, Volume 236, Issue 3, pp 887–900 | Cite as

AtGSNOR1 function is required for multiple developmental programs in Arabidopsis

  • Eunjung Kwon
  • Angela Feechan
  • Byung-Wook Yun
  • Byung-Ho Hwang
  • Jacqueline A. Pallas
  • Jeong-Gu Kang
  • Gary J. LoakeEmail author
Original Article


Nitric oxide (NO) has been proposed to regulate a diverse array of activities during plant growth, development and immune function. S-nitrosylation, the addition of an NO moiety to a reactive cysteine thiol, to form an S-nitrosothiol (SNO), is emerging as a prototypic redox-based post-translational modification. An ARABIDOPSIS THALIANA S-NITROSOGLUTATHIONE (GSNO) REDUCTASE (AtGSNOR1) is thought to be the major regulator of total cellular SNO levels in this plant species. Here, we report on the impact of loss- and gain-of-function mutations in AtGSNOR1 upon plant growth and development. Loss of AtGSNOR1 function in atgsnor1-3 plants increased the number of initiated higher order axillary shoots that remain active, resulting in a loss of apical dominance relative to wild type. In addition atgsnor1-3 affected leaf shape, germination, 2,4-D sensitivity and reduced hypocotyl elongation in both light and dark grown seedlings. Silique size and seed production were also decreased in atgsnor1-3 plants and the latter was reduced in atgsnor1-1 plants, which overexpress AtGSNOR1. Overexpression of AtGSNOR1 slightly delayed flowering time in both long and short days, whereas atgsnor1-3 showed early flowering compared to wild type. In the atgsnor1-3 line, FLOWERING LOCUS C (FLC) expression was reduced, whereas transcription of CONSTANS (CO) was enhanced. Therefore, AtGSNOR1 may negatively regulate the autonomous and photoperiod flowering time pathways. Both overexpression and loss of AtGSNOR1 function also reduced primary root growth, while root hair development was increased in atgsnor1-1 and reduced in atgsnor1-3 plants. Collectively, our findings imply that AtGSNOR1 controls multiple genetic networks integral to plant growth and development.


Nitric oxide S-nitrosylation AtGSNOR1 S-nitrosothiols Plant development 



Nitric oxide




S-nitrosoglutathione reductase


Flowering locus C






Gibberellic acid


Salicylic acid


2,4-Dichlorophenoxyacetic acid



AF was the recipient of a BBSRC CASE studentship. EK and BW were funded by BBSRC grant BB/D0118091/1 to the Loake lab. BW was supported by a grant from the Next-Generation BioGreen 21 Program (SSAC, grant# : PJ009011), Rural Development Administration, Republic of Korea. JK was the recipient of a Staff Scholarship from the University of Edinburgh.


  1. Belenghi B, Romero-Puertas MC, Vercammen D, Brackenier A, Inze D, Delledonne M, Van Breusegem F (2007) Metacaspase activity of Arabidopsis thaliana is regulated by S-nitrosylation of a critical cysteine residue. J Biol Chem 282:1352–1358PubMedCrossRefGoogle Scholar
  2. Beligni MV, Lamattina L (2000) Nitric oxide stimulates seed germination and de-etiolation, and inhibits hypocotyl elongation, three light-inducible responses in plants. Planta 210:215–221PubMedCrossRefGoogle Scholar
  3. Beligni MV, Lamattina L (2001) Nitric oxide: a non-traditional regulator of plant growth. Trends Plant Sci 6:508–509PubMedCrossRefGoogle Scholar
  4. Beveridge CA, Weller JL, Singer SR, Hofer JM (2003) Axillary meristem development. Budding relationships between networks controlling flowering, branching, and photoperiod responsiveness. Plant Physiol 131:927–934PubMedCrossRefGoogle Scholar
  5. Blazquez MA, Weigel D (2000) Integration of floral inductive signals in Arabidopsis. Nature 404:889–892PubMedCrossRefGoogle Scholar
  6. Blazquez MA, Soowal LN, Lee I, Weigel D (1997) LEAFY expression and flower initiation in Arabidopsis. Development 124:3835–3844PubMedGoogle Scholar
  7. Blazquez MA, Green R, Nilsson O, Sussman MR, Weigel D (1998) Gibberellins promote flowering of Arabidopsis by activating the LEAFY promoter. Plant Cell 10:791–800PubMedGoogle Scholar
  8. Chen R, Sun S, Wang C, Li Y, Liang Y, An F, Li C, Dong H, Yang X, Zhang J, Zuo J (2009) The Arabidopsis PARAQUAT RESISTANT2 gene encodes an S-nitrosoglutathione reductase that is a key regulator of cell death. Cell Res 19:1377–1387PubMedCrossRefGoogle Scholar
  9. Chou ML, Haung MD, Yang CH (2001) EMF genes interact with late-flowering genes in regulating floral initiation genes during shoot development in Arabidopsis thaliana. Plant Cell Physiol 42:499–507PubMedCrossRefGoogle Scholar
  10. Cline MG (1994) The role of hormones in apical dominance. New approaches to an old problem in plant development. Physiol Plant 90:230–237CrossRefGoogle Scholar
  11. Corpas FJ, Barroso JB, Carreras A, Valderrama R, Palma JM, Leon AM, Sandalio LM, del Rio LA (2006) Constitutive arginine-dependent nitric oxide synthase activity in different organs of pea seedlings during plant development. Planta 224:246–254PubMedCrossRefGoogle Scholar
  12. Correa-Aragunde N, Graziano M, Lamattina L (2004) Nitric oxide plays a central role in determining lateral root development in tomato. Planta 218:900–905PubMedCrossRefGoogle Scholar
  13. Delledonne M, Xia Y, Dixon RA, Lamb C (1998) Nitric oxide functions as a signal in plant disease resistance. Nature 394:585–588PubMedCrossRefGoogle Scholar
  14. 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 USA 98:13454–13459PubMedCrossRefGoogle Scholar
  15. Durner J, Wendehenne D, Klessig DF (1998) Defense gene induction in tobacco by nitric oxide, cyclic GMP, and cyclic ADP-ribose. Proc Natl Acad Sci USA 95:10328–10333PubMedCrossRefGoogle Scholar
  16. Eriksson S, Bohlenius H, Moritz T, Nilsson O (2006) Gas is the active gibberellin in the regulation of LEAFY transcription and Arabidopsis floral initiation. Plant Cell 18:2172–2181PubMedCrossRefGoogle Scholar
  17. Feechan A, Kwon E, Yun BW, Wang Y, Pallas JA, Loake GJ (2005) A central role for S-nitrosothiols in plant disease resistance. Proc Natl Acad Sci USA 102:8054–8059PubMedCrossRefGoogle Scholar
  18. Fernàndez-Marcos M, Sanz L, Lorenzo O (2012) Nitric oxide: an emerging regulator of cell elongation during primary root growth. Plant Signal Behav 2:1–7Google Scholar
  19. Foreman J, Demidchik V, Bothwell JH, Mylona P, Miedema H, Torres MA, Linstead P, Costa S, Brownlee C, Jones JD, Davies JM, Dolan L (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422:442–446PubMedCrossRefGoogle Scholar
  20. 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 22:3816–3830PubMedCrossRefGoogle Scholar
  21. Furchgott RF (1995) Special topics: nitric oxide. Annu Rev Physiol 57:659–682CrossRefGoogle Scholar
  22. Gendall AR, Levy YY, Wilson A, Dean C (2001) The VERNALIZATION 2 gene mediates the epigenetic regulation of vernalization in Arabidopsis. Cell 107:525–535PubMedCrossRefGoogle Scholar
  23. Goretski J, Hollocher TC (1988) Trapping of nitric oxide produced during denitrification by extracellular hemoglobin. J Biol Chem 263:2316–2323PubMedGoogle Scholar
  24. Grant JJ, Loake GJ (2000) Role of reactive oxygen intermediates and cognate redox signaling in disease resistance. Plant Physiol 124:21–29PubMedCrossRefGoogle Scholar
  25. Grant JJ, Chini A, Basu D, Loake GJ (2003) Targeted activation tagging of the Arabidopsis NBS-LRR gene, ADR1, conveys resistance to virulent pathogens. Mol Plant Microbe Interact 16:669–680PubMedCrossRefGoogle Scholar
  26. Guo FQ, Crawford NM (2005) Arabidopsis nitric oxide synthase1 is targeted to mitochondria and protects against oxidative damage and dark-induced senescence. Plant Cell 17:3436–3450PubMedCrossRefGoogle Scholar
  27. Guo FQ, Okamoto M, Crawford NM (2003) Identification of a plant nitric oxide synthase gene involved in hormonal signaling. Science 302:100–103PubMedCrossRefGoogle Scholar
  28. Gupta KJ, lgamberdiev AU, Manjunatha G, Segu S, Moran JF, Neelawarne B, Bauwe H, Kaiser WM (2011) The emerging roles of nitric oxide (NO) in plant mitochondria. Plant Sci 181:520–526Google Scholar
  29. Hay A, Kaur H, Phillips A, Hedden P, Hake S, Tsiantis M (2002) The gibberellin pathway mediates KNOTTED1-type homeobox function in plants with different body plans. Curr Biol 12:1557–1565PubMedCrossRefGoogle Scholar
  30. He Y, Tang RH, Hao Y, Stevens RD, Cook CW, Ahn SM, Jing L, Yang Z, Chen L, Guo F, Fiorani F, Jackson RB, Crawford NM, Pei ZM (2004) Nitric oxide represses the Arabidopsis floral transition. Science 305:1968–1971PubMedCrossRefGoogle Scholar
  31. Hempel FD, Feldman LJ (1994) Bi-directional inflorescence development in Arabidopsis thaliana: acropetal initiation of flowers and basipetal initiation of paraclades. Planta 192:276–286CrossRefGoogle Scholar
  32. Jacobsen SE, Olszewski NE (1993) Mutations at the SPINDLY locus of Arabidopsis alter gibberellin signal transduction. Plant Cell 5:887–896PubMedGoogle Scholar
  33. Klepper L (1979) Nitric-oxide (NO) and nitrogen-dioxide (NO2) emissions from herbicide-treated soybean plants. Atmos Environ 13:537–542CrossRefGoogle Scholar
  34. Kolbert Z, Ortega L, Erdei L (2010) Involvement of nitrate reductase (NR) in osmotic stress-induced NO generation of Arabidopsis thaliana. J Plant Physiol 167:77–80PubMedCrossRefGoogle Scholar
  35. Lamattina L, Garcia-Mata C, Graziano M, Pagnussat G (2003) Nitric oxide: the versatility of an extensive signal molecule. Annu Rev Plant Biol 54:109–136PubMedCrossRefGoogle Scholar
  36. Lee U, Wie C, Fernandez BO, Feelisch M, Vierling E (2008) Modulation of nitrosative stress by S-nitrosoglutathione reductase is critical for thermotolerance and plant growth in Arabidopsis. Plant Cell 20:786–802PubMedCrossRefGoogle Scholar
  37. Lincoln C, Britton JH, Estelle M (1990) Growth and development of the axr1 mutants of Arabidopsis. Plant Cell 2:1071–1080PubMedGoogle Scholar
  38. Lindermayr C, Saalbach G, Durner J (2005) Proteomic identification of S-nitrosylated proteins in Arabidopsis. Plant Physiol 137:921–930PubMedCrossRefGoogle Scholar
  39. Lindermayr C, Saalbach G, Bahnweg G, Durner J (2006) Differential inhibition of Arabidopsis methionine adenosyltransferases by protein S-nitrosylation. J Biol Chem 281:4285–4291PubMedCrossRefGoogle Scholar
  40. 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 410:490–494PubMedCrossRefGoogle Scholar
  41. Liu L, Yan Y, Zeng M, Zhang J, Hanes MA, Ahearn G, McMahon TJ, Dickfeld T, Marshall HE, Que LG, Stamler JS (2004) Essential roles of S-nitrosothiols in vascular homeostasis and endotoxic shock. Cell 116:617–628PubMedCrossRefGoogle Scholar
  42. Lombardo MC, Graziano M, Polacco JC, Lamattina L (2006) Nitric oxide function as a positive regulator of root hair development. Plant Signal Behav 1:28–33PubMedCrossRefGoogle Scholar
  43. Malik SI, Hussain A, Yun B-W, Spoel SH, Loake GJ (2011) GSNOR-mediated de-nitrosylation in the plant defence response. Plant Sci 181:540–544PubMedCrossRefGoogle Scholar
  44. Mannick JB, Hausladen A, Liu L, Hess DT, Zeng M, Miao QX, Kane LS, Gow AJ, Stamler JS (1999) Fas-induced caspase denitrosylation. Science 284:651–654PubMedCrossRefGoogle Scholar
  45. Melino G, Bernassola F, Knight RA, Corasaniti MT, Nistico G, Finazzi-Agro A (1997) S-nitrosylation regulates apoptosis. Nature 388:432–433PubMedCrossRefGoogle Scholar
  46. Michaels SD, Amasino RM (1999) FLOWERING LOCUS C encodes a novel MADS domain protein that acts as a repressor of flowering. Plant Cell 11:949–956PubMedGoogle Scholar
  47. Michaels JE, Shiba K, Miller WT (1999) Autonomous folding of a C-terminal inhibitory fragment of Escherichia coli isoleucine-tRNA synthetase. Biochim Biophys Acta 1433:103–109PubMedCrossRefGoogle Scholar
  48. Morris DA (1977) Transport of exogenous auxin in two-branched dwarf pea seedlings (Pisum sativum L.). Planta 136:91–96CrossRefGoogle Scholar
  49. Mutasa-Göttgens E, Hedden P (2009) Gibberellin as a factor in floral regulatory networks. J Exp Bot 60:1979–1989PubMedCrossRefGoogle Scholar
  50. Onouchi H, Igeno MI, Perilleux C, Graves K, Coupland G (2000) Mutagenesis of plants overexpressing CONSTANS demonstrates novel interactions among Arabidopsis flowering-time genes. Plant Cell 12:885–900PubMedGoogle Scholar
  51. Ötvös K, Pasternak TP, Miskolczi P, Domoki M, Dorjgotov D, Szucs A, Bottka S, Dudits D, Feher A (2005) Nitric oxide is required for, and promotes auxin-mediated activation of, cell division and embryogenic cell formation but does not influence cell cycle progression in alfalfa cell cultures. Plant J 43:849–860PubMedCrossRefGoogle Scholar
  52. Pagnussat GC, Simontacchi M, Puntarulo S, Lamattina L (2002) Nitric oxide is required for root organogenesis. Plant Physiol 129:954–956PubMedCrossRefGoogle Scholar
  53. Pagnussat GC, Lanteri ML, Lamattina L (2003) Nitric oxide and cyclic GMP are messengers in the indole acetic acid-induced adventitious rooting process. Plant Physiol 132:1241–1248PubMedCrossRefGoogle Scholar
  54. Palmer RM, Hickery MS, Charles IG, Moncada S, Bayliss MT (1993) Induction of nitric oxide synthase in human chondrocytes. Biochem Biophys Res Commun 193:398–405PubMedCrossRefGoogle Scholar
  55. Piñeiro M, Coupland G (1998) The control of flowering time and floral identity in Arabidopsis. Plant Physiol 117:1–8PubMedCrossRefGoogle Scholar
  56. Putterill J, Robson F, Lee K, Simon R, Coupland G (1995) The CONSTANS gene of Arabidopsis promotes flowering and encodes a protein showing similarities to zinc finger transcription factors. Cell 80:847–857PubMedCrossRefGoogle Scholar
  57. Reeves PH, Coupland G (2001) Analysis of flowering time control in Arabidopsis by comparison of double and triple mutants. Plant Physiol 126:1085–1091PubMedCrossRefGoogle Scholar
  58. Reintanz B, Lehnen M, Reichelt M, Gershenzon J, Kowalczyk M, Sandberg G, Godde M, Uhl R, Palme K (2001) Bus, a bushy Arabidopsis CYP79F1 knockout mutant with abolished synthesis of short-chain aliphatic glucosinolates. Plant Cell 13:351–367PubMedGoogle Scholar
  59. 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 53:103–110PubMedCrossRefGoogle Scholar
  60. Sheldon CC, Burn JE, Perez PP, Metzger J, Edwards JA, Peacock WJ, Dennis ES (1999) The FLF MADS box gene: a repressor of flowering in Arabidopsis regulated by vernalization and methylation. Plant Cell 11:445–458PubMedGoogle Scholar
  61. Sheldon CC, Rouse DT, Finnegan EJ, Peacock WJ, Dennis ES (2000) The molecular basis of vernalization: the central role of FLOWERING LOCUS (FLC). Proc Natl Acad Sci USA 97:3753–3758PubMedCrossRefGoogle Scholar
  62. Simpson GG, Dean C (2002) Arabidopsis, the Rosetta stone of flowering time? Science 296:285–289PubMedCrossRefGoogle Scholar
  63. Singh DP, Jermakow AM, Swain SM (2002) Gibberellins are required for seed development and pollen tube growth in Arabidopsis. Plant Cell 14:3133–3147PubMedCrossRefGoogle Scholar
  64. Sorefan K, Booker J, Haurogne K, Goussot M, Bainbridge K, Foo E, Chatfield S, Ward S, Beveridge C, Rameau C, Leyser O (2003) MAX4 and RMS1 are orthologous dioxygenase-like genes that regulate shoot branching in Arabidopsis and pea. Genes Dev 17:1469–1474PubMedCrossRefGoogle Scholar
  65. Stamler JS, Toone EJ, Lipton SA, Sucher NJ (1997) (S)NO signals: translocation, regulation, and consensus motif. Neuron 18:691–696PubMedCrossRefGoogle Scholar
  66. Stirnberg P, Chatfield SP, Leyser HM (1999) AXR1 acts after lateral bud formation to inhibit lateral bud growth in Arabidopsis. Plant Physiol 121:839–847PubMedCrossRefGoogle Scholar
  67. Stirnberg P, Van De Sande K, Leyser HM (2002) MAX1 and MAX2 control shoot lateral branching in Arabidopsis. Development 129:1131–1141PubMedGoogle Scholar
  68. Suarez-Lopez P, Wheatley K, Robson F, Onouchi H, Valverde F, Coupland G (2001) CONSTANS mediates between the circadian clock and the control of flowering in Arabidopsis. Nature 410:1116–1120PubMedCrossRefGoogle Scholar
  69. Sussex IM, Kerk NM (2001) The evolution of plant architecture. Curr Opin Plant Biol 4:33–37PubMedCrossRefGoogle Scholar
  70. Terrile MC, Parı′s R, Caldero′ n-Villalobos LA, Lglesias MJ, Lamattina L, Estelle M, Casalongue CA (2012) Nitric oxide influences auxin signaling through S-nitrosylation of the Arabidopsis TRANSPORT INHIBITOR RESPONSE 1 auxin receptor. Plant J 70:492–500Google Scholar
  71. Thimann KV, Skoog F (1933) Studies on the growth hormone of plants: III. The inhibiting action of the growth substance on bud development. Proc Natl Acad Sci USA 19:714–716PubMedCrossRefGoogle Scholar
  72. Vanacker H, Lu H, Rate DN, Greenberg JT (2001) A role for salicylic acid and NPR1 in regulating cell growth in Arabidopsis. Plant J 28:209–216PubMedCrossRefGoogle Scholar
  73. Wang ZY, Tobin EM (1998) Constitutive expression of the CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) gene disrupts circadian rhythms and suppresses its own expression. Cell 93:1207–1217PubMedCrossRefGoogle Scholar
  74. Wang Y, Yun BW, Kwon E, Hong JK, Yoon J, Loake GJ (2006) S-nitrosylation: an emerging redox-based post-translational modification in plants. J Exp Bot 57:1777–1784PubMedCrossRefGoogle Scholar
  75. Wilcox D, Dove B, McDavid D, Greer D (1995) UTHSCSA image tool for windows. The University of Texas Health Science Center, San AntonioGoogle Scholar
  76. Wilson RN, Heckma JW, Somerville CR (1992) Gibberellin is required for flowering in Arabidopsis thaliana under short days. Plant Physiol 100:403–408PubMedCrossRefGoogle Scholar
  77. Wilson ID, Neill SJ, Hancock JT (2008) Nitric oxide synthesis and signaling in plants. Plant Cell Environ 31:622–631PubMedCrossRefGoogle Scholar
  78. Yamasaki H (2000) Nitrite-dependent nitric oxide production pathway: implications for involvement of active nitrogen species in photoinhibition in vivo. Phil Trans R Soc Lond B 355:1477–1488CrossRefGoogle Scholar
  79. Yanovsky MJ, Kay SA (2002) Molecular basis of seasonal time measurement in Arabidopsis. Nature 419:308–312PubMedCrossRefGoogle Scholar
  80. Yun B-W, Feechan A, Yin M, Saidi NBB, Bihan TL, Yu M, Moore JW, Kang J-G, Kwon E, Spoel SH, Pallas JA, Loake GJ (2011) S-nitrosylation of NADPH oxidase regulates cell death in plant immunity. Nature 478:264–268PubMedCrossRefGoogle Scholar
  81. Zago E, Morsa S, Dat JF, Alard P, Ferrarini A, Inzé D, Delledonne M, Van Breusegem F (2006) Nitric oxide- and hydrogen peroxide-responsive gene regulation during cell death induction in tobacco. Plant Physiol 141:404–411PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Eunjung Kwon
    • 1
  • Angela Feechan
    • 1
    • 2
  • Byung-Wook Yun
    • 1
    • 3
  • Byung-Ho Hwang
    • 1
  • Jacqueline A. Pallas
    • 4
    • 5
  • Jeong-Gu Kang
    • 1
  • Gary J. Loake
    • 1
    Email author
  1. 1.Institute of Molecular Plant Sciences, School of Biological SciencesUniversity of EdinburghEdinburghUK
  2. 2.CSIRO Plant IndustryGlen OsmondAustralia
  3. 3.School of Applied Biosciences, College of Agriculture and Life SciencesKyungpook National UniversityDaeguSouth Korea
  4. 4.Trait Research, SyngentaBerkshireUK
  5. 5.Bloomsbury Centre for BioinformaticsUniversity College LondonLondonUK

Personalised recommendations