Pi Uptake Research Articles

Vascular bundle cell-specific expression of a phosphate transporter improves phosphate use efficiency of transgenic Arabidopsis without detrimental effects

Constitutive overexpression of phosphate (Pi) transporter family 1 often results in the accumulation of toxic levels of Pi, which causes growth retardation in plants. In contrast, we had previously reported that root epidermis-specific overexpression of the phosphate transporter TaPT2 in Arabidopsis leads to improved growth and Pi use efficiency. In the present study, we used promoters AtHKT1;1 and SKOR, which are predominantly expressed in the vascular bundle tissues, to overexpress TaPT2. Transgenic lines exhibited increased shoot growth compared to wild type plants under normal- and low-Pi conditions, along with elevated root Pi and total P content, and higher xylem sap Pi concentration, specifically under low-Pi conditions. This was attributed to moderate Pi accumulation in the xylem parenchyma cells, enhancing the Pi uploading capacity to the xylem. SKOR-TaPT2, however, did not complement pho1 mutant, which was defective in uploading Pi to the xylem. The transcriptional levels of VPT1 and VPT3, which are responsible for transporting excess Pi into a vacuole, were upregulated in SKOR promoter lines under normal-Pi conditions. Our results suggested that root vascular bundle-specific expression of TaPT2 is another promising strategy for increasing biomass production, Pi uptake, and Pi use efficiency while preventing growth retardation in transgenic plants.

A plastidial lipoyl synthase LIP1p plays a crucial role in phosphate homeostasis in Arabidopsis.

Phosphate (Pi) homeostasis is important for plant growth and adaptation to the dynamic environment, which requires the precise regulation of phosphate transporter (PHT) trafficking from the endoplasmic reticulum to the plasma membrane. LIPOYL SYNTHASE 1p (LIP1p) is known as a key enzyme in plastids to catalyze lipoylation of pyruvate dehydrogenase complex for de novo fatty acid synthesis. It is unknown whether this process is involved in regulating Pi homeostasis. Here, we demonstrate a new role of LIP1p in controlling Pi homeostasis by regulating PHT1 trafficking. We recovered a weak mutant allele of LIP1p in Arabidopsis that accumulates much less Pi and has enhanced expression of phosphate starvation-induced genes. LIP1p mutation alters the lipid profile and compromises vesicle trafficking of PHT1 to the plasma membrane to impair Pi uptake. Beside phosphorus, the homeostasis of a series of mineral nutrients was also perturbed in lip1p mutant. Our findings provide powerful genetic evidence to support the linkage between lipoylation and ion homeostasis in plants.

Downregulation of the rice HRS1 HOMOLOG3 transcriptional repressor gene due to N deficiency directly co-activates ammonium and phosphate transporter genes.

Rice HRS1 HOMOLOG3 (OsHHO3) acts as a transcriptional repressor of AMMONIUM TRANSPORTER1 (OsAMT1) genes in rice; thus, reduced OsHHO3 expression in nitrogen (N)-deficient environments promotes ammonium uptake. In this study, we show that OsHHO3 also functions as a repressor of a specific subset of phosphate (Pi) transporter (PT) genes involved in the uptake and root-to-shoot translocation of Pi, including OsPT2, OsPT4, and OsPHO1;1. Consistently, disruption of OsHHO3 increased Pi uptake and Pi contents in shoots and roots, while overexpression of OsHHO3 generated the opposite effects. Furthermore, phosphorus (P) deficiency slightly decreased OsHHO3 expression, upregulating a specific subset of PT genes. However, N deficiency was more effective than P deficiency in suppressing OsHHO3 expression in roots, and unlike N deficiency-dependent activation of PT genes under the control of OsHHO3, the P deficiency-dependent activation of OsAMT1 genes was minimal. Interestingly, the simultaneous deficiency of both N and P promoted the OsHHO3-regulated expression of PT genes more significantly than the deficiency of either N or P, but diminished the expression of genes regulated by OsPHR2, a master regulator of Pi starvation-responsive transcriptional activation. Phenotypic analysis revealed that the inactivation and overexpression of OsHHO3 improved and reduced plant growth, respectively, under N-deficient and P-deficient conditions. These results suggest that OsHHO3 regulates a specific subset of PT genes independently of OsPHR2-mediated regulation and plays a critical role in the adaptation to diverse N and P environments.

The microRNA399d-PHOSPHATE2 module alters rice sensitivity to rice ragged stunt virus by manipulating phosphate uptake.

Rice (Oryza sativa L.) production frequently faces threats from biotic and abiotic stressors, with rice ragged stunt virus (RRSV) as a substantial biotic factor. The relationship between inorganic phosphorus (Pi) content and susceptibility to RRSV is crucial yet poorly understood. This study investigates how phosphorus metabolism influences rice resistance to RRSV, focusing on genetic manipulations that modulate this relationship. The RRSV infection increased phosphate (Pi) content in the aerial parts of rice plants by enhancing Pi uptake and transport. Furthermore, the upregulation of microRNA399d (miR399d) and the suppression of its target gene OsPHOSPHATE2 (OsPHO2) enhanced Pi accumulation, increasing rice susceptibility to RRSV infection. Additionally, elevated Pi levels, which are associated with altered ROS dynamics, reduced ROS activity and potentially dampened the plant's innate immune response to viral infection. The miR399d-PHO2 module was identified as pivotal in mediating phosphate uptake and influencing susceptibility to RRSV through modulations in the phosphorus and ROS pathways. This study shed light on the regulatory mechanisms of phosphorus nutrition in rice, revealing a critical interaction between phosphorus metabolism, ROS dynamics, and viral defense. The findings suggest potential strategies for manipulating Pi levels to enhance plant resistance against viruses, opening avenues for agricultural improvements and disease management in rice.

Developmental responses of roots to limited phosphate availability: research progress and application in cereals.

Phosphorus (P), an essential macronutrient, is crucial for plant growth and development. However, available inorganic phosphate (Pi) is often scarce in soil, and its limited mobility exacerbates P deficiency in plants. Plants have developed complex mechanisms to adapt to Pi-limited soils. The root, the primary interface of the plant with soil, plays an essential role in plant adaptation to Pi-limited soil environments. Root system architecture significantly influences Pi acquisition via the dynamic modulation of primary root and/or crown root length, lateral root proliferation and length, root hair development, and root growth angle in response to Pi availability. This review focuses on the physiological, anatomical, and molecular mechanisms underpinning changes in root development in response to Pi starvation in cereals, mainly focusing on the model monocot plant rice (Oryza sativa). We also review recent efforts to modify root architecture to enhance P uptake efficiency in crops and propose future research directions aimed at the genetic improvement of Pi uptake and use efficiency in crops based on root system architecture.

In Vitro Antimicrobial Synergistic Activity and the Mechanism of the Combination of Naringenin and Amikacin Against Antibiotic-Resistant Escherichia coli.

Bacterial drug resistance is becoming an increasingly serious problem, and the development of antibacterial synergists is urgently needed. Combining existing antibiotics with promising nonantibiotic agents is one strategy that has been shown to be effective at overcoming the widespread emergence of antibiotic-resistant pathogens. In this study, we investigated the antibacterial activities and mechanism of naringenin (NG) combined with amikacin (AMK) against multidrug-resistant Escherichia coli (E. coli). We first measured the fractional inhibitory concentration (FIC) of NG combined with antibiotics via the checkerboard method. The results indicated that the combination of NG and AMK had a synergistic effect on E. coli ATCC 25922 and E. coli C7F3. In addition, this synergistic effect was verified by time-kill assays. Moreover, scanning electron microscopy (SEM) was used to observe cell morphology. The results showed that the cell wall of E. coli was destroyed. Furthermore, we assessed the leakage of alkaline phosphatase (AKP), K+, and protein. The extracellular AKP activity increased after the combinational group of 1/2MIC NG and 1/2MIC AMK, suggesting an impairment in cell wall permeability. An increase in the leakage of intracellular K+ and protein indicated an increase in cell inner membrane permeability. These results revealed that NG and AMK inhibited E. coli by damaging cell walls and membranes. In addition, PI uptake rapidly increased after treatment with NG and AMK. Confocal laser scanning microscopy (CLSM) revealed that NG caused cell wall and cell membrane damage in E. coli. In summary, our results provide a new strategy for responding to the development of E. coli drug resistance.

A lipid synthase maintains metabolic flux for jasmonate synthesis to regulate root growth and phosphate homeostasis.

Plants require phosphate (Pi) for proper growth and development but often face scarcity of this vital nutrient in the soil. Pi-starvation triggers membrane lipid remodeling to utilize the membrane phospholipid-bound Pi in plants. In this process, phospholipids are replaced by non-Pi-containing galactolipids (MGDG, DGDG) and sulfolipids. The galactolipids ratio (MGDG:DGDG) is suggested to influence jasmonic acid (JA) biosynthesis. However, how the MGDG:DGDG ratio, JA levels, and root growth are coordinated under Pi deficiency in rice (Oryza sativa) remains unknown. Here, we characterized DGDG synthase 1 (OsDGD1) for its role in regulating root development by maintaining metabolic flux for JA biosynthesis. We showed that OsDGD1 is responsive under low Pi and is under the direct control of Phosphate Starvation Response 2 (OsPHR2), the master regulator of low Pi adaptations. Further, OsDGD1 knockout (KO) lines showed marked phenotypic differences compared to the wild type (WT), including a significant reduction in root length and biomass, leading to reduced Pi uptake. Further, lipidome analyses revealed reduced DGDG levels in the KO line, leading to reduced membrane remodeling, thus affecting P utilization efficiency. We also observed an increase in the MGDG: DGDG ratio in KO lines, which enhanced the endogenous JA levels and signaling. This imbalance of JA in KO plants led to changes in auxin levels, causing drastic root growth inhibition. These findings indicate the critical role of OsDGD1 in maintaining optimum levels of JA during Pi deficiency for conducive root growth. Besides acting as signaling molecules and structural components, our study widens the role of lipids as metabolic flux controllers for phytohormone biosynthesis.

Orthophosphate uptake onto woodchips in denitrifying bioreactors is enhanced by anoxic-(anaerobic-)oxic cycling

Woodchips are widely used as a low-cost and renewable organic carbon source for denitrifying biofilms in passive nutrient removal systems. One limitation of wood-based biofiltration systems is their relatively poor removal of phosphorus (P) from subsurface drainage and stormwaters, necessitating the use of additional filter media when co-treatment of nitrogen (N) and P is required. Here, we show that anoxic-oxic cycling of woodchip media, which enhances nitrate (NO3−) removal by increasing the mobilization of organic carbon from wood, also improves orthophosphate (Pi) uptake onto woodchips. Orthophosphate removal rates in flow-through woodchip columns ranged from 0 to 34.9 μg PO43− L−1 h−1 under continuously-saturated (anoxic) conditions, and increased to 17.5 to 71.9 μg PO43− L−1 h−1 in columns undergoing drying-rewetting (oxic-anoxic) cycles. The highest Pi removal efficiencies were observed in the first 20 h after reactors were re-flooded, and were concurrent with maxima in polyphosphate kinase (ppk) gene expression by the polyphosphate accumulating organisms (PAOs) Accumulibacter spp. and Pseudomonas spp. Batch experiments confirmed that anoxic-anaerobic-oxic pre-incubation conditions led to orthophosphate uptake onto woodchips as high as 74.9 ± 0.8 mg PO43−/kg woodchip, and batch tests with autoclaved woodchips demonstrated that Pi uptake was due to biological processes and not adsorption. NO3− removal in batch tests was also greatest under oxic incubation conditions, attributed to greater carbon availability in hypoxic to anoxic zones in woodchip biofilms. While further research is needed to elucidate the mechanisms controlling enhanced Pi uptake by woodchip biofilms under anoxic-(anaerobic-)oxic cycling, these results suggest a role for enhanced Pi uptake by PAOs in a nature-based system for treatment of nonpoint source nutrients.

Identification and expression analysis of Phosphate Transporter 1 (PHT1) genes in the highly phosphorus-use-efficient Hakea prostrata (Proteaceae).

Heavy and costly use of phosphorus (P) fertiliser is often needed to achieve high crop yields, but only a small amount of applied P fertiliser is available to most crop plants. Hakea prostrata (Proteaceae) is endemic to the P-impoverished landscape of southwest Australia and has several P-saving traits. We identified 16 members of the Phosphate Transporter 1 (PHT1) gene family (HpPHT1;1-HpPHT1;12d) in a long-read genome assembly of H. prostrata. Based on phylogenetics, sequence structureand expression patterns, we classified HpPHT1;1 as potentially involved in Pi uptake from soil and HpPHT1;8 and HpPHT1;9 as potentially involved in Pi uptake and root-to-shoot translocation. Three genes, HpPHT1;4, HpPHT1;6and HpPHT1;8, lacked regulatory PHR1-binding sites (P1BS) in the promoter regions. Available expression data for HpPHT1;6 and HpPHT1;8 indicated they are not responsive to changes in P supply, potentially contributing to the high P sensitivity of H. prostrata. We also discovered a Proteaceae-specific clade of closely-spaced PHT1 genes that lacked conserved genetic architecture among genera, indicating an evolutionary hot spot within the genome. Overall, the genome assembly of H. prostrata provides a much-needed foundation for understanding the genetic mechanisms of novel adaptations to low P soils in southwest Australian plants.

Candida albicans' inorganic phosphate transport and evolutionary adaptation to phosphate scarcity.

Phosphorus is essential in all cells' structural, metabolic and regulatory functions. For fungal cells that import inorganic phosphate (Pi) up a steep concentration gradient, surface Pi transporters are critical capacitators of growth. Fungi must deploy Pi transporters that enable optimal Pi uptake in pH and Pi concentration ranges prevalent in their environments. Single, triple and quadruple mutants were used to characterize the four Pi transporters we identified for the human fungal pathogen Candida albicans, which must adapt to alkaline conditions during invasion of the host bloodstream and deep organs. A high-affinity Pi transporter, Pho84, was most efficient across the widest pH range while another, Pho89, showed high-affinity characteristics only within one pH unit of neutral. Two low-affinity Pi transporters, Pho87 and Fgr2, were active only in acidic conditions. Only Pho84 among the Pi transporters was clearly required in previously identified Pi-related functions including Target of Rapamycin Complex 1 signaling, oxidative stress resistance and hyphal growth. We used in vitro evolution and whole genome sequencing as an unbiased forward genetic approach to probe adaptation to prolonged Pi scarcity of two quadruple mutant lineages lacking all 4 Pi transporters. Lineage-specific genomic changes corresponded to divergent success of the two lineages in fitness recovery during Pi limitation. Initial, large-scale genomic alterations like aneuploidies and loss of heterozygosity eventually resolved, as populations gained small-scale mutations. Severity of some phenotypes linked to Pi starvation, like cell wall stress hypersensitivity, decreased in parallel to evolving populations' fitness recovery in Pi scarcity, while severity of others like membrane stress responses diverged from Pi scarcity fitness. Among preliminary candidate genes for contributors to fitness recovery, those with links to TORC1 were overrepresented. Since Pi homeostasis differs substantially between fungi and humans, adaptive processes to Pi deprivation may harbor small-molecule targets that impact fungal growth, stress resistance and virulence.

Metabolic modelling identifies mitochondrial Pi uptake and pyruvate efflux as key aspects of daytime metabolism and proton homeostasis in crassulacean acid metabolism leaves.

Crassulacean acid metabolism (CAM) leaves are characterized by nocturnal acidification and diurnal deacidification processes related with the timed actions of phosphoenolpyruvate carboxylase and Rubisco, respectively. How CAM leaves manage cytosolic proton homeostasis, particularly when facing massive diurnal proton effluxes from the vacuole, remains unclear. A 12-phase flux balance analysis (FBA) model was constructed for a mature malic enzyme-type CAM mesophyll cell in order to predict diel kinetics of intracellular proton fluxes. The charge- and proton-balanced FBA model identified the mitochondrial phosphate carrier (PiC, Pi/H+ symport), which provides Pi to the matrix to sustain ATP biosynthesis, as a major consumer of cytosolic protons during daytime (> 50%). The delivery of Pi to the mitochondrion, co-transported with protons, is required for oxidative phosphorylation and allows sufficient ATP to be synthesized to meet the high energy demand during CAM Phase III. Additionally, the model predicts that mitochondrial pyruvate originating from decarboxylation of malate is exclusively exported to the cytosol, probably via a pyruvate channel mechanism, to fuel gluconeogenesis. In this biochemical cycle, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) acts as another important cytosolic proton consumer. Overall, our findings emphasize the importance of mitochondria in CAM and uncover a hitherto unappreciated role in metabolic proton homeostasis.

Characterization and stress-responsive regulation of CmPHT1 genes involved in phosphate uptake and transport in Melon (Cucumis melo L.)

BackgroundPhosphorus (P) deficiency, a major nutrient stress, greatly hinders plant growth. Phosphate (Pi) uptake in plant roots relies on PHT1 family transporters. However, melon (Cucumis melo L.) lacks comprehensive identification and characterization of PHT1 genes, particularly their response patterns under diverse stresses.ResultsThis study identified and analyzed seven putative CmPHT1 genes on chromosomes 3, 4, 5, 6, and 7 using the melon genome. Phylogenetic analysis revealed shared motifs, domain compositions, and evolutionary relationships among genes with close histories. Exon number varied from 1 to 3. Collinearity analysis suggested segmental and tandem duplications as the primary mechanisms for CmPHT1 gene family expansion. CmPHT1;4 and CmPHT1;5 emerged as a tandemly duplicated pair. Analysis of cis-elements in CmPHT1 promoters identified 14 functional categories, including putative PHR1-binding sites (P1BS) in CmPHT1;4, CmPHT1;6, and CmPHT1;7. We identified that three WRKY transcription factors regulated CmPHT1;5 expression by binding to its W-box element. Notably, CmPHT1 promoters harbored cis-elements responsive to hormones and abiotic factors. Different stresses regulated CmPHT1 expression differently, suggesting that the adjusted expression patterns might contribute to plant adaptation.ConclusionsThis study unveils the characteristics, evolutionary diversity, and stress responsiveness of CmPHT1 genes in melon. These findings lay the foundation for in-depth investigations into their functional mechanisms in Cucurbitaceae crops.

Astrocytes modulate brain phosphate homeostasis via polarized distribution of phosphate uptake transporter PiT2 and exporter XPR1

Aberrant inorganic phosphate (Pi) homeostasis causes brain calcification and aggravates neurodegeneration, but the underlying mechanism remains unclear. Here, we found that primary familial brain calcification (PFBC)-associated Pi transporter genes Pit2 and Xpr1 were highly expressed in astrocytes, with importer PiT2 distributed over the entire astrocyte processes and exporter XPR1 localized to astrocyte end-feet on blood vessels. This polarized PiT2 and XPR1 distribution endowed astrocyte with Pi transport capacity competent for brain Pi homeostasis, which was disrupted in mice with astrocyte-specific knockout (KO) of either Pit2 or Xpr1. Moreover, we found that Pi uptake by PiT2, and its facilitation by PFBC-associated galactosidase MYORG, were required for the high Pi transport capacity of astrocytes. Finally, brain calcification was suppressed by astrocyte-specific PiT2 re-expression in Pit2-KO mice. Thus, astrocyte-mediated Pi transport is pivotal for brain Pi homeostasis, and elevating astrocytic Pi transporter function represents a potential therapeutic strategy for reducing brain calcification.

Mode of Action of Antimicrobial Potential Protease SH21 Derived from Bacillus siamensis.

Global public health is facing a major issue with emerging resistance to antimicrobial agents. Antimicrobial agents that are currently on the market are strong and efficient, but it has not been ruled out that these medications will eventually cause resistance to bacteria. Exploring novel bioactive compounds derived from natural sources is therefore, crucial to meet future demands. The present study evaluated the mode of action of the antimicrobial potential protease enzyme SH21. Protease SH21 exhibited antimicrobial activity, strong heat stability (up to 100 °C), and pH stability (pH 3.0 to 9.0). In terms of mode of action, we found that protease SH21 was able to disrupt the bacterial cell membrane as the results of the nucleotide leakage and cell membrane permeability assay. In addition, we also checked inner membrane permeability by PI uptake assay which suggested that protease SH21 has the ability to enter the bacterial cell membrane. Our results revealed that the antimicrobial protease SH21 might be a promising candidate for treating microbial infections.

Phosphate overplus response in Chlamydomonas reinhardtii: Polyphosphate dynamics to monitor phosphate uptake and turnover

There is widespread interest in using microalgae to sustainably control and recover nutrients from wastewater to meet environmental discharge consents and recycle nutrients into agriculture or other applications. Like bacteria and yeasts, microalgae exhibit a phosphate (Pi) overplus response when Pi-deprived cells are resupplied with this nutrient. Quantitative and qualitative methods were used to follow the dynamics of polyP synthesis and turnover in four strains of Chlamydomonas reinhardtii during Pi deprivation followed by nutrient resupply (either as Pi or complete media). The lowest level of in-cell polyP during Pi deprivation, which correlates with the cessation of growth, is the key parameter for timing Pi resupply to maximise the Pi overplus response. Pi deprivation beyond this point leads to a reduced Pi uptake and reduced overplus response. Additional nutrients do not affect the magnitude of the overplus response but are important for continued growth and maximal Pi removal from the media. One strain tested shows enhanced Pi uptake and increased polyP and total in-cell P, suggesting that strain selection is also important. Although polyP levels are maintained after Pi resupply, the polymer is dynamically remodelled. Inositol hexakisphosphate (IP6) increases during this time but does not precede polyP synthesis as predicted by a model where inositol phosphates switch on polyP synthesis. Tracking polyP allows the correct time for nutrient resupply to be determined and therefore a reproducible Pi overplus response to be achieved. Depending on whether maximum cellular P content or maximum Pi removal is desired different strategies may be required. Pi deprivation until growth cessation then resupplying complete nutrients gives the best trade-off between high in-cell P accumulation, high Pi removal and algal biomass growth. This work provides robust measurements of quantitative physiological parameters, which allows reproducibility in laboratory studies and provides design parameters for algal-based nutrient recovery systems from waste waters.

Homeostatic coordination of cellular phosphate uptake and efflux requires an organelle-based receptor for the inositol pyrophosphate IP8

Phosphate (Pi) serves countless metabolic pathways and is involved in macromolecule synthesis, energy storage, cellular signaling, and bone maintenance. Herein, we describe the coordination of Pi uptake and efflux pathways to maintain mammalian cell Pi homeostasis. We discover that XPR1, the presumed Pi efflux transporter, separately supervises rates of Pi uptake. This direct, regulatory interplay arises from XPR1 being a binding partner for the Pi uptake transporter PiT1, involving a predicted transmembrane helix/extramembrane loop in XPR1, and its hitherto unknown localization in a subset of intracellular LAMP1-positive puncta (named "XLPVs"). A pharmacological mimic of Pi homeostatic challenge is sensed by the inositol pyrophosphate IP8, which functionalizes XPR1 to respond in a temporally hierarchal manner, initially adjusting the rate of Pi efflux, followed subsequently by independent modulation of PiT1 turnover to reset the rate of Pi uptake. These observations generate a unifying model of mammalian cellular Pi homeostasis, expanding opportunities for therapeutic intervention.

#1825 Treating hyperphosphatemia in CKD via targeting Npt2a in the kidney

Abstract Background and Aims Targeting the renal sodium phosphate cotransporter 2a (Npt2a) offers a novel strategy for treating hyperphosphatemia in chronic kidney disease (CKD). In patients with and without reduced kidney function, hyperphosphatemia is associated with cardiovascular complications and currently treatment options are limited. Method To study the role of a novel Npt2a inhibitor (PF-06869206) we used in vitro (Opossum kidney [OK] cells) and in vivo approaches (C57Bl/6 and Npt2a knockout [Npt2a−/−] mice). As models of CKD, 5/6 nephrectomy (5/6 Nx) as well as novel Alport/Npt2a knockout mice (Alport/Npt2a−/−) were studied. Results OK cell experiments showed that the Npt2a inhibitor caused dose-dependent reductions of Na+-dependent Pi uptake (IC50: ∼1.4 μmol/L) and Michaelis-Menten kinetics identified a ∼2.4-fold higher Km for Pi in response to Npt2a inhibition with no significant change in apparent Vmax, indicating a competitive mode of action. In vivo, Npt2a inhibition enhanced urinary excretions of Pi, Ca2+, and Na+ dose-dependently, which is recapitulated in animal models with reduced kidney function (5/6 Nx and Alport mice). Reductions in plasma Pi occur within 30 minutes and reach a maximum (40-50%) after 2 hours, this coincided with ∼50% reductions in PTH levels. No effects on FGF23 were observed in this time frame. In C57Bl/6 mice qualitative immunohistochemistry after 24 hours showed reductions in Npt2a labelling in apical microvilli compared to vehicle treatment. Providing proof of concept, Npt2a−/− and Alport/Npt2a−/− mice confirmed specificity of the drug, effects on urinary and plasma Pi were absent. Further electrophysiological studies in acutely isolated connecting tubule/cortical collecting duct showed that open probability of the epithelial Na+ channel (ENaC) was reduced in response to Npt2a inhibition, suggesting that the natriuretic effect is mediated, at least in part, by inhibition of ENaC. Conclusion Npt2a inhibition offers a promising therapeutic approach for treating hyperphosphatemia and possibly offers the opportunity of reducing cardiovascular complications in CKD.

Attenuated down-regulation of PHOSPHATE TRANSPORTER1 genes as a mechanism for phosphorus sensitivity in phosphorus-efficient Hakea prostrata (Proteaceae)

Abstract Background and aims Phosphorus (P) is an essential plant nutrient and integral for crop yield. However, plants adapted to P-impoverished environments, such as Hakea prostrata (Proteaceae), are often sensitive to P supplies that would be beneficial to other plants. The strategies for phosphate uptake and transport in P-sensitive species have received little attention. Methods Using a recently-assembled transcriptome of H. prostrata, we identified 10 putative members of the PHOSPHATE TRANSPORTER1 (PHT1) gene family, which is responsible for inorganic phosphate (Pi) uptake and transport in plants. We examined plant growth, organ P concentrations and the transcript levels for the eight PHT1 members that were expressed in roots of H. prostrata at Pi supplies ranging from P-impoverished to P-excess. Key results Hakea prostrata plants suppressed cluster root growth above ecologically-relevant Pi supplies, whilst non-cluster root mass ratios were constant. Root P concentrations increased with increasing Pi supply. Of the eight H. prostrata PHT1 genes tested, four had relatively high transcript amounts in young roots suggesting important roles in Pi uptake; however, a maximum five-fold difference in expression between P-impoverished and P-excess conditions indicated a low P-responsiveness for these genes. The HpPHT1;8 and HpPHT1;9 genes were paralogous to Pi-responsive Arabidopsis thaliana PHT1;8 and PHT1;9 orthologues involved in root-to-shoot translocation of P, but only HpPHT1;9 was P responsive. Conclusions An attenuated ability of H. prostrata to regulate PHT1 expression in response to Pi supply is likely responsible for its low capacity to control P uptake and contributes to its high P sensitivity.

Synergistic antibacterial activity of cell wall hydrolase Lys14579 combined with cinnamaldehyde against emetic Bacillus cereus and their application in foods

Synergistic antibacterial effect as a promising strategy against foodborne pathogens is compelling since the development of novel antibacterial agents is not so easy. Here we investigated the synergistic effect of cell wall hydrolase Lys14579 and cinnamaldehyde (CAL) against emetic Bacillus cereus. The MICs of Lys14579 and CAL against emetic B. cereus were 25 and 200 μg/mL, respectively. The combination of Lys14579 and CAL displayed a synergistic antimicrobial effect with a Fractional Inhibitory Concentration Index (FICI) of 0.375. Bactericidal assays revealed that this combination had better bactericidal performance than either substance alone. Furthermore, the combination of Lys14579 and CAL effectively inhibited the biofilm formation of emetic B. cereus at the concentrations of 6.25 and 50 μg/mL, respectively. Additionally, it down-regulated the cereulide synthetase gene (cesB) expression at the transcription level and reduced swarming motility by 55%. Scanning electron microscope and PI uptake assays further demonstrated that the combination accelerated the disruption of emetic B. cereus cells. Finally, the synergistic preservative effect of this combinational was observed in boiled rice and fresh lean beef. Thus, the combination of Lys14579 and CAL represents a potential synergistic strategy for controlling emetic B. cereus contamination in the food industry.

Silicon regulates phosphate deficiency through involvement of auxin and nitric oxide in barley roots.

Silicon application mitigates phosphate deficiency in barley through an interplay with auxin and nitric oxide, enhancing growth, photosynthesis, and redox balance, highlighting the potential of silicon as a fertilizer for overcoming nutritional stresses. Silicon (Si) is reported to attenuate nutritional stresses in plants, but studies on the effect of Si application to plants grown under phosphate (Pi) deficiency are still very scarce, especially in barley. Therefore, the present work was undertaken to investigate the potential role of Si in mitigating the adverse impacts of Pi deficiency in barleyHordeum vulgareL. (var. BH902). Further, the involvement of two key regulatory signaling molecules--auxin and nitric oxide (NO)--in Si-induced tolerance against Pi deficiency in barley was tested. Morphological attributes, photosynthetic parameters, oxidative stress markers (O2·-, H2O2, and MDA), antioxidant system (enzymatic--APX, CAT, SOD, GR, DHAR, MDHAR as well as non-enzymatic--AsA and GSH), NO content, and proline metabolism were the key traits that were assessed under different treatments. The P deficiency distinctly declined growth of barley seedlings, which was due to enhancement in oxidative stress leading to inhibition of photosynthesis. These results were also in parallel with an enhancement in antioxidant activity, particularly SOD and CAT, and endogenous proline level and its biosynthetic enzyme (P5CS). The addition of Si exhibited beneficial effects on barley plants grown in Pi-deficient medium as reflected in increased growth, photosynthetic activity, and redox balance through the regulation of antioxidant machinery particularly ascorbate-glutathione cycle. We noticed that auxin and NO were also found to be independently participating in Si-mediated improvement of growth and other parameters in barley roots under Pi deficiency. Data of gene expression analysis for PHOSPHATE TRANSPORTER1 (HvPHT1) indicate that Si helps in increasing Pi uptake as per the need of Pi-deficient barley seedlings, and also auxin and NO both appear to help Si in accomplishing this task probably by inducing lateral root formation. These results are suggestive of possible application of Si as a fertilizer to correct the negative effects of nutritional stresses in plants. Further research at genetic level to understand Si-induced mechanisms for mitigating Pi deficiency can be helpful in the development of new varieties with improved tolerance against Pi deficiency, especially for cultivation in areas with Pi-deficient soils.