ATPase Gene Research Articles

The A subunit of vacuolar H+-ATPase gene (CmVHA-A) plays opposite roles in plant growth and drought tolerance of chrysanthemum under different growing conditions

As the most prominent proton pumps in plants, vacuolar H+-ATPases (VHAs) comprise multiple subunits that are important for physiological processes and stress tolerance in plants. However, few studies on the roles of subunit genes of VHAs in chrysanthemum have been reported to date. In this study, the gene of A subunit of V-ATPase in chrysanthemum (CmVHA-A) was cloned and identified. CmVHA-A was conserved with VHA-A proteins from other plants. Expression analysis showed that CmVHA-A was highly expressed in most tissues of chrysanthemum except for the flower bud, and was readily induced by polyethylene glycol (PEG) treatment. Functional analysis demonstrated that CmVHA-A exerted a negative influence on the growth and development of shoot and root of chrysanthemum under normal conditions. RNA-sequencing (RNA-seq) analysis revealed the possible explanations for phenotypic differences between transgenic and wild-type (WT) plants. Under drought conditions, CmVHA-A positively affected the drought tolerance of chrysanthemum by enhancing antioxidase activity and alleviating photosynthetic disruption. Overall, CmVHA-A plays opposite roles in plant growth and drought tolerance of chrysanthemums under different growing conditions.

Characterization of Genomic, Physiological, and Probiotic Features of Lactiplantibacillus plantarum JS21 Strain Isolated from Traditional Fermented Jiangshui.

This study aimed to understand the genetic and metabolic traits of a Lactiplantibacillus plantarum JS21 strain and its probiotic abilities through laboratory tests and computer analysis. L. plantarum JS21 was isolated from a traditional fermented food known as "Jiangshui" in Hanzhong city. In this research, the complete genetic makeup of JS21 was determined using Illumina and PacBio technologies. The JS21 genome consisted of a 3.423 Mb circular chromosome and five plasmids. It was found to contain 3023 protein-coding genes, 16 tRNA genes, 64 rRNA operons, 40 non-coding RNA genes, 264 pseudogenes, and six CRISPR array regions. The GC content of the genome was 44.53%. Additionally, the genome harbored three complete prophages. The evolutionary relationship and the genome collinearity of JS21 were compared with other L. plantarum strains. The resistance genes identified in JS21 were inherent. Enzyme genes involved in the Embden-Meyerhof-Parnas (EMP) and phosphoketolase (PK) pathways were detected, indicating potential for facultative heterofermentative pathways. JS21 possessed bacteriocins plnE/plnF genes and genes for polyketide and terpenoid assembly, possibly contributing to its antibacterial properties against Escherichia coli (ATCC 25922), Escherichia coli (K88), Staphylococcus aureus (CMCC 26003), and Listeria monocytogenes (CICC 21635). Furthermore, JS21 carried genes for Na+/H+ antiporters, F0F1 ATPase, and other stress resistance genes, which may account for its ability to withstand simulated conditions of the human gastrointestinal tract in vitro. The high hydrophobicity of its cell surface suggested the potential for intestinal colonization. Overall, L. plantarum JS21 exhibited probiotic traits as evidenced by laboratory experiments and computational analysis, suggesting its suitability as a dietary supplement.

Functional identification of CCR1 gene in apple (Malus halliana) demonstrates that it enhances saline-alkali stress tolerance

BackgroundLignin is a complex aromatic polymer that plays an important biological role in maintaining plant structure and defending plants against biotic and abiotic stresses. Cinnamoyl-CoA reductase (CCR) is a key enzyme involved in the lignin synthesis-specific pathway and regulates lignin biosynthesis and accumulation.MethodsBased on transcriptome data, MhCCR1, which was significantly induced by saline-alkali stress, was cloned from Malus halliana. The physicochemical properties, evolutionary relationships and cis-acting elements were analyzed. We obtained transgenic materials by heterologous transformation of MhCCR1 into Arabidopsis thaliana and tobacco, as well as homologous transformation into apple callus, and characterized their saline-alkali stress resistance by a series of physiological and biochemical experiments. And yeast two-hybridization technique was applied to screen and validate the interacting proteins.ResultsWe found that overexpression of MhCCR1 enhanced the tolerance of A. thaliana, tobacco and apple calli under saline-alkali stress, and caused a variety of physiological and biochemical changes. As compared to the wild type, the transgenic plants showed better growth, higher lignin, chlorophyll and proline contents, lower conductivity and MDA content, and significant increase in antioxidant enzyme activities (SOD, POD, CAT) in the transgenic lines under stress condition. In addition, expression of saline-alkali stress-related genes in overexpressed A. thaliana were also higher than in WT, including the antioxidant genes, the Na+ transporter genes, and the H+-ATPase genes, while expression of the K+ transporter genes displayed opposite changes. Meanwhile, the expression levels of genes related to lignin synthesis, AtPAL1, AtCOMT, AtC4H, At4CL1, and AtCCOAOMT, were also significantly up-regulated. At last, the Y2H experiment confirmed the interaction between MhCCR1 and MhMYB4, MhMYB1R1, MhHXK, and MhbZIP23 proteins.ConclusionsThese results suggest that MhCCR1 may play a positive regulatory role in saline-alkali tolerance of transgenic lines by regulating the lignin content, osmoregulatory substances, chlorophyll content, antioxidant enzyme activities, and genes related to saline-alkali stress, thus providing excellent resistance genes for the stress-responsive regulatory network of apples, and providing a theoretical basis for the cultivation of saline and alkali-resistant apple varieties.Graphical

Identification and characterization of the plasma membrane H+-ATPase genes in Brassica napus and functional analysis of BnHA9 in salt tolerance

As a primary proton pump, plasma membrane (PM) H+-ATPase plays critical roles in regulating plant growth, development, and stress responses. PM H+-ATPases have been well characterized in many plant species. However, no comprehensive study of PM H+-ATPase genes has been performed in Brassica napus (rapeseed). In this study, we identified 32 PM H+-ATPase genes (BnHAs) in the rapeseed genome, and they were distributed on 16 chromosomes. Phylogenetical and gene duplication analyses showed that the BnHA genes were classified into five subfamilies, and the segmental duplication mainly contributed to the expansion of the rapeseed PM H+-ATPase gene family. The conserved domain and subcellular analyses indicated that BnHAs encoded canonical PM H+-ATPase proteins with 14 highly conserved domains and localized on PM. Cis-acting regulatory element and expression pattern analyses indicated that the expression of BnHAs possessed tissue developmental stage specificity. The 25 upstream open reading frames with the canonical initiation codon ATG were predicted in the 5’ untranslated regions of 11 BnHA genes and could be used as potential target sites for improving rapeseed traits. Protein interaction analysis showed that BnBRI1.c associated with BnHA2 and BnHA17, indicating that the conserved activity regulation mechanism of BnHAs may be present in rapeseed. BnHA9 overexpression in Arabidopsis enhanced the salt tolerance of the transgenic plants. Thus, our results lay a foundation for further research exploring the biological functions of PM H+-ATPases in rapeseed.

Melatonin reduces cadmium accumulation through cell wall fraction fixation capacity in cotton seedlings

Melatonin (MT) participates in plants' responses in plants’ response to cadmium (Cd) tolerance, although its work model remains elusive. Here, the function of MT in adjusting Cd accumulation in Gossypium hirsutum (CCRI12) was investigated. In this research we observed that exogenous 50 μM MT significantly reduced the Cd accumulation under 100 μМ Cd stress for 7 days of treatment in cotton seedlings. Cd markedly induced the ROS (H2O2, O2-) accumulation and elevated the expression of transporters for Cd uptake and translocation. After application of MT reduced ROS and MDA accumulation in response to high induction of antioxidant defense system; the genes encoding superoxide dismutase (GhSOD), catalase (GhCAT), and peroxidase (GhPOD), ascorbate peroxidase (GhAPX), and glutathione S-transferase (GhGST) were significantly upregulated accompanied by their level of content. Furthermore, MT down-regulated transporters for Cd uptake include Natural Resistance-Associated Macrophage Protein 1 (GhNRAMP3), Zinc/Iron-regulated transporter-like protein (GhZIP1/4/6), Heavy Metal ATPase gene (GhHMA2), and ATP-binding Cassette sub family C (GhABCC3/2). Exogenous application of MT raised the hemicelluloses, cellulose and pectin content accompanied by their expression levels, xyloglucan endotransglucosylase/hydrolase (GhXTH28/6/9/B), cellulose synthase like (GhCSLG1/2) and COBRA-like (GhCOBL4), pectin acetylesterases (GhPAE8/7), pectin methylesterases (GhPME31) which in turn increased the cell wall's binding capacity to Cd.

Seasonal drought promotes citrate accumulation in citrus fruit through the CsABF3-activated CsAN1-CsPH8 pathway.

Plenty of rainfall but unevenly seasonal distribution happens regularly in southern China. Seasonal drought from summer to early autumn leads to citrus fruit acidification, but how seasonal drought regulates citrate accumulation remains unknown. Herein, we employed a set of physiological, biochemical, and molecular approaches to reveal that CsABF3 responds to seasonal drought stress and modulates citrate accumulation in citrus fruits by directly regulating CsAN1 and CsPH8. Here, we demonstrated that irreversible acidification of citrus fruits is caused by drought lasting for > 30 d during the fruit enlargement stage. We investigated the transcriptome characteristics of fruits affected by drought and corroborated the pivotal roles of a bHLH transcription factor (CsAN1) and a P3A-ATPase gene (CsPH8) in regulating citrate accumulation in response to drought. Abscisic acid (ABA)-responsive element binding factor 3 (CsABF3) was upregulated by drought in an ABA-dependent manner. CsABF3 activated CsAN1 and CsPH8 expression by directly and specifically binding to the ABA-responsive elements (ABREs) in the promoters and positively regulated citrate accumulation. Taken together, this study sheds new light on the regulatory module ABA-CsABF3-CsAN1-CsPH8 responsible for citrate accumulation under drought stress, which advances our understanding of quality formation of citrus fruit.

Impacts of electric field coupled membrane bioreactor on phenol wastewater with high salinity: Performance, membrane fouling and eco-friendly strategy

Electric field coupled membrane bioreactor (EMBR) enhanced the treatment performance of phenol wastewater with high salinity, improved sludge settleability, and alleviated membrane fouling. The sludge volume index (SVI) and fouling rate were positively correlated with the extracellular polymeric substance (EPS) content, indicating that the electric field improved sludge settleability by improving the properties of EPS, successfully reducing the fouling rate by 42.86 %. The phenol-degrading and salt-tolerant genera were enriched, and the EPS secretion genera were reduced in EMBR. Moreover, EMBR effectively resisted salt stress by increasing the relative abundance of betaine synthesis (betA) and transport genes (proV and proW), glutamate synthesis genes (gltB and gltD), salt stress response gene (rpoS), and F-type H+/Na+-transport ATPase genes (atpA and atpD), as well as reducing the relative abundance of genes related to cell membrane permeability. Meanwhile, EMBR decreased the production of intracellular reactive oxygen species (ROS), activated the antioxidant system and inhibited the ROS-mediated Save our Soul (SOS) response, which further improved salt tolerance of microorganisms. Furthermore, the inhibition of the SOS response reduced the dissemination of antibiotic resistance genes (ARGs). The co-occurrence patterns of ARGs and genera indicated that EMBR could also suppress the dissemination of ARGs by reducing the relative abundance of ARGs hosts.

Chitosan oligosaccharide accelerates the dissemination of antibiotic resistance genes through promoting conjugative plasmid transfer

The dissemination of antibiotic resistance genes (ARGs), especially via plasmid-mediated horizontal gene transfer, poses a pervasive threat to global health. Chitosan-oligosaccharide (COS) is extensively utilized in medicine, plant and animal husbandry. However, their impact on microflora implies the potential to exert selective pressure on plasmid transfer. To explore the role of COS in facilitating the dissemination of ARGs via plasmid conjugation, we established in vitro mating models. The addition of COS to conjugation mixtures significantly enhanced the transfer of RP4 plasmid and mcr-1 positive IncX4 plasmid in both intra- and inter-specific. Phenotypic and transcriptome analysis revealed that COS enhanced intercellular contact by neutralizing cell surface charge and increasing cell surface hydrophobicity. Additionally, COS increased membrane permeability by inhibiting the Tol-Pal system, thereby facilitating plasmid conjugative transfer. Furthermore, COS served as the carbon source and was metabolized by E. coli, providing energy for plasmid conjugation through regulating the expression of ATPase and global repressor factor-related genes in RP4 plasmid. Overall, these findings improve our awareness of the potential risks associated with the presence of COS and the spread of bacterial antibiotic resistance, emphasizing the need to establish guidelines for the prudent use of COS and its discharge into the environment.

Improvement of Na+-K+ ATPase Activity and Gene Expression Associated with Glomerular Ultrastructural Protection in Diabetic Nephropathy by Acacia senegal: Role of Oxidative Stress Disturbances

Improvement of Na+-K+ ATPase Activity and Gene Expression Associated with Glomerular Ultrastructural Protection in Diabetic Nephropathy by Acacia senegal: Role of Oxidative Stress Disturbances

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.

A Case of Wilson's Disease Suspected to Involve a Novel Genetic Mutation of ATP7B Gene, Requiring a Differential Diagnosis with Non-alcoholic Steatohepatitis.

A 19-year-old Japanese man was referred for a further evaluation of liver dysfunction. Despite the absence of symptoms or obesity, the liver biopsy results were consistent with non-alcoholic steatohepatitis. Subsequent investigations revealed low serum ceruloplasmin, increased urinary copper excretion, and a known mutation c.3809A>G (p.Asn1270Ser) in the copper-transporting enzyme P-type ATPase (ATP7B) gene, leading to a diagnosis of Wilson's disease. A previously unreported variant, i.e., c.3866A>T (p.Asp1289Val) was detected on the patient's other allele and was considered a novel mutation, classified as 'likely pathogenic' according to the American College of Medical Genetics guidelines.

Identification and pathogenicity of Alternaria species causing leaf blotch and fruit spot of apple in California

In late summer 2020, symptoms of leaf blotch and fruit spot were observed in two different commercial apple orchards (cultivars ‘Pink Lady’ and ‘Modi’) in San Joaquin County, California, USA. Ninety Alternaria isolates were obtained from symptomatic leaves and fruits collected from the orchards. Based on morphological characteristics of the colonies, sporulation patterns, and conidia, the isolates were preliminarily separated into three morphogroups, tentatively identified as A. alternata, A. tenuissima and A. arborescens. Multi-locus phylogenetic analyses, using nucleotide sequences of plasma membrane ATPase, calmodulin, and Alternaria major allergen genes, showed that the isolates initially identified as A. tenuissima clustered with strains of A. alternata, following the current taxonomical arrangement of the genus. Pathogenicity tests on detached wounded apple leaves and fruits, using representative isolates of the three morphogroups, fulfilled Koch’s postulates. This is the first report of A. alternata and A. arborescens as causal agents of leaf blotch and fruit spot of apple in California.

LcVHA2 and LcVHA3 positively modulate energy metabolism and reduce pericarp browning in litchi fruit after application of Bacillus siamensis

Pericarp browning, postharvest disease, and a short storage life are the leading causes of economic losses in the litchi industry globally. Energy metabolism plays an integral role in regulating browning, disease resistance, and the shelf-life of litchi fruit. The ability of Bacillus siamensis (N-1) to control postharvest diseases and maintain the quality of a variety of tropical fruits has previously been reported, however its effect on litchi fruit has not been explored. In the present study, we investigated the effect of Bacillus siamensis (N-1) on the postharvest energy metabolism of litchi fruit and its relation to pericarp browning, as well as the expression of energy-related genes. Results indicated that litchi fruit treated with Bacillus siamensis (N-1) exhibited a significant improvement in storability as evidenced by a decrease in the rate of the browning index (BI), color change, rate of weight loss, and laccase (LAC) and polyphenol oxidase (PPO) activity. There was also an increase in the energy status of litchi fruit due to the enhanced activity of Ca2+-ATPase, H+-ATPase, oxidase (CCO), cytochrome C, and succinic dehydrogenase (SDH), as well as the expression of their corresponding genes. LcVHA2 and LcVHA3, two major vacuolar-type H+-ATPase genes, were cloned and characterized. Phylogenetic analysis revealed that these genes are members of the V-ATPase subunit D and E gene family, respectively. Both heterologous (in tomato) and homologous (in litchi) transient overexpression (OE) assays indicated that both LcVHA2 and LcVHA3 have the potential to delay changes in fruit coloration and pericarp browning, but also increase ADP, ATP, and V-H+-ATPase activity, as well as the energy charge (EC) of fruit. Collectively, results indicated that the LcVHA2 and LcVHA3 play a critical role in preventing the browning and senescence of litchi fruit. This study provides new information on the ability of Bacillus siamensis (N-1), which has been previously demonstrated to be an effective postharvest biocontrol agent, to also improve the storage life of litchi fruit.

Enhanced Rice Resistance to Sheath Blight through Nitrate Transporter 1.1B Mutation without Yield Loss under NH4+ Fertilization.

Nitrogen fertilization can promote rice yield but decrease resistance to sheath blight (ShB). In this study, the nitrate transporter 1.1b (nrt1.1b) mutant that exhibited less susceptibility to ShB but without compromising yield under NH4+ fertilization was screened. NRT1.1B's regulation of ShB resistance was independent of the total nitrogen concentration in rice under NH4+ conditions. In nrt1.1b mutant plants, the NH4+ application modulated auxin signaling, chlorophyll content, and phosphate signaling to promote ShB resistance. Furthermore, the findings indicated that NRT1.1B negatively regulated ShB resistance by positively modulating the expression of H+-ATPase gene OSA3 and phosphate transport gene PT8. The mutation of OSA3 and PT8 promoted ShB resistance by increasing the apoplastic pH in rice. Our study identified the ShB resistance mutant nrt1.1b, which maintained normal nitrogen use efficiency without compromising yield.

Response of Fibroblasts from Menkes' and Wilson's Copper Metabolism-Related Disorders to Ionizing Radiation: Influence of the Nucleo-Shuttling of the ATM Protein Kinase.

Menkes' disease (MD) and Wilson's disease (WD) are two major copper (Cu) metabolism-related disorders caused by mutations of the ATP7A and ATP7B ATPase gene, respectively. While Cu is involved in DNA strand breaks signaling and repair, the response of cells from both diseases to ionizing radiation, a common DNA strand breaks inducer, has not been investigated yet. To this aim, three MD and two WD skin fibroblasts lines were irradiated at two Gy X-rays and clonogenic cell survival, micronuclei, anti-γH2AX, -pATM, and -MRE11 immunofluorescence assays were applied to evaluate the DNA double-strand breaks (DSB) recognition and repair. MD and WD cells appeared moderately radiosensitive with a delay in the radiation-induced ATM nucleo-shuttling (RIANS) associated with impairments in the DSB recognition. Such delayed RIANS was notably caused in both MD and WD cells by a highly expressed ATP7B protein that forms complexes with ATM monomers in cytoplasm. Interestingly, a Cu pre-treatment of cells may influence the activity of the MRE11 nuclease and modulate the radiobiological phenotype. Lastly, some high-passage MD cells cultured in routine may transform spontaneously becoming immortalized. Altogether, our findings suggest that exposure to ionizing radiation may impact on clinical features of MD and WD, which requires cautiousness when affected patients are submitted to radiodiagnosis and, eventually, radiotherapy.

Systematic investigation of the emerging pathogen of Tsukamurella species in a Chinese tertiary teaching hospital.

Tsukamurella species have been clinically regarded as rare but emerging opportunistic pathogens causing various infections in humans. Tsukamurella pneumonia has often been misdiagnosed as pulmonary tuberculosis due to its clinical presentation resembling tuberculosis-like syndromes. Tsukamurella species have also been confused in the laboratory with other phylogenetic bacteria, such as Gordonia. This study aimed to investigate the clinical, microbiological, and molecular characteristics; species distribution; and antimicrobial susceptibility of Tsukamurella species. Immunodeficiency and chronic pulmonary disease appeared to be risk factors for Tsukamurella pneumonia, and the presence of bronchiectasis and pulmonary nodules on imaging was highly correlated with this infection. The study confirmed that groEL (heat shock protein 60) and secA (the secretion ATPase) genes are reliable for identifying Tsukamurella species. Additionally, the ssrA (stable small RNA) gene showed promise as a tool for discriminating between different Tsukamurella species with the shortest sequence length. In terms of antimicrobial susceptibility, quinolones, trimethoprim/sulfamethoxazole, amikacin, minocycline, linezolid, and tigecycline demonstrated potent in vitro activity against Tsukamurella isolates in our study. The study also proposed a resistance mechanism involving a substitution (S91R) within the quinolone-resistance-determining region of the gyrA gene, which confers resistance to levofloxacin and ciprofloxacin. Furthermore, we found that disk diffusion testing is not suitable for testing the susceptibilities of Tsukamurella isolates to ciprofloxacin, imipenem, and minocycline. In conclusion, our systematic investigation may contribute to a better understanding of this rare pathogen. IMPORTANCE Tsukamurella species are rare but emerging human pathogens that share remarkable similarities with other mycolic acid-containing genera of the order Actinomycetales, especially Mycobacterium tuberculosis. Consequently, misdiagnosis and therapeutic failures can occur in clinical settings. Despite the significance of accurate identification, antimicrobial susceptibility, and understanding the resistance mechanism of this important genus, our knowledge in these areas remains fragmentary and incomplete. In this study, we aimed to address these gaps by investigating promising identification methods, the antimicrobial susceptibility patterns, and a novel quinolone resistance mechanism in Tsukamurella species, utilizing a collection of clinical isolates. The findings of our study will contribute to improve diagnosis and successful management of infections caused by Tsukamurella species, as well as establishing well-defined performance and interpretive criteria for antimicrobial susceptibility testing.

Improvement of ethanol and 2,3-butanediol production in Saccharomyces cerevisiae by ATP wasting

Background“ATP wasting” has been observed in 13C metabolic flux analyses of Saccharomyces cerevisiae, a yeast strain commonly used to produce ethanol. Some strains of S. cerevisiae, such as the sake strain Kyokai 7, consume approximately two-fold as much ATP as laboratory strains. Increased ATP consumption may be linked to the production of ethanol, which helps regenerate ATP.ResultsThis study was conducted to enhance ethanol and 2,3-butanediol (2,3-BDO) production in the S. cerevisiae strains, ethanol-producing strain BY318 and 2,3-BDO-producing strain YHI030, by expressing the fructose-1,6-bisphosphatase (FBPase) and ATP synthase (ATPase) genes to induce ATP dissipation. The introduction of a futile cycle for ATP consumption in the pathway was achieved by expressing various FBPase and ATPase genes from Escherichia coli and S. cerevisiae in the yeast strains. The production of ethanol and 2,3-BDO was evaluated using high-performance liquid chromatography and gas chromatography, and fermentation tests were performed on synthetic media under aerobic conditions in batch culture. The results showed that in the BY318-opt_ecoFBPase (expressing opt_ecoFBPase) and BY318-ATPase (expressing ATPase) strains, specific glucose consumption was increased by 30% and 42%, respectively, and the ethanol production rate was increased by 24% and 45%, respectively. In contrast, the YHI030-opt_ecoFBPase (expressing opt_ecoFBPase) and YHI030-ATPase (expressing ATPase) strains showed increased 2,3-BDO yields of 26% and 18%, respectively, and the specific production rate of 2,3-BDO was increased by 36%. Metabolomic analysis confirmed the introduction of the futile cycle.ConclusionATP wasting may be an effective strategy for improving the fermentative biosynthetic capacity of S. cerevisiae, and increased ATP consumption may be a useful tool in some alcohol-producing strains.

Mechanosensation in leaf veins.

Whether the plant vasculature has the capacity to sense touch is unknown. We developed a quantitative assay to investigate touch-response electrical signals in the leaves and veins of Arabidopsis thaliana. Mechanostimulated electrical signaling in leaves displayed strong diel regulation. Signals of full amplitude could be generated by repeated stimulation at the same site after approximately 90 minutes. However, the signals showed intermediate amplitudes when repeatedly stimulated in shorter timeframes. Using intracellular electrodes, we detected touch-response membrane depolarizations in the phloem. On the basis of this, we mutated multiple Arabidopsis H+-ATPase (AHA) genes expressed in companion cells. We found that aha1 aha3 double mutants attenuated touch-responses, and this was coupled to growth rate reduction. Moreover, propagating membrane depolarizations could be triggered by mechanostimulating the exposed primary vasculature of wild-type plants but not of aha1 aha3 mutants. Primary veins have autonomous mechanosensory properties which depend on P-type proton pumps.

CaHMA1 promotes Cd accumulation in pepper fruit

The main planting areas for pepper (Capsicum sp.) are high in cadmium (Cd), which is the most prevalent heavy metal pollutant worldwide. Breeding pepper cultivars with low Cd levels can promote sustainable agricultural production and ensure the safety of pepper products. To identify breeding targets for reducing Cd accumulation in pepper fruits, we performed a genome-wide association study on 186 accessions. Polymorphisms were associated with fruit Cd content in a genomic region containing a homolog of Arabidopsis (Arabidopsis thaliana) Heavy metal-transporting ATPase 1 (HMA1) encoding a P-type ATPase. In two cultivars with contrasting Cd accumulation, transcriptome analysis revealed differentially expressed genes enriched for carbohydrate metabolism and photosynthesis in fruits with high Cd accumulation, and a Cd2+/Zn2+-exporting ATPase gene (HMA). Heterologous expression of CaHMA1 in yeast increases Cd sensitivity. Overexpression of CaHMA1 conferred a severe increase in Cd content in Arabidopsis plants, whereas reduced CaHMA1 expression in pepper fruits decreased Cd content. We propose that CaHMA1 expression may be an important component of the high Cd accumulation in pepper plants.

The Na<sup>+</sup>/K<sup>+</sup>-ATPase Inhibitor Ouabain Has Different Effects on the Electrophysiological Properties of Excitatory and Inhibitory Neurons in the Entorhinal Cortex E

Na+/K+-ATPase maintains the neuron’s resting potential and the transmembrane gradient of K+ and Na+ cations, thus regulating ion transport and cellular volume. Mutations in Na+/K+-ATPase genes that impair its function can cause significant impairments in the nervous system function, including the development of epilepsy, if not lethal. Different forms of Na+/K+-ATPase are expressed in various classes of neurons and exhibit different characteristics. Thus, the impaired function of Na+/K+-ATPase may differentially affect the functioning of inhibitory and excitatory neurons. This study aims to determine the effects of the Na+/K+-ATPase antagonist ouabain on the electrophysiological characteristics of pyramidal cells and fast-spiking interneurons, as well as its impact on synaptic transmission. The results indicate that exposure to 5 µM ouabain results in depolarization of the resting membrane potential by 5 mV, as well as decreased amplitude and increased duration of the action potential of pyramidal neurons. Furthermore, ouabain caused a decrease in the amplitude of afterhyperpolarization in fast-spiking i-nterneurons. Moreover, both types of neurons exhibited a decrease in the threshold of action potential generation and the current at which depolarization block occurs. The addition of ouabain did not alter other electrophysiological characteristics of neurons. Furthermore, ouabain rapidly attenuates GABAergic transmission without affecting e-xcitatory synaptic transmission. These new findings on the effects of ouabain on excitatory pyramidal neurons and inhibitory interneurons contribute to the understanding of the mechanism underlying changes in the balance of excitation and inhibition in neural networks under Na+/K+-ATPase function impairment.