Abstract
Main conclusion
The phosphate transporters LpPHT1;1 and LpPHT1;4 have different roles in phosphate uptake and translocation in ryegrass under P stress condition.
Abstract
The phosphate transporter 1 (PHT1) family are integral membrane proteins that operate in phosphate uptake, distribution and remobilization within plants. In this study, we report on the functional characterization and expression of two PHT1 family members from ryegrass plants (Lolium perenne L.) and determine their roles in the specificity of Pi transport. The expression level of LpPHT1;4 was strongly influenced by phosphorus (P) status, being higher under P-starvation condition. In contrast, the expression level of LpPHT1;1 was not correlated with P supply. Yeast mutant complementation assays showed that LpPHT1;4 can complement the growth defect of the yeast mutant Δpho84 under Pi-deficient conditions, whereas the yeast mutant expressing LpPHT1;1 was not able to restore growth. Phylogenetic and molecular analyses indicated high sequence similarity to previously identified PHT1s from other species in the Poaceae. These results suggest that LpPHT1;1 may function as a low-affinity Pi transporter, whereas LpPHT1;4 could acts as a high-affinity Pi transporter to maintain Pi homeostasis under stress conditions in ryegrass plants. This study will form the basis for the long-term goal of improving the phosphate use efficiency of ryegrass plants.
Similar content being viewed by others
Abbreviations
- PHT1:
-
Phosphate transporter 1
- PAE:
-
Phosphorus acquisition efficiency
- PUE:
-
Phosphorus utilization efficiency
References
Ai P, Sun S, Zhao J, Fan X, Xin W, Guo Q et al (2009) Two rice phosphate transporters, OsPht1;2 and OsPht1;6, have different functions and kinetic properties in uptake and translocation. Plant J 57:798–809. https://doi.org/10.1111/j.1365-313X.2008.03726.x
Ashkenazy H, Erez E, Martz E, Pupko T, Ben-Tal N (2010) ConSurf 2010: calculating evolutionary conservation in sequence and structure of proteins and nucleic acids. Nucleic Acids Res 38:529–533. https://doi.org/10.1093/nar/gkq399
Ayadi A, David P, Arrighi JF, Chiarenza S, Thibaud MC, Nussaume L et al (2015) Reducing the genetic redundancy of Arabidopsis PHOSPHATE TRANSPORTER1 transporters to study phosphate uptake and signaling. Plant Physiol 167:1511–1526. https://doi.org/10.1104/pp.114.252338
Baker A, Ceasar SA, Palmer AJ, Paterson JB, Qi W, Muench SP et al (2015) Replace, reuse, recycle: improving the sustainable use of phosphorus by plants. J Exp Bot 66:3523–3540. https://doi.org/10.1093/jxb/erv210
Bun-Ya M, Nishimura M, Harashima S, Oshima Y (1991) The PHO84 gene of Saccharomyces cerevisiae encodes an inorganic phosphate transporter. Mol Cell Biol 11:3229–3238. https://doi.org/10.1128/mcb.11.6.3229
Byrne SL, Foito A, Hedley PE, Morris JA, Stewart D, Barth S (2011) Early response mechanisms of perennial ryegrass (Lolium perenne) to phosphorus deficiency. Ann Bot 107:243–254. https://doi.org/10.1093/aob/mcq234
Ceasar SA, Hodge A, Baker A, Baldwin SA (2014) Phosphate concentration and arbuscular mycorrhizal colonisation influence the growth, yield and expression of twelve PHT1 family phosphate transporters in foxtail millet (Setaria italica). PLoS One 9:e108459. https://doi.org/10.1371/journal.pone.0108459
Ceasar SA, Baker A, Muench SP, Ignacimuthu S, Baldwin SA (2016) The conservation of phosphate-binding residues among PHT1 transporters suggests that distinct transport affinities are unlikely to result from differences in the phosphate-binding site. Biochem Soc Trans 44:1541–1548. https://doi.org/10.1042/BST20160016
Ceasar SA, Baker A, Ignacimuthu S (2017) Functional characterization of the PHT1 family transporters of foxtail millet with development of a novel Agrobacterium-mediated transformation procedure. Sci Rep 7:1–16. https://doi.org/10.1038/s41598-017-14447-0
Chien PS, Chiang CP, Leong SJ, Chiou TJ (2018) Sensing and signaling of phosphate starvation: from local to long distance. Plant Cell Physiol 59:1714–1722. https://doi.org/10.1093/pcp/pcy148
Cordell D, White S (2014) Life’s bottleneck: sustaining the world’s phosphorus for a food secure future. Annu Rev Environ Resour 39:161–188. https://doi.org/10.1146/annurev-environ-010213-113300
Felsenstein J (1985) Phylogenies and the comparative method. Am Nat 125:1–15. https://doi.org/10.1086/284325
Ghillebert R, Swinnen E, De Snijder P, Smets B, Winderickx J (2011) Differential roles for the low-affinity phosphate transporters Pho87 and Pho90 in Saccharomyces cerevisiae. Biochem J 434:243–251. https://doi.org/10.1042/BJ20101118
Gietz RD, Schiestl RH (2007) High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2:31–34. https://doi.org/10.1038/nprot.2007.13
Gu M, Chen A, Sun S, Xu G (2016) Complex regulation of plant phosphate transporters and the gap between molecular mechanisms and practical application: what is missing? Mol Plant 9:396–416. https://doi.org/10.1016/j.molp.2015.12.012
Ham BK, Chen J, Yan Y, Lucas WJ (2018) Insights into plant phosphate sensing and signaling. Curr Opin Biotechnol 49:1–9. https://doi.org/10.1016/j.copbio.2017.07.005
Huang CY, Shirley N, Genc Y, Shi B, Langridge P (2011) Phosphate utilization efficiency correlates with expression of low-affinity phosphate transporters and noncoding RNA, IPS1, in barley. Plant Physiol 156:1217–1229. https://doi.org/10.1104/pp.111.178459
Jain A, Nagarajan VK, Raghothama KG (2012) Transcriptional regulation of phosphate acquisition by higher plants. Cell Mol Life Sci 69:3207–3224. https://doi.org/10.1007/s00018-012-1090-6
Julia CC, Rose TJ, Pariasca-Tanaka J, Jeong K, Masuda T, Wissuwa M (2018) Phosphorus uptake commences at the earliest stages of seedling development in rice (Oryza sativa L.). J Exp Bot 69:5233–5240. https://doi.org/10.1093/jxb/ery267
Kochian LV, Hoekenga OA, Piñeros MA (2004) How do crop plants tolerate acid soils? Mechanisms of aluminum tolerance and phosphorous efficiency. Annu Rev Plant Biol 55:459–493. https://doi.org/10.1146/annurev.arplant.55.031903.141655
Lapis-Gaza HR, Jost R, Finnegan PM (2014) Arabidopsis PHOSPHATE TRANSPORTER1 genes PHT1; 8 and PHT1; 9 are involved in root-to-shoot translocation of orthophosphate. BMC Plant Biol 14:334. https://doi.org/10.1186/s12870-014-0334-z
Liu X, Zhao X, Zhang L, Lu W, Li X, Xiao K (2013) TaPht1;4, a high-affinity phosphate transporter gene in wheat (Triticum aestivum), plays an important role in plant phosphate acquisition under phosphorus deprivation. Funct Plant Biol 40:329–341. https://doi.org/10.1071/FP12242
Liu P, Chen S, Song A, Zhao S, Fang W, Guan Z et al (2014) A putative high affinity phosphate transporter, CmPT1, enhances tolerance to Pi deficiency of chrysanthemum. BMC Plant Biol 14:18. https://doi.org/10.1186/1471-2229-14-18
Liu F, Xu Y, Jiang H, Jiang C, Du Y, Gong C et al (2016) Systematic identification, evolution and expression analysis of the Zea mays PHT1 gene family reveals several new members involved in root colonization by arbuscular mycorrhizal fungi. Int J Mol Sci 17:930. https://doi.org/10.3390/ijms17060930
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔct method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
López-Arredondo DL, Leyva-González MA, González-Morales SI, López-Bucio J, Herrera-Estrella L (2014) Phosphate nutrition: improving low-phosphate tolerance in crops. Annu Rev Plant Biol 65:95–123. https://doi.org/10.1146/annurev-arplant-050213-035949
López-Arredondo DL, Sánchez-Calderón L, Yong-Villalobos L (2017) Molecular and genetic basis of plant macronutrient use efficiency: concepts, opportunities, and challenges. In: Hossain M, Kamiya T, Burritt DJ, Tran LP, Fujiwara T (eds) Plant macronutrient use efficiency. Academic Press, New York, pp 1–29
Maharajan T, Ceasar SA, Ajeesh Krishna TP, Ramakrishnan M, Duraipandiyan V, Abdulla Naif et al (2018) Utilization of molecular markers for improving the phosphorus efficiency in crop plants. Plant Breed 137:10–26. https://doi.org/10.1111/pbr.12537
Mayrose I, Graur D, Ben-Tal N, Pupko A (2004) Comparison of site specific rate-inference methods for protein sequences: empirical bayesian methods are superior. Mol Biol Evol 21:1781–1791. https://doi.org/10.1093/molbev/msh194
Mora ML, Alfaro MA, Jarvis SC, Demanet R, Cartes P (2006) Soil aluminium availability in Andisols of southern Chile and its effect on forage production and animal metabolism. Soil Use Manag 22:95–101. https://doi.org/10.1111/j.1475-2743.2006.00011.x
Muchhal US, Raghothama KG (1999) Transcriptional regulation of plant phosphate transporters. Proc Natl Acad Sci USA 96:5868–5872. https://doi.org/10.1073/pnas.96.10.5868
Nussaume L, Kanno S, Javot H, Marin E, Pochon N, Ayadi A et al (2011) Phosphate import in plants: focus on the PHT1 transporters. Front Plant Sci 2:83. https://doi.org/10.3389/fpls.2011.00083
Ohno T, Zibilske LM (1991) Determination of low concentrations of phosphorus in soil extracts using malachite green. Soil Sci Soc Am J 55:892–895. https://doi.org/10.2136/sssaj1991.03615995005500030046x
Parra-Almuna L, Diaz-Cortez A, Ferrol N, de la Mora ML (2018) Aluminium toxicity and phosphate deficiency activates antioxidant systems and up-regulates expression of phosphate transporters gene in ryegrass (Lolium perenne L.) plants. Plant Physiol Biochem 130:445–454. https://doi.org/10.1016/j.plaphy.2018.07.031
Pedersen BP, Kumar H, Waight AB, Risenmay AJ, Roe-Zurz Z, Chau BH et al (2013) Crystal structure of a eukaryotic phosphate transporter. Nature 496:533–536. https://doi.org/10.1038/nature12042
Peret B, Desnos T, Jost R, Kanno S, Berkowitz O, Nussaume L (2014) Root architecture responses: in search of phosphate. Plant Physiol 166:1713–1723. https://doi.org/10.1104/pp.114.244541
Plaxton WC, Tran HT (2011) Metabolic adaptations of phosphate-starved plants. Plant Physiol 156:1006–1015. https://doi.org/10.1104/pp.111.175281
Qin L, Guo Y, Chen L, Liang R, Gu M, Xu G et al (2012) Functional characterization of 14 Pht1 family genes in yeast and their expressions in response to nutrient starvation in soybean. PLoS One 7:e47726. https://doi.org/10.1371/journal.pone.0047726
Raghothama KG, Karthikeyan AS (2005) Phosphate acquisition. Plant Soil 274:37–49. https://doi.org/10.1146/annurev.arplant.50.1.665
Rasmussen S, Liu Q, Parsons AJ, Jones CS, Xue H (2014) Transcriptional regulation of phosphate transporters from Lolium perenne and its mycorrhizal symbionts in response to phosphorus supply. Funct Plant Biol 42:1–8. https://doi.org/10.1071/FP14043
Sadzawka A, Grez R, Carrasco MA, Mora ML (2004) Métodos de Análisis de Tejidos Vegetales. Comisión de Normalización y Acreditación (CNA), Sociedad Chilena de la Ciencia del Suelo, Chile
Saitou N, Nei M (1987) The neighbor-joining method—a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454
Sattari SZ, Bouwman AF, Martinez-Rodríguez R, Beusen AH, Van Ittersum MK (2016) Negative global phosphorus budgets challenge sustainable intensification of grasslands. Nat Commun. https://doi.org/10.1038/ncomms10696
Smith FW, Mudge SR, Rae AL, Glassop D (2003) Phosphate transport in plants. Plant Soil 248:71–83. https://doi.org/10.1023/A:1022376332180
Sun S, Gu M, Cao Y, Huang X, Zhang X, Ai P et al (2012) A constitutive expressed phosphate transporter, OsPht1;1, modulates phosphate uptake and translocation in phosphate-replete rice. Plant Physiol 159:1571–1581. https://doi.org/10.1104/pp.112.196345
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Taylor GJ, Foy CD (1985) Mechanisms of aluminum tolerance in Triticum aestivum L. (wheat). II. Differential pH induced by spring cultivars in nutrient solutions. Am J Bot 72:702–706. https://doi.org/10.1080/01904168509363372
Teng W, Zhao YY, Zhao XQ, He X, Ma W et al (2017) Genome-wide identification, characterization, and expression analysis of PHT1 phosphate transporters in wheat. Front Plant Sci 8:543. https://doi.org/10.3389/fpls.2017.00543
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. https://doi.org/10.1093/nar/22.22.4673
Vance CP, Uhde-Stone C, Allan DL (2003) Phosphorus acquisition and use: critical adaptations by plants for securing a nonrenewable resource. New Phytol 157:423–447. https://doi.org/10.1046/j.1469-8137.2003.00695.x
Veneklaas EJ, Lambers H, Bragg J, Finnegan PM, Lovelock CE, Plaxton WC et al (2012) Opportunities for improving phosphorus-use efficiency in crop plants. New Phytol 195:306–320. https://doi.org/10.1111/j.1469-8137.2012.04190.x
Versaw WK, Garcia LR (2017) Intracellular transport and compartmentation of phosphate in plants. Curr Opin Plant Biol 39:25–30. https://doi.org/10.1016/j.pbi.2017.04.015
Wang X, Shen J, Liao H (2010) Acquisition or utilization, which is more critical for enhancing phosphorus efficiency in modern crops? Plant Sci 179:302–306. https://doi.org/10.1016/j.plantsci.2010.06.007
Wang D, Lv S, Jiang P, Li Y (2017) Roles, regulation, and agricultural application of plant phosphate transporters. Front Plant Sci 8:817. https://doi.org/10.3389/fpls.2017.00817
Wang F, Deng M, Xu J, Zhu X, Mao C (2018) Molecular mechanisms of phosphate transport and signaling in higher plants. Semin Cell Dev Biol 74:114–122. https://doi.org/10.1016/j.semcdb.2017.06.013
Waterhouse A, Bertoni M, Bienert S, Studer G, Tauriello G, Gumienny R et al (2018) SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 46:296–303. https://doi.org/10.1093/nar/gky427
Ye Y, Yuan J, Chang X, Yang M, Zhang L, Lu K et al (2015) The phosphate transporter gene OsPht1;4 is involved in phosphate homeostasis in rice. PLoS One 10:e0126186. https://doi.org/10.1371/journal.pone.0126186
Zhang Z, Liao H, Lucas WJ (2014) Molecular mechanisms underlying phosphate sensing, signaling, and adaptation in plants. J Integr Plant Biol 56:192–220. https://doi.org/10.1111/jipb.12163
Zheng R, Wang J, Liu M, Duan G, Gao X, Bai S et al (2016) Molecular cloning and functional analysis of two phosphate transporter genes from Rhizopogon luteolus and Leucocortinarius bulbiger, two ectomycorrhizal fungi of Pinus tabulaeformis. Mycorrhiza 26:633–644. https://doi.org/10.1007/s00572-016-0702-7
Zuckerkandl E, Pauling L (1965) Evolutionary divergence and convergence in proteins. In: Bryson V, Vogel H (eds) Evolving genes and proteins. Academic Press, New York, pp 97–166
Acknowledgements
We would like to thank the Scientific and Technological Bioresource Nucleus of Universidad de La Frontera (BIOREN-UFRO) for providing access to specialized equipment. This work was supported by Project (FONDECYT 1181050), CONICYT Doctoral scholarship (No. 21151320) and the Spanish Ministry of Economy, Industry and Competitivity Project (AGL2015-67098-R).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
No conflicts of interest declared.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Parra-Almuna, L., Pontigo, S., Larama, G. et al. Expression analysis and functional characterization of two PHT1 family phosphate transporters in ryegrass. Planta 251, 6 (2020). https://doi.org/10.1007/s00425-019-03313-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00425-019-03313-0