In vitro gastrointestinal simulated digestion of three plant proteins: determination of digestion rate, free amino acids and peptide contents.

Cassava protein (CP), barley protein (BP) and yellow pea protein (YPP) are important nutrient and integral constituent of staple in pet foods. It is known that the digestion of proteins directly influences their absorption and utilisation. In the present work, we performed in vitro simulated gastrointestinal digestion of three plant proteins as a staple for dog and cat food. The digestion rate of CP, BP and YPP in dog food was 56.33 ± 0.90%, 48.53 ± 0.91%, and 66.96 ± 0.37%, respectively, whereas the digestion rate of CP, BP, and YPP in cat food was 66.25 ± 0.72%, 43.42 ± 0.83%, and 58.05 ± 0.85%, respectively. Using SDS-polyacrylamide gel electrophoresis to determine the molecular weight (MW) of each protein and the products of their digestion, it was revealed that MW of digestion samples decreased, and MW during the small intestine phase was lower than that during the gastric phase. Peptide sequences of digested products were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and it was found that the total number of peptides in the small intestine digestion samples was higher than that in the gastric phase samples. The MW of peptides obtained from CP was within the range of 1000-1500 Da, while MW of peptides derived from BP and YPP was within the range of 400-2000 Da. In addition, free amino acids were mainly produced in the small intestine phase. Furthermore, the percentage of essential amino acids in the small intestine phase (63 ~ 82%) was higher than that in the gastric phase (37 ~ 63%). Taken together, these findings contribute to the current understanding of the utilisation of plant proteins in dog and cat foods and provide important insights into the selection and application of plant proteins as a staple in dog and cat foods.

Biosynthesis of the allelopathic alkaloid gramine in barley by a cryptic oxidative rearrangement.

The defensive alkaloid gramine not only protects barley and other grasses from insects but also negatively affects their palatability to ruminants. The key gene for gramine formation has remained elusive, hampering breeding initiatives. In this work, we report that a gene encoding cytochrome P450 monooxygenase CYP76M57, which we name AMI synthase (AMIS), enables the production of gramine in Nicotiana benthamiana, Arabidopsis thaliana, and Saccharomyces cerevisiae. We reconstituted gramine production in the gramine-free barley (Hordeum vulgare) variety Golden Promise and eliminated it from cultivar Tafeno by Cas-mediated gene editing. In vitro experiments unraveled that an unexpected cryptic oxidative rearrangement underlies this noncanonical conversion of an amino acid to a chain-shortened biogenic amine. The discovery of the genetic basis of gramine formation now permits tailor-made optimization of gramine-linked traits in barley by plant breeding.

Contrasting cytosolic glutathione redox dynamics under abiotic and biotic stress in barley as revealed by the biosensor Grx1-roGFP2.

Barley is a staple crop of major global importance and relatively resilient to a wide range of stress factors in the field. Transgenic reporter lines to investigate physiological parameters during stress treatments remain scarce. We generated and characterized transgenic homozygous barley lines (cv. Golden Promise Fast) expressing the genetically encoded biosensor Grx1-roGFP2, which indicates the redox potential of the major antioxidant glutathione in the cytosol. Our results demonstrated functionality of the sensor in living barley plants. We determined the glutathione redox potential (EGSH) of the cytosol to be in the range of -308 mV to -320 mV. EGSH was robust against a combined NaCl (150 mM) and water deficit treatment (-0.8 MPa) but responded with oxidation to infiltration with the phytotoxic secretome of the necrotrophic fungus Botrytis cinerea. The generated reporter lines are a novel resource to study biotic and abiotic stress resilience in barley, pinpointing that even severe abiotic stress leading to a growth delay does not automatically induce cytosolic EGSH oxidation, while necrotrophic pathogens can undermine this robustness.

Uptake and bioaccumulation of iron oxide nanoparticles (Fe3O4) in barley (Hordeum vulgare L.): effect of particle-size.

Root-to-shoot translocation of nanoparticles (NPs) is a matter of interest due to their possible unprecedented effects on biota. Properties of NPs, such as structure, surface charge or coating, and size, determine their uptake by cells. This study investigates the size effect of iron oxide (Fe3O4) NPs on plant uptake, translocation, and physiology. For this purpose, Fe3O4 NPs having about 10 and 100 nm in average sizes (namely NP10 and NP100) were hydroponically subjected to barley (Hordeum vulgare L.) in different doses (50, 100, and 200 mg/L) at germination (5 days) and seedling (3 weeks) stages. Results revealed that particle size does not significantly influence the seedlings' growth but improves germination. The iron content in root and leaf tissues gradually increased with increasing NP10 and NP100 concentrations, revealing their root-to-shoot translocation. This result was confirmed by vibrating sample magnetometry analysis, where the magnetic signals increased with increasing NP doses. The translocation of NPs enhanced chlorophyll and carotenoid contents, suggesting their contribution to plant pigmentation. On the other hand, catalase activity and H2O2 production were higher in NP10-treated roots compared to NP100-treated ones. Besides, confocal microscopy revealed that NP10 leads to cell membrane damages. These findings showed that Fe3O4 NPs were efficiently taken up by the roots and transported to the leaves regardless of the size factor. However, small-sized Fe3O4 NPs may be more reactive due to their size properties and may cause cell stress and membrane damage. This study may help us better understand the size effect of NPs in nanoparticle-plant interaction.

Bacterial crude oil and polyaromatic hydrocarbon degraders from Kazakh oil fields as barley growth support.

Bacterial strains of the genera Arthrobacter, Bacillus, Dietzia, Kocuria, and Micrococcus were isolated from oil-contaminated soils of the Balgimbaev, Dossor, and Zaburunye oil fields in Kazakhstan. They were selected from 1376 isolated strains based on their unique ability to use crude oil and polyaromatic hydrocarbons (PAHs) as sole source of carbon and energy in growth experiments. The isolated strains degraded a wide range of aliphatic and aromatic components from crude oil to generate a total of 170 acid metabolites. Eight metabolites were detected during the degradation of anthracene and of phenanthrene, two of which led to the description of a new degradation pathway. The selected bacterial strains Arthrobacter bussei/agilis SBUG 2290, Bacillus atrophaeus SBUG 2291, Bacillus subtilis SBUG 2285, Dietzia kunjamensis SBUG 2289, Kocuria rosea SBUG 2287, Kocuria polaris SBUG 2288, and Micrococcus luteus SBUG 2286 promoted the growth of barley shoots and roots in oil-contaminated soil, demonstrating the enormous potential of isolatable and cultivable soil bacteria in soil remediation. KEY POINTS: • Special powerful bacterial strains as potential crude oil and PAH degraders. • Growth on crude oil or PAHs as sole source of carbon and energy. • Bacterial support of barley growth as resource for soil remediation.

Jasmonic acid improves barley photosynthetic efficiency through a possible regulatory module, MYC2-RcaA, under combined drought and salinity stress.

The combined stress of drought and salinity is prevalent in various regions of the world, affects several physiological and biochemical processes in crops, and causes their yield to decrease. Photosynthesis is one of the main processes that are disturbed by combined stress. Therefore, improving the photosynthetic efficiency of crops is one of the most promising strategies to overcome environmental stresses, making studying the molecular basis of regulation of photosynthesis a necessity. In this study, we sought a potential mechanism that regulated a major component of the combined stress response in the important crop barley (Hordeum vulgare L.), namely the Rubisco activase A (RcaA) gene. Promoter analysis of the RcaA gene led to identifying Jasmonic acid (JA)-responsive elements with a high occurrence. Specifically, a Myelocytomatosis oncogenes 2 (MYC2) transcription factor binding site was highlighted as a plausible functional promoter motif. We conducted a controlled greenhouse experiment with an abiotic stress-susceptible barley genotype and evaluated expression profiling of the RcaA and MYC2 genes, photosynthetic parameters, plant water status, and cell membrane damages under JA, combined drought and salinity stress (CS) and JA + CS treatments. Our results showed that applying JA enhances barley's photosynthetic efficiency and water relations and considerably compensates for the adverse effects of combined stress. Significant association was observed among gene expression profiles and evaluated physiochemical characteristics. The results showed a plausible regulatory route through the JA-dependent MYC2-RcaA module involved in photosynthesis regulation and combined stress tolerance. These findings provide valuable knowledge for further functional studies of the regulation of photosynthesis under abiotic stresses toward the development of multiple-stress-tolerant crops.

A two-sequence motif-based method for the inventory of gene families in fragmented and poorly annotated genome sequences.

Databases of genome sequences are growing exponentially, but, in some cases, assembly is incomplete and genes are poorly annotated. For evolutionary studies, it is important to identify all members of a given gene family in a genome. We developed a method for identifying most, if not all, members of a gene family from raw genomes in which assembly is of low quality, using the P-type ATPase superfamily as an example. The method is based on the translation of an entire genome in all six reading frames and the co-occurrence of two family-specific sequence motifs that are in close proximity to each other. To test the method's usability, we first used it to identify P-type ATPase members in the high-quality annotated genome of barley (Hordeum vulgare). Subsequently, after successfully identifying plasma membrane H+-ATPase family members (P3A ATPases) in various plant genomes of varying quality, we tested the hypothesis that the number of P3A ATPases correlates with the ability of the plant to tolerate saline conditions. In 19 genomes of glycophytes and halophytes, the total number of P3A ATPase genes was found to vary from 7 to 22, but no significant difference was found between the two groups. The method successfully identified P-type ATPase family members in raw genomes that are poorly assembled.

Phytosiderophore pathway response in barley exposed to iron, zinc or copper starvation.

Efficient micronutrient acquisition is a critical factor in selecting micronutrient dense crops for human consumption. Enhanced exudation and re-uptake of metal chelators, so-called phytosiderophores, by roots of graminaceous plants has been implicated in efficient micronutrient acquisition. We compared PS biosynthesis and exudation as a response mechanism to either Fe, Zn or Cu starvation. Two barley (Hordeum vulgare L.) lines with contrasting micronutrient grain yields were grown hydroponically and PS exudation (LC-MS) and root gene expression (RNAseq) were determined after either Fe, Zn, or Cu starvation. The response strength of the PS pathway was micronutrient dependent and decreased in the order Fe > Zn > Cu deficiency. We observed a stronger expression of PS pathway genes and greater PS exudation in the barley line with large micronutrient grain yield suggesting that a highly expressed PS pathway might be an important trait involved in high micronutrient accumulation. In addition to several metal specific transporters, we also found that the expression of IRO2 and bHLH156 transcription factors was not only induced under Fe but also under Zn and Cu deficiency. Our study delivers important insights into the role of the PS pathway in the acquisition of different micronutrients.

Synthesis and properties of nano-cadmium oxide and its size-dependent responses by barley plant.

Present study included technological methods that made it possible to synthesize CdO nanoparticles and carry out their qualitative and quantitative diagnostics, confirming the as-prepared CdO nanoparticles (NPs) were spherical and had a size of 25 nm. Then, under the conditions of the model experiment the effect of CdO in macro and nanosized particles on absorption, transformation, and structural and functional changes occurring in cells and tissues of Hordeum vulgare L. (spring barley) during its ontogenesis was analyzed. Different analytical techniques were used to detect the transformation of CdO forms: Fourier-transform infrared spectroscopy (FTIR), Dynamic light scattering (DLS), X-ray fluorescence analysis (XRF), Scanning electron microscopy (SEM-EDXMA and TEM), X-ray diffraction (XRD), and X-ray absorption fine structure, consists of XANES - X-ray absorption near edge structure, and EXAFS - Extended X-ray absorption fine structure. Quantitative differences in the elemental chemical composition of barley root and leaf samples were observed. The predominant root uptake of Cd was revealed. CdO-NPs were found to penetrate deeply into barley plant tissues, where they accumulated and formed new mineral phases such as Cd5(PO4)3Cl and CdSO4 according to XRD analysis. The molecular-structural state of the local Cd environment in plant samples corresponding to Cd-O and Cd-Cd. The toxicity of CdO-NPs was found to significantly affect the morphology of intracellular structures are the main organelles of photosynthesis therefore, destructive changes in them obviously reduce the level of metabolic processes ensuring the growth of plants. This study is an attempt to show results how it is possible to combine some instrumental techniques to characterize and behavior of NPs in complex matrices of living organisms.

Mechanism of selenite tolerance during barley germination: A combination of tissue selenium metabolism alterations and ascorbate-glutathione cycle modulation.

Selenite is widely used to increase Selenium (Se) content in cereals, however excessive selenite may be toxic to plant growth. In this study, barley was malted to elucidate the action mechanism of selenite in the generation and detoxification of oxidative toxicity. The results showed that high doses (600 µM) of selenite radically increased oxidative stress by the elevated accumulation of superoxide and malondialdehyde, leading to phenotypic symptoms of selenite-induced toxicity like stunted growth. Barley tolerates selenite through a combination of mechanisms, including altering Se distribution in barley, accelerating Se efflux, and increasing the activity of some essential antioxidant enzymes. Low doses (150 µM) of selenite improved barley biomass, respiratory rate, root vigor, and maintained the steady-state equilibrium between reactive oxygen species (ROS) and antioxidant enzyme. Selenite-induced proline may act as a biosignal to mediate the response of barley to Se stress. Furthermore, low doses of selenite increased the glutathione (GSH) and ascorbate (AsA) concentrations by mediating the ascorbate-glutathione cycle (AsA-GSH cycle). GSH intervention and dimethyl selenide volatilization appear to be the primary mechanisms of selenite tolerance in barley. Thus, results from this study will provide a better understanding of the mechanisms of selenite tolerance in crops.

Response of salt stress resistance in highland barley (Hordeum vulgare L. var. nudum) through phenylpropane metabolic pathway.

Highland barley (Hordeum vulgare L. var. nudum) is a grain crop that grows on the plateau under poor and high salt conditions. Therefore, to cultivate high-quality highland barley varieties, it is necessary to study the molecular mechanism of strong resistance in highland barley, which has not been clearly explained. In this study, a high concentration of NaCl (240 mmol/L), simulating the unfavorable environment, was used to spray the treated highland barley seeds. Transcriptomic analysis revealed that the expression of more than 8,000 genes in highland barley seed cells was significantly altered, suggesting that the metabolic landscape of the cells was deeply changed under salt stress. Through the KEGG analysis, the phenylpropane metabolic pathway was significantly up-regulated under salt stress, resulting in the accumulation of polyphenols, flavonoids, and lignin, the metabolites for improving the stress resistance of highland barley seed cells, being increased 2.71, 1.22, and 1.17 times, respectively. This study discovered that the phenylpropane metabolic pathway was a significant step forward in understanding the stress resistance of highland barley, and provided new insights into the roles of molecular mechanisms in plant defense.

Effect of 3-nitrooxypropanol on enteric methane emissions of feedlot cattle fed with a tempered barley-based diet with canola oil.

A dose-response experiment was designed to examine the effect of 3-nitrooxypropanol (3-NOP) on methane (CH4) emissions, rumen function and performance of feedlot cattle fed a tempered barley-based diet with canola oil. Twenty Angus steers of initial body weight (BW) of 356 ±â€…14.4 kg were allocated in a randomized complete block design. Initial BW was used as the blocking criterion. Cattle were housed in individual indoor pens for 112 d, including the first 21 d of adaptation followed by a 90-d finishing period when five different 3-NOP inclusion rates were compared: 0 mg/kg dry matter (DM; control), 50 mg/kg DM, 75 mg/kg DM, 100 mg/kg DM, and 125 mg/kg DM. Daily CH4 production was measured on day 7 (last day of starter diet), day 14 (last day of the first intermediate diet), and day 21 (last day of the second intermediate diet) of the adaptation period and on days 28, 49, 70, 91, and 112 of the finisher period using open circuit respiration chambers. Rumen digesta samples were collected from each steer on the day prior to chamber measurement postfeeding, and prefeeding on the day after the chamber measurement, for determination of rumen volatile fatty acids (VFA), ammonium-N, protozoa enumeration, pH, and reduction potential. Dry matter intake (DMI) was recorded daily and BW weekly. Data were analyzed in a mixed model including period, 3-NOP dose and their interaction as fixed effects, and block as a random effect. Our results demonstrated both a linear and quadratic (decreasing rate of change) effect on CH4 production (g/d) and CH4 yield (g/kg DMI) as 3-NOP dose increased (P < 0.01). The achieved mitigation for CH4 yield in our study ranged from approximately 65.5% up to 87.6% relative to control steers fed a finishing feedlot diet. Our results revealed that 3-NOP dose did not alter rumen fermentation parameters such as ammonium-N, VFA concentration nor VFA molar proportions. Although this experimental design was not focused on the effect of 3-NOP dose on feedlot performance, no negative effects of any 3-NOP dose were detected on animal production parameters. Ultimately, the knowledge on the CH4 suppression pattern of 3-NOP may facilitate sustainable pathways for the feedlot industry to lower its carbon footprint.

Livestock methane (CH4) is the main source of greenhouse gases (GHGs) in agriculture, contributing to 11.6% of global GHGs emissions from human-related activities. Therefore, mitigating CH4 emissions from ruminant animals is a great opportunity for meeting the current climate targets. In this experiment, increasing inclusion rates of a promising CH4-mitigating compound, 3-nitrooxypropanol (3-NOP, from 50 to 125 mg of 3-NOP/kg of dry matter [DM]), were added to a barley-based feedlot diet containing 25 ppm of monensin and 7% fat (DM-basis) and fed to Angus steers. Under these conditions, increasing inclusion rate of 3-NOP reduced both production and yield of CH4 by up to 90%. Rumen fermentation, feed intake, and average daily gain were not affected by the 3-NOP dose. Our results on the potential CH4 suppression of 3-NOP may assist the feedlot industry towards sustainability by lowering its GHG output.

In vitro gastrointestinal digestion of highland barley protein: identification and characterization of novel bioactive peptides involved in gut cholecystokinin secretion.

BACKGROUND: The satiety hormone cholecystokinin (CCK) plays an important role in food intake inhibition. Its secretion is regulated by dietary components. The search for bioactive compounds that induce CCK secretion is currently an active area of research. The objective of this study was to evaluate the ability of highland barley protein digest (HBPD) to stimulate CCK secretion in vitro and in vivo and identify the responsible peptide sequences. RESULTS: HBPD was prepared by in vitro gastrointestinal digestion model. Peptides of <1000 Da in HBPD accounted for 82%. HBPD was rich in essential amino acids Leu, Phe and Val, but lack in sulfur amino acids Met and Cys. HBPD treatment at a concentration of 5 mg mL-1 significantly stimulated CCK secretion from enteroendocrine STC-1 cells (P < 0.05). Moreover, oral gavage with HBPD in mice significantly increased plasma CCK level. Chromatographic separation was performed to isolate peptide fractions involved in CCK secretion and peptide sequence was determined by ultra-performance liquid chromatography-tandem mass spectrometry. Two novel CCK-releasing peptides, PDLP and YRIVPL, were pointed out for their outstanding CCK secretagogue activity. CONCLUSION: This study demonstrated for the first time that HBPD had an ability to stimulate CCK secretion in vitro and in vivo and determined the bioactive peptides exerting CCK secretagogue activity in HBPD. © 2023 Society of Chemical Industry.

Molecular identification of a laccase that catalyzes the oxidative coupling of a hydroxycinnamic acid amide for hordatine biosynthesis in barley.

Plants produce dimerized phenolic compounds as secondary metabolites. Hordatine A (HA), a dehydrodimer of p-coumaroylagmatine (pCA), is an antifungal compound accumulated at high levels in young barley (Hordeum vulgare) seedlings. The enzyme responsible for the oxidative dimerization of pCA, which is the final step of the hordatine biosynthetic pathway, has not been identified. In this study, we first verified the presence of this enzyme activity in the crude extract of barley seedlings. Because the enzyme activity was not dependent on H2 O2 , the responsible enzyme was not peroxidase, which was previously implicated in HA biosynthesis. The analysis of the dissection lines of wheat (Triticum aestivum) carrying aberrant barley 2H chromosomes detected HA in the wheat lines carrying the distal part of the 2H short arm. This chromosomal region contains two laccase genes (HvLAC1 and HvLAC2) that are highly expressed at the seedling stage and may encode enzymes that oxidize pCA during the formation of HA. Changes in the HvLAC transcript levels coincided with the changes in the HA biosynthesis-related enzyme activities in the crude extract and the HA content in barley seedlings. Moreover, HvLAC genes were heterologously expressed in Nicotiana benthamiana leaves and in bamboo (Phyllostachys nigra) suspension cells and HA biosynthetic activities were detected in the crude extract of transformed N. benthamiana leaves and bamboo suspension cells. The HA formed by the enzymatic reaction had the same stereo-configuration as the naturally occurring HA. These results demonstrate that HvLAC enzymes mediate the oxidative coupling of pCA during HA biosynthesis.

Enhancement of Polyphenols and Antioxidant Activity in Germinated Black Highland Barley by Ultrasonication.

The aim of this study was to investigate the effect of ultrasonic stress germination (USG) on total phenolic contents (TPC), total flavonoid contents (TFC), the phenolic compositions, and antioxidant activities of black highland barley (BHB). The USG processing parameters, polyphenol profile, phenolic compositions, and antioxidant activities were explored after USG. Results showed that the optimal USG parameters were as follows: 350 W ultrasonic pretreatment power, 30 °C ultrasonication temperature, 25 min ultrasonication time, and 64 h germination time. Under these conditions, the total phenolic content (688.84 ± 5.30 mg/100 g) and total flavonoid content (59.23 ± 0.45 mg/100 g) of BHB were increased by 28.55% and 10.15%, respectively, compared to the untreated samples. In addition, the USG treatment could more effectively enrich bound phenolic acids and free flavonoids, among which the content of catechin was significantly increased by USG and was the main characteristic substance. Moreover, the USG treatment could improve the antioxidant activity and had a higher antioxidant potency composite index (APC index) (97.91%) of BHB. These results indicate that USG might be an effective method to enrich polyphenols and improve antioxidant activity in BHB.

The association between hordein polypeptide banding and agronomic traits in partitioning genetic diversity in six-rowed Ethiopian barley lines (Hordeum vulgare L.).

BACKGROUND: Evaluation of the extent of genetic variation within and between the populations of crop genetic resources are of paramount importance in any breeding program. An experiment aimed at assessing the extent of variation among barley lines and the degree of association between hordein polypeptide and agronomic traits was hence executed. METHODS: Field experiment was conducted in six environments between 2017-2019 involving 19 barley lines. Hordein bands were separated using vertical Sodium Dodecyl Sulphate Poly- acrylamide Gel Electrophoresis (SDS-PAGE). RESULTS: The analysis of variance revealed significant variation among lines and wider range units were observed for the agronomic traits. The line (Acc# 16,811-6) was superior, producing the highest grain yield (2.97 ton ha-1) across environments, 3.6 ton ha-1 at Holleta, and 1.93 ton ha-1 at Chefedonsa. At Arsi Negelle a different line Acc# 17146-9 was the highest yielding (3.15ton ha-1). SDS-PAGE-based analysis of barley lines separated 12 hordein bands between C (four bands) and B (eight bands) subunits. Interestingly bands 52, 46a, and 46b were uniquely conserved in the four naked barley lines (Acc#16809-14,16956-11, 17240-3, 17244-19). A considerably high proportion of genetic diversity within the populations than among the populations could be a repercussion of high gene flow which substantiates the longstanding and dominant informal seed exchange system among the farmers. The significant positive association between grain yield and band 50 evocates the expression of this allele may code for higher grain yield. The negative association between days to maturity and band 52 perhaps stipulates earliness in barely lines upon the manifestation of the band. Band 52 and 60 appeared to be associated with more than one agronomic trait (days to maturity and thousand kernel weight; grain filling period and grain yield respectively) and could be the result of pleiotropic characteristics of the genes residing in these banding regions. CONCLUSION: The barley lines exhibited substantial variation for hordein protein and agronomic traits. However, imparted the need for the implementation of decentralized breeding as a consequence of genotype-by-environment interaction. Significant hordein polypeptide and agronomic traits association advocated the utilization of hordein as a protein marker and perhaps consider them in the parental line selection.

Bacillus mycoides PM35 in combination with titanium dioxide (TiO2)⎯nanoparticles enhanced morpho-physio-biochemical attributes in Barley (Hordeum vulgare L.) under cadmium stress.

Plant growth-promoting rhizobacteria (PGPR) are naturally occurring soil bacteria and are known to induce plant growth promotion and titanium dioxide (TiO2)⎯nanoparticles (NPs) used in a range of applications that need increased whiteness, improved corrosion resistance and photocatalytic activity. Keeping in view the stress mitigation potential of TiO2⎯NPS and B. mycoides PM35, the existing research work was premeditated to inspect the beneficial role of seed priming with using different levels of TiO2⎯NPs i.e., [(0 no TiO2⎯NPs), 25 and 50 µg/ml] and soil incubation plant growth promoting rhizobacteria (B. mycoides PM35) i.e., [(0 no B. mycoides PM35), 10 and 20 µL] on biochemical, morphological and physiological characteristics of Barley (Hordeum vulgare L.) plants under different levels of Cd in the soil i.e., [(0 Cd), 50 and 100 mg kg-1]. Results from the present study showed that the increasing levels of Cd in the soil significantly (P < 0.05) decreased plant growth and biomass, photosynthetic pigments, gas exchange attributes, sugars, and nutritional contents from the roots and shoots of the plants. In contrast, increasing levels of Cd in the soil significantly (P < 0.05) increased oxidative stress indicators in term of malondialdehyde, hydrogen peroxide, and electrolyte leakage, and also increased organic acid exudation patter in the roots of H. vulgare. Although, the activities of enzymatic antioxidants and the response of their gene expressions in the roots and shoots of the plants and non-enzymatic such as phenolic, flavonoid, ascorbic acid, and anthocyanin contents were initially increased with the exposure of 50 mg kg-1 Cd, but decreased by the increasing the Cd concentration 100 mg kg-1 in the soil. The negative impact of Cd toxicity can overcome the application of PGPR (B. mycoides PM35) and TiO2⎯NPs, which ultimately increased plant growth and biomass by capturing the reactive oxygen species, and decreased oxidative stress in H. vulgare by decreasing the Cd contents in the roots and shoots of the plants. Our results also showed that the TiO2⎯NPs were more sever and showed better results when we compared with PGPR (B. mycoides PM35) under the same treatment of Cd in the soil. Research findings, therefore, suggest that the combined application of PGPR (B. mycoides PM35) and TiO2⎯NPs can ameliorate Cd toxicity in H. vulgare, resulting in improved plant growth and composition under metal stress, as depicted by balanced exudation of organic acids.

Highland Barley Tea Polyphenols Extract Alleviates Skeletal Muscle Fibrosis in Mice by Reducing Oxidative Stress, Inflammation, and Cell Senescence.

The tea of roasted Highland barley is a cereal-based drink rich in polyphenols. A model of skeletal muscle senescence and fibrosis was constructed using d-galactose-induced C2C12 myotubes, and Highland barley tea Polyphenols (HBP) were extracted for the intervention. We found that HBP effectively alleviated oxidative stress, inflammation, and fibrosis induced by d-galactose-induced skeletal muscle senescence. Also, HBP treatment significantly down-regulated pro-fibrotic genes, inflammation, and oxidative stress levels in a contusion model of senescent mice. Reduced levels of SIRT3 protein was found to be an essential factor in skeletal muscle senescence and fibrosis in both cellular and animal models, while HBP treatment significantly increased SIRT3 protein levels and viability in skeletal muscle. The ability of HBP to mitigate skeletal muscle fibrosis and oxidative stress was significantly reduced after SIRT3 silencing. Together, these results suggest that HBP intervention can significantly alleviate aging-induced oxidative stress, inflammation, and skeletal muscle fibrosis, with the activation of SIRT3 as the underlying mechanism of action.

Effects of bulk and nano-ZnO particles on functioning of photosynthetic apparatus in barley (Hordeum vulgare L.).

The functioning of the photosynthetic apparatus in barley (Hordeum vulgare L.) after 7-days of exposure to bulk (b-ZnO) and nanosized ZnO (n-ZnO) (300, 2000, and 10,000 mg/l) has been investigated. An impact on the amount of chlorophylls, photosynthetic efficiency, as well as the zinc accumulation in chloroplasts was demonstrated. Violation of the chloroplast fine structure was revealed. These changes were generally more pronounced with n-ZnO exposure, especially at high concentrations. For instance, the chlorophyll deficiency under 10,000 mg/l b-ZnO treatment was 31% and with exposure to 10,000 mg/l n-ZnO, the chlorophyll deficiency was already 52%. The expression analysis of the photosynthetic genes revealed their different sensitivity to b-ZnO and n-ZnO exposure. The genes encoding subunits of photosystem II (PSII) and, to a slightly lesser extent, photosystem I (PSI) showed the highest suppression of transcriptional levels. The mRNA levels of the subunits of cytochrome-b6f, NADH dehydrogenase, ribulose-1,5-bisphosphate carboxylase and ATP synthase, which, in addition to linear electron flow (LEF), participate in cyclic electron flow (CEF) and autotrophic CO2 fixation, were more stable or increased under b-ZnO and n-ZnO treatments. At the same time, CEF was increased. It was assumed that under the action of b-ZnO and n-ZnO, the processes of LEF are disrupted, and CEF is activated. This allows the plant to prevent photo-oxidation and compensate for the lack of ATP for the CO2 fixation process, thereby ensuring the stability of photosynthetic function in the initial stages of stress factor exposure. The study of photosynthetic structures of crops is important from the point of view of understanding the risks of reducing the production potential and the level of food security due to the growing use of nanoparticles in agriculture.

Cadmium distribution and transformation in leaf cells involved in detoxification and tolerance in barley.

Barley is a diagnostic plant that often used in the research of soil pollution by heavy metals, our research explored the detoxification and tolerance mechanism of cadmium(Cd) in barley through pot experiment. We investigated subcellular distribution, chemical forms and oxidative damage of Cd in barley leaves, combing with the transmission electron microscopy and Fourier-transform infrared spectroscopy(FT-IR) to further understand the translocation, transformation characteristics and toxic effect of Cd in cells. The results showed that, the bioaccumulation factors in roots and shoots of barley were ranged of 4.03-7.48 and 0.51-1.30, respectively. Barley reduces the toxic effects by storing Cd in the roots and reducing its transport to the shoots. Compared to the control treatment (0 mg/kg), the percentage of Cd in the cell wall fractions of leaves in 300 mg/kg Cd treatment increased from 34.74 % to 38.41 %; the percentage of the organelle fractions increased from 24.47 % to 56.02 %; and the percentage of soluble fraction decreased from 40.80 % to 5.57 %. We found that 69.13 % of the highly toxic inorganic Cd and water-soluble Cd were converted to less toxic pectates and protein-integrated Cd (50.20 %) and undissolved Cd phosphates (18.93 %). This conversion of Cd was mainly due to its combination with -OH, -NH, -CN, -C-O-C, and -C-O-P groups. Excessive Cd induced a significant (P < 0.05) increase in the levels of peroxidase, malondialdehyde, and cell membrane permeability, which damaged the cell membrane and allowed Cd to enter the organelles. The chloroplasts and mitochondria were destroyed, and eventually the metabolism of intracellular substances was affected, resulting in symptoms of toxicity. Our research provides cellular-scale insight into the mechanisms of Cd tolerance in barley.