, Volume 256, Issue 1, pp 147–159 | Cite as

Elevated CO2-induced production of nitric oxide differentially modulates nitrate assimilation and root growth of wheat seedlings in a nitrate dose-dependent manner

  • Sandeep B. Adavi
  • Lekshmy SatheeEmail author
Original Article


Wheat is a major staple food crop worldwide contributing approximately 20% of total protein consumed by mankind. The nitrogen and protein concentration of wheat crop and grain often decline as a result of exposure of the crop to elevated CO2 (EC). The changes in nitrogen (N) assimilation, root system architecture, and nitric oxide (NO)-mediated N signaling and expression of genes involved in N assimilation and high affinity nitrate uptake were examined in response to different nitrate levels and EC in wheat. Activity of enzyme nitrate reductase (NRA) was downregulated under EC both in leaf and root tissues. Plants grown under EC displayed enhanced production of NO and more so when nitrate supply was high. Based on exogenous supply of NO, inhibitors of NO production, and NO scavenger, regulatory role of NO on EC mediated changes in root morphology and NRA was revealed. The enhanced NO production under EC and high N levels negatively regulated the transcript abundance of NR and high affinity nitrate transporters (HATS).


Nitric oxide (NO) Elevated CO2 High affinity transport system (HATS) 



SAB acknowledges ICAR for the junior research fellowship support received during the course of the study.


ICAR-Indian Agricultural Research Institute funded (institute project-CRSCIARISIL20144047279) and provided the necessary facilities.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

709_2018_1285_MOESM1_ESM.docx (12 kb)
ESM 1 (DOCX 11.5 kb)
709_2018_1285_MOESM2_ESM.docx (13 kb)
ESM 2 (DOCX 12.7 kb)
709_2018_1285_MOESM3_ESM.docx (487 kb)
ESM 3 (DOCX 486 kb)


  1. Balotf S, Islam S, Kavoos G, Kholdebarin B, Juhasz A, Ma W (2018) How exogenous nitric oxide regulates nitrogen assimilation in wheat seedlings under different nitrogen sources and levels. PLoS One 13:0190269CrossRefGoogle Scholar
  2. BassiriRad H, Gutschick VP, Lussenhop J (2001) Root system adjustments: regulation of plant nutrient uptake and growth responses to elevated CO2. Oecologia 126:305–320CrossRefGoogle Scholar
  3. Bethke PC, Gubler F, Jacobsen JV, Jones RL (2004) Dormancy of Arabidopsis seeds and barley grains can be broken by nitric oxide. Planta 219:847–855CrossRefGoogle Scholar
  4. Bloom AJ (2015) Photorespiration and nitrate assimilation: a major intersection between plant carbon and nitrogen. Photosynth Res 123:117–128CrossRefGoogle Scholar
  5. Bloom AJ, Smart DR, Nguyen DT, Searles PS (2002) Nitrogen assimilation and growth of wheat under elevated carbon dioxide. Proc Natl Acad Sci 99:1730–1735CrossRefGoogle Scholar
  6. Bloom AJ, Burger M, Asensio JSR, Cousins AB (2010) Carbon dioxide enrichment inhibits nitrate assimilation in wheat and Arabidopsis. Science 328:899–903CrossRefGoogle Scholar
  7. Bloom AJ, Asensio JSR, Randall L, Rachmilevitch S, Cousins AB, Carlisle EA (2012) CO2 enrichment inhibits shoot nitrate assimilation in C3 but not C4 plants and slows growth under nitrate in C3 plants. Ecology 93:355–367Google Scholar
  8. Bloom AJ, Burger M, Kimball BA, Pinter PJ (2014) Nitrate assimilation is inhibited by elevated CO2 in field-grown wheat. Nat Clim Chang 4:477–480CrossRefGoogle Scholar
  9. Bogdan C, Rollinghoff M, Diefenbach A (2000) The role of nitric oxide in innate immunity. Immunol Rev 173:17–26CrossRefGoogle Scholar
  10. Bowes G (1993) Facing the inevitable: plants and increasing atmospheric CO2. Annu Rev Plant Physiol Plant Mol Biol 44:309–332CrossRefGoogle Scholar
  11. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  12. Cheng T, Chen J, Abd Allah EF, Wang P, Wang G, Hu X, Shi J (2015) Quantitative proteomics analysis reveals that S-nitrosoglutathione reductase (GSNOR) and nitric oxide signaling enhance poplar defense against chilling stress. Planta 242:1361–1390CrossRefGoogle Scholar
  13. Correa-Aragunde N, Graziano M, Lamattina L (2004) Nitric oxide plays a central role in determining lateral root development in tomato. Planta 218:900–905CrossRefGoogle Scholar
  14. Correa-Aragunde N, Lombardo C, Lamattina L (2008) Nitric oxide: an active nitrogen molecule that modulates cellulose synthesis in tomato roots. New Phytol 179:386–396CrossRefGoogle Scholar
  15. Courtois C, Besson A, Dahan J, Bourque S, Dobrowolska G, Pugin A, Wendehenne D (2008) Nitric oxide signalling in plants: interplays with Ca2+ and protein kinases. J Exp Bot 59:155–163Google Scholar
  16. de Pinto MC, Tommasi F, De Gara L (2002) Changes in the antioxidant systems as part of the signaling pathway responsible for the programmed cell death activated by nitric oxide and reactive oxygen species in tobacco bright-yellow 2 cells. Plant Physiol 130:698–708CrossRefGoogle Scholar
  17. Delledonne M, Zeier J, Marocco A, Lamb C (2001) Signal interactions between nitric oxide and reactive oxygen intermediates in the plant hypersensitive disease resistance response. Proc Natl Acad Sci 98:13454–13459CrossRefGoogle Scholar
  18. 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 3:195–204Google Scholar
  19. Du S, Zhang R, Zhang P, Liu H, Yan M, Chen N, Ke S (2016) Elevated CO2-induced production of nitric oxide (NO) by NO synthase differentially affects nitrate reductase activity in Arabidopsis plants under different nitrate supplies. J Exp Bot 67:893–904CrossRefGoogle Scholar
  20. Evans J, Nason A (1953) Pyridine nucleotide-nitrate reductase from extracts of higher plants. Plant Physiol 28:233–254CrossRefGoogle Scholar
  21. Feechan A, Kwon E, Yuri B, Wang Y, Pallas J, Loake G (2005) A central role for S-nitrosothiols in plant disease resistance. Proc Natl Acad Sci 102:8054–8059CrossRefGoogle Scholar
  22. Flores T, Todd CD, Tovar-Mendez A, Dhanoa PK, Correa-Aragunde N, Hoyos ME, Brownfield DM, Mullen RT, Lamattina L Polacco JC (2008) Arginase-negative mutants of Arabidopsis exhibit increased nitric oxide signaling in root development. Plant Physiol 147:1936–1946Google Scholar
  23. Foresi N, Correa-Aragunde N, Parisi G, Calo G, Salerno G, Lamattina L (2010) Characterization of a nitric oxide synthase from the plant kingdom: NO generation from the green alga Ostreococcus tauri is light irradiance and growth phase dependent. Plant Cell 22:3816–3830CrossRefGoogle Scholar
  24. Frungillo L, Skelly MJ, Loake GJ, Spoel SH, Salgado I (2014) S-nitrosothiols regulate nitric oxide production and storage in plants through the nitrogen assimilation pathway. Nat Commun 5:5401CrossRefGoogle Scholar
  25. Geiger M, Haake V, Ludewig F, Sonnewald U, Stitt M (1999) The nitrate and ammonium nitrate supply have a major influence on the response of photosynthesis, carbon metabolism, nitrogen metabolism and growth to elevated carbon dioxide in tobacco. Plant Cell Environ 22:1177–1199CrossRefGoogle Scholar
  26. Gonzalez AM, de los AC, Henriquez MJ, Contreras RA, Morales B, Moenne A (2012) Cross talk among calcium, hydrogen peroxide, and nitric oxide and activation of gene expression involving calmodulins and calcium-dependent protein kinases in Ulva compressa exposed to copper excess. Plant Physiol 158:1451–1462CrossRefGoogle Scholar
  27. Gupta KJ, Fernie AR, Kaiser WM, van Dongen JT (2011) On the origins of nitric oxide. Trends Plant Sci 16:160–168CrossRefGoogle Scholar
  28. Hageman RH, Hucklesby DP (1971) Nitrate reductase from higher plants. In Methods in enzymology Academic Press 23:491–503Google Scholar
  29. Hung KT, Chang CJ, Kao CH (2002) Paraquat toxicity is reduced by nitric oxide in rice leaves. J Plant Physiol 159:159–166CrossRefGoogle Scholar
  30. IPCC (2014) Summary for Policymakers. In: Field CB, Barros VR, Dokken DJ, Mach KJ, Mastrandrea MD, Bilir TE, Chatterjee M, Ebi KL, Estrada YO, Genova RC, Girma B, Kissel ES, Levy AN, MacCracken S, Mastrandrea PR, White LL (eds) Climate change 2014: impacts, adaptation, and vulnerability. Part a: global and sectoral aspects. Contribution of working group II to the fifth assessment report of the intergovernmental panel on climate change. Cambridge University Press, Cambridge, pp 1–32Google Scholar
  31. Jackson RB, Reynolds HL (1996) Nitrate and ammonium uptake for single-and mixed-species communities grown at elevated CO2. Oecologia 105:74–80Google Scholar
  32. Jeandroz S, Lamotte O, Astier J, Rasul S, Trapet P, Besson-Bard A, Bourque S, Nicolas-Francès V, Ma W, Berkowitz G, Wendehenne D (2013) There's more to the picture than meets the eye: Nitric oxide cross-talk with Ca2+ signaling. Plant Physiol 163:459–470Google Scholar
  33. 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 104:9–17CrossRefGoogle Scholar
  34. Kaur G, Singh HP, Batish DR, Mahajan P, Kohli RK, Rishi V (2015) Exogenous nitric oxide (NO) interferes with lead (Pb)-induced toxicity by detoxifying reactive oxygen species in hydroponically grown wheat (Triticum aestivum) roots. PLoS One 24:0138713Google Scholar
  35. Klepper L, Flesher D, Hageman RH (1971) Generation of reduced nicotinamide adenine dinucleotide for nitrate reduction in green leaves. Plant Physiol 48:580–590CrossRefGoogle Scholar
  36. Kolbert Z, Ortega L, Erdei L (2010) Involvement of nitrate reductase (NR) in osmotic stress-induced NO generation of Arabidopsis thaliana L. roots. J Plant Physiol 167:77–80CrossRefGoogle Scholar
  37. Larios B, Aguera E, Cabello P, Maldonado JM, De La Haba P (2004) The rate of CO2 assimilation controls the expression and activity of glutamine synthetase through sugar formation in sunflower (Helianthus annuus L.) leaves. J Exp Bot 55:69–75CrossRefGoogle Scholar
  38. Lekshmy S, Jha SK (2017) Selection of reference genes suitable for qRT-PCR expression profiling of biotic stress, nutrient deficiency and plant hormone responsive genes in bread wheat. Ind J Plant Physiol 22:101–106CrossRefGoogle Scholar
  39. Lekshmy S, Jain V, Kheterpal S, Pandey R, Singh R (2009) Effect of elevated CO2 on kinetics of nitrate uptake in wheat roots. Ind J Plant Physiol 14:16–22Google Scholar
  40. Lekshmy S, Jain V, Khetarpal S, Pandey R (2013) Inhibition of nitrate uptake and assimilation in wheat seedlings grown under elevated CO2. Ind J Plant Physiol 18:23–29CrossRefGoogle Scholar
  41. Liu L, Hausladen A, Zeng M, Que L, Heitman J, Stamler JS (2001) A metabolic enzyme for S-nitrosothiol conserved from bacteria to humans. Nature 410:490–494CrossRefGoogle Scholar
  42. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408CrossRefGoogle Scholar
  43. Loladze I (2014) Hidden shift of the ionome of plants exposed to elevated CO2 depletes minerals at the base of human nutrition. elife 3:02245CrossRefGoogle Scholar
  44. Nair TVR, Abrol YP (1973) Nitrate reductase activity in developing wheat ears. Cell Mol Life Sci 29:1480–1481CrossRefGoogle Scholar
  45. Niu Y, Jin C, Jin G, Zhou Q, Lin X, Tang C, Zhang Y (2011) Auxin modulates the enhanced development of root hairs in Arabidopsis thaliana (L.) Heynh under elevated CO2. Plant Cell Environ 34:1304–1317CrossRefGoogle Scholar
  46. 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 53:103–110CrossRefGoogle Scholar
  47. Rosales EP, Iannone MF, Groppa MD, Benavides MP (2011) Nitric oxide inhibits nitrate reductase activity in wheat leaves. Plant Physiol Biochem 49:124–130CrossRefGoogle Scholar
  48. Rusterucci C, Espunya MC, Diaz M, Chabannes M, Martinez MC (2007) S-nitrosoglutathione reductase affords protection against pathogens in Arabidopsis, both locally and systemically. Plant Physiol 143:1282–1292CrossRefGoogle Scholar
  49. Sakamoto A, Ueda M, Morikawa H (2002) Arabidopsis glutathione- dependent formaldehyde dehydrogenase is an S-nitrosoglutathione reductase. FEBS Lett 515:20–24CrossRefGoogle Scholar
  50. Scheible WR, Morcuende R, Czechowski T, Fritz C, Osuna D, Palacios-Rojas N, Schindelasch D, Thimm O, Udvardi MK, Stitt M (2004) Genome-wide reprogramming of primary and secondary metabolism, protein synthesis, cellular growth processes, and the regulatory infrastructure of Arabidopsis in response to nitrogen. Plant Physiol 136:2483–2499CrossRefGoogle Scholar
  51. Shapiro AD (2005) Nitric oxide signalling in plants. Vitam Horm 72:339–398CrossRefGoogle Scholar
  52. Shimono H, Bunce JA (2008) Acclimation of nitrogen uptake capacity of rice to elevated atmospheric CO2 concentration. Ann Bot 103:87–94CrossRefGoogle Scholar
  53. Simontacchi M, Galatro A, Ramos-Artuso F, Santa-Maria GE (2015) Plant survival in a changing environment: the role of nitric oxide in plant responses to abiotic stress. Front Plant Sci 6:977CrossRefGoogle Scholar
  54. Singh HP, Bati DR, Kaur G, Arora K, Kohli RK (2008) Nitric oxide (as sodium nitroprusside) supplementation ameliorates Cd toxicity in hydroponically grown wheat roots. Environ Exp Bot 63:158–167CrossRefGoogle Scholar
  55. Stitt M, Muller C, Matt P, Gibon Y, Carillo P, Morcuende R, Scheible WR, Krapp A (2002) Steps towards an integrated view of nitrogen metabolism. J Exp Bot 53:959–970CrossRefGoogle Scholar
  56. Sun H, Li J, Song W, Tao J, Huang S, Chen S, Hou M, Xu G, Zhang Y (2015) Nitric oxide generated by nitrate reductase increases nitrogen uptake capacity by inducing lateral root formation and inorganic nitrogen uptake under partial nitrate nutrition in rice. J Exp Bot 66:2449–2459CrossRefGoogle Scholar
  57. Teng N, Wang J, Chen T, Wu X, Wang Y, Lin J (2006) Elevated CO2 induces physiological, biochemical and structural changes in leaves of Arabidopsis thaliana. New Phytol 172:92–103CrossRefGoogle Scholar
  58. Undurraga SF, Ibarra-Henriquez C, Fredes I, Alvarez JM, Gutierrez RA (2017) Nitrate signaling and early responses in Arabidopsis roots. J Exp Bot 68:2541–2551CrossRefGoogle Scholar
  59. Wang X, Hargrove MS (2013) Nitric oxide in plants: the roles of ascorbate and hemoglobin. PLoS One 8:2611CrossRefGoogle Scholar
  60. Wang YS, Yang ZM (2005) Nitric oxide reduces aluminum toxicity by preventing oxidative stress in the roots of Cassia tora L. Plant Cell Physiol 46:1915–1923Google Scholar
  61. Wang Y, Li K, Li X (2009) Auxin redistribution modulates plastic development of root system architecture under salt stress in Arabidopsis thaliana. J Plant Physiol 166:1637–1645CrossRefGoogle Scholar
  62. Wang H, Xiao W, Niu Y, Jin C, Chai R, Tang C, Zhang Y (2013) Nitric oxide enhances development of lateral roots in tomato (Solanum lycopersicum L.) under elevated carbon dioxide. Planta 237:137–144CrossRefGoogle Scholar
  63. Xiong J, Lu H, Lu K, Duan Y, An L, Zhu C (2009) Cadmium decreases crown root number by decreasing endogenous nitric oxide, which is indispensable for crown root primordia initiation in rice seedlings. Planta 230:599–610CrossRefGoogle Scholar
  64. Yaacov YL, Wills RB, Ku VVV (1998) Evidence for the function of the free radical gas nitric oxide (NO•) as an endogenous maturation and senescence regulating factor in higher plants. Plant Physiol Biochem 36:825–833CrossRefGoogle Scholar
  65. Yamasaki H, Sakihama Y, Takahashi S (2000) An alternative pathway for nitric oxide production in plants: new features of an old enzyme. Trends Plant Sci 4:128–129CrossRefGoogle Scholar
  66. Ye YQ, Jin CW, Fan SK, Mao QQ, Sun CL, Yu Y, Lin XY (2015) Elevation of NO production increases Fe immobilization in the Fe-deficiency roots apoplast by decreasing pectin methylation of cell wall. Sci Rep 5:10746CrossRefGoogle Scholar
  67. Zhang H, Shen WB, Xu LL (2003) Effects of nitric oxide on the germination of wheat seeds and its reactive oxygen species metabolisms under osmotic stress. Acta Bot Sin 45:901–905Google Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Division of Plant PhysiologyIndian Agricultural Research InstituteNew DelhiIndia

Personalised recommendations