Search Results (36)

This study investigates the effects of γ-aminobutyric acid (GABA) and proline, both individually and in combination, on the growth of oilseed rape under drought stress and following the resumption of irrigation. The goal was to determine whether the exogenous application of these compounds enhances the plants response to prolonged water deficit and, if so, to identify the biochemical processes involved in the plant tissue. The experiment was conducted under controlled laboratory conditions. After 21 days of plant cultivation, at the 3–4 leaf stage, seedlings were sprayed with aqueous solutions of GABA (0.1 mM) and proline (0.1 mM). The plants were then subjected to 8 days of severe drought stress, after which irrigation was resumed, and recovery was assessed over 4 days. The results showed that both amino acids alleviated the drought-induced stress as indicated by higher relative water content (RWC), increased levels of endogenous proline and photosynthetic pigments in leaves, and enhanced survival and growth recovery after drought. GABA-treated plants maintained membrane integrity and preserved plasma membrane (PM) ATPase activity during prolonged drought stress while reducing ethylene, H₂O₂, and MDA levels. Proline also influenced these biochemical responses, though to a lesser extent. The combination of GABA and proline facilitated better recovery of oilseed rape compared to the drought control group following rewatering. Notably, GABA treatment resulted in a significant increase in gene expression compared to the untreated control. Molecular analysis of drought-responsive genes revealed that the gene expression in plants treated with both proline and GABA was typically intermediate between those treated with proline alone and those treated with GABA alone. Based on these findings, we propose that GABA application could serve as an alternative to proline for improving oilseed rape’s drought tolerance, potentially increasing both crop yield and quality. Full article

Plant plasma membrane (PM) H⁺-ATPase functions as a proton-motive force by exporting cellular protons to establish a transmembrane chemical gradient of H⁺ ions and an accompanying electrical gradient. These gradients are crucial for plant growth and development and for plant responses to abiotic and biotic stresses. In this study, a comprehensive identification of the PM H⁺-ATPase gene family was conducted across four cotton species. Specifically, 14 genes were identified in the diploid species Gossypium arboreum and Gossypium raimondii, whereas 39 and 43 genes were identified in the tetraploid species Gossypium hirsutum and Gossypium barbadense, respectively. The characteristics of this gene family were subsequently compared and analyzed using bioinformatics. Chromosomal localization and collinearity analyses elucidated the distribution characteristics of this gene family within the cotton genomes. Gene structure and phylogenetic analyses demonstrated the conservation of this family across cotton species, whereas the examination of cis-acting elements in gene promoters highlighted their involvement in environmental stress and hormone response categories. An expression profile analysis revealed eight genes whose expression was upregulated under salt stress conditions, and quantitative real-time PCR results suggested that the cotton PM H⁺-ATPase genes may play crucial roles in conferring resistance to salt stress. These findings establish a robust foundation for subsequent investigations into the functions of cotton PM H⁺-ATPase genes and may offer valuable insights for selecting genes for resistance breeding programs. Full article

Human lactoferrin (hLf) is an innate host defense protein that inhibits microbial H⁺-ATPases. This protein includes an ancestral structural motif (i.e., γ-core motif) intimately associated with the antimicrobial activity of many natural Cys-rich peptides. Peptides containing a complete γ-core motif from hLf or other phylogenetically diverse antimicrobial peptides (i.e., afnA, SolyC, PA1b, PvD₁, thanatin) showed microbicidal activity with similar features to those previously reported for hLf and defensins. Common mechanistic characteristics included (1) cell death independent of plasma membrane (PM) lysis, (2) loss of intracellular K⁺ (mediated by Tok1p K⁺ channels in yeast), (3) inhibition of microbicidal activity by high extracellular K⁺, (4) influence of cellular respiration on microbicidal activity, (5) involvement of mitochondrial ATP synthase in yeast cell death processes, and (6) increment of intracellular ATP. Similar features were also observed with the BM2 peptide, a fungal PM H⁺-ATPase inhibitor. Collectively, these findings suggest host defense peptides containing a homologous γ-core motif inhibit PM H⁺-ATPases. Based on this discovery, we propose that the γ-core motif is an archetypal effector involved in the inhibition of PM H⁺-ATPases across kingdoms of life and contributes to the in vitro microbicidal activity of Cys-rich antimicrobial peptides. Full article

Xanthomonas campestris pathovar campestris (Xcc) is a significant phytopathogen causing black rot disease in crucifers. Xcc injects a variety of type III effectors (T3Es) into the host cell to assist infection or propagation. A number of T3Es inhibit plant immunity, but the biochemical basis for a vast majority of them remains unknown. Previous research has revealed that the evolutionarily conserved XopL-family effector XopL_Xcc inhibits plant immunity, although the underlying mechanisms remain incompletely elucidated. In this study, we identified proton pump interactor (PPI1) as a specific virulence target of XopL_Xcc in Arabidopsis. Notably, the C-terminus of PPI1 and the Leucine-rich repeat (LRR) domains of XopL_Xcc are pivotal for facilitating this interaction. Our findings indicate that PPI1 plays a role in the immune response of Arabidopsis to Xcc. These results propose a model in which XopL_Xcc binds to PPI1, disrupting the early defense responses activated in Arabidopsis during Xcc infection and providing valuable insights into potential strategies for regulating plasma membrane (PM) H⁺-ATPase activity during infection. These novel insights enhance our understanding of the pathogenic mechanisms of T3Es and contribute to the development of effective strategies for controlling bacterial diseases. Full article

Human defensins are cysteine-rich peptides (Cys-rich peptides) of the innate immune system. Defensins contain an ancestral structural motif (i.e., γ-core motif) associated with the antimicrobial activity of natural Cys-rich peptides. In this study, low concentrations of human α- and β-defensins showed microbicidal activity that was not associated with cell membrane permeabilization. The cell death pathway was similar to that previously described for human lactoferrin, also an immunoprotein containing a γ-core motif. The common features were (1) cell death not related to plasma membrane (PM) disruption, (2) the inhibition of microbicidal activity via extracellular potassium, (3) the influence of cellular respiration on microbicidal activity, and (4) the influence of intracellular pH on bactericidal activity. In addition, in yeast, we also observed (1) partial K⁺-efflux mediated via Tok1p K⁺-channels, (2) the essential role of mitochondrial ATP synthase in cell death, (3) the increment of intracellular ATP, (4) plasma membrane depolarization, and (5) the inhibition of external acidification mediated via PM Pma1p H⁺-ATPase. Similar features were also observed with BM2, an antifungal peptide that inhibits Pma1p H⁺-ATPase, showing that the above coincident characteristics were a consequence of PM H⁺-ATPase inhibition. These findings suggest, for the first time, that human defensins inhibit PM H⁺-ATPases at physiological concentrations, and that the subsequent cytosolic acidification is responsible for the in vitro microbicidal activity. This mechanism of action is shared with human lactoferrin and probably other antimicrobial peptides containing γ-core motifs. Full article

Chickens are sensitive to heat stress because their capacity to dissipate body heat is low. Hence, in chickens, excessive ambient temperature negatively influences their reproductive performance and health. Heat stress induces inflammation and oxidative stress, thereby rendering many reproductive organs dysfunctional. In this study, we evaluated the effects of the supplementation of dietary quercetin and vitamin E on the uterine function, eggshell quality via estrogen concentration, calcium metabolism, and antioxidant status of the uterus of laying hens under heat stress. The ambient temperature transformation was set at 34 ± 2 °C for 8 h/d (9:00 am–5:00 pm), which was followed by 22 °C to 28 °C for 16 h/d. Throughout the experiment, the relative humidity in the chicken’s pen was at 50 to 65%. A total of 400 Tianfu breeder hens (120-days-old) were randomly divided into four dietary experimental groups, including basal diet (Control); basal diet + 0.4 g/kg quercetin; basal diet + 0.2 g/kg vitamin E; and basal diet + the combination of quercetin (0.4 g/kg) and vitamin E (0.2 g/kg). The results show that the combination of quercetin and vitamin E significantly increased the serum alkaline phosphatase levels and the antioxidant status of the uterus (p < 0.05). In addition, the combination of quercetin and vitamin E significantly increased the concentrations of serum estrogen and progesterone, as well as elevated the expression of hypothalamic gonadotropin-releasing hormone-1 and follicular cytochrome P450 family 19 subfamily A member-1 (p < 0.05). We also found that the calcium levels of the serum and uterus were significantly increased by the synergistic effects of quercetin and vitamin E (p < 0.05), and they also increased the expression of Ca²⁺-ATPase and the mRNA expression of calcium-binding-related genes in the uterus (p < 0.05). These results are consistent with the increased eggshell quality of the laying hens under heat stress. Further, the combination of quercetin and vitamin E significantly increased the uterine morphological characteristics, such as the height of the uterine mucosal fold and the length of the uterine mucosa villus of the heat-stressed laying hens. These results collectively improve the uterine function, serum and uterine calcium concentration, eggshell strength, and eggshell thickness (p < 0.05) in heat-stressed laying hens. Taken together, we demonstrated in the present study that supplementing the combination of dietary quercetin and vitamin E alleviated the effects of heat stress and improved calcium metabolism, hormone synthesis, and uterine function in the heat-stressed laying hens. Thus, the supplementation of the combination of quercetin and vitamin E alleviates oxidative stress in the eggshell gland of heat-stressed laying hens, thereby promoting calcium concentration in the serum and eggshell gland, etc., in laying hens. Hence, the combination of quercetin and vitamin E promotes the reproductive performance of the laying hens under heat stress and can also be used as a potent anti-stressor in laying hens. Full article

Poor tenderness of camel meat has seriously hampered the development of the camel meat industry. This study investigated the effects of muscle fiber composition and ageing time on meat quality, glycolytic potential, and glycolysis-related enzyme activities. Muscle samples of the longissimus thoracis (LT), psoas major (PM), and semitendinosus (ST) were collected from eight 8–10 year old Sonid Bactrian camels (females). Muscle fiber composition was examined by ATPase staining and immunohistochemistry. Meat quality indexes, glycolytic potential, and activities of major glycolytic enzymes were examined at 4 °C aging for 1, 6, 24, 72, and 120 h. The results showed that LT was mainly composed of type IIb muscle fibers, whereas PM and ST were mainly composed of type I muscle fibers. The PCR results of the myosin heavy chain (MyHC) were consistent with the ATPase staining results. During aging, the shear force of LT muscle was always greater than that of PM and ST, and its glycolysis was the strongest; type IIa, IIb, and IIx muscle fibers were positively correlated with muscle shear force and glycolysis rate, and type I muscle fibers were significantly and negatively correlated with the activities of the key enzymes of glycolysis within 6 h. The results showed that the muscle fibers of LT muscle had the greatest glycolysis capacity. These results suggest that an excessive type IIb muscle fiber number percentage and area in camel meat accelerated the glycolysis process, but seriously affected the sensory profile of the camel meat. The results of this study provide directions for the camel industry when addressing the poor tenderness of camel meat. Full article

Bafilomycin A1 inhibits V-type H⁺ ATPases on the molecular level, which acidifies endo-lysosomes. The main objective of the study was to assess the effect of bafilomycin A1 on Ca²⁺ content, NAADP-induced Ca²⁺ release, and ATPase activity in rat hepatocytes and human colon cancer samples. Chlortetracycline (CTC) was used for a quantitative measure of stored calcium in permeabilized rat hepatocytes. ATPase activity was determined by orthophosphate content released after ATP hydrolysis in subcellular post-mitochondrial fraction obtained from rat liver as well as from patients’ samples of colon mucosa and colorectal cancer samples. In rat hepatocytes, bafilomycin A1 decreased stored Ca²⁺ and prevented the effect of NAADP on stored Ca²⁺. This effect was dependent on EGTA–Ca²⁺ buffers in the medium. Bafilomycin A1 significantly increased the activity of Ca²⁺ ATPases of endoplasmic reticulum (EPR), but not plasma membrane (PM) Ca²⁺ ATPases in rat liver. Bafilomycin A1 also prevented the effect of NAADP on these pumps. In addition, bafilomycin A1 reduced Na⁺/K⁺ ATPase activity and increased basal Mg²⁺ ATPase activity in the subcellular fraction of rat liver. Concomitant administration of bafilomycin A1 and NAADP enhanced these effects. Bafilomycin A1 increased the activity of the Ca²⁺ ATPase of EPR in the subcellular fraction of normal human colon mucosa and also in colon cancer tissue samples. In contrast, it decreased Ca²⁺ ATPase PM activity in samples of normal human colon mucosa and caused no changes in colon cancer. Bafilomycin A1 decreased Na⁺/K⁺ ATPase activity and increased basal Mg²⁺ ATPase activity in normal colon mucosa samples and in human colon cancer samples. It can be concluded that bafilomycin A1 targets NAADP-sensitive acidic Ca²⁺ stores, effectively modulates ATPase activity, and assumes the link between acidic stores and EPR. Bafilomycin A1 may be useful for cancer therapy. Full article

When plants are exposed to environmental stress, their growth is inhibited. Under such conditions, controlled inhibition of growth is beneficial for plant survival. Jasmonic acid (JA) is a well-known phytohormone that limits plant growth, which has been confirmed in several species. However, its role in cucumber seedlings has not yet been comprehensively investigated. For this reason, we aimed to determine the involvement of JA in the regulation of proteins crucial for growth including plasma membrane proton pump (PM H⁺-ATPase), PM nitrate transporters, and nitrate reductase (NR). Treatment of cucumber seedlings with JA not only limited their growth but also increased the H₂O₂ content in their roots. The main sources of ROS generated for signalling purposes are PM NADPH oxidase (RBOH) and superoxide dismutase (SOD). Exposure of seedlings to JA induced the expression of some CsRBOH and SOD encoding genes, suggesting that ROS signalling can be activated by JA. As a consequence of JA exposure, the activity of all analysed proteins was inhibited and the expression of their genes was modified. The results indicate that reduction of PM H⁺-ATPase activity and the related decrease in nitrate uptake and assimilation are responsible for the root growth retardation of JA-treated plants. Full article

Soil salinity is a worldwide problem that adversely affects plant growth and development. Soil salinization in Xinjiang of China is very serious. Ping’ou hybrid hazelnut, as an important ecological and economic tree species, as well as a salt-tolerant plant, has been grown in Xinjiang for over 20 years. Understanding the salt-tolerance mechanism of Ping’ou hybrid hazelnut is of great significance for the breeding of salt-tolerant varieties and the rational utilization of salinized land. In this study, ‘Liaozhen 7’, a fine variety of Ping’ou hybrid hazelnut, was selected as test material, and seedlings were treated with 0 (control), 50, 100 and 200 mM NaCl. Subsequently, the pattern of NaCl-induced fluxes of Na⁺, K⁺ and H⁺ in the root meristematic zone and their response to ion transport inhibitors were studied using non-invasive micro-test technology (NMT). Different concentrations of NaCl stress significantly increased the Na⁺ concentration in roots, while K⁺ concentration decreased first and then increased with the increase of NaCl concentration. Meanwhile, NaCl stress induced a significant decline in K⁺/Na⁺ ratio. Control and 200 mM NaCl-induced Na⁺ and K⁺ fluxes in roots exhibited an outward efflux, whereas an inward flux was observed for H⁺. Under 200 mM NaCl stress, the average rates of net Na⁺ and K⁺ efflux, as well as H⁺ influx in roots were significantly increased, which were 11.6, 6.7 and 2.3 times higher than that of control, respectively. Furthermore, pharmacological experiments showed that 200 mM NaCl-induced Na⁺ efflux; H⁺ influx was significantly suppressed by amiloride, an inhibitor of plasma membrane (PM) Na⁺/H⁺ antiporter, and sodium vanadate, an inhibitor of PM H⁺-ATPase. Net Na⁺ efflux and H⁺ influx induced by NaCl decreased by 89.9% and 135.0%, respectively. The NaCl-induced Na⁺ efflux was mediated by a Na⁺/H⁺ antiporter using energy provided by PM H⁺-ATPase. The NaCl-induced K⁺ efflux was significantly restricted by tetraethylamine chloride, a K⁺ channel inhibitor, and promoted by sodium vanadate, which decreased by 111.2% and increased by 80.8%, respectively, indicating that K⁺ efflux was regulated by depolarization-activated outward-rectifying K⁺ channels and non-selective cation channels (NSCCs). In conclusion, NMT data revealed that NaCl stress up regulated the root Na⁺/H⁺ antiporter and H⁺ pump (an activity of PM Na⁺/H⁺ antiport system) of ‘Liaozhen 7’, which compelled the Na⁺/H⁺ exchange across the PM and restricted K⁺ loss via depolarization-activated K⁺ channels and NSCCs simultaneously, thereby maintaining the K⁺/Na⁺ homeostasis and higher salt tolerance. Full article

The geomagnetic field (GMF) is a natural component of the biosphere, and, during evolution, all organisms experienced its presence while some evolved the ability to perceive magnetic fields (MF). We studied the response of 14-3-3 proteins and the plasma membrane (PM) proton pump H⁺-ATPase to reduced GMF values by lowering the GMF intensity to a near-null magnetic field (NNMF). Seedling morphology, H⁺-ATPase activity and content, 14-3-3 protein content, binding to PM and phosphorylation, gene expression, and ROS quantification were assessed in maize (Zea mays) dark-grown seedlings. Phytohormone and melatonin quantification were also assessed by LG-MS/MS. Our results suggest that the GMF regulates the PM H⁺-ATPase, and that NNMF conditions alter the proton pump activity by reducing the binding of 14-3-3 proteins. This effect was associated with both a reduction in H₂O₂ and downregulation of genes coding for enzymes involved in ROS production and scavenging, as well as calcium homeostasis. These early events were followed by the downregulation of IAA synthesis and gene expression and the increase in both cytokinin and ABA, which were associated with a reduction in root growth. The expression of the homolog of the MagR gene, ZmISCA2, paralleled that of CRY1, suggesting a possible role of ISCA in maize magnetic induction. Interestingly, melatonin, a widespread molecule present in many kingdoms, was increased by the GMF reduction, suggesting a still unknown role of this molecule in magnetoreception. Full article

Two ATP-dependent proton pumps function in plant cells. Plasma membrane H⁺-ATPase (PM H⁺-ATPase) transfers protons from the cytoplasm to the apoplast, while vacuolar H⁺-ATPase (V-ATPase), located in tonoplasts and other endomembranes, is responsible for proton pumping into the organelle lumen. Both enzymes belong to two different families of proteins and, therefore, differ significantly in their structure and mechanism of action. The plasma membrane H⁺-ATPase is a member of the P-ATPases that undergo conformational changes, associated with two distinct E1 and E2 states, and autophosphorylation during the catalytic cycle. The vacuolar H⁺-ATPase represents rotary enzymes functioning as a molecular motor. The plant V-ATPase consists of thirteen different subunits organized into two subcomplexes, the peripheral V₁ and the membrane-embedded V₀, in which the stator and rotor parts have been distinguished. In contrast, the plant plasma membrane proton pump is a functional single polypeptide chain. However, when the enzyme is active, it transforms into a large twelve-protein complex of six H⁺-ATPase molecules and six 14-3-3 proteins. Despite these differences, both proton pumps can be regulated by the same mechanisms (such as reversible phosphorylation) and, in some processes, such as cytosolic pH regulation, may act in a coordinated way. Full article

The length of sorghum mesocotyl plays a vital role in seed emergence from the soil, which is the foundation of healthy growth. In this study, we aimed to understand how exogenous auxin (IAA) promoted mesocotyl elongation of sorghum and its physiology mechanism. The results presented that exogenous IAA significantly promoted mesocotyl elongation in MS24B (short mesocotyl inbred line) by increasing the cell length, while with extra exogenous NPA (IAA inhibitor) application, the mesocotyl length presented a significant short phenotype. In Z210 (long mesocotyl inbred line), exogenous IAA had a slight effect on mesocotyl length elongation, while the NPA treatment decreased the mesocotyl length considerably. In MS24B, IAA treatment increased the activity of amylase to degrade starch to soluble sugar, and the activity of hexokinase was improved to consume the increased soluble sugar to offer more energy. The energy will help to increase the activity of PM H⁺-ATPase and the expression of expansin-related genes, which ultimately will promote the acidification of the plasma membrane in MS24B for cell elongation. Overall, the exogenous IAA functioned on the activation of energy metabolism, which in turn, inducted the acidification of the plasma membrane for mesocotyl elongation. Full article

Low nitrogen stress severely impedes crop growth and productivity. There has been substantial research on root adaptation to low nitrogen conditions in many plant species. However, the mechanism underlying the morphological response of the strawberry (Fragaria × ananassa Duch.) root to low-NO₃⁻ or low-NH₄⁺ stress remains poorly understood. Strawberry plants were hydroponically cultivated under 1 mM NO₃⁻, 1 mM NH₄⁺, and control (15 mM NO₃⁻) conditions to assess the physiological responses of their roots to low nitrogen stress. As a result, low nitrogen stresses increased the fresh weight of root, lateral root density, and root surface area, as well as enhanced the accumulation of indole-3-acetic acid and jasmonic acid while significantly reducing salicylic acid in the roots. Correspondingly, low nitrogen stresses increased PM H⁺-ATPase activity. Low-NO₃⁻ stress enhanced the activities of nitrate reductase and glutamine synthetase, whereas low-NH₄⁺ treatment led to higher glutamine synthetase and glutamate synthase activities. Collectively, the present results demonstrate that low nitrogen stresses enhance nitrogen uptake of strawberry roots by regulating hormones (indole-3-acetic acid, jasmonic acid, and salicylic acid) and thereby mediating PM H⁺-ATPase activity, while promoting nitrogen metabolism by upregulating the activities of nitrate reductase, glutamine synthetase, and glutamate synthase. In conclusion, low nitrogen conditions may facilitate more efficient acquisition of available N from the soil by strawberry root system. Full article

Hydrogen gas (H₂) is considered as a signaling molecule and plays multiple roles in plant growth. However, the effect of H₂ on postharvest physiology in lily scales during storage has not been reported. In this study, the regulatory roles of hydrogen-rich water (HRW, a H₂ donor, a concentration of 0.45 mM for 100% HRW) in water status, ion balance, and nutrients in Lanzhou lily (Lilium davidii var. unicolor) scales were investigated. The scales were soaked in HRW for 12 d, and sampling was performed every 3 d for a total of 5 times. The results show that HRW (0, 10, 50, and 100%) increased the fresh weight, dry weight, relative water content, and water loss rate in lily scales, with maximum biological response at 50% HRW. Treatment with 50% HRW significantly increased the K⁺ content and K⁺/Na⁺ ratio in lily scales and decreased Na⁺ content. The Na⁺ K⁺-ATPase, and PM H⁺-ATPase activities were also increased by 50% HRW treatment. Meanwhile, 50% HRW up-regulated the expression of AKT1 and HA3 genes and down-regulated the expression of NHX2 and SOS1 genes. In addition, 50% HRW treatment significantly increased the expression level of PIP1;5, PIP2A, TIP1;3, and TIP2;2 genes. Treatment with 50% HRW significantly increased the content of water-soluble carbohydrate, sucrose, glucose, and fructose in lily scales, and decreased the content of starch. In addition, 50% HRW treatment significantly increased the activity of α-amylase, β-amylase, total amylase, sucrose synthase, and sucrose phosphate synthase. Collectively, H₂ might enhance the water retention capacity and nutrient content in lily scales by maintaining ion balance, regulating aquaporin, and increasing sugar-metabolizing enzyme activity, thereby prolonging the storage period of postharvest scales of Lanzhou lily. Full article