, Volume 248, Issue 1, pp 185–196 | Cite as

Nitrate inhibits the remobilization of cell wall phosphorus under phosphorus-starvation conditions in rice (Oryza sativa)

  • Chun Quan Zhu
  • Xiao Fang Zhu
  • Chao Wang
  • Xiao Ying Dong
  • Ren Fang Shen
Original Article


Main conclusion

NO3 not only inhibited the reutilization of cell wall P via decreasing root cell wall pectin content and PME activity, but also hampered the P translocation from root to shoot.

The rice cultivars ‘Kasalath’ (Kas) and ‘Nipponbare’ (Nip) were used to demonstrate that the nitrogen source NO3 inhibits internal phosphorus (P) reutilization in rice under P-absence conditions. Analysis using Kas showed that the expression of − P-induced marker genes OsIPS1/2 and OsSPX1/2/3/5 are significantly higher under 1 mM NO 3 −  − P (1N − P) treatment than 0 mM NO 3 −  − P (0N − P) treatment. The absence of NO3 from the nutrient solution significantly increased cell wall P release by increasing pectin synthesis and increasing the activity of pectin methylesterase (PME), and also significantly improved the translocation of soluble P from the root to the shoot by increasing xylem sap P content under P-absence conditions. The rice seedlings grown in 0 mM NO3 accumulated significantly higher nitric oxide (NO) in the roots than those grown in 1 mM NO3. Exogenously applying the NO donor sodium nitroprusside (SNP) revealed that NO is a major contributor to differential cell wall P remobilization in rice by mediating pectin synthesis and demethylation under different NO3 concentrations (0 and 1 mM) under P-deprived conditions.


Cell wall Nitrate (NO3Nitric oxide (NO) Phosphorus (P) Remobilization Rice Translocation 



Nitric oxide


Nitrate reductase


Pectin methylesterase


Sodium nitroprusside



This work was funded by the National Key Basic Research Program of China (Grant number 2014CB441000), the Natural Science Foundation of China (Grant number 31501825), and the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant numbers XDB15030302 and XDB15030202).

Supplementary material

425_2018_2892_MOESM1_ESM.docx (56 kb)
Supplemental Fig. S1 The contents of NH4+ and NO3 in Kas and Nip rice cultivars under P-sufficient and P-deficient conditions. Seedlings were grown in P-sufficient or P-deficient nutrient solutions for 1 week. The roots and shoots were collected to measure the NH4+ and NO3 contents. Columns with asterisks are significantly different at P < 0.05. Data are mean ± SD (n = 4) (DOCX 55 kb)


  1. Arnaud N, Murgia I, Boucherez J, Briat JF, Cellier F, Gaymard F (2006) An iron-induced nitric oxide burst precedes ubiquitin-dependent protein degradation for Arabidopsis AtFer1 ferritin gene expression. J Biol Chem 281:23579–23588CrossRefPubMedGoogle Scholar
  2. Besson-Bard A, Gravot A, Richaud P, Auroy P, Duc C, Gaymard F, Taconnat L, Renou JP, Pugin A, Wendehenne D (2009) Nitric oxide contributes to cadmium toxicity in Arabidopsis by promoting cadmium accumulation in roots and by up-regulating genes related to iron uptake. Plant Physiol 149:1302–1315CrossRefPubMedPubMedCentralGoogle Scholar
  3. Blumenkrantz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem 54:484–489CrossRefPubMedGoogle Scholar
  4. Chang YC, Yamamoto Y, Matsumoto H (1999) Accumulation of aluminium in the cell wall pectin in cultured tobacco (Nicotiana tabacum L.) cells treated with a combination of aluminium and iron. Plant Cell Environ 22:1009–1017CrossRefGoogle Scholar
  5. Che J, Yamaji N, Shao JF, Ma JF, Shen RF (2016) Silicon decreases both uptake and root-to-shoot translocation of manganese in rice. J Exp Bot 67:1535–1544CrossRefPubMedPubMedCentralGoogle Scholar
  6. Du S, Zhang Y, Lin X, Wang Y, Tang C (2008) Regulation of nitrate reductase by nitric oxide in Chinese cabbage pakchoi (Brassica chinensis L.). Plant Cell Environ 31:195–204PubMedGoogle Scholar
  7. Duan K, Yi K, Dang L, Huang H, Wu W, Wu P (2008) Characterization of a sub-family of Arabidopsis genes with the SPX domain reveals their diverse functions in plant tolerance to phosphorus starvation. Plant J 54:965–1128CrossRefPubMedGoogle Scholar
  8. Fan X, Jia L, Li Y, Smith SJ, Miller AJ, Shen Q (2007) Comparing nitrate storage and remobilization in two rice cultivars that differ in their nitrogen use efficiency. J Exp Bot 58:1729–1740CrossRefPubMedGoogle Scholar
  9. Fitter A, Williamson L, Linkohr B, Leyser O (2002) Root system architecture determines fitness in an Arabidopsis mutant in competition for immobile phosphate ions but not for nitrate ions. Proc R Soc Lond B Biol Sci 269:2017–2022CrossRefGoogle Scholar
  10. Gamuyao R, Chin JH, Pariasca-Tanaka J, Pesaresi P, Catausan S, Dalid C, Slamet-Loedin I, Tecson-Mendoza EM, Wissuwa M, Heuer S (2012) The protein kinase Pstol1 from traditional rice confers tolerance of phosphorus deficiency. Nature 488:535–539CrossRefPubMedGoogle Scholar
  11. Graziano M, Lamattina L (2007) Nitric oxide accumulation is required for molecular and physiological responses to iron deficiency in tomato roots. Plant J 52:949–960CrossRefPubMedGoogle Scholar
  12. Hinsinger P (2001) Bioavailability of soil inorganic P in the rhizosphere as affected by root-induced chemical changes: a review. Plant Soil 237:173–195CrossRefGoogle Scholar
  13. Hou XL, Wu P, Jiao FC, Jia QJ, Chen HM, Yu J, Song XW, Yi KK (2005) Regulation of the expression of OsIPS1 and OsIPS2 in rice via systemic and local Pi signalling and hormones. Plant Cell Environ 28:353–364CrossRefGoogle Scholar
  14. Hu B, Wang W, Ou S, Tang J, Li H, Che R, Zhang Z, Chai X, Wang H, Wang Y (2015) Variation in NRT1.1B contributes to nitrate-use divergence between rice subspecies. Nat Genet 47:834–838CrossRefPubMedGoogle Scholar
  15. Husted S, Hebbern CA, Mattsson M, Schjoerring JK (2000) A critical experimental evaluation of methods for determination of NH4 + in plant tissue, xylem sap and apoplastic fluid. Physiol Plantarum 109:167–179CrossRefGoogle Scholar
  16. Jin CW, Du ST, Zhang YS, Lin XY, Tang CX (2009) Differential regulatory role of nitric oxide in mediating nitrate reductase activity in roots of tomato (Solanum lycocarpum). Ann Bot 104:9–17CrossRefPubMedPubMedCentralGoogle Scholar
  17. Jing J, Rui Y, Zhang F, Rengel Z, Shen J (2010) Localized application of phosphorus and ammonium improves growth of maize seedlings by stimulating root proliferation and rhizosphere acidification. Field Crop Res 119:355–364CrossRefGoogle Scholar
  18. Kaiser WM, Huber SC (2001) Post-translational regulation of nitrate reductase: mechanism, physiological relevance and environmental triggers. J Exp Bot 52:1981–1989CrossRefPubMedGoogle Scholar
  19. Kant S, Peng M, Rothstein SJ (2011) Genetic regulation by NLA and microRNA827 for maintaining nitrate-dependent phosphate homeostasis in Arabidopsis. PLoS Genet 7:e1002021CrossRefPubMedPubMedCentralGoogle Scholar
  20. Kirk G, Kronzucker H (2005) The potential for nitrification and nitrate uptake in the rhizosphere of wetland plants: a modelling study. Ann Bot 96:639–646CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kopyra M, Gwóźdź EA (2003) Nitric oxide stimulates seed germination and counteracts the inhibitory effect of heavy metals and salinity on root growth of Lupinus luteus. Plant Physiol Biochem 41:1011–1017CrossRefGoogle Scholar
  22. Kronzucker H, Glass A, Siddiqi M, Kirk G (2000) Comparative kinetic analysis of ammonium and nitrate acquisition by tropical lowland rice: implications for rice cultivation and yield potential. New Phytol 145:471–476CrossRefGoogle Scholar
  23. Lawlor DW, Lemaire G, Gastal F (2001) Nitrogen, plant growth and crop yield. In: Lea PJ, Morot-Gaudry JF (eds) Plant nitrogen. Springer, Berlin, pp 343–367CrossRefGoogle Scholar
  24. Li YL, Fan XR, Shen QR (2008) The relationship between rhizosphere nitrification and nitrogen-use efficiency in rice plants. Plant Cell Environ 31:73–85PubMedGoogle Scholar
  25. Liang C, Wang J, Zhao J, Tian J, Liao H (2014) Control of phosphate homeostasis through gene regulation in crops. Curr Opin Plant Biol 21:59–66CrossRefPubMedGoogle Scholar
  26. Lin Y, Hwang CF, Cheng CL (1994) 5′ proximal regions of Arabidopsis nitrate reductase genes direct nitrate-induced transcription in transgenic tobacco. Plant Physiol 106:477–484CrossRefPubMedPubMedCentralGoogle Scholar
  27. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408CrossRefPubMedPubMedCentralGoogle Scholar
  28. Lynch JP, Brown KM (2001) Topsoil foraging—an architectural adaptation of plants to low phosphorus availability. Plant Soil 237:225–237CrossRefGoogle Scholar
  29. Mackerness AHS, John CF, Jordan B, Thomas B (2001) Early signaling components in ultraviolet-B responses: distinct roles for different reactive oxygen species and nitric oxide. FEBS Lett 489:237–242CrossRefGoogle Scholar
  30. Manoli A, Begheldo M, Genre A, Lanfranco L, Trevisan S, Quaggiotti S (2014) NO homeostasis is a key regulator of early nitrate perception and root elongation in maize. J Exp Bot 65:185–200CrossRefPubMedGoogle Scholar
  31. Marschner H (1995) Mineral nutrition of higher plants, 2nd edn. Academic Press, LondonGoogle Scholar
  32. Meng ZB, Chen LQ, Suo D, Li GX, Tang CX, Zheng SJ (2012) Nitric oxide is the shared signalling molecule in phosphorus-and iron-deficiency-induced formation of cluster roots in white lupin (Lupinus albus). Ann Bot 109:1055–1064CrossRefPubMedPubMedCentralGoogle Scholar
  33. Meyer C, Lea US, Provan F, Kaiser WM, Lillo C (2005) Is nitrate reductase a major player in the plant NO (nitric oxide) game? Photosynth Res 83:181–189CrossRefPubMedGoogle Scholar
  34. Noriharu A, Arihara J, Okada K, Yoshihara T, Johansen C (1990) Phosphorus uptake by pigeon pea and its role in cropping systems of the Indian subcontinent. Science 248:477–480CrossRefGoogle Scholar
  35. Pagnussat GC, Simontacchi M, Puntarulo S, Lamattina L (2002) Nitric oxide is required for root organogenesis. Plant Physiol 129:954–956CrossRefPubMedPubMedCentralGoogle Scholar
  36. Pedroso MC, Magalhaes JR, Durzan D (2000) Nitric oxide induces cell death in Taxus cells. Plant Sci 157:173–180CrossRefPubMedGoogle Scholar
  37. Raghothama K (1999) Phosphate acquisition. Annu Rev Plant Biol 50:665–693CrossRefGoogle Scholar
  38. Rockel P, Strube F, Rockel A, Wildt J, Kaiser WM (2002) Regulation of nitric oxide (NO) production by plant nitrate reductase in vivo and in vitro. J Exp Bot 53:103–110CrossRefPubMedGoogle Scholar
  39. Smith FW, Jackson WA (1987) Nitrogen enhancement of phosphate transport in roots of Zea mays L. I. Effects of ammonium and nitrate pretreatment. Plant Physiol 84:1314–1318CrossRefPubMedPubMedCentralGoogle Scholar
  40. Smoleń S, Sady W (2007) The effect of nitrogen fertilizer form and foliar application on Cd, Cu and Zn concentrations in carrot. Folia Hort 19:87–96Google Scholar
  41. Song L, Ding W, Zhao M, Sun B, Zhang L (2006) Nitric oxide protects against oxidative stress under heat stress in the calluses from two ecotypes of reed. Plant Sci 171:449–458CrossRefPubMedGoogle Scholar
  42. Sun CL, Lu LL, Yu Y, Liu LJ, Hu Y, Ye YQ, Jin CW, Lin XY (2016) Decreasing methylation of pectin caused by nitric oxide leads to higher aluminium binding in cell walls and greater aluminium sensitivity of wheat roots. J Exp Bot 67:979–989CrossRefPubMedGoogle Scholar
  43. Szczerba MW, Britto DT, Balkos KD, Kronzucker HJ (2008) Alleviation of rapid, futile ammonium cycling at the plasma membrane by potassium reveals K+-sensitive and-insensitive components of NH4 + transport. J Exp Bot 59:303–313CrossRefPubMedGoogle Scholar
  44. Tiessen H (2008) Phosphorus in the global environment. In: White PJ, Hammond JP (eds) The ecophysiology of plant–phosphorus interactions. Springer, Dordrecht, pp 1–7Google Scholar
  45. Trevisan S, Manoli A, Quaggiotti S (2014) NO signaling is a key component of the root growth response to nitrate in Zea mays L. Plant Signal Behav 9:1–6CrossRefGoogle Scholar
  46. Uchida A, Jagendorf AT, Hibino T, Takabe T, Takabe T (2002) Effects of hydrogen peroxide and nitric oxide on both salt and heat stress tolerance in rice. Plant Sci 163:515–523CrossRefGoogle Scholar
  47. Vance CP (2008) Plants without arbuscular mycorrhizae. In: White PJ, Hammond JP (eds) The ecophysiology of plant–phosphorus interactions. Springer, Dordrecht, pp 117–142CrossRefGoogle Scholar
  48. Wang Y, Rezzonico E, Poirier Y (2004) Structure and expression profile of the Arabidopsis PHO1 gene family indicates a broad role in inorganic phosphate homeostasis. Plant Physiol 135:400–411CrossRefPubMedPubMedCentralGoogle Scholar
  49. Wang Z, Hu H, Huang H, Duan K, Wu Z, Wu P (2009) Regulation of OsSPX1 and OsSPX3 on expression of OsSPX domain genes and Pi-starvation signaling in rice. J Integr Plant Biol 51:663–674CrossRefPubMedGoogle Scholar
  50. Wang B, Tang X, Cheng L, Zhang A, Zhang W, Zhang F, Liu J, Cao Y, Allan D, Vance C, Shen B (2010) Nitric oxide is involved in phosphorus deficiency-induced cluster-root development and citrate exudation in white lupin. New Phytol 187:1112–1123CrossRefPubMedGoogle Scholar
  51. Xiong J, An LY, Lu H, Zhu C (2009) Exogenous nitric oxide enhances cadmium tolerance of rice by increasing pectin and hemicellulose contents in root cell wall. Planta 230:755–765CrossRefPubMedGoogle Scholar
  52. Yamasaki H, Sakihama Y (2000) Simultaneous production of nitric oxide and peroxynitrite by plant nitrate reductase: in vitro evidence for the NR-dependent formation of active nitrogen species. FEBS Lett 468:89–92CrossRefPubMedGoogle Scholar
  53. Yin XM, Luo W, Wang SW, Shen QR, Long XH (2014) Effect of nitrogen starvation on the responses of two rice cultivars to nitrate uptake and utilization. Pedosphere 24:690–698CrossRefGoogle Scholar
  54. Zeng H, Liu G, Kinoshita T, Zhang R, Zhu Y, Shen Q, Xu G (2012) Stimulation of phosphorus uptake by ammonium nutrition involves plasma membrane H+ ATPase in rice roots. Plant Soil 357:205–214CrossRefGoogle Scholar
  55. Zhao DY, Tian QY, Li LH, Zhang WH (2007) Nitric oxide is involved in nitrate-induced inhibition of root elongation in Zea mays. Ann Bot 100:497–503CrossRefPubMedPubMedCentralGoogle Scholar
  56. Zhao XQ, Guo SW, Shinmachi F, Sunairi M, Noguchi A, Hasegawa I, Shen RF (2013) Aluminium tolerance in rice is antagonistic with nitrate preference and synergistic with ammonium preference. Ann Bot 111:69–77CrossRefPubMedGoogle Scholar
  57. Zheng C, Jiang D, Liu F, Dai T, Liu W, Jing Q, Cao W (2009a) Exogenous nitric oxide improves seed germination in wheat against mitochondrial oxidative damage induced by high salinity. Environ Exp Bot 67:222–227CrossRefGoogle Scholar
  58. Zheng L, Huang F, Narsai R, Wu J, Giraud E, He F, Cheng L, Wang F, Wu P, Whelan J (2009b) Physiological and transcriptome analysis of iron and phosphorus interaction in rice seedlings. Plant Physiol 151:262–274CrossRefPubMedPubMedCentralGoogle Scholar
  59. Zhu XF, Wang ZW, Wan JX, Sun Y, Wu YR, Li GX, Shen RF, Zheng SJ (2015) Pectin enhances rice (Oryza sativa) root phosphorus remobilization. J Exp Bot 66:1017–1024CrossRefPubMedGoogle Scholar
  60. Zhu CQ, Zhu XF, Hu AY, Wang C, Wang B, Dong XY, Shen RF (2016a) Differential effects of nitrogen forms on cell wall phosphorus remobilization are mediated by nitric oxide, pectin content, and phosphate transporter expression. Plant Physiol 171:1407–1417CrossRefPubMedPubMedCentralGoogle Scholar
  61. Zhu XF, Wang B, Song WF, Zheng SJ, Shen RF (2016b) Putrescine alleviates iron deficiency via NO-dependent reutilization of root cell wall Fe in Arabidopsis. Plant Physiol 170:558–567CrossRefPubMedGoogle Scholar
  62. Zhu XF, Zhu CQ, Zhao XS, Zheng SJ, Shen RF (2016c) Ethylene is involved in root phosphorus remobilization in rice (Oryza sativa) by regulating cell-wall pectin and enhancing phosphate translocation to shoots. Ann Bot 118:645–653CrossRefPubMedCentralGoogle Scholar
  63. Zhu XF, Wu Q, Zheng L, Shen RF (2017) NaCl alleviates iron deficiency through facilitating root cell wall iron reutilization and its translocation to the shoot in Arabidopsis thaliana. Plant Soil 417:155–167CrossRefGoogle Scholar
  64. Zou C, Shen J, Zhang F, Guo S, Rengel Z, Tang C (2001) Impact of nitrogen form on iron uptake and distribution in maize seedlings in solution culture. Plant Soil 235:143–149CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Chun Quan Zhu
    • 1
    • 2
    • 3
  • Xiao Fang Zhu
    • 1
  • Chao Wang
    • 1
  • Xiao Ying Dong
    • 1
  • Ren Fang Shen
    • 1
    • 2
  1. 1.State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil ScienceChinese Academy of ScienceNanjingChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.National Key Laboratory of Rice BiologyChina National Rice Research InstituteHangzhouChina

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