliable indicators in screening barley germplasm for drought tolerance.

Assessing Crop Water Demand and Deficit for the Growth of Spring Highland Barley in Tibet, China

The aim of this study was to assess the crop water demand and deficit of spring highland barley and discuss suitable irrigation systems for different regions in Tibet, China. Long-term trends in reference crop evapotranspiration and crop water demand were analyzed in different regions, together with crop water demand and deficit of spring highland barley under different precipitation frequencies. Results showed that precipitation trends during growth stages did not benefit the growth of spring highland barley. The crop coefficient of spring highland barley in Tibet was 0.87 and crop water demand was 389.0 ram. In general, a water deficit was found in Tibet, because precipitation was lower than water consumption of spring highland barley. The most severe water deficit were in the jointing to heading stage and the heading to wax ripeness stage, which are the most important growth stages for spring highland barley; water deficit in these two stages would be harmful to the yield. Water deficit showed different characteristics in different regions. In conclusion, irrigation systems may be more successful if based on an analysis of water deficit within different growth stages and in different regions.

Differences in the Efficiency of Potassium (K) Uptake and Use in Barley Varieties

Screening of crop genotypes with high K efficiency plays a fundamental role in understanding their physiological mechanisms. In this study, pot culture experiment was conducted to reveal the differences in uptake and use of K among 56 barley varieties. The coefficient of variation （CV） of K uptake （Ku） and K dry matter production index （KDMPI） of these varieties were higher than other indices and both Ku and KDMPI were significantly different among the barley varieties. Furthermore, Ku and KDMPI were positively correlated with dry matter weight （DMW） during the whole growth period; K use efficiency （KUE） and K harvesting index （KHI） were significantly related to dry grain weight （DGW） at maturing stage. Through cluster of these indices, Sandrime and AC Westech were found to be the most K-efficient and K-inefficient genotypes of barley, respectively. Significant differences were observed in uptake and use capacity of K between the two genotypes. During the whole growth period, Ku, K dry matter production efficiency （KDMPE）, KDMPI and DMW of the K- efficient genotype were found to be 1.5-1.8, 1.4-2.3, 2.1-3.9, and 1.7-2.1 times higher than those of the K-inefficient genotype, respectively, and at maturing stage, DWG, KUE, and KHI of efficient genotype were higher than those of the inefficient one. The results also showed that Sandrime was the most efficient candidate among the tested varieties for K efficiency and further studies should be conducted to investigate its physiological and biochemical characteristics.

Nitrogen(N) metabolism related enzyme activities,cell uitrastructure and nutrient contents as affected by N level and barley genotype

Development of the new crop cultivars with high yield under low nitrogen （N） input is a fundamental approach to enhance agricultural sustainability, which is dependent on the exploitation of the elite germplasm. In the present study, four barley genotypes （two Tibetan wild and two cultivated）, differing in N use efficiency （NUE）, were characterized for their physiolog- ical and biochemical responses to different N levels. Higher N levels significantly increased the contents of other essential nutrients （P, K, Ca, Fe, Cu and Mn）, and the increase was more obvious for the N-efficient genotypes （ZD9 and XZ149）. The observation of ultrastructure showed that chloroplast structure was severely damaged under low nitrogen, and the two high N efficient genotypes were relatively less affected. The activities of the five N metabolism related enzymes, i.e., nitrate reductase （NR）, glutamine sy＇nthetase （GS）, nitrite reductase （NiR）, glutamate synthase （GOGAT） and glutamate dehydrogenase （GDH） all showed the substantial increase with the increased N level in the culture medium. However the increased extent differed among the four genotypes, with the two N efficient genotypes showing more increase in com- parison with the other two genotypes with relative N inefficiency （HXRL and XZ56）. The current findings showed that a huge difference exists in low N tolerance among barley genotypes, and improvement of some physiological traits （such as enzymes） could be helpful for increasing N utilization efficiency.

Mapping of Mutant Gene prbs Controlling Poly-Row-and-Branched Spike in Barley (Hordeum vulgare L.)

A row-type mutant of barley named poly-row-and-branched spike （prbs） was previously obtained from a two-rowed cultivar Pudamai-2 after treated by inflorescence soaking in maize total DNA solution. The mutant produces branched spikes with irregular multiple rows. Genetic analysis indicated that the mutant phenotype was caused by a recessive gene prbs, and the PRBS locus had a recessive epistatic effect on an independent locus （denoted as Vrsx） conferring the variation of two-rowed spike vs. six-rowed spike. This study aimed to map PRBS as well as Vrsx using simple sequence repeats （SSR） markers. We developed an F2 population from a cross between the prbs mutant and a six-rowed cultivar Putianwudu for the gene mapping. As the two target loci interacted to result in a segregation ratio of two-rowed type：six-rowed type：prbs=9：3：4 in the population, we adopted a special strategy to map the two loci. PRBS was mapped between SSR markers HvLTPPB and Bmag0508A on the short arm of chromosome 3H, with distances of 24.7 and 14.3 cM to the two markers, respectively. Vrsx was mapped between SSR markers Bmag0125 and Bmag0378 on chromosome 2H, with distances of 6.9 and 15.3 cM to the two markers, respectively. This suggests that Vrsx should be the known locus Vrs1, which predominantly controls row-type variation in barley cultivars, and PRBS is a new locus related to the row type of spikes in barley.

Transformation of TrxS Gene into Barley by Particle Bombardment

The production of malting barley in China can＇t meet the demand of beer industries because of poor quality and it becomes a bottleneck problem in beer manufacture industry. In this paper, TrxS gene cloned from Phalaris coerulescens was transferred into barley cultivar Yupi 1 （YP1） via biolistic bombardment. 1206 immature embryos were bombarded and seven transgenic plants carrying TrxS gene were confirmed by PCR and PCR-Southern blotting analysis. TrxS gene was expressed in transgenic plants by RT-PCR analysis. The activity of Trxh and α-amylase of transgenic line were higher than that of non-transgenic line, which is helpful to improve malting quality of barley.

Alleviation of Al Toxicity in Barley by Addition of Calcium

The potential mechanism by which Ca alleviates Al toxicity was investigated in barley seedlings. It was found that 100 Al-alone treatment inhibited barley plant growth and thereby reduced shoot height and root length, and dry weights of root, shoot and leaf; promoted Al accumulation but inhibited Ca absorption in plant tissues; and induced an increase in the activities of superoxide dismutase （SOD）, catalase （CAT）, and peroxidase （POD） and in the level of lipid peroxidation （MDA content） in leaves. Except for the increase in Ca concentration in plant tissues, treatment with 0.5 mM Ca in the absence of Al had less effect on the above-mentioned parameters, compared with the control. Addition of Ca efficiently reduced Al toxicity, which is reflected by the promotion of plant growth, reduction in Al concentration and MDA content, increase in Ca concentration and in SOD, POD, and CAT activities compared with the Al-alone-treatment; with increase in Ca level （3.0 raM）, the ameliorative effect became more dominant. This indicated that the alleviation of aluminum toxicity in barley seedlings with Ca supplementation could be associated with less absorption of Al and the enhancement of the protective ability of the cell because of increased activity of the antioxidative enzyme.

Fast mapping of a chlorophyll b synthesis-deficiency gene in barley(Hordeum vulgare L.) via bulked-segregant analysis with reduced-representation sequencing

Bulked-segregant analysis coupled with next-generation sequencing(BSA-seq) has emerged as an efficient tool for genetic mapping of single genes or major quantitative trait loci controlling(agronomic) traits of interest. However, such a mapping-by-sequencing approach usually relies on deep sequencing and advanced statistical methods. Application of BSA-Seq based on construction of reduced-representation libraries and allele frequency analysis permitted anchoring the barley pale-green(pg) gene on chromosome 3 HL. With further marker-assisted validation, pg was mapped to a 3.9 Mb physical-map interval. In the pg mutant a complete deletion of chlorophyllide a oxygenase(HvCAO) gene was identified.Because the product of this gene converts Chl a to Chl b, the pg mutant is deficient in Chl b.An independent Chl b-less mutant line M4437_2 carried a nonsynonymous substitution(F263 L) in the C domain of HvCAO. The study demonstrates an optimized pooling strategy for fast mapping of agronomically important genes using a segregating population.

Systemic acquired resistance, NPR1, and pathogenesis-related genes in wheat and barley

In Arabidopsis, systemic acquired resistance （SAR） is established beyond the initial infection by a pathogen or is directly induced by treatment with salicylic acid （SA） or its functional analogs, 2,6-dichloroisonicotinic acid （INA） and benzothiadiazole （BTH）. NPR1 protein is considered the master regulator of SAR in both SA signal sensing and transduction. In wheat （Triticum aesfivum） and barley （Hordeum vulgare）, both pathogen infection and BTH treatment can induce broad-spectrum resistance to various diseases, including powdery mildew, leaf rust, Fusarium head blight, etc. However, three different types of SAR-like responses including acquired resistance （AR）, systemic immunity （SI）, and BTH-induced resistance （BIR） seem to be achieved by activating different gene pathways. Recent research on wheat and barley NPR1 homologs in AR and SI has provided the initial clue for understanding the mechanism of SAR in these two plant species. In this review, the specific features ofAR, Si, and BIR in wheat and barley were summarized and compared with that of SAR in model plants of Arabidopsis and rice. Research updates on downstream genes of SAR, including pathogenesis-related （PR） and BTH-induced genes, were highlighted.

Identification of SNPs in barley(Hordeum vulgare L.)by deep sequencing of six reduced representation libraries

High-density genetic markers are required for genotyping and linkage mapping in identifying genes from crops with complex genomes, such as barley. As the most common variation, single nucleotide polymorphisms(SNPs) are suitable for accurate genotyping by using the next-generation sequencing(NGS) technology. Reduced representation libraries(RRLs) of five barley accessions and one mutant were sequenced using NGS technology for SNP discovery. Twenty million short reads were generated and the proportion of repetitive sequences was reduced by more than 56%. A total of 6061 SNPs were identified, and 451 were mapped to the draft sequence of the barley genome with pairing reads. Eleven SNPs were validated using length polymorphic allele-specific PCR markers.

Supplementation of Fermented Barley Extracts with Lactobacillus Plantarum dy-1 Inhibits Obesity via a UCP1-dependent Mechanism

Objective We aimed to explore how fermented barley extracts with Lactobacillus plantarum dy-1(LFBE) affected the browning in adipocytes and obese rats.Methods In vitro, 3T3-L1 cells were induced by LFBE, raw barley extraction(RBE) and polyphenol compounds(PC) from LFBE to evaluate the adipocyte differentiation.In vivo, obese SD rats induced by high fat diet(HFD) were randomly divided into three groups treated with oral gavage:(a) normal control diet with distilled water,(b) HFD with distilled water,(c) HFD with 800 mg LFBE/kg body weight(bw).Results In vitro, LFBE and the PC in the extraction significantly inhibited adipogenesis and potentiated browning of 3T3-L1 preadipocytes, rather than RBE.In vivo, we observed remarkable decreases in the body weight, serum lipid levels, white adipose tissue(WAT) weights and cell sizes of brown adipose tissues(BAT) in the LFBE group after 10 weeks.LFBE group could gain more mass of interscapular BAT(IBAT) and promote the dehydrogenase activity in the mitochondria.And LFBE may potentiate process of the IBAT thermogenesis and epididymis adipose tissue(EAT) browning via activating the uncoupling protein 1(UCP1)-dependent mechanism to suppress the obesity.Conclusion These results demonstrated that LFBE decreased obesity partly by increasing the BAT mass and the energy expenditure by activating BAT thermogenesis and WAT browning in a UCP1-dependent mechanism.