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.

Improvement of glucose metabolism in middle-aged mice on a high-fat diet by whole-grain highland barley is related to low methionine levels

Methionine restriction(MR)is an effective dietary strategy to regulate energy metabolism and alleviate oxidative stress and inflammation in the body,especially in the middle-aged and elderly population.However,the high methionine content of meat products makes this dietary strategy impossible to combine with protein supplementation and MR.Highland barley(HB),a low-methionine cereal,not only provides the body with protein but also has improved glucose metabolism and antioxidant and anti-inflammatory properties.Therefore,this study evaluated the feasibility of HB as a source of methionine-restricted dietary protein and the potential mechanisms.Middle-aged C57BL/6J mice were fed a control diet(CON),a high-fat diet(HFD),a whole-grain HB high-fat diet(HBHF),or a HBHF+methionine diet(HBHFmet)for 25 weeks.The results showed that the HBHF could keep the body weight,fasting glucose,insulin,homeostasis model assessment of insulin resistance(HOMA-IR),blood lipids,inflammation,and oxidative stress of HFD mice at normal levels.Compared with the HFD groups,HBHF inhibited pancreatic cell apoptosis and improved insulin secretion while improving hepatic and skeletal muscle glucose metabolism.However,these efficacies were attenuated in HBHFmet group mice.These findings suggest that HBHF has an MR strategy.

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.

Utilization of Barley Functional Foods for Preventing Chronic Diseases in China

Chronic diseases are the leading global causes of death in China and the world, especially hypertension ＇and diabetes, and the main reasons are heredity, abnormal metabolism and an unhealthy lifestyle for dietary. Barley is taditional medicine in China. There is an very important effect for functional foods for preventing chronic diseases of barley grains and barley grass powder as well as its products because of its high contents of minerals, vitamins, amino acids, antioxidant enzymes, chlorophyll and bioactive compounds such as β-glucans, phenolic compounds, GABA, alkaloid, tocopherols and tocotrienols, dietary fiber and so on. There are a lot of challenges to promote health for functional food with barley grains and its grass in China. Barley grain will produce pearled barley, grits, flakes and flour, and it adds texture, flavor, aroma, nutritional and medicinal value to product. Barley grass powder can be produced in barley green, barley grass rice noodle, barley grass ersi, barley grass noodles and barley green beer and other new functional foods. The future for barley use in food products is improving and very promising.

Ornithine Pathway in Proline Biosynthesis Activated by Salt Stress in Barley Seedlings

C-14-glutamate and C-14-arginine were spreaded on leaves of six-day old barley (Hordeum vulgare L.) seedlings that were treated with NaCl 200 mmol/L. The result showed that the pathway of arginine-->ornithine-->proline existed in the six-day old barley seedlings and was provoked remarkably by NaCl treatment. After seven days, proline accumulation contributed via the arginine-->ornithine-->proline pathway was 1.0 - 1.5 folds of that via the glutamate-->proline pathway. The activation of arginine-->ornithine-->proline pathway by salt stress in the salt-tolerant cultivar 'Jian 4' was 1.7 - 2.0 folds of that in the salt-sensitive cultivar 'KP 7', which suggested that the activation of arginine-->ornithine-->proline pathway in barley seedlings played an important role in improving salt tolerance of plants.

Effects of Salinity Stress on the Levels of Covalently and Noncovalently Conjugated Polyamines in Plasma Membrane and Tonoplast Isolated from Barley Seedlings

When the 7_d old barley ( Hordeum vulgare L.) seedlings were treated with different concentrations of NaCl for 3 d, the levels of the noncovalently conjugated polyamines (PAs) in the plasma membrane and tonoplast vesicles and the covalently conjugated PAs in the membrane proteins were promoted by NaCl of low concentrations and suppressed by NaCl of high concentrations. Among the noncovalently conjugated PAs in the vesicles, spermidine (Spd) level was the most abundant, while putrescine (Put) content was predominant among the covalently conjugated PAs, accounted for 40%-70%, 35%-60%, respectively. In addition, the TLC (thin_layer chromatography) profiles of the benzoylated PAs presented an unknown polyamine with Rf =0.92 (X 0.92 ), which conjugated covalently and noncovalently in root tonoplast and its content changed as well as Spd with NaCl treatment. The total PA contents in the roots were higher than that in the leaves, and the types and contents of covalently and noncovalently conjugated PAs in the tonoplast were higher than those in the plasma membrane. The results showed that the above two PAs associated with the membrane might be essential in salt adaption of cells and the maintenance of membrane function.

Relationship Between Polyamine Metabolism in Roots and Salt Tolerance of Barley Seedlings

Plant growth rate (GR), contents of free polyamines (fPAs) and bound polyamines (bPAs) and activities of some key enzymes involved in polyamine (PA) metabolism in the roots of two barley (Hordeum valgare L.) cultivars differing in salt sensitivity were investigated with 0-300 mmol/L NaCl treatments. With 0-200 mmol/L NaCl treatments, activities of arginine decarboxylase (ADC) and transglutaminase (TGase) and PA oxidase (PAO) in the roots of barley seedlings all increased, while TGase and PAO activities decreased slightly at 300 mmol/L NaCl. As a result, free Put (fPut) content increased continuously with increasing concentrations of NaCl, while levels of free Spd (fSpd) and an unknown PA (fPAx) and bPAs (bPut, bSpd and bPAx), as well as (fSpd + fPAx)/fPut ratio rose at 50-200 mmol/L NaCl and reduced at 300 mmol/L NaCl. However, no significant change in the tetra-amine spermine (Spin) content was observed. Statistical analysis showed that GR was very significantly positively correlated with (fSpd + fPAx)/fPut ratios and the contents of bPAs, whereas a significant inverse correlation existed between GR and the ratios of fPA contents to bPA levels. These results showed that, under salt stress, the balance between fSpd, fPAx and fPut levels and an equipoise between fPA and bPA contents in roots were important to salt tolerance of barley seedlings.

Relationship Between H^+-ATPase Activity and Fluidity of Tonoplast in Barley Roots Under NaCl Stress

H +_ATPase activity of tonoplast in roots of Hordeum vulgare L. cv. 'Tanyin 2' (salt_tolerant cultivar) increased when the roots were exposed to 50-200 mmol/L NaCl for 2 d, and decreased when NaCl concentration was increased to 600 mmol/L. In 'Kepin 7' (salt_sensitive cultivar), tonoplast H +_ATPase activity in roots also increased at lower levels of NaCl (50-100 mmol/L), but decreased at higher levels of NaCl (200-600 mmol/L). Tonoplast fluidity in roots of 'Tanyin 2' decreased at 50-200 mmol/L NaCl, and increased significantly at 600 mmol/L NaCl. Under salt stress, the change of tonoplast fluidity was identical with that of the ratio of unsaturated fatty acids to saturated fatty acids in tonoplast lipid of barley roots. It is proposed that the increase of tonoplast fluidity due to increased degree of unsaturation of fatty acids is one of the reasons leading to the decrease of H +_ATPase activity under higher level of NaCl stress.

Mechanism of the Effect of Polyamines on the Activity of Tonoplasts of Barley Roots Under Salt Stress

The seeds of barley Hordeum vulgare L. cv. Jian 4) were soaked with 0.1 mmol/L putrescine (Put) and 0.5 mmol/L spermidine (Spd), and then the seedlings were treated with 200 mmol/L NaCl. The growth rate (GR), dry matter accumulation, distribution of ions, the amount of polyamines (PAs) bound to tonoplast proteins as well as lipid composition and the activity of tonoplast vesicles isolated from roots were investigated. The results showed that soaking with Put or Spd could retard salt injury, promote GR and dry matter accumulation, and increase K+/Na+ in the roots. Compared with NaCl_treated plants, phospholipid content in root tonoplast rose by soaking with Put and Spd, while the level of galactose in lipids was decreased. Moreover, the ratio in noncovalently conjugated PA contents of (Spd+PAx (an unknown PA)) to (Put+Dap (diaminopropane)), and the total contents of covalently and noncovalently conjugated PAs were all increased. Statistical analysis indicated that the ratio of (Spd+PAx) to (Put+Dap) was significantly and positively correlated with the activities of membrane associated enzymes H+_ATPase and H+_PPase.

Identification of Wheat-Barley 2H Alien Substitution Lines

The genetic constitution of fifteen materials derived from the cross wheat (Triticum aestivum L. cv. 'Chinese Spring') X barley (Hordeum vulgare L. cv. 'Betzes') was analyzed, and six disomic alien substitution lines were screened by GISH. The chromosome configurations in pollen mother cells at meiotic metaphase I (PMCs M I) of F, from each disomic substitution line respectively crossed with double ditelocentric lines 2A, 2B and 2D of 'Chinese Spring' were observed, and a set of wheat-barley disomic alien substitution lines 2H(A), 2H(B) and 2H(D) were obtained. The RFLP analysis with the probe psr131 on the short arm of wheat homeologous group 2 combining with four restriction enzymes were carried out. The results indicated that the probe psr131 could be used as molecular marker to tag the barley chromosome 2H. The barley chromosome 2H had good genetic compensation ability for wheat chromosomes 2B and 2D in vitality and other agronomic characters. The result of testing seed was that the wheat appearance starch quality had been changed from the half-farinaceous of 'Chinese Spring' to the half-cutin of substitution lines by transferring the barley chromosome 2H to wheat.

Genetic Variation of Functional Components in Grains of Improved Barley Lines from Four Continents

[Objective] The study was to analyze functional components in improved barley grains. [Method] Genetic variations of functional components in grains among 629 barley improved lines from four continents were determined with DU640-type ultraviolet spectrophotometry (BECKMAN). [Result] The contents (mg/100 g) of total flavones and GABA in grains of Asian barley lines (123.09±29.56, 9.49±4.34) were significantly higher than that of American barley lines (103.85±22.33, 7.38±3.59), while no significant difference was observed between Asia/Americas and Europe (115.47±11.41, 9.66±3.98) and Australia (104.20±4.76, 8.83±3.41); furthermore, there was no significant difference of resistant starch content (%) in barley grains among four continents [Asia (1.63±1.44), America (1.54±1.13), Europe (1.20±0.85) and Australia (0.27±0.26)]. The contents (%) of resistant starch in grains of two-rowed barley (ssp. Hordeum distichon Koern., 1.45±1.20) was significantly lower than that of poly-rowed barley (ssp. Hordeum vulgare Orlov., 1.95±1.24). On the contrary, the content of total flavones in two-rowed lines (111.43±27.79 mg/100 g) was significantly higher than that of poly-rowed lines (102.15±14.95 mg/100 g), and the content of GABA in two-rowed lines (8.55±3.73 mg/100 g) was also significantly higher than that of poly-rowed lines (5.96±3.95 mg/100 g). There was the most significant correlation between GABA content and resistant starch (-0.21**)/total flavones content (0.12**, P<0.01, n=629). There were great genotype differences among the functional components in barley grains. The coefficient of variation (78.60%) and range (0-9.29%) of resistant starch (1.56±1.22%) were relatively large, including 11 high-resistant starch lines above 5%; the coefficient of variation (49.00%) and range (0-30.67 mg/100g) of GABA (8.00±3.92 mg/100 g) were also relatively large, including 26 lines with GABA higher than 15 mg/100 g; the coefficient of variation (23.63%) and range (58.44-236.91 mg/100 g) of total flavones (109.44±25.85 mg/100 g) was relatively small, including 14 lines with total flavones higher than 176 mg/100 g. [Conclusion] There are zonal and genotypic differences in the contents of of functional components in barley grains.

Identification of Molecular Markers for the Thinopyrum intermedium Chromosome 2Ai-2 with Resistance to Barley Yellow Dwarf Virus

The wheat_ Thinopyrum intermedium addition lines Z1,Z2 contain a pair of Th. intermedium chromosomes 2Ai_2 carrying the gene with resistance to barley yellow dwarf virus (BYDV). Genomic in situ hybridization (GISH) was used to analyze the chromosome constitution of Z1,Z2 by using genomic DNA probes from Th. intermedium and Pseudoroegneria strigosa . The results showed that the chromosome constitution of either Z1 or Z2 composes of 42 wheat chromosomes and two Th. intermedium chromosomes (2Ai_2). The 2Ai_2 chromosome is St_E intercalary translocation, in which the E genomic chromosome segment translocated into the middle region of the long arm of chromosome belonging to St genome. With the genomic DNA probe of Ps. strigosa , the GISH pattern specific to the 2Ai_2 chromosome may be used as a molecular cytogenetic marker. A detailed RFLP analysis on Z1, Z2 and their parents was carried out by using 12 probes on the wheat group 2 chromosomes. Twenty RFLP markers specific to the 2Ai_2 chromosome were identified. Two RAPD markers of OPR16 -350 and OPH09 -1580 , specific to the 2Ai_2 chromosome, were identified from 280 RAPD primers. These molecular markers could be used to assisted_select translocation lines with small segment of the 2Ai_2 chromosome and provide tools to localize the BYDV resistance.

Relationship Between Tonoplast H^+-ATPase Activity, Ion Uptake and Calcium in Barley Roots Under NaCl Stress

Under NaCl stress for 2 d, H+-ATPase activity increased, and H+-PPase activity decreased in the tonoplast of salt-tolerant barley ( Hordeum vulgare L. cv. 'Tanyin 2') roots. La3+ (1 mmol/L), an inhibitor of Ca2+ channel in plasma membrane, and EGTA (5 mmol/L), a Ca2+ chelator, inhibited this NaCl-induced increase in H+-ATPase activity but stimulated the H+-PPase activity. Treatment of barley roots with CaM antagonist (trifluoperazine, TFP, 20 mumol/L) also diminished the increase of H+-ATPase activity induced by NaCl. La3+, TFP or La3+ + TFP increased Na+ uptake and decreased K+ and Ca2+ uptake in barley roots under NaCl stress. These results suggested that the activation of tonoplast H+-ATPase and the regulation of Na+ and K+ uptake under NaCl stress may be related to Ca2+-CaM system.