Abstract
Plasma membrane H+-ATPase (PM H+-ATPase, EC 3.6.1.3.) is a proton pump that is necessary to promote cell growth and ion fluxes across the plasma membrane. The main goal of this study was to evaluate the role of PM H+-ATPase isoform OsA7 expression in rice growth and nitrogen (N) accumulation using three genetically engineered lineages with artificial micro RNA (amiRNA) targeting OsA7 (osa7.1, osa7.2, and osa7.3). PM H+-ATPase isoform expression in rice shoots and roots (wild-type) revealed that OsA7 is highly expressed in roots and is the most highly expressed PM H+-ATPase isoform. The three osa7 lineages had lower fresh weight, grain yield, height, and 1000-grain weight compared to control IRS plants. The hydroponic experiment comprised three NO3− levels over 30 days: 0.2 mM NO3−–N, 2.0 mM NO3−–N, and NO3− starvation for 3 days. The three osa7 lineages had lower PM H+-ATPase and V-H+-PPase activity as compared to the IRS plants. The root and shoot fresh weights were lower in osa7 lineages. The root/shoot ratio was lower in the osa7 lineages cultivated without nitrogen for 3 days and with 0.2 mM of NO3−–N as compared to IRS, and did not change in plants cultivated with 2.0 mM NO3−–N. The total N concentration did not change in the three osa7 lineages as compared to IRS. Overall, the results indicate that OsA7 is important for rice growth, grain production, and root growth, but does not affect N accumulation, highlighting the importance of other PM H+-ATPase isoforms in N uptake.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arango M, Gévaudant F, Oufattol M, Boutry M (2003) The plasma membrane proton pump ATPase: the significance of gene subfamilies. Planta 216:355–365. https://doi.org/10.1007/s00425-002-0856-8
Baker RF, Leach KA, Braun DM (2012) SWEET as sugar: new sucrose effluxers in plants. Mol Plant 5:766–768. https://doi.org/10.1093/mp/SSS054
Baxter I, Tchieu J, Sussman MR, Boutry M, Palmgren MG, Gribskov M, Harper JF, Axelsen KB (2003) Genomic comparison of P-type ATPase ion pump in Arabidopsis and rice. Plant Physiol 132:618–628. https://doi.org/10.1104/pp.103.021923
Bobik K, Duby G, Nizet Y, Vandermeeren C, Stiernet P, Kanczewska J, Boutry M (2010) Two widely expressed plasma membrane H+-ATPase isoforms of Nicotiana tabacum are differentially regulated by phosphorylation of their penultimate threonine. Plant J 62:291–301. https://doi.org/10.1111/j.1365-313X.2010.04147.x
Bradford MM (1976) Rapid and sensitive method for quantification of microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Chang C, Hu Y, Sun S, Zhu Y, Ma G, Xu G (2009) Proton pump OsA8 is linked to phosphorus uptake and translocation in rice. J Exp Bot 60:557–565. https://doi.org/10.1093/jxb/ern298
Chen M, Wei X, Shao G, Tang S, Luo J, Hu P (2012) Fragrance of the rice grain achieved via artificial microRNA-induced down-regulation of OsBADH2. Plant Breed 131:584–590. https://doi.org/10.1111/j.1439-0523.2012.01989.x
Chen H, Zhang Q, Cai H, Xu F (2017) Ethylene mediates alkaline-induced rice growth inhibition by negatively regulating plasma membrane H+-ATPase activity in roots. Front Plant Sci 8:1839. https://doi.org/10.3389/fpls.2017.01839
Facanha AR, De Meis L (1995) Inhibition of maize root H+-ATPase by fluoride and fluoroaluminate complexes. Plant Physiol 108:241–246. https://doi.org/10.1104/pp.108.1.241
Fageria NK, Baligar VC (2001) Low land rice response to nitrogen fertilization. Commun Soil Sci Plant Anal 32:1405–1428. https://doi.org/10.1081/CSS-100104202
Falhof J, Pedersen JT, Fuglsang AT, Palmgren M (2016) Plasma membrane H+-ATPase regulation in the center of plant physiology. Mol Plant 9:323–337. https://doi.org/10.1016/j.molp.2015.11.002
Fuglsang AT, Paez-Valencia J, Gaxiola RA (2011) Plant proton pumps: regulatory circuits involving H+-ATPase and H+-PPase. In: Geisler M, Venema K (eds) Transporters and pumps in plant signaling. Springer, Berlin, pp 39–64. https://doi.org/10.1007/978-3-642-14369-4_2.
Gao J, Liu J, Li B, Li Z (2001) Isolation and purification of functional total RNA from blue-grained wheat endosperm tissues contaning high levels of starches and flavonoids. Plant Mol Biol Rep 19:185a–185i. https://doi.org/10.1007/BF02772163
Gaxiola RA, Palmgren MG, Schumacher K (2007) Plant proton pumps. FEBS Lett 581:2204–2214. https://doi.org/10.1016/j.febslet.2007.03.050
Gaxiola RA, Sanchez CA, Paez-Valencia J, Ayre BG, Elser JJ (2012) Genetic manipulation of a “vacuolar” H+-PPase: from salt tolerance to yield enhancement under phosphorus-deficient soils. Plant Physiol 159:3–11. https://doi.org/10.1104/pp.112.195701
Glass AD (2003) Nitrogen use efficiency of crop plants: physiological constraints upon nitrogen absorption. Crit Rev Plant Sci 22:453–470. https://doi.org/10.1080/07352680390243512
Hager A (2003) Role of the plasma membrane H+-ATPase in auxin-induced elongation growth: historical and new aspects. J Plant Res 116:483–505. https://doi.org/10.1007/s10265-003-0110-x
Haruta M, Burch HL, Nelson RB, Barrett-Wilt G, Kline KG, Mohsin SB, Young JC, Otegui MS, Sussman MR (2010) Molecular characterization of mutant arabidopsis plants with reduced plasma membrane proton pump activity. J Biol Chem 285:17918–17929. https://doi.org/10.1074/jbc.M110.101733
Hayashi Y, Takahashi K, Inoue S, Kinoshita T (2014) Abscisic acid suppresses hypocotyl elongation by dephosphorylating plasma membrane H+-ATPase in Arabidopsis thaliana. Plant Cell Physiol 55:845–853. https://doi.org/10.1093/pcp/pcu028
Hiei Y, Komari T (2008) Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed. Nat Protoc 3:824–834. https://doi.org/10.1038/nprot.2008.46
Ho CH, Lin SH, Hu HC, Tsay YF (2009) CHL1 functions as a nitrate sensor in plants. Cell 138:1184–1194. https://doi.org/10.1016/j.cell.2009.07.004
Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. University of California, Berkeley, pp 1–32
Hoffmann RD, Olsen LI, Ezike CV, Pedersen JT, Manstretta R, López-Marqués RL, Palmgren M (2018) Roles of plasma membrane proton ATPases AHA2 and AHA7 in normal growth of roots and root hairs in Arabidopsis thaliana. Physiol Plant. https://doi.org/10.1111/ppl.12842
Jain M, Nijhawan A, Tyagi AK, Khurana JP (2006) Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. Biochem Biophys Res Commun 345:646–651. https://doi.org/10.1016/j.bbrc.2006.04.140
Jiang S, Sun J, Tian Z, Hu H, Michel EJS, Gao J, Jiang D, Cao W, Dai T (2017) Root extension and nitrate transporter up-regulation induced by nitrogen deficiency improves nitrogen status and plant growth at the seedling stage of winter wheat (Triticum aestivum L.). Environ Exp Bot 141:28–40. https://doi.org/10.1016/j.envexpbot.2017.06.006
Kiba T, Kudo T, Kojima M, Sakakibara H (2011) Hormonal control of nitrogen acquisition: roles of auxin, abscisic acid, and cytokinin. J Exp Bot 62:1399–1409. https://doi.org/10.1093/jxb/erq410
Kinoshita T, Shimazaki K (2002) Biochemical evidence for the requirement of 14–3-3 protein binding in activation of the guard-cell plasma membrane H+-ATPase by blue light. Plant Cell Physiol 43:1359–1365. https://doi.org/10.1093/pcp/pcf167
Krouk G (2016) Hormones and nitrate: a two-way connection. Plant Mol Biol 91:599–606. https://doi.org/10.1007/s11103-016-0463-x
Krouk G, Lacombe B, Bielach A, Perrine-Walker F, Malinska K, Mounier E, Hoyerova K, Tillard P, Leon S, Ljung K, Zazimalova E, Benkova E, Nacry P, Gojon A (2010) Nitrate-regulated auxin transport by NRT1.1 defines a mechanism for nutrient sensing in plants. Dev Cell 18(927):937. https://doi.org/10.1016/j.devcel.2010.05.008
Li J, Yang H, Peer WA, Richter G, Blaskelee J, Bandyopadhyay A, Titapiwantakun B, Undurraga S, Khodakovskaya M, Richards EL, Krizek B, Murphy AS, Gilroy S, Gaxiola R (2005) Arabidopsis H+-PPase AVP1 regulates auxin-mediated organ development. Science 310:121–125. https://doi.org/10.1126/science.1115711
Li X, Guo C, Gu J, Duan W, Zhao M, Ma C, Du X, Lu W, Xiao K (2014) Overexpression of VP, a vacuolar H+-pyrophosphatase gene in wheat (Triticum aestivum L.), improves tobacco plant growth under Pi and N deprivation, high salinity, and drought. J Exp Bot 65:683–696. https://doi.org/10.1093/jxb/ert442
Li Z, Zhang X, Zhao Y, Li Y, Zhang G, Peng Z, Zhang J (2018) Enhancing auxin accumulation in maize root tips improves root growth and dwarfs plant height. Plant Biotechnol J 16:86–99. https://doi.org/10.1111/pbi.12751
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCт method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Młodzińska E, Kłobus G, Christensen MD, Fuglsang AT (2015) The plasma membrane H+ATPase AHA2 contributes to the root architecture in response to different nitrogen supply. Physiol Plant 154:270–282. https://doi.org/10.1111/ppl.12305
Paez-Valencia J, Sanchez-Lares J, Marsh E, Dorneles LT, Santos MP, Sanchez D, Winter D, Murphy S, Cox J, Trzaska M, Metler J, Kozic A, Facanha AR, Schachtman D, Sanchez CA, Gaxiola RA (2013) Enhanced proton translocating pyrophosphatase activity improves nitrogen use efficiency in romaine lettuce. Plant Physiol 161:1557–1569. https://doi.org/10.1104/pp.112.212852
Quaggiotti S, Ruperti B, Borsa P, Destro T, Malagoli M (2003) Expression of a putative high-affinity NO3- transporter and of an H+-ATPase in relation to whole plant nitrate transport physiology in two maize genotypes differently responsive to low nitrogen availability. J Exp Bot 54:1023–1031. https://doi.org/10.1093/jxb/erg106
Sambrook J, Russel DW (2001) Molecular cloning. A laboratory manual, 3rd ed. CSHL Press, New York, p 999. https://doi.org/10.1016/0092-8674(90)90210-6
Santi S, Schmidt W (2009) Dissecting iron deficiency-induced proton extrusion in Arabidopsis roots. New Phytol 183:1072–1084. https://doi.org/10.1111/j.1469-8137.2009.02908.x
Santi S, Locci G, Monte R, Pinton R, Varanini Z (2003) Induction of nitrate uptake in maize roots: expression of a putative high-affinity nitrate transporter and plasma membrane H+-ATPase isoforms. J Exp Bot 54:1851–1864. https://doi.org/10.1093/jxb/erg208
Santos LA, Bucher CA, Souza SR, Fernandes MS (2009) Effects of nitrogen stress on proton-pumping and nitrogen metabolism in rice. J Plant Nutr 32:549–564. https://doi.org/10.1080/01904160802714953
Schilling RK, Tester M, Marschner P, Plett DC, Roy SJ (2017) AVP1: one protein, many roles. Trends Plant Sci 22:154–162. https://doi.org/10.1016/j.tplants.2016.11.012
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 7:671–675. https://doi.org/10.1038/nmeth.2089
Sperandio MVL, Santos LA, Bucher CA, Fernandes MS, Souza SR (2011) Isoforms of plasma membrane H+-ATPase in rice root and shoot are differentially induced by starvation and resupply of NO3- or NH4+. Plant Sci 180:251–258. https://doi.org/10.1016/j.plantsci.2010.08.018
Sperandio MVL, Santos LA, Araújo OJL, Braga RP, Coelho CP, Nogueira EM, Fernandes MS, Souza SR (2014) Response of nitrate transporters and PM H+-ATPase expression to nitrogen flush in two upland rice varieties contrasting in nitrate uptake kinetics. Aust J Crop Sci 8:568–576
Sun L, Pehlivan N, Esmaeili N, Jiang W, Yang X, Jarret P, Mishra N, Zhu X, Ca Y, Herath M, Shen G, Zhang H (2018) Co-overexpression of AVP1 and PP2A-C5 in Arabidopsis makes plants tolerant to multiple abiotic stresses. Plant Sci 274:271–283. https://doi.org/10.1016/j.plantsci.2018.05.026
Takahashi K, Hayashi K, Kinoshita T (2012) Auxin activates the plasma membrane H+-ATPase by phosphorylation during hypocotyl elongation in Arabidopsis. Plant Physiol 159:632–641. https://doi.org/10.1104/pp.112.196428
Toda Y, Wang Y, Takahashi A, Kawai Y, Tada Y, Yamaji N, Ma JM, Ashikari M, Kinoshita T (2016) Oryza sativa H+-ATPase (OSA) is involved in the regulation of dumbbell-shaped guard cells of rice. Plant Cell Physiol 57:1220–1230. https://doi.org/10.1093/pcp/pcw070
Toki S, Hara N, Ono K, Onodera H, Tagiri A, Oka S, Tanaka H (2006) Early infection of scutellum tissue with Agrobacterium allows high-speed transformation of rice. Plant J 47:969–976. https://doi.org/10.1111/j.1365-313X.2006.02836.x
Undurraga SF, Santos MP, Paez-Valencia J, Yang H, Hepler PK, Facanha AR, Hirschi KD, Gaxiola RA (2012) Arabidopsis sodium dependent and independent phenotypes triggered by H+-PPase up-regulation are SOS1 dependent. Plant Sci 183:96–105. https://doi.org/10.1016/j.plantsci.2011.11.011
Vidal EA, Araus V, Lu C, Parry G, Green PJ, Coruzzi GM, Gutiérrez RA (2010) Nitrate-responsive miR393/AFB3 regulatory module controls root system architecture in Arabidopsis thaliana. Proc Natl Acad Sci 107:4477–4482. https://doi.org/10.1073/pnas.0909571107
Warthmann N, Chen H, Ossowski S, Weigel D, Hervé P (2008) Highly specific gene silencing by artificial miRNAs in rice. PLoS ONE 3:e1829. https://doi.org/10.1371/journal.pone.0001829
Wu D, Shen H, Yokawa K, Baluška F (2014) Alleviation of aluminium-induced cell rigidity by overexpression of OsPIN2 in rice roots. J Exp Bot 65:5305–5315. https://doi.org/10.1093/jxb/eru292
Yang H, Zhang X, Gaxiola RA, Xu G, Peer WA, Murphy AS (2014) Over-expression of the Arabidopsis proton-pyrophosphatase AVP1 enhances transplant survival, root mass, and fruit development under limiting phosphorus conditions. J Exp Bot 65:3045–3053. https://doi.org/10.1093/jxb/eru149
Yuan Z, Cao Q, Zhang K, Ata-UI-Karim ST, Tian Y, Zhu Y, Cao W, Liu X (2016) Optimal leaf positions for SPAD meter measurement in rice. Front Plant Sci 7:719. https://doi.org/10.3389/fpls.2016.00719
Zandonadi DB, Canellas LP, Façanha AR (2007) Indolacetic and humic acids induce lateral root development through a concerted plasmalemma and tonoplast H+ pumps activation. Planta 225:1583–1589. https://doi.org/10.1007/s00425-006-0454-2
Acknowledgements
We would like to thank Emmanuel Guiderdoni and Martine Bes (Centre de coopération internationale en recherche agronomique pour le développement; CIRAD, France) for providing the IRS154 vector, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and the Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) for the financial support.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
On behalf of all authors, the corresponding author states that there is no conflict of interest.
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
Sperandio, M.V.L., Santos, L.A., Tavares, O.C.H. et al. Reduced Plasma Membrane H+-ATPase Isoform OsA7 Expression and Proton Pump Activity Decrease Growth Without Affecting Nitrogen Accumulation in Rice. J Plant Growth Regul 40, 67–77 (2021). https://doi.org/10.1007/s00344-020-10081-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00344-020-10081-9