Low Pi Stress Research Articles

Multi-omics-based identification of purple acid phosphatases and metabolites involved in phosphorus recycling in stylo root exudates

Stylo (Stylosanthes guianensis) is a tropical forage and cover crop that possesses low phosphate (Pi) tolerance traits. However, the mechanisms underlying its tolerance to low-Pi stress, particularly the role of root exudates, remain unclear. This study employed an integrated approach using physiological, biochemical, multi-omics, and gene function analyses to investigate the role of stylo root exudates in response to low-Pi stress. Widely targeted metabolomic analysis revealed that eight organic acids and one amino acid (L-cysteine) were significantly increased in the root exudates of Pi-deficient seedlings, among which tartaric acid and L-cysteine had strong abilities to dissolve insoluble-P. Furthermore, flavonoid-targeted metabolomic analysis identified 18 flavonoids that were significantly increased in root exudates under low-Pi conditions, mainly belonging to the isoflavonoid and flavanone subclasses. Additionally, transcriptomic analysis revealed that 15 genes encoding purple acid phosphatases (PAPs) had upregulated expression in roots under low-Pi conditions. Among them, SgPAP10 was characterized as a root-secreted phosphatase, and overexpression of SgPAP10 enhanced organic-P utilization by transgenic Arabidopsis. Overall, these findings provide detailed information regarding the importance of stylo root exudates in adaptation to low-Pi stress, highlighting the plant's ability to release Pi from organic-P and insoluble-P sources through root-secreted organic acids, amino acids, flavonoids, and PAPs.

Comprehensive Evolution and Expression anaLysis of PHOSPHATE 1 Gene Family in Allotetraploid Brassica napus and Its Diploid Ancestors.

The members of PHOSPHATE 1 (PHO1) family play important roles in plant phosphate (Pi) transport and adaptation to Pi deficiency. The functions of PHO1 family proteins have been reported in several plant species, with the exception of Brassica species. Here, we identified 23, 23, and 44 putative PHO1 family genes in Brassica rapa, Brassica oleracea, and Brassica napus by whole genome analysis, respectively. The phylogenetic analysis divided PHO1 family proteins into eight groups, which represented the orthologous relationships among PHO1 members. The gene structure and the conserved motif analysis indicated that the most PHO1 family genes had similar gene structures and the PHO1 proteins shared mutual conserved motifs. The chromosome distribution analysis showed that the majority of BnPHO1 family genes distributed analogously at chromosomes with BrPHO1 and BoPHO1 family genes. The data showed that PHO1 family genes were highly conserved during evolution from diploid to tetraploid. Furthermore, the expression analysis showed that PHO1 family genes had different expression patterns in plant tissues, suggesting the diversity of gene functions in Brassica species. Meanwhile, the expression analysis also revealed that some PHO1 family genes were significantly responsive to Pi deficiency, suggesting that PHO1 family genes play critical roles in Pi uptake and homeostasis under low Pi stress. Altogether, the characteristics of PHO1 family genes provide a reliable groundwork for further dissecting their functions in Brassica species.

Genome-wide association study identifies a gene conferring high physiological phosphorus use efficiency in rice.

Phosphate (Pi) is indispensable for the growth and development of plant, and low-Pi stress is a major limitation for crop growth and yield worldwide. The tolerance to low-Pi stress varied among rice germplasm resources. However, the mechanisms underlying the tolerance of rice to low-Pi stress, as a complex quantitative trait, are not clear. We performed a genome-wide association study (GWAS) through a diverse worldwide collection of 191 rice accessions in the field under normal-Pi and low-Pi supply in two years. Twenty and three significant association loci were identified for biomass and grain yield per plant under low-Pi supply respectively. The expression level of OsAAD as a candidate gene from a associated locus was significantly up-regulated after low-Pi stress treatment for five days and tended to return to normal levels after Pi re-supply in shoots. Suppression of OsAAD expression could improve the physiological phosphorus use efficiency (PPUE) and grain yields through affecting the expression of several genes associated with GA biosynthesis and metabolism. OsAAD would be a promising gene for increasing PPUE and grain yield in rice under normal- and low-Pi supply via genome editing.

NtPIN3 positively regulates low phosphorus tolerance by changing root elongation, Pi concentration and antioxidant capacity in tobacco

Phosphorus (P) is a critical nutrient for plants, and inorganic orthophosphate (Pi) deficiency results in declining crop quality and yields. Auxin plays a key regulatory effect on plant growth in response to Pi deficiency, yet the regulatory mechanisms remain poorly understood. In this study, we cloned and identified an auxin efflux transporter, NtPIN3, involved in the low phosphate response of tobacco (Nicotiana tabacum). NtPIN3 is located on the cytoplasmic membrane, and high expression levels of NtPIN3 were observed under phosphorus deficiency conditions. Moreover, NtPIN3 overexpression in tobacco could dramatically enhance low Pi tolerance by increasing the root length, which was consistent with the GUS activity and the auxin concentration in the root. Notably, Pi concentration and antioxidant capacity in overexpression transgenic tobacco were significantly increased compared with the wild type (WT) under low Pi stress conditions due to Pi uptake (NtPT1 and NtPT2) and antioxidant capacity (superoxide dismutase, peroxidase, and catalase). In contrast, compared with WT, the ntpin3 mutant significantly reduced root length, auxin concentration, Pi concentration, and antioxidant capacity under low Pi stress conditions, suggesting that NtPIN3 is a positive regulator of low phosphate in tobacco. In addition, the yeast-two-hybrid and luciferase complementation imaging assay confirmed that NtPIN3 interacted with NtPIN2. Altogether, the NtPIN3 involved in low Pi response and overexpression of NtPIN3 contributed to enhanced tolerance to low Pi stress, which proved an underlying gene for breeding low Pi-stress tolerant plants.

Mutation of OsLPR3 Enhances Tolerance to Phosphate Starvation in Rice.

Low Phosphate Root (LPR) encodes a protein localized to the endoplasmic reticulum (ER) and cell wall. This gene plays a key role in responding to phosphate (Pi) deprivation, especially in remodeling the root system architecture (RSA). An identification and expression analysis of the OsLPR family in rice (Oryza sativa) has been previously reported, and OsLPR5, functioning in Pi uptake and translocation, is required for the normal growth and development of rice. However, the role of OsLPR3, one of the five members of this family in rice, in response to Pi deficiency and/or in the regulation of plant growth and development is unknown. Therefore, in this study, the roles of OsLPR3 in these processes were investigated, and some functions were found to differ between OsLPR3 and OsLPR5. OsLPR3 was found to be induced in the leaf blades, leaf sheaths, and roots under Pi deprivation. OsLPR3 overexpression strongly inhibited the growth and development of the rice but did not affect the Pi homeostasis of the plant. However, oslpr3 mutants improved RSA and Pi utilization, and they exhibited a higher tolerance to low Pi stress in rice. The agronomic traits of the oslpr3 mutants, such as 1000-grain weight and seed length, were stimulated under Pi-sufficient conditions, indicating that OsLPR3 plays roles different from those of OsLPR5 during plant growth and development, as well as in the maintenance of the Pi status of rice.

Functional Analysis of Bna-miR399c-PHO2 Regulatory Module Involved in Phosphorus Stress in Brassica napus

Phosphorus stress is one of the important factors restricting plant growth and development, and the microRNA (miRNA) family is involved in the regulation of the response to plant nutrient stress by repressing the expression of target genes at the post-transcriptional or translational level. miR399 is involved in the transportation of phosphate in multiple plants by improving tolerance to low Pi conditions. However, the effect of miR399 on the response of low Pi stress in rapeseed (Brassica napus L.) is unclear. The present study showed a significant increase in taproot length and lateral root number of plants overexpressing Bna-miR399c, while the biomass and Pi accumulation in shoots and roots increased, and the anthocyanin content decreased and chlorophyll content improved under low Pi stress. The results illustrate that Bna-miR399c could enhance the uptake and transportation of Pi in soil, thus making B. napus more tolerant to low Pi stress. Furthermore, we confirmed that BnPHO2 is one of the targets of Bna-miR399c, and the rejection of Pi in rapeseed seedlings increased due to the overexpression of BnPHO2. Hence, we suggest that miR399c-PHO2 module can effectively regulate the homeostasis of Pi in B. napus. Our study can also provide the theoretical basis for germplasm innovation and the design of intelligent crops with low nutrient input and high yield to achieve the dual objectives of income and yield increase and environmental protection in B. napus.

Genome wide association studies for acid phosphatase activity at varying phosphorous levels in Brassica juncea L.

Acid phosphatases (Apases) are an important group of enzymes that hydrolyze soil and plant phosphoesters and anhydrides to release Pi (inorganic phosphate) for plant acquisition. Their activity is strongly correlated to the phosphorus use efficiency (PUE) of plants. Indian mustard (Brassica juncea L. Czern & Coss) is a major oilseed crop that also provides protein for the animal feed industry. It exhibits low PUE. Understanding the genetics of PUE and its component traits, especially Apase activity, will help to reduce Pi fertilizer application in the crop. In the present study, we evaluated 280 genotypes of the diversity fixed foundation set of Indian mustard for Apase activity in the root (RApase) and leaf (LApase) tissues at three- low (5µM), normal (250µM) and high (1mM) Pi levels in a hydroponic system. Substantial effects of genotype and Pi level were observed for Apase activity in both tissues of the evaluated lines. Low Pi stress induced higher mean RApase and LApase activities. However, mean LApase activity was relatively more than mean RApase at all three Pi levels. JM06016, IM70 and Kranti were identified as promising genotypes with higher LApase activity and increased R/S at low Pi. Genome-wide association study revealed 10 and 4 genomic regions associated with RApase and LApase, respectively. Annotation of genomic regions in the vicinity of peak associated SNPs allowed prediction of 15 candidates, including genes encoding different family members of the acid phosphatase such as PAP10 (purple acid phosphatase 10), PAP16, PNP (polynucleotide phosphorylase) and AT5G51260 (HAD superfamily gene, subfamily IIIB acid phosphatase) genes. Our studies provide an understanding of molecular mechanism of the Apase response of B. juncea at varying Pi levels. The identified SNPs and candidate genes will support marker-assisted breeding program for improving PUE in Indian mustard. This will redeem the crop with enhanced productivity under restricted Pi reserves and degrading agro-environments.

Physiological and Transcriptomic Analyses Uncover the Reason for the Inhibition of Photosynthesis by Phosphate Deficiency in Cucumis melo L.

Phosphate (Pi) deficiency is a common phenomenon in agricultural production and limits plant growth. Recent work showed that long-term Pi deficiency caused the inhibition of photosynthesis and inefficient electron transport. However, the underlying mechanisms are still unknown. In this study, we used the physiological, histochemical, and transcriptomic methods to investigate the effect of low-Pi stress on photosynthetic gas exchange parameters, cell membrane lipid, chloroplast ultrastructure, and transcriptional regulation of key genes in melon seedlings. The results showed that Pi deficiency significantly downregulated the expression of aquaporin genes, induced an increase in ABA levels, and reduced the water content and free water content of melon leaves, which caused physiological drought in melon leaves. Therefore, gas exchange was disturbed. Pi deficiency also reduced the phospholipid contents in leaf cell membranes, caused the peroxidation of membrane lipids, and destroyed the ultrastructure of chloroplasts. The transcriptomic analysis showed that 822 differentially expressed genes (DEGs) were upregulated and 1254 downregulated by Pi deficiency in leaves. GO and KEGG enrichment analysis showed that DEGs significantly enriched in chloroplast thylakoid membrane composition (GO:0009535), photosynthesis-antenna proteins (map00196), and photosynthesis pathways (map00195) were downregulated by Pi deficiency. It indicated that Pi deficiency regulated photosynthesis-related genes at the transcriptional level, thereby affecting the histochemical properties and physiological functions, and consequently causing the reduced light assimilation ability and photosynthesis efficiency. It enriches the mechanism of photosynthesis inhibition by Pi deficiency.

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.

Integration of small RNA, degradome, and transcriptome sequencing data illustrates the mechanism of low phosphorus adaptation in Camellia oleifera.

Phosphorus (P) is an indispensable macronutrient for plant growth and development, and it is involved in various cellular biological activities in plants. Camellia oleifera is a unique high-quality woody oil plant that grows in the hills and mountains of southern China. However, the available P content is deficient in southern woodland soil. Until now, few studies focused on the regulatory functions of microRNAs (miRNAs) and their target genes under low inorganic phosphate (Pi) stress. In this study, we integrated small RNA, degradome, and transcriptome sequencing data to investigate the mechanism of low Pi adaptation in C. oleifera. We identified 40,689 unigenes and 386 miRNAs by the deep sequencing technology and divided the miRNAs into four different groups. We found 32 miRNAs which were differentially expressed under low Pi treatment. A total of 414 target genes of 108 miRNAs were verified by degradome sequencing. Gene ontology (GO) functional analysis of target genes found that they were related to the signal response to the stimulus and transporter activity, indicating that they may respond to low Pi stress. The integrated analysis revealed that 31 miRNA–target pairs had negatively correlated expression patterns. A co-expression regulatory network was established based on the profiles of differentially expressed genes. In total, three hub genes (ARF22, WRKY53, and SCL6), which were the targets of differentially expressed miRNAs, were discovered. Our results showed that integrated analyses of the small RNA, degradome, and transcriptome sequencing data provided a valuable basis for investigating low Pi in C. oleifera and offer new perspectives on the mechanism of low Pi tolerance in woody oil plants.

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.

TaMPK2B, a member of the MAPK family in T. aestivum, enhances plant low-Pi stress tolerance through modulating physiological processes associated with phosphorus starvation defensiveness

The mitogen-activated protein kinase (MAPK) cascades are present in plant species and modulate plant growth and stress responses. This study characterizes TaMPK2B, a MAPK family gene in T. aestivum that regulates plant adaptation to low-Pi stress. TaMPK2B harbors the conserved domains involving protein phosphorylation and protein-protein interaction. A yeast two-hybrid assay reveals an interaction between TaMPK2B and TaMPKK2 and between the latter and TaMPKKK;A, suggesting that all comprise a MAPK signaling cascade TaMPKKK;A-TaMPKK2-TaMPK2B. TaMPK2B expression levels were elevated in roots and leaves under a Pi starvation (PS) condition. Additionally, the induced TaMPK2B transcripts under PS in tissues were gradually restored following the Pi normal recovery condition. TaMPK2B overexpression conferred on plants improved PS adaptation; the tobacco lines with TaMPK2B overexpression enhanced the plant’s dry mass production, Pi uptake capacity, root system architecture (RSA) establishment, and ROS homeostasis relative to wild type under PS treatment. Moreover, the transcripts of genes in phosphate transporter (PT), PIN-FORMED, and antioxidant enzyme (AE) families, including NtPT3 and NtPT4, NtPIN9, and NtMnSOD1 and NtPOD1;7, were elevated in Pi-deprived lines overexpressing TaMPK2B. Transgene analyses validated their functions in regulating Pi uptake, RSA establishment, and AE activities of plants treated by PS. These results suggest that TaMPK2B-mediated plant PS adaptation is correlated with the modified transcription of distinct PT, PIN, and AE genes. Our investigation suggests that TaMPK2B is one of the crucial regulators in plant low-Pi adaptation by improving Pi uptake, RSA formation, and ROS homeostasis via transcriptionally regulating genes associated with the above physiological processes.

Organ specific transcriptome analysis of upland cotton (Gossypium hirsutum) in response to low phosphorus stress during early stage of growth

ABSTRACT In order to systematically identify the critical genes involved in the response to low inorganic phosphorus (Pi) in cotton, we performed RNA sequencing (RNA-seq) analyses of root, stem, and leaf from Gossypium hirsutum seedlings grown under both Pi replete and Pi deplete conditions. In total, 1679, 1611, and 841 differentially expressed genes (DEGs) were observed in the roots, stems, and leaves, respectively. Several Pi-starvation-induced genes homologous to Arabidopsis were identified, such as SPX, PHO1/2, and PHF1, as well as several members of the PHT1 family, which showed different expression levels in specific organs. Subsequently, 25 randomly selected DEGs were validated by quantitative real-time PCR, the result of which was consistent with the RNA sequencing data, indicating the reliability of RNA-seq data. By analyzing the differentially expressed transcription factors, we found that the MYB family was one of the three most abundant transcription factor families in all three tissues. Moreover, some phytohormone-related DEGs were also found in G. hirsutum seedlings in this study. In particular, some auxin-responsive genes, such as IAA24, IAA11/14/16, and IAA4/14/16/29 were differentially expressed in the roots, stems and leaves of G. hirsutum seedlings respectively. Finally, homologous genes for some important phosphatases such as PPCK1 and VTC4 were also found to be differentially expressed. On analyzing the protein interaction network of 121 shared DEGs, five interaction networks – two of which contained 10 and 6 DEGs, respectively – were identified. These findings provided a theoretical basis for studying the gene function of different cotton tissues in response to low Pi stress.

The involvement of abscisic acid-insensitive mutants in low phosphate stress responses during rhizosphere acidification, anthocyanin accumulation and Pi homeostasis in Arabidopsis

Phosphorus is an essential plant nutrient, used in the formation of macromolecules such as nucleic acids and phospholipids. Abscisic acid (ABA) may be involved in the process of low inorganic phosphate (Pi) responses. The phenotypes of ABA-insensitive Arabidopsis mutants (abi1/2/3/4/5) under low Pi stress were investigated to identify possible low Pi response mutant genes. The results showed enhanced rhizosphere acidification in the abi1–1/abi2–1/abi5–1 mutants under low Pi stress compared with wild-type (WT) seedlings. The abi1–1/abi2–1/ abi3–1/abi5–1 mutants accumulated less anthocyanin than the WT, while the abi4–1 mutant showed greater accumulation, implicating all the ABA-insensitive mutants in anthocyanin deposition under Pi deficiency. Alterations in the Pi contents of roots or shoots were also observed in the mutants in response to both Pi sufficiency and deficiency, indicating that the mutants were involved in Pi uptake or transportation. The primary root length and root-shoot ratio of abi3–1 and abi4–1 mutants decreased compared with WT seedlings under low Pi condition. Further research showed that ABI5 could regulate PHT1;5 and WRKY42 expression by combining with ACGT cis-acting elements of the PHT1;5 and WRKY42 promoters.

High transcriptome plasticity drives phosphate starvation responses in tomato

Tomato is an important vegetable crop and fluctuating available soil phosphate (Pi) level elicits several morpho-physiological responses driven by underlying molecular responses. Therefore, understanding these molecular responses at the gene and isoform levels has become critical in the quest for developing crops with improved Pi use efficiency. A quantitative time-series RNA-seq analysis was performed to decipher the global transcriptomic changes that accompany Pi starvation in tomato. Apart from changes in the expression levels of genes, there were also alterations in the expression of alternatively-spliced transcripts. Physiological responses such as anthocyanin accumulation, reactive oxygen species generation and cell death are obvious 7 days after Pi deprivation accompanied with the maximum amount of transcriptional change in the genome making it an important stage for in-depth study while studying Pi stress responses (PSR). Our study demonstrates that transcriptomic changes under Pi deficiency are dynamic and complex in tomato. Overall, our study dwells on the dynamism of the transcriptome in eliciting a response to adapt to low Pi stress and lays it bare. Findings from this study will prove to be an invaluable resource for researchers using tomato as a model for understanding nutrient deficiency.

Genome-wide evolution and expression analysis of the MYB-CC gene family in Brassica spp.

The MYB-CC family is a subtype within the MYB superfamily. This family contains an MYB domain and a predicted coiled-coil (CC) domain. Several MYB-CC transcription factors are involved in the plant’s adaptability to low phosphate (Pi) stress. We identified 30, 34, and 55 MYB-CC genes in Brassica rapa, Brassica oleracea, and Brassica napus, respectively. The MYB-CC genes were divided into nine groups based on phylogenetic analysis. The analysis of the chromosome distribution and gene structure revealed that most MYB-CC genes retained the same relative position on the chromosomes and had similar gene structures during allotetraploidy. Evolutionary analysis showed that the ancestral whole-genome triplication (WGT) and the recent allopolyploidy are critical for the expansion of the MYB-CC gene family. The expression patterns of MYB-CC genes were found to be diverse in different tissues of the three Brassica species. Furthermore, the gene expression analysis under low Pi stress revealed that MYB-CC genes may be related to low Pi stress responses. These results may increase our understanding of MYB-CC gene family diversification and provide the basis for further analysis of the specific functions of MYB-CC genes in Brassica species.

The Ubiquitin E3 Ligase PRU2 Modulates Phosphate Uptake in Arabidopsis.

Phosphorus is an essential macronutrient for plants. The phosphate (Pi) concentration in soil solutions is typically low, and plants always suffer from low-Pi stress. During Pi starvation, a number of adaptive mechanisms in plants have evolved to increase Pi uptake, whereas the mechanisms are not very clear. Here, we report that an ubiquitin E3 ligase, PRU2, modulates Pi acquisition in Arabidopsis response to the low-Pi stress. The mutant pru2 showed arsenate-resistant phenotypes and reduced Pi content and Pi uptake rate. The complementation with PRU2 restored these to wild-type plants. PRU2 functioned as an ubiquitin E3 ligase, and the protein accumulation of PRU2 was elevated during Pi starvation. PRU2 interacted with a kinase CK2α1 and a ribosomal protein RPL10 and degraded CK2α1 and RPL10 under low-Pi stress. The in vitro phosphorylation assay showed that CK2α1 phosphorylated PHT1;1 at Ser-514, and prior reports demonstrated that the phosphorylation of PHT1;1 Ser-514 resulted in PHT1;1 retention in the endoplasmic reticulum. Then, the degradation of CK2α1 by PRU2 under low-Pi stress facilitated PHT1;1 to move to the plasma membrane to increase Arabidopsis Pi uptake. Taken together, this study demonstrated that the ubiquitin E3 ligase—PRU2—was an important positive regulator in modulating Pi acquisition in Arabidopsis response to low-Pi stress.

Integrated multi-omics analysis provides insights into genome evolution and phosphorus deficiency adaptation in pigeonpea (Cajanus cajan).

Pigeonpea (Cajanus cajan) is an important legume food crop and plays a crucial role in a secure food supply in many developing countries. Several previous studies have suggested that pigeonpea has great potential for phosphorus (P) deficiency tolerance, but little is known about the underlying mechanism. In this study, the physiological and molecular responses of pigeonpea roots to phosphate (Pi) starvation were investigated through integrating phenotypic, genomic, transcriptomic, metabolomic, and lipidomic analyses. The results showed that low-Pi treatment increased total root length, root surface area, and root acid phosphatase activity, and promoted the secretion of organic acids (e.g. citric acids, piscidic acids, and protocatechuic acids) and the degradation of phospholipids and other P-containing metabolites in the roots of pigeonpea. Consistent with the morphological, physiological, and biochemical changes, a large number of genes involved in these Pi-starvation responses were significantly upregulated in Pi-deficient pigeonpea roots. Among these Pi-starvation response genes upregulated by low-Pi treatment, four gene families were expanded through recent tandem duplication in the pigeonpea genome, namely phosphate transporter 1 (PHT1), phosphoethanolamine/phosphocholine phosphatase (PECP), fasciclin-like arabinogalactan protein (FLA), and glutamate decarboxylase (GAD). These gene families may be associated with Pi uptake from the soil, phospholipid recycling, root morphological remodeling, and regulation of organic acid exudation. Taken together, our results suggest that pigeonpea employs complex Pi-starvation responses to strengthen P acquisition and utilization during low-Pi stress. This study provides new insights into the genome evolution and P deficiency adaptation mechanism of pigeonpea.

Tampk2b, a Member of the Mapk Family in T. Aestivum, Enhances Plant Low-Pi Stress Tolerance Through Modulating Physiological Processes Associated with Low-P Stress Defensiveness

Tampk2b, a Member of the Mapk Family in T. Aestivum, Enhances Plant Low-Pi Stress Tolerance Through Modulating Physiological Processes Associated with Low-P Stress Defensiveness

TaMADS2-3D, a MADS transcription factor gene, regulates phosphate starvation responses in plants

Soil inorganic phosphate (Pi) levels are frequently suboptimal for the growth and development of crop plants. Although MADS-box genes participate in diverse plant developmental processes, their involvement in phosphate starvation responses (PSRs) remains unclear. We identified a type I MADS-box transcription factor gene, TaMADS2-3D, which was rapidly induced under low-Pi stress in roots of wheat (Triticum aestivum). A TaMADS2-3D-GFP fusion protein was found located in the nucleus. Transgenic Arabidopsis plants overexpressing TaMADS2-3D (TaMADS2-3DOE) showed shortened primary roots, increased lateral root density, and retarded seedling growth under high-Pi (HP) conditions, accompanied by increased Pi contents in their shoots and roots. The Arabidopsis TaMADS2-3DOE plants showed similar PSR phenotypes under low Pi (LP) conditions. These results indicate constitutive activation of PSRs by overexpression of TaMADS2-3D in Arabidopsis. Reactive oxygen species (ROS), H2O2 and O2−, levels were increased in root tips of Arabidopsis TaMADS2-3DOE plants under HP conditions. Transcriptome analysis of Arabidopsis TaMADS2-3DOE plants under different Pi regimes revealed expression changes for a variety of PSR genes including AtZAT6. Overexpression of TaMADS2-3D in wheat also led to constitutive activation of PSRs. We propose that TaMADS2-3D regulates plant PSRs probably by modulating ROS homeostasis, root development, PSR gene expression, and Pi uptake. This study increases our understanding of plant PSR regulation and provides a valuable gene for improving phosphorus-use efficiency in wheat and other crops.