Pi Starvation Signaling Research Articles

Recent advances in research on phosphate starvation signaling in plants

Phosphorus is indispensable for plant growth and development, with its status crucial for determining crop productivity. Plants have evolved various biochemical, morphological, and developmental responses to thrive under conditions of low P availability, as inorganic phosphate (Pi), the primary form of P uptake, is often insoluble in soils. Over the past 25 years, extensive research has focused on understanding these responses, collectively forming the Pi starvation response system. This effort has not only expanded our knowledge of strategies to cope with Pi starvation (PS) but also confirmed their adaptive significance. Moreover, it has identified and characterized numerous components of the intricate regulatory network governing P homeostasis. This review emphasizes recent advances in PS signaling, particularly highlighting the physiological importance of local PS signaling in inhibiting primary root growth and uncovering the role of TORC1 signaling in this process. Additionally, advancements in understanding shoot-root Pi allocation and a novel technique for studying Pi distribution in plants are discussed. Furthermore, emerging data on the regulation of plant-microorganism interactions by the PS regulatory system, crosstalk between the signaling pathways of phosphate starvation, phytohormones and immunity, and recent studies on natural variation in Pi homeostasis are addressed.

OsMYB58 Negatively Regulates Plant Growth and Development by Regulating Phosphate Homeostasis.

Phosphate (Pi) starvation is a critical factor limiting crop growth, development, and productivity. Rice (Oryza sativa) R2R3-MYB transcription factors function in the transcriptional regulation of plant responses to various abiotic stresses and micronutrient deprivation, but little is known about their roles in Pi starvation signaling and Pi homeostasis. Here, we identified the R2R3-MYB transcription factor gene OsMYB58, which shares high sequence similarity with AtMYB58. OsMYB58 expression was induced more strongly by Pi starvation than by other micronutrient deficiencies. Overexpressing OsMYB58 in Arabidopsis thaliana and rice inhibited plant growth and development under Pi-deficient conditions. In addition, the overexpression of OsMYB58 in plants exposed to Pi deficiency strongly affected root development, including seminal root, lateral root, and root hair formation. Overexpressing OsMYB58 strongly decreased the expression of the rice microRNAs OsmiR399a and OsmiR399j. By contrast, overexpressing OsMYB58 strongly increased the expression of rice PHOSPHATE 2 (OsPHO2), whose expression is repressed by miR399 during Pi starvation signaling. OsMYB58 functions as a transcriptional repressor of the expression of its target genes, as determined by a transcriptional activity assay. These results demonstrate that OsMYB58 negatively regulates OsmiR399-dependent Pi starvation signaling by enhancing OsmiR399s expression.

Overexpressing Ugp1 promotes phosphate uptake and accumulation in rice (Oryza sativa)

Plant responses to phosphate (Pi) starvation stress involve an array of adaptive strategies including enhanced accumulation and shoot-to-root transport of carbohydrates required for activating the plant Pi starvation signaling. However, the contribution of carbohydrate biosynthesis genes to maintaining phosphorus (P) homeostasis remains unknown, and the functional characterization of sugar metabolism genes is often impeded or compromised due to the loss of fertility of the null mutants. Here, a highly expressed gene encoding UDP-glucose pyrophosphorylase in rice (Oryza sativa), Ugp1, was functionally characterized. Ugp1 was transcriptionally induced in leaf blades by Pi starvation. As a link between sucrose (Suc) and P, we assumed that overexpression of Upg1 would alter the Suc concentration. Given that Suc is a signaling molecule, such concentration changes would affect the P deficiency signal transduction, thereby altering the homeostasis of endogenous P. The results showed that, overexpression of Ugp1 decreased plant biomass, increased sucrose content, and promoted Pi accumulation. The elevated Pi accumulation in Ugp1-OX plants was accompanied by the up-regulation of Pi-starvation-induced genes. Taken together, these results demonstrate that Ugp1 is a positive regulator of sucrose accumulation, and is required for maintaining P homeostasis in rice. Ugp1 thus represents a novel entry point to dissect the mechanisms underlying the carbon-P crosstalk.

PHT1;5 Repressed by ANT Mediates Pi Acquisition andDistribution under Low Pi andSalinity inSalt Cress.

Salinity and phosphate (Pi) starvation are the most common abiotic stresses that threaten crop productivity. Salt cress (Eutrema salsugineum) displays good tolerance to both salinity and Pi limitation. Previously, we found several Phosphate Transporter (PHT) genes in salt cress upregulated under salinity. Here, EsPHT1;5 induced by both low Pi (LP) and salinity was further characterized. Overexpression of EsPHT1;5 in salt cress enhanced plant tolerance to LP and salinity, while the knock-down lines exhibited growth retardation. The analysis of phosphorus (P) content and shoot/root ratio of total P in EsPHT1;5-overexpressing salt cress seedlings and the knock-down lines as well as arsenate uptake assays suggested the role of EsPHT1;5 in Pi acquisition and root-shoot translocation under Pi limitation. In addition, overexpression of EsPHT1;5 driven by the native promoter in salt cress enhanced Pi mobilization from rosettes to siliques upon a long-term salt treatment. Particularly, the promoter of EsPHT1;5 outperformed that of AtPHT1;5 in driving gene expression under salinity. We further identified a transcription factor EsANT, which negatively regulated EsPHT1;5 expression and plant tolerance to LP and salinity. Taken together, EsPHT1;5 plays an integral role in Pi acquisition and distribution in plant response to LP and salt stress. Further, EsANT may be involved in the cross-talk between Pi starvation and salinity signaling pathways. This work provides further insight into the mechanism underlying high P use efficiency in salt cress in its natural habitat, and evidence for a link between Pi and salt signaling.

NIGT1 represses plant growth and mitigates phosphate starvation signaling to balance the growth response tradeoff in rice.

Inorganic phosphate (Pi) availability is an important factor that affects the growth and yield of crops, thus an appropriate and effective response to Pi-fluctuation is critical. However, how crops orchestrate Pi-signaling and growth under Pi-starvation conditions to optimize the growth defense tradeoff remains unclear. Here we show that a Pi-starvation induced transcription factor NIGT1 (NITRATE-INDUCIBLE GARP-TYPE TRANSCRIPTIONAL REPRESSOR 1) controls plant growth and prevents a hyperresponse to Pi-starvation by directly repressing the expression of growth-related and Pi-signaling genes to achieve a balance between growth and response under a varying Pi environment. NIGT1 directly binds to the promoters of Pi-starvation signaling marker genes, like IPS1, miR827 and SPX2, under Pi-deficient conditions to mitigate the Pi-starvation responsive (PSR). It also directly represses the expression of vacuolar Pi efflux transporter genes VPE1/2 to regulate plant Pi-homeostasis. We further demonstrate that NIGT1 constrains shoot growth by repressing the expression of growth-related regulatory genes, including brassinolide signal transduction master regulator BZR1, cell division regulator CYCB1;1, and DNA replication regulator PSF3. Our findings reveal the function of NIGT1 in orchestrating plant growth and Pi-starvation signaling, and also provide evidence that NIGT1 acts as a safeguard to avoid hyperresponse during Pi-starvation stress in rice. This article is protected by copyright. All rights reserved.

SlPHL1 is involved in low phosphate stress promoting anthocyanin biosynthesis by directly upregulation of genes SlF3H, SlF3′H, and SlLDOX in tomato

Phosphate (Pi) deficiency is a common stress that limits plant growth and development. Plants exhibit a variety of Pi starvation responses (PSRs), including anthocyanin accumulation. The transcription factors of the PHOSPHATE STARVATION RESPONSE (PHR) family, such as AtPHR1 in Arabidopsis, play central roles in the regulation of Pi starvation signaling. Solanum lycopersicum PHR1-like 1 (SlPHL1) is a recently identified PHR involved in PSR regulation in tomato, but the detailed mechanism of its participation in Pi starvation-inducing anthocyanin accumulation remains unclear. Here we found that overexpression of SlPHL1 in tomato increases the expression of genes associated with anthocyanin biosynthesis, thereby promoting anthocyanin biosynthesis, but silencing SlPHL1 with Virus Induced Gene Silencing (VIGS) attenuated low phosphate (LP) stress-induced anthocyanin accumulation and expression of the biosynthesis-related genes. Notably, SlPHL1 is able to bind the promoters of genes Flavanone 3-Hydroxylase (SlF3H), Flavanone 3′-Hydroxylase (SlF3′H), and Leucoanthocyanidin Dioxygenase (SlLDOX) by yeast one-hybrid (Y1H) analysis. Furthermore, Electrophoretic Mobility Shift Assay (EMSA) and transient transcript expression assay showed that PHR1 binding t (sequence (P1BS) motifs located on the promoters of these three genes are critical for SlPHL1 binding and enhancing the gene transcription. Additionally, allogenic overexpression of SlPHL1 could promote anthocyanin biosynthesis in Arabidopsis under LP conditions through the similar mechanism to AtPHR1, suggesting that SlPHL1 might be functionally conserved with AtPHR1 in this process. Taken together, SlPHL1 positively regulates LP-induced anthocyanin accumulation by directly promoting the transcription of SlF3H, SlF3′H and SlLDOX. These findings will contribute to understanding the molecular mechanism of PSR in tomato.

PtoWRKY40 interacts with PtoPHR1-LIKE3 while regulating the phosphate starvation response in poplar.

Plants usually suffer from phosphorus starvation because of the low inorganic phosphate (Pi) status of most soils. To cope with this, plants have evolved an adaptive phosphate starvation response (PSR) which involves both developmental and metabolic changes regulated mainly by PHOSPHATE STARVATION RESPONSE1 (PHR1) and its homologs. Here, we elucidated how perennial woody plants, such as poplars (Populus spp.), respond to low-Pi stress. We first performed RNA-seq analysis of low-Pi-treated poplars and identified PtoWRKY40 is rapidly downregulated and protein degraded after stress. Overexpressing and knocking-down PtoWRKY40 downregulated and upregulated the expression of Pi starvation signaling genes, respectively, such as PHOSPHATE TRANSPORTER1 (PHT1)-type genes and PURPLE ACID PHOSPHATASE genes. PtoWRKY40 bound to the W box in the promoter of several PtoPHT1s and repressed their expression. Moreover, PtoWRKY40 interacted with PtoPHR1-LIKE3 (PtoPHL3), a PHR1 homolog in poplar, to inhibit the latter binding to the P1BS element and thus reduced PtoPHT1s' transcription under Pi-sufficient conditions. However, Pi deficiency decreased PtoWRKY40 abundance and therefore released its inhibition on PHT1s. In conclusion, we have uncovered a PSR mechanism mediated by PtoWRKY40 and PtoPHL3 which regulates Pi content in poplars, deepening our understanding of how poplars adapt to diverse Pi conditions and regulate appropriate responses to maintain Pi homeostasis.

PHR1 positively regulates phosphate starvation-induced anthocyanin accumulation through direct upregulation of genes F3'H and LDOX in Arabidopsis.

Phosphate deficiency promotes anthocyanin accumulation in Arabidopsis through direct binding of PHR1 to the P1BS motifs on the promoters of F3'H and LDOX and thereby upregulating their expression. Phosphorus is one of the essential elements for plants, and plants mainly absorb inorganic phosphate (Pi) from soil. But Pi deficiency is a common factor limiting plant growth and development. Anthocyanin accumulation in green tissues (such as leaves) is one of the characteristics of many plants in response to Pi starvation. However, little is known about the mechanism by which Pi starvation induces anthocyanin accumulation. Here, we found that the mutation of the gene PHOSPHATE STARVATION RESPONSE1 (PHR1), which encodes a key factor involved in Pi starvation signaling in Arabidopsis, significantly attenuates anthocyanin accumulation under Pi-limiting conditions. Moreover, the expression of several Pi deficiency-upregulated genes that are involved in anthocyanin biosyntheses, such as flavanone 3'-hydroxylase (F3'H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX), and production of anthocyanin pigment 1 (PAP1), was significantly lower in the phr1-1 mutant than in the wild type (WT). Both yeast one-hybrid (Y1H) analysis and chromatin immunoprecipitation quantitative PCR (ChIP-qPCR) showed that PHR1 can interact with the promoters of F3'H and LDOX, but not DFR and PAP1. By electrophoretic mobility shift assay (EMSA), it was further confirmed that the PHR1-binding sequence (P1BS) motifs located on the F3'H and LDOX promoters are required for the PHR1 bindings. Also, in Arabidopsis protoplasts, PHR1 enhanced the transcriptional activity of the F3'H and LDOX promoters, but these effects were markedly impaired when the P1BS motifs were mutated. Taken together, these results indicate that PHR1 positively regulates Pi starvation-induced anthocyanin accumulation in Arabidopsis, at least in part, by directly binding the P1BS motifs located on the promoters to upregulate the transcription of anthocyanin biosynthetic genes F3'H and LDOX.

Alternative splicing of REGULATOR OF LEAF INCLINATION 1 modulates phosphate starvation signaling and growth in plants.

Phosphate (Pi) limitation represents a primary constraint on crop production. To better cope with Pi deficiency stress, plants have evolved multiple adaptive mechanisms for phosphorus acquisition and utilization, including the alteration of growth and the activation of Pi starvation signaling. However, how these strategies are coordinated remains largely unknown. Here, we found that the alternative splicing (AS) of REGULATOR OF LEAF INCLINATION 1 (RLI1) in rice (Oryza sativa) produces two protein isoforms: RLI1a, containing MYB DNA binding domain and RLI1b, containing both MYB and coiled-coil (CC) domains. The absence of a CC domain in RLI1a enables it to activate broader target genes than RLI1b. RLI1a, but not RLI1b, regulates both brassinolide (BL) biosynthesis and signaling by directly activating BL-biosynthesis and signaling genes. Both RLI1a and RLI1b modulate Pi starvation signaling. RLI1 and PHOSPHATE STARVATION RESPONSE 2 function redundantly to regulate Pi starvation signaling and growth in response to Pi deficiency. Furthermore, the AS of RLI1-related genes to produce two isoforms for growth and Pi signaling is widely present in both dicots and monocots. Together, these findings indicate that the AS of RLI1 is an important and functionally conserved strategy to orchestrate Pi starvation signaling and growth to help plants adapt to Pi-limitation stress.

A reciprocal inhibitory module for Pi and iron signaling

Phosphorous (P) and iron (Fe), two essential nutrients for plant growth and development, are highly abundant elements in the earth's crust but often display low availability to plants. Due to the ability to form insoluble complexes, the antagonistic interaction between P and Fe nutrition in plants has been noticed for decades. However, the underlying molecular mechanism modulating the signaling and homeostasis between them remains obscure. Here, we show that the possible iron sensors HRZs, the iron deficiency–induced E3 ligases, could interact with the central regulator of phosphate (Pi) signaling, PHR2, and prompt its ubiquitination at lysine residues K319 and K328, leading to its degradation in rice. Consistent with this, the hrzs mutants displayed a high Pi accumulation phenotype. Furthermore, we found that iron deficiency could attenuate Pi starvation signaling by inducing the expression of HRZs, which in turn trigger PHR2 protein degradation. Interestingly, on the other hand, rice PHRs could negatively regulate the expression of HRZs to modulate iron deficiency responses. Therefore, PHR2 and HRZs form a reciprocal inhibitory module to coordinate Pi and iron signaling and homeostasis in rice. Taken together, our results uncover a molecular link between Pi and iron master regulators, which fine-tunes plant adaptation to Pi and iron availability in rice.

SlPHL1, a MYB-CC transcription factor identified from tomato, positively regulates the phosphate starvation response.

Inorganic phosphate (Pi) deficiency is a major limiting factor for plant growth and development. Previous reports have demonstrated that PHOSPHATE STARVATION RESPONSE 1 (PHR1) and OsPHR2 play central roles in Pi-starvation signaling in Arabidopsis and rice, respectively. However, the Pi-starvation signaling network in tomato (Solanum lycopersicum) is still not fully understood. In this work, SlPHL1, a homolog of AtPHR1 and OsPHR2, was identified from tomato. It was found that SlPHL1 contains the MYB and coiled-coil (CC) domains, localizes in the nucleus, and has transcriptional activity, indicating that it is a typical MYB-CC transcription factor (TF). Overexpression of SlPHL1 enhanced Pi-starvation responses both in Arabidopsis Col-0 and in tomato Micro-Tom, including elevated root hair growth, promoted APase activity, favored Pi uptake, and increased transcription of Pi starvation-inducing (PSI) genes. Besides, overexpressing SlPHL1 was able to compensate for the Pi-starvation response weakened by the AtPHR1 mutation. Notably, electrophoretic mobility shift assay (EMSA) showed that SlPHL1 could bind to the PHR1-binding sequence (P1BS, GNATATNC)-containing DNA fragments. Furthermore, SlPHL1 specifically interacted with the promoters of the tomato PSI genes SlPht1;2 and SlPht1;8 through the P1BS cis-elements. Taken these results together, SlPHL1 is a newly identified MYB-CC TF from tomato, which participates in Pi-starvation signaling by directly upregulating the PSI genes. These findings might contribute to the understanding of the Pi-starvation signaling in tomato.

Modulation of Phosphate Deficiency-Induced Metabolic Changes by Iron Availability in Arabidopsis thaliana.

Concurrent suboptimal supply of several nutrients requires the coordination of nutrient-specific transcriptional, phenotypic, and metabolic changes in plants in order to optimize growth and development in most agricultural and natural ecosystems. Phosphate (Pi) and iron (Fe) deficiency induce overlapping but mostly opposing transcriptional and root growth responses in Arabidopsis thaliana. On the metabolite level, Pi deficiency negatively modulates Fe deficiency-induced coumarin accumulation, which is controlled by Fe as well as Pi deficiency response regulators. Here, we report the impact of Fe availability on seedling growth under Pi limiting conditions and on Pi deficiency-induced accumulation of amino acids and organic acids, which play important roles in Pi use efficiency. Fe deficiency in Pi replete conditions hardly changed growth and metabolite profiles in roots and shoots of Arabidopsis thaliana, but partially rescued growth under conditions of Pi starvation and severely modulated Pi deficiency-induced metabolic adjustments. Analysis of T-DNA insertion lines revealed the concerted coordination of metabolic profiles by regulators of Fe (FIT, bHLH104, BRUTUS, PYE) as well as of Pi (SPX1, PHR1, PHL1, bHLH32) starvation responses. The results show the interdependency of Pi and Fe availability and the interplay between Pi and Fe starvation signaling on the generation of plant metabolite profiles.

Two ADP-glucose pyrophosphorylase subunits, OsAGPL1 and OsAGPS1, modulate phosphorus homeostasis in rice.

Plant acclimatory responses to phosphate (Pi) starvation stress include the accumulation of carbohydrates, namely sugar and starch. However, whether altered endogenous carbohydrate profile could in turn affect plant Pi starvation responses remains widely unexplored. Here, two genes encoding the large and small subunits of an ADP-glucose pyrophosphorylase (AGP) in rice (Oryza sativa), AGP Large Subunit 1 (AGPL1) and AGP Small Subunit 1 (AGPS1), were functionally characterized with regard to maintenance of phosphorus (P) homeostasis and regulation of Pi starvation signaling. AGPL1 and AGPS1 were both positively responsive to nitrogen (N) or Pi deprivation, and expressed in almost all the tissues except in the meristem and mature zones of root. AGPL1 and AGPS1 physically interacted in chloroplast, and catalyzed the rate-limiting step of starch biosynthesis. Low-N- (LN) and low-Pi (LP)-triggered starch accumulation in leaves was impaired in agpl1, agps1 and apgl1 agps1 mutants compared with the wild-type plants. By contrast, mutation of AGPL1 and/or AGPS1 led to an increase in the content of the major sugar, sucrose, in leaf sheath and root under control and LN conditions. Moreover, the Pi accumulation was enhanced in the mutants under control and LN conditions, but not LP conditions. Notably, the LN-induced suppression of Pi accumulation was compromised attributed to the mutation of AGPL1 and/or AGPS1. Furthermore, the increased Pi accumulation was accompanied by the specific suppression of OsSPX2 and activation of several Pi transporter genes. These results indicate that a balanced level of carbohydrates is vital for maintaining plant P homeostasis.

SmMYB98b positive regulation to tanshinones in Salvia miltiorrhiza Bunge hairy roots

Tanshinones are major secondary metabolites in Salvia miltiorrhiza Bunge, the traditional Chinese medicinal plant Danshen. Increasing the production of tanshinones is important because of their significant economic value in human medicine, for the treatment of cardiovascular and cerebrovascular diseases and for their anti-tumor properties. Phosphate (Pi) starvation can change secondary metabolism in plants. MYB transcription factors (TFs) have been reported to be involved in both Pi starvation signaling and secondary metabolism regulation, enabling plants to build a relationship between the Pi starvation signal and secondary metabolism. In the present study, a nucleus-located R2R3 MYB TF, SmMYB98b, was found in S. militorrhiza. SmMYB98b was shown to belong to subgroup 20 of R2R3-MYB TFs, and expression could be induced by low-Pi conditions. The overexpression of SmMYB98b in S. militorrhiza hairy roots stimulated tanshinone accumulation. Plants overexpressing SmMYB98b exhibited Pi-starvation responses, such as decreased Pi concentration and activation of genes inducible by phosphate starvation, whereas transgenic plants with the SmMYB98b-RNA interference (RNAi) construct showed the opposite phenotype. In summary, the present study highlighted the role of SmMYB98b as a regulator of tanshinone biosynthesis and its potential function in Pi-starvation signaling. These results might have wider implications for studying the crosstalk between Pi stress responses and secondary metabolism pathways in plants. This study highlights the role of SmMYB98b as a positive regulator of tanshinone biosynthesis and its potential function in Pi-starvation signaling in Salvia miltiorrhiza.

Internal phosphate starvation signaling and external phosphate availability have no obvious effect on the accumulation of cadmium in rice

Phosphorus (P) is a limiting nutrient element for crop. To obtain maximum crop yield, P fertilizer is often over-applied, which leads to accelerating exhaustion of phosphate resources and serious environmental problems. Reducing the application of P fertilizer and enhancing the P utilization efficiency of crops are significant for the sustainable development of agriculture. Cadmium (Cd) contamination in rice is another serious agricultural issue. However, whether reducing the application of P fertilizer and enhancing the P utilization efficiency of crops will increase the risk of Cd accumulation in crops remains obscure. In this study, we are aiming to elucidate the relationship between Cd and P in rice from physiological and genetic perspectives. For this purpose, the wild type (WT) rice plants and phosphate (Pi)-starvation signaling repressed mutant phr2 were used to analyze the relationship between Cd and P. Here, we found that Cd stress could promote P accumulation and induce Pi-starvation signaling in WT and phr2 shoots under Pi-sufficient condition in a PHOSPHATE STARVATION RESPONSE 2 (PHR2) independent manner. Besides, the expression level of Cd transporter of OsNramp5 and the uptake speed of Cd2+ were not obviously changed under Pi-sufficient and Pi-deficient conditions. Furthermore, our Cd determination results showed that the Cd concentrations in WT and phr2 were not obviously changed under Pi-sufficient and Pi-deficient conditions. These results indicate that the external P availability and internal Pi-starvation signaling cannot obviously affect the accumulation of Cd in rice seedling.

Two RING-Finger Ubiquitin E3 Ligases Regulate the Degradation of SPX4, An Internal Phosphate Sensor, for Phosphate Homeostasis and Signaling in Rice

SPX-domain-containing proteins (SPXs) play an important role in inorganic phosphate (Pi) sensing, signaling, and transport in eukaryotes. In plants, SPXs are known to integrate cellular Pi status and negatively regulate the activity of Pi central regulators, the PHOSPATE STARVATION RESPONSE proteins (PHRs). The stability of SPXs, such as SPX4, is reduced under Pi-deficient conditions. However, the mechanisms by which SPXs are degraded remain unclear. In this study, using a yeast-two-hybrid screen we identified two RING-finger ubiquitin E3 ligases regulating SPX4 degradation, designated SDEL1 and SDEL2, which were post-transcriptionally induced by Pi starvation. We found that both SDELs were located in the nucleus and cytoplasm, had ubiquitin E3 ligase activity, and directly ubiquitinated the K213 and K299 lysine residues in SPX4 to regulate its stability. Furthermore, we found that PHR2, a Pi central regulator in rice, could compete with SDELs by interacting with SPX4 under Pi-sufficient conditions, which protected SPX4 from ubiquitination and degradation. Consistent with the biochemical function of SDEL1 and SDEL2, overexpression of SDEL1 or SDEL2 resulted in Pi overaccumulation and induced Pi-starvation signaling even under Pi-sufficient conditions. Conversely, their loss-of-function mutants displayed decreased Pi accumulation and reduced Pi-starvation signaling. Collectively, our study revealed that SDEL1 and SDEL2 facilitate the degradation of SPX4 to modulate PHR2 activity and regulate Pi homeostasis and Pi signaling in response to external Pi availability in rice.

AtSPX1-mediated transcriptional regulation during leaf senescence in Arabidopsis thaliana

Leaf senescence is the final stage of leaf growth, a highly coordinated and complicated process. Phosphorus as an essential macronutrient for plant growth is remobilized from senescing leaves to other vigorous parts of the plant. In this study, through data mining, we found some phosphate starvation induced genes such as AtSPX1, were significantly induced in aging leaves in Arabidopsis. We applied a reverse genetics approach to investigate the phenotypes of transgenic plants and mutant plants, and the results showed that the overexpression of AtSPX1 accelerated leaf senescence, suppressed Pi accumulation, promoted SA production and H2O2 levels in leaves, while the mutant lines of AtSPX1 showed slightly delayed leaf senescence. We conducted RNA-seq-based transcriptome analysis together with GO and GSEA enrichment analyses for transgenic vs. wild-type plants to elucidate the possible underlying regulatory mechanism. The 558 genes that were up-regulated in the overexpression plants 35S::AtSPX1/WT, were significantly enriched in the process of leaf senescence, Pi starvation responses and SA signaling pathways, as were the target genes of some transcription factors such as WRKYs and NACs. In a word, we characterized AtSPX1 as a key regulator, which mediated the crosstalks among leaf senescence, Pi starvation and SA signaling pathways in Arabidopsis thaliana.

The FvPHR1 transcription factor control phosphate homeostasis by transcriptionally regulating miR399a in woodland strawberry

Plants have evolved phosphate (Pi) starvation response to adapt the low-Pi environment. The regulation of adaptive responses to phosphorus deficiency by the PHR1-miR399-PHO2 module has been well studied in Arabidopsis thaliana but not in strawberry. Transcription factor PHR1 as the central regulator in the Pi starvation signaling has been revealed in a few plant species. However, the function of PHR1 homologues in strawberry is still unknown. In this study, a total of 13 MYB-CC genes were identified in the woodland strawberry (Fragaria vesca) genome and the FvPHR1 gene was characterized. FvPHR1 contains MYB domain and coiled–coil (CC) domain and is localized in the nucleus. FvPHR1 also exhibits trans-activation ability. Furthermore, the P content in leaves of FvPHR1-overexpressing woodland strawberries was significantly increased by 1.38-fold to 1.78-fold compared with that in the wild type. FvPHR1 was also demonstrated to directly bind to the FvMIR399a promoter and positively regulate the expression of FvmiR399a in woodland strawberry. These results showed that PHR1-miR399 module is involved in the regulation of phosphate-signaling pathway and phosphate homeostasis in woodland strawberry.

Rice SPX6 negatively regulates the phosphate starvation response through suppression of the transcription factor PHR2.

Phosphorus (P) is an essential macronutrient for plant growth and development, but the molecular mechanism determining how plants sense external inorganic phosphate (Pi) levels and reprogram transcriptional and adaptive responses is incompletely understood. In this study, we investigated the function of OsSPX6 (hereafter SPX6), an uncharacterized member of SPX domain (SYG1, Pho81 and XPR1)-containing proteins in rice, using reverse genetics and biochemical approaches. Transgenic plants overexpressing SPX6 exhibited decreased Pi concentrations and suppression of phosphate starvation-induced (PSI) genes. By contrast, transgenic lines with decreased SPX6 transcript levels or spx6 mutant showed significant Pi accumulation in the leaf and upregulation of PSI genes. Overexpression of SPX6 genetically suppressed the overexpression of PHOSPHATE STARVATION RESPONSE REGULATOR 2 (PHR2) in terms of the accumulation of high Pi content. Moreover, direct interaction of SPX6 with PHR2 impeded PHR2 translocation into the nucleus, and inhibited PHR2 binding to the P1BS (PHR1 binding sequence) element. SPX6 protein was degraded in leaves under Pi-deficient conditions, whereas it accumulated in roots. We conclude that rice SPX6 is another important negative regulator in Pi starvation signaling through the interaction with PHR2. SPX6 shows different responses to Pi starvation in shoot and root, which differ from those of other SPX proteins.

Maintenance of phosphate homeostasis and root development are coordinately regulated by MYB1, an R2R3-type MYB transcription factor in rice

The adaptive responses of plants to phosphate (Pi) starvation stress are fine-tuned by an elaborate regulatory network. In this study, we identified and characterized a novel Pi starvation-responsive gene, MYB1, encoding an R2R3-type transcription factor in rice. MYB1 was transcriptionally induced in leaf sheaths and old leaf blades. It was localized to the nucleus and expressed mainly in vascular tissues. Mutation of MYB1 led to an increase in Pi uptake and accumulation, accompanied by altered expression of a subset of Pi transporters and several genes involved in Pi starvation signaling. Furthermore, MYB1 affected the elongation of the primary root in a Pi-dependent manner and lateral roots in a Pi-independent manner. Moreover, gibberellic acid (GA)-triggered lateral root elongation was largely suppressed in wild-type plants under Pi starvation conditions, whereas this suppression was partially rescued in myb1 mutant lines, correlating with the up-regulation of a GA biosynthetic gene upon MYB1 mutation. Taken together, the findings of this study highlight the role of MYB1 as a regulator involved in both Pi starvation signaling and GA biosynthesis. Such a co-regulator might have broad implications for the study of cross-talk between nutrient stress and other signaling pathways.