High-affinity Phosphate Transporter Research Articles

Pi-efficient wheat cultivars screened by using both functional marker CAPS-799 and field experiment

ContextIn our previous study, we identified TaPHT1;9–4B as a critical high-affinity phosphate (Pi) transporter that plays a significant role in Pi acquisition and transport. Subsequently, we developed the functional marker CAPS-799 to facilitate the identification of its Pi-efficient elite haplotype. ObjectiveThis study aimed to screen a variety of wheat cultivars using the CAPS-799 marker to identify those exhibiting Pi-efficient elite haplotypes. Additionally, we sought to elucidate the physiological mechanisms underlying these traits through both field and hydroponic experiments. MethodsField experiments over two successive years without phosphorus (P) fertilizer, along with a hydroponic experiment using low Pi (10 μM) were conducted. P concentration, biomass, grain yield, yield components, root growth parameters, and transcript levels of TaPHT1;9–4B were measured. Total P accumulation, P transport and utilization efficiency, and relative parameters were also calculated. ResultsEight Pi-efficient wheat cultivars with the Hap3 haplotype were identified using CAPS-799 from a group of 80 modern major cultivars. In two-year field experiments without P fertilizer, these Hap3 cultivars exhibited significantly higher grain yield, spike number, P uptake, and transport efficiency compared to Non-Hap3 haplotypes (Hap1, Hap2, and Hap4). As a result, these eight cultivars were classified as Pi-efficient elite haplotypes. In field experiments, TaPHT1;9–4B transcript levels in the roots during the early grain-filling stage, as well as root surface area, volume, and mean root diameter in hydroponic experiments, were significantly greater in Hap3 cultivars than in Non-Hap3 cultivars. ConclusionsOur field and hydroponic experiments demonstrated that CAPS-799 is a valuable functional marker for identifying Pi-efficient wheat cultivars. These Pi-efficient cultivars were characterized by higher TaPHT1;9–4B transcript levels and increased root growth. ImplicationsCAPS-799 can be used to screen or develop Pi-efficient wheat cultivars.

Rhizophagus intraradices mediated mitigation of arsenic toxicity in wheat involves differential distribution of arsenic in subcellular fractions and modulated expression of Phts and ABCCs

Biomagnification of arsenic in food chain through wheat consumption poses a serious threat to human health. Therefore, it is necessary to elucidate mechanism of arsenic tolerance and detoxification in wheat. The study aimed to unravel the strategies adopted by arbuscular mycorrhizal fungi to alleviate arsenic toxicity in wheat. To accomplish this, independent and interactive effects of arsenic and Rhizophagus intraradices were assessed. Colonization by R. intraradices resulted in lower expression of high-affinity phosphate transporters (Phts) in comparison with non-mycorrhizal (NM) plants, thereby lowering arsenic concentrations in mycorrhizal (M) plants. Additionally, the subcellular fractionation analysis indicated differential distribution of arsenic. In NM plants, arsenic accumulated primarily in cell wall and organelle fractions. Conversely, in M plants arsenic was more concentrated in the cell wall and vacuolar fractions. This was related to higher levels of hydroxyl and aldehyde groups in cell wall fraction of root along with increased expression of C-type ATP-binding cassette transporters in root and leaves. These factors enabled effective sequestration of arsenic in the cell wall and vacuoles of M plants, thereby reducing its toxicity. Furthermore, the proportion of inorganic arsenic was lower in M plant, as it transformed it into less toxic organic forms.

Phosphate availability conditions caspofungin tolerance, capsule attachment and titan cell formation in Cryptococcus neoformans.

There is an urgent need for new antifungal drugs to treat invasive fungal diseases. Unfortunately, the echinocandin drugs that are fungicidal against other important fungal pathogens are ineffective against Cryptococcus neoformans, the causative agent of life-threatening meningoencephalitis in immunocompromised people. Contributing mechanisms for echinocandin tolerance are emerging with connections to calcineurin signaling, the cell wall, and membrane composition. In this context, we discovered that a defect in phosphate uptake impairs the tolerance of C. neoformans to the echinocandin caspofungin. Our previous analysis of mutants lacking three high affinity phosphate transporters revealed reduced elaboration of the polysaccharide capsule and attenuated virulence in mice. We investigated the underlying mechanisms and found that loss of the transporters and altered phosphate availability influences the cell wall and membrane composition. These changes contribute to the shedding of capsule polysaccharide thus explaining the reduced size of capsules on mutants lacking the phosphate transporters. We also found an influence of the calcineurin pathway including calcium sensitivity and an involvement of the endoplasmic reticulum in the response to phosphate limitation. Furthermore, we identified membrane and lipid composition changes consistent with the role of phosphate in phospholipid biosynthesis and with previous studies implicating membrane integrity in caspofungin tolerance. Finally, we discovered a contribution of phosphate to titan cell formation, a cell type that displays modified cell wall and capsule composition. Overall, our analysis reinforces the importance of phosphate as a regulator of cell wall and membrane composition with implications for capsule attachment and antifungal drug susceptibility.

Discovery of Candidatus Nitrosomaritimum as a New Genus of Ammonia-Oxidizing Archaea Widespread in Anoxic Saltmarsh Intertidal Aquifers.

Ammonia-oxidizing archaea (AOA) are widely distributed in marine and terrestrial habitats, contributing significantly to global nitrogen and carbon cycles. However, their genomic diversity, ecological niches, and metabolic potentials in the anoxic intertidal aquifers remain poorly understood. Here, we discovered and named a novel AOA genus, Candidatus Nitrosomaritimum, from the intertidal aquifers of Yancheng Wetland, showing close metagenomic abundance to the previously acknowledged dominant Nitrosopumilus AOA. Further construction of ammonia monooxygenase-based phylogeny demonstrated the widespread distribution of Nitrosomaritimum AOA in global estuarine-coastal niches and marine sediment. Niche differentiation among sublineages of this new genus in anoxic intertidal aquifers is driven by salinity and dissolved oxygen gradients. Comparative genomics revealed that Candidatus Nitrosomaritimum has the genetic capacity to utilize urea and possesses high-affinity phosphate transporter systems (phnCDE) for surviving phosphorus-limited conditions. Additionally, it contains putative nosZ genes encoding nitrous-oxide (N2O) reductase for reducing N2O to nitrogen gas. Furthermore, we gained first genomic insights into the archaeal phylum Hydrothermarchaeota populations residing in intertidal aquifers and revealed their potential hydroxylamine-detoxification mutualism with AOA through utilizing the AOA-released extracellular hydroxylamine using hydroxylamine oxidoreductase. Together, this study unravels the overlooked role of priorly unknown but abundant AOA lineages of the newly discovered genus Candidatus Nitrosomaritimum in biological nitrogen transformation and their potential for nitrogen pollution mitigation in coastal environments.

Biochemical characterization of a high affinity phosphate transporter (PiPT) from root endophyte fungus Piriformospora indica

We have functionally characterized the high-affinity phosphate transporter (PiPT) from the root endophyte fungus Piriformospora indica. PiPT belongs to the major facilitator superfamily (MFS). PiPT protein was purified by affinity chromatography (Ni-NTA) and Size Exclusion Chromatography (SEC). The functionality of solubilized PiPT was determined in detergent-solubilized state by fluorescence quenching and in proteoliposomes. In the fluorescence quenching assay, PiPT exhibited a saturation concentration of approximately 2 μM, at a pH of 4.5. Proteoliposomes of size 121.6 nm radius, showed transportation of radioactive phosphate. Vmax was measured to be 232.2 ± 11 pmol/min/mg protein. We have found Km to be 45.8 ± 6.2 μM suggesting high affinity towards phosphate.

Whole Genome Analysis of Streptomyces spp. Strains Isolated from the Rhizosphere of Vitis vinifera L. Reveals Their Role in Nitrogen and Phosphorus Metabolism

The rhizospheric microorganisms of agricultural crops play a crucial role in plant growth and nutrient cycling. In this study, we isolated two Streptomyces strains, Streptomyces sp. LM32 and Streptomyces sp. LM65, from the rhizosphere of Vitis vinifera L. We then conducted genomic analysis by assembling, annotating, and inferring phylogenomic information from the whole genome sequences. Streptomyces sp. strain LM32 had a genome size of 8.1 Mb and a GC content of 72.14%, while Streptomyces sp. strain LM65 had a genome size of 7.3 Mb and a GC content of 71%. Through ANI results, as well as phylogenomic, pan-, and core-genome analysis, we found that strain LM32 was closely related to the species S. coelicoflavus, while strain LM65 was closely related to the species S. achromogenes subsp. achromogenes. We annotated the functional categories of genes encoded in both strains, which revealed genes involved in nitrogen and phosphorus metabolism. This suggests that these strains have the potential to enhance nutrient availability in the soil, promoting agricultural sustainability. Additionally, we identified gene clusters associated with nitrate and nitrite ammonification, nitrosative stress, allantoin utilization, ammonia assimilation, denitrifying reductase gene clusters, high-affinity phosphate transporter and control of PHO regulon, polyphosphate, and phosphate metabolism. These findings highlight the ecological roles of these strains in sustainable agriculture, particularly in grapevine and other agricultural crop systems.

Kinetic network modeling with molecular simulation inputs: A proton-coupled phosphate symporter

Phosphate, an essential metabolite involved in numerous cellular functions, is taken up by proton-coupled phosphate transporters of plants and fungi within the major facilitator family. Similar phosphate transporters have been identified across a diverse range of biological entities, including various protozoan parasites linked to human diseases, breast cancer cells with increased phosphate requirements, and osteoclast-like cells engaged in bone resorption. Prior studies have proposed an overview of the functional cycle of a proton-driven phosphate transporter (PiPT), yet a comprehensive understanding of the proposed reaction pathways necessitates a closer examination of each elementary reaction step within an overall kinetic framework. In this work, we leverage kinetic network modeling in conjunction with a “bottom-up” molecular dynamics approach to show how such an approach can characterize the proton-phosphate co-transport behavior of PiPT under different pH and phosphate concentration conditions. In turn, this allows us to reveal the prevailing reaction pathway within a high-affinity phosphate transporter under different experimental conditions and to uncover the molecular origin of the optimal pH condition of this transporter.

Integrated genomics provides insights into the evolution of the polyphosphate accumulation trait of Ca. Accumulibacter

Candidatus Accumulibacter, a prominent polyphosphate-accumulating organism (PAO) in wastewater treatment, plays a crucial role in enhanced biological phosphorus removal (EBPR). The genetic underpinnings of its polyphosphate accumulation capabilities, however, remain largely unknown. Here, we conducted a comprehensive genomic analysis of Ca. Accumulibacter-PAOs and their relatives within the Rhodocyclaceae family, identifying 124 core genes acquired via horizontal gene transfer (HGT) at its least common ancestor. Metatranscriptomic analysis of an enrichment culture of Ca. Accumulibacter revealed active transcription of 44 of these genes during an EBPR cycle, notably including the polyphosphate kinase 2 (PPK2) gene instead of the commonly recognized polyphosphate kinase 1 (PPK1) gene. Intriguingly, the phosphate regulon (Pho) genes showed minimal transcriptions, pointing to a distinctive fact of Pho dysregulation, where PhoU, the phosphate signaling complex protein, was not regulating the high-affinity phosphate transport (Pst) system, resulting in continuous phosphate uptake. To prevent phosphate toxicity, Ca. Accumulibacter utilized the laterally acquired PPK2 to condense phosphate into polyphosphate, resulting in the polyphosphate-accumulating feature. This study provides novel insights into the evolutionary emergence of the polyphosphate-accumulating trait in Ca. Accumulibacter, offering potential advancements in understanding the PAO phenotype in the EBPR process.

Phosphate Transporter OsPT4, Ubiquitinated by E3 Ligase OsAIRP2, Plays a Crucial Role in Phosphorus and Nitrogen Translocation and Consumption in Germinating Seed

Phosphorus (P) and nitrogen (N) are essential macronutrients necessary for plant growth and development. OsPT4 is a high-affinity phosphate (Pi) transporter that has a positive impact on nutrient uptake and seed development. In this study, the expression patterns of different Pi transporter genes in germinating seeds were determined, and the relative expression of OsPT4 was induced in Pi-deficient seeds and gradually increased with the passage of germination time. The analysis of P, N, Pi, and amino acid concentrations in germinating seeds of OsPT4 mutants showed that the OsPT4 mutation caused P and N retention and a continuous reduction in multiple amino acid concentrations in germinating seeds. Transcriptome analysis and qRT-PCR results also indicated that the OsPT4 mutation inhibits the expression of genes related to P and N transportation and amino acid synthesis in germinating seeds. In addition, the paraffin section and TUNEL assay of OsPT4 mutant germinating seeds suggests that OsPT4 mutation causes programmed cell death (PCD) delayed in the aleurone layer and inhibition of leaf outgrowth. Moreover, we also found that OsPT4 was ubiquitinated by OsAIRP2, which is a C3HC4-type RING E3 Ub ligase. Our studies illustrate that OsPT4 plays a crucial role in P and N collaborative translocation and consumption in germinating seeds. It also provides a theoretical basis for the molecules and physiological mechanisms of P and N cross-talk under suppressed Pi uptake conditions.

Interaction of calcium responsive proteins and transcriptional factors with the PHO regulon in yeasts and fungi.

Phosphate and calcium ions are nutrients that play key roles in growth, differentiation and the production of bioactive secondary metabolites in filamentous fungi. Phosphate concentration regulates the biosynthesis of hundreds of fungal metabolites. The central mechanisms of phosphate transport and regulation, mediated by the master Pho4 transcriptional factor are known, but many aspects of the control of gene expression need further research. High ATP concentration in the cells leads to inositol pyrophosphate molecules formation, such as IP3 and IP7, that act as phosphorylation status reporters. Calcium ions are intracellular messengers in eukaryotic organisms and calcium homeostasis follows elaborated patterns in response to different nutritional and environmental factors, including cross-talking with phosphate concentrations. A large part of the intracellular calcium is stored in vacuoles and other organelles forming complexes with polyphosphate. The free cytosolic calcium concentration is maintained by transport from the external medium or by release from the store organelles through calcium permeable transient receptor potential (TRP) ion channels. Calcium ions, particularly the free cytosolic calcium levels, control the biosynthesis of fungal metabolites by two mechanisms, 1) direct interaction of calcium-bound calmodulin with antibiotic synthesizing enzymes, and 2) by the calmodulin-calcineurin signaling cascade. Control of very different secondary metabolites, including pathogenicity determinants, are mediated by calcium through the Crz1 factor. Several interactions between calcium homeostasis and phosphate have been demonstrated in the last decade: 1) The inositol pyrophosphate IP3 triggers the release of calcium ions from internal stores into the cytosol, 2) Expression of the high affinity phosphate transporter Pho89, a Na+/phosphate symporter, is controlled by Crz1. Also, mutants defective in the calcium permeable TRPCa7-like of Saccharomyces cerevisiae shown impaired expression of Pho89. This information suggests that CrzA and Pho89 play key roles in the interaction of phosphate and calcium regulatory pathways, 3) Finally, acidocalcisomes organelles have been found in mycorrhiza and in some melanin producing fungi that show similar characteristics as protozoa calcisomes. In these organelles there is a close interaction between orthophosphate, pyrophosphate and polyphosphate and calcium ions that are absorbed in the polyanionic polyphosphate matrix. These advances open new perspectives for the control of fungal metabolism.

RNAseq analysis of mutants in coding and non-coding transcription of phosphate genes in the yeast Saccharomyces cerevisiae

In the yeast Saccharomyces cerevisiae phosphate starvation induces the expression of PHO genes, including PHO84, encoding an high-affinity phosphate transporter, and SPL2, encoding a regulatory protein. PHO84 is down-regulated by antisense transcription. Here, using strand-specific RNAseq the effect is studied of mutations related to sense and antisense transcription of phosphate genes. Replacement of the transcriptional terminator of PHO84 by that of CYC1 resulted, unexpectedly, in an increased antisense transcription and a strongly reduced sense transcription of PHO84 and a strongly reduced SPL2 expression. The expression of unrelated genes was altered as well. The data suggest that antisense transcription of PHO84 and not the Pho84 transporter affects the expression of SPL2. Deletion of the two putative binding sites for Ume6 in the SPL2 promoter or deletion of UME6 differently affected SPL2 expression, suggesting that Ume6 regulates SPL2 by a mechanism different from a simple binding to the putative Ume6 binding sites.

Wheat PHT1;9 acts as one candidate arsenate absorption transporter for phytoremediation

Arsenate (AsV) is one of the most common forms of arsenic (As) in environment and plant high-affinity phosphate transporters (PHT1s) are the primary plant AsV transporters. However, few PHT1s involved in AsV absorption have been identified in crops. In our previous study, TaPHT1;3, TaPHT1;6 and TaPHT1;9 were identified to function in phosphate absorption. Here, their AsV absorption capacities were evaluated using several experiments. Ectopic expression in yeast mutants indicated that TaPHT1;9 had the highest AsV absorption rates, followed by TaPHT1;6, while not for TaPHT1;3. Under AsV stress, further, BSMV-VIGS-mediated TaPHT1;9-silencing wheat plants exhibited higher AsV tolerance and lower As concentrations than TaPHT1;6-silenced plants, whereas TaPHT1;3-silencing plants had similar phenotype and AsV concentrations to control. These suggested that TaPHT1;9 and TaPHT1;6 possessed AsV absorption capacity with the former showing higher activities. Under hydroponic condition, furthermore, CRISPR-edited TaPHT1;9 wheat mutants showed the enhanced tolerance to AsV with decreased As distributions and concentrations, whereas TaPHT1;9 ectopic expression transgenic rice plants had the opposite results. Also, under AsV-contaminated soil condition, TaPHT1;9 transgenic rice plants exhibited depressed AsV tolerance with increased As concentrations in roots, straws and grains. Moreover, Pi addition alleviated the AsV toxicity. These suggested that TaPHT1;9 should be a candidate target gene for AsV phytoremediation.

The nutrient-responsive CDK Pho85 primes the Sch9 kinase for its activation by TORC1.

Yeast cells maintain an intricate network of nutrient signaling pathways enabling them to integrate information on the availability of different nutrients and adjust their metabolism and growth accordingly. Cells that are no longer capable of integrating this information, or that are unable to make the necessary adaptations, will cease growth and eventually die. Here, we studied the molecular basis underlying the synthetic lethality caused by loss of the protein kinase Sch9, a key player in amino acid signaling and proximal effector of the conserved growth-regulatory TORC1 complex, when combined with either loss of the cyclin-dependent kinase (CDK) Pho85 or loss of its inhibitor Pho81, which both have pivotal roles in phosphate sensing and cell cycle regulation. We demonstrate that it is specifically the CDK-cyclin pair Pho85-Pho80 or the partially redundant CDK-cyclin pairs Pho85-Pcl6/Pcl7 that become essential for growth when Sch9 is absent. Interestingly, the respective three CDK-cyclin pairs regulate the activity and distribution of the phosphatidylinositol-3 phosphate 5-kinase Fab1 on endosomes and vacuoles, where it generates phosphatidylinositol-3,5 bisphosphate that serves to recruit both TORC1 and its substrate Sch9. In addition, Pho85-Pho80 directly phosphorylates Sch9 at Ser726, and to a lesser extent at Thr723, thereby priming Sch9 for its subsequent phosphorylation and activation by TORC1. The TORC1-Sch9 signaling branch therefore integrates Pho85-mediated information at different levels. In this context, we also discovered that loss of the transcription factor Pho4 rescued the synthetic lethality caused by loss of Pho85 and Sch9, indicating that both signaling pathways also converge on Pho4, which appears to be wired to a feedback loop involving the high-affinity phosphate transporter Pho84 that fine-tunes Sch9-mediated responses.

Genetic linkage map construction and QTL mapping of blade length and width in Saccharina japonica using SSR and SNP markers

MAS (marker assisted selection) is useful for cultivar development in the lower plant Saccharina japonica. In this study, 167 screened SSR markers and 125 individuals were used to construct a S.japonica genetic linkage map with 27 linkage groups and a total Haldane length of 2069 cM. Constructed linkage groups were aligned with assembled pseudo-chromosomes of S. japonica genome sequences (NCBI: MEHQ00000000) and anchored to 23 of 31 pseudo-chromosomes. A genetic linkage map including 33 linkage groups was constructed with combined 3397 SSR and SNP markers. In the integrated genetic map used for QTL mapping, SSR marker M1895 was closely associated with the major QTL for blade length and width with LOD threshold of >5.0. Annotation indicated that SjPT, coded as a high affinity phosphate transporter, was mapped to this major QTL with cDNA sequence of 1422 bp, which coded 473 aa for 8 transmembrane helics. qRT-PCR and the transcriptomic detection confirmed that SjPT transcription was consistent with increases in kelp blade length and width. Marker M1895 appears to be particularly useful for MAS (marker assisted selection) in S. japonica breeding.

Systematic Identification and Expression Analysis of the Sorghum Pht1 Gene Family Reveals Several New Members Encoding High-Affinity Phosphate Transporters.

Sorghum (Sorghum bicolor) is known to have a more robust capability of phosphorus uptake than many other cereal plants, which could be attributed to its phosphate transporter 1 (Pht1) that has a high phosphorus affinity. There are eleven SbPht1 genes in the sorghum genome, nine of which are expressed in sorghum roots or shoots in response to phosphorus deficiency (low-P). The molecular features of these nine genes were investigated by gene expression analysis, subcellular localization, and a yeast mutant complementation growth assay. They were found to be induced in response to low-P stress in root or shoot. All these SbPht1 proteins were found to be localized on the cell membrane, and SbPht1;8 was also detected in the endoplasmic reticulum. These SbPht1s were able to complement the yeast mutant EY917 that lacks all the functional phosphate transporters, and, among them, SbPht1;5, SbPht1;6 and SbPht1;8 could partially complement the yeast mutant strain EY917 in low-P conditions. Overall, these findings demonstrate that SbPht1;5, SbPht1;6, and SbPht1;8 are high-affinity phosphate transporters. SbPht1;5, in particular, is specifically involved in phosphorus uptake in the roots, whilst SbPht1;6 and SbPht1;8 are key players in both P uptake and P transport in response to low-P stress in sorghum.

Root Morphological and Physiological Adaptations to Low Phosphate Enhance Phosphorus Efficiency at Melon (Cucumis melo L.) Seedling Stage

The high phosphorus (P) acquisition ability of crops can reduce their dependence on artificial inorganic phosphate (Pi) supplementation under Pi-limited conditions. Melon (Cucumis melo L.) is vulnerable to Pi deficiency. This study was carried out to explore the morphological and physiological responses of melon to low-Pi stress under a hydroponic system. The results show that low-Pi stress significantly disturbed nutrient homeostasis, reduced P content, and resulted in iron accumulation in melon seedlings and brown iron plaque formation on the root surface. A nutrient pool of P and Fe formed on the roots to forage for more Pi under low-Pi conditions. Severe long-term low-Pi stress promoted primary root elongation and inhibited lateral root growth, which increased the longitudinal absorption zone of the roots. The decrease in P content of the roots upregulated the expression of the acid phosphatase (APase) gene and increased APase activity. The high-affinity phosphate transporter (Pht1) genes were also upregulated significantly. These morphological and physiological responses significantly increased Pi uptake rate and P utilization efficiency at the melon seedling stage. These findings will be useful for screening low-Pi-tolerant varieties and sustaining melon production in P-limited environments.

Abscisic acid facilitates phosphate acquisition through the transcription factor ABA INSENSITIVE5 in Arabidopsis

Low phosphate (LP) in soil is a common nutrient stress that severely restricts agricultural production, but the role, if any, of the major stress phytohormone abscisic acid (ABA) in plant phosphate (Pi) starvation responses remains elusive. Here, we report that LP-induced ABA accumulation promotes Pi uptake in an ABA INSENSITIVE5 (ABI5)-dependent manner in Arabidopsis thaliana. LP significantly activated plant ABA biosynthesis, metabolism, and stress responses, suggesting a role of ABA in the plant response to Pi availability. LP-induced ABA accumulation and expression of two major high-affinity phosphate transporter genes PHOSPHATE TRANSPORTER1;1/1;4 (PHT1;1/1;4) were severely impaired in a mutant lacking BETA-GLUCOSIDASE1 (BG1), which converts conjugated ABA to active ABA, and the mutant had shorter roots and less Pi content than wild-type plants under LP conditions. Moreover, a mutant of ABI5, which encodes a central transcription factor in ABA signaling, also exhibited suppressed root elongation and had reduced Pi content under LP conditions. ABI5 facilitated Pi acquisition by activating the expression of PHT1;1 by directly binding to its promoter, while overexpression of PHT1;1 completely rescued its Pi content under LP conditions. Together, our findings illustrate a molecular mechanism by which ABA positively modulates phosphate acquisition through ABI5 in the Arabidopsis response to phosphate deficiency.

Two Pathways Diverge: Glycerophosphocholine Impacts Both Phosphate and Lipid Metabolism in Candida albicans

Candida albicans (C. albicans)is a leading source of fungal infections in humans, resulting in over 250,000 deaths annually. C. albicanscolonizes multiple host niches and causes invasive disease, requiring its adaptation to wide variations in nutrient availability. Phosphate is an essential nutrient, and several genes associated with phosphate acquisition are differentially expressed during infection. In addition to inorganic phosphate (Pi), C. albicans utilizes the abundant lipid metabolite, glycerophosphocholine (GPC), as a phosphate source. GPC is transported into the cell via the Git3 and Git4 transporters. The Git transporters are required for full virulence in a mouse model of disseminated candidiasis, but the mechanism for that virulence defect is unknown. Here, we probe the importance of GPC uptake to cellular metabolism by examining two aspects of its metabolic fate: i) its catabolism to liberate Pi), and ii) its potential conversion to phosphatidylcholine (PC) via reacylation. To query its role in phosphate homeostasis, we employed a knockout strain of the high affinity phosphate transporter, Pho84. Cells lacking Pho84 display sensitivity to several host‐related stressors and decreased virulence. Our results show that provision of cells with GPC as phosphate source alleviates hypersensitivity of pho84∆/∆ mutants to osmotic, oxidative, and cell wall stress. The rate at which GPC and Pi are transported into the cell varies depending upon the total level of phosphate provided, with Pitransport exceeding GPC transport by a maximum of two‐fold at 500 µM total phosphate. The effect of exogenous GPC on induction of the PHO regulon, the cellular system of phosphate homeostasis, is complex and concentration dependent. Beyond its role in phosphate metabolism, GPC has other physiologically relevant metabolic fates, namely its reacylation to form a phosphatidylcholine (PC) molecule. In Saccharomyces cerevisiae, the first step of this novel reacylation pathway is catalyzed by the acyltransferase, Gpc1, which converts GPC to lyso‐PC. Initial in vivo labeling studies under phosphate replete conditions indicate that the C. albicans homolog of Gpc1 is a GPC acyltransferase and that the reacylation pathway is functional in C. albicans. In vitro assays in strains lacking or containing Gpc1 are underway. Taken together, our results indicate that C. albicans can utilize the abundant lipid metabolite, GPC, via at least two metabolic pathways to promote its growth and survival in the human host.

Sorting Nexin1 negatively modulates phosphate uptake by facilitating Phosphate Transporter1;1 degradation in Arabidopsis.

High-affinity phosphate (Pi) transporters (PHTs) PHT1;1 and PHT1;4 are necessary for plant root Pi uptake especially under Pi-deficient conditions, but how their protein stability is modulated remains elusive. Here, we identified a Ttransfer DNA insertion mutant of Sorting Nexin1 (SNX1), which had more Pi content and less anthocyanin accumulation than the wild type under deficient Pi. By contrast, the snx1-2 mutant displayed higher sensitivity to exogenous arsenate in terms of seed germination and root elongation, revealing higher Pi uptake rates. Further study showed that SNX1 could co-localize and interact with PHT1;1 and PHT1;4 in vesicles and at the plasma membrane. Genetic analysis showed that increased Pi content in the snx1-2 mutant under low Pi conditions could be extensively compromised by mutating PHT1;1 in the double mutant snx1-2 pht1;1, revealing that SNX1 is epistatic to PHT1;1. In addition, SNX1 negatively controls PHT1;1 protein stability; therefore, PHT1;1 protein abundance in the plasma membrane was increased in the snx1-2 mutant compared with the wild type under either sufficient or deficient Pi. Together, our study (i) identifies SNX1 as a key modulator of the plant response to low Pi and (ii) unravels its role in the modulation of PHT1;1 protein stability, PHT1;1 accumulation at the plasma membrane, and root Pi uptake.

Nitrogen Fixation Influenced by Phosphorus and Nitrogen Availability in the Benthic Bloom-forming Cyanobacterium Hydrocoleum sp. Identified in a Temperate Marine Lagoon.

The nitrogen-fixing, non-heterocystous cyanobacterium Hydrocoleum sp. (Oscillatoriales) is a common epiphytic and benthic bloom-former in tropical and subtropical shallow water systems but shares high phylogenetic similarity with the planktonic, globally important diazotroph Trichodesmium. Multiphasic observations in this study resulted in unexpected identification of Hydrocoleum sp. in mass accumulations in a coastal lagoon in the Western temperate North Atlantic Ocean. Hydrocoleum physiology was examined in situ through measurements of N2 and CO2 fixation rates and expression of genes involved with N2 fixation, CO2 fixation, and phosphorus (P) stress. Bulk N2 fixation rates and Hydrocoleum nifH expression peaked at night and were strongly suppressed by dissolved inorganic nitrogen (DIN). The expression of high affinity phosphate transporter (pstS) and alkaline phosphatase (phoA) genes of Hydrocoleum was elevated during the night and negatively responded to phosphate amendments, as evidence that these mechanisms contribute to P acquisition during diazotrophic growth of Hydrocoleum in situ. This discovery at the edge of the previously known Hydrocoleum habitat range in the warming oceans raises intriguing questions about diazotrophic cyanobacterial adaptations and transitions on the benthic-pelagic continuum.