, Volume 237, Issue 1, pp 65–75 | Cite as

Shoot to root communication is necessary to control the expression of iron-acquisition genes in Strategy I plants

  • María J. García
  • Francisco J. RomeraEmail author
  • Minviluz G. Stacey
  • Gary Stacey
  • Eduardo Villar
  • Esteban Alcántara
  • Rafael Pérez-Vicente
Original Article


Previous research showed that auxin, ethylene, and nitric oxide (NO) can activate the expression of iron (Fe)-acquisition genes in the roots of Strategy I plants grown with low levels of Fe, but not in plants grown with high levels of Fe. However, it is still an open question as to how Fe acts as an inhibitor and which pool of Fe (e.g., root, phloem, etc.) in the plant acts as the key regulator for gene expression control. To further clarify this, we studied the effect of the foliar application of Fe on the expression of Fe-acquisition genes in several Strategy I plants, including wild-type cultivars of Arabidopsis [Arabidopsis thaliana (L.) Heynh], pea [Pisum sativum L.], tomato [Solanum lycopersicon Mill.], and cucumber [Cucumis sativus L.], as well as mutants showing constitutive expression of Fe-acquisition genes when grown under Fe-sufficient conditions [Arabidopsis opt3-2 and frd3-3, pea dgl and brz, and tomato chln (chloronerva)]. The results showed that the foliar application of Fe blocked the expression of Fe-acquisition genes in the wild-type cultivars and in the frd3-3, brz, and chln mutants, but not in the opt3-2 and dgl mutants, probably affected in the transport of a Fe-related repressive signal in the phloem. Moreover, the addition of either ACC (ethylene precursor) or GSNO (NO donor) to Fe-deficient plants up-regulated the expression of Fe-acquisition genes, but this effect did not occur in Fe-deficient plants sprayed with foliar Fe, again suggesting the existence of a Fe-related repressive signal moving from leaves to roots.


dgl Ethylene Iron Nitric oxide opt3 Peptide Phloem 



1-Aminocyclopropane-1-carboxylic acid




3-(2-Pyridyl)-5,6-bis(4-phenyl-sulfonic acid)-1,2,4-triazine







We thank Dr Michael Grusak (USDA/ARS, Houston, Texas, USA), Dr Ross Welch (USDA/ARS, Ithaca, New York, USA), and Dr Petra Bauer (Saarland University, Saarbrücken, Germany) for kindly providing seeds of the Sparkle [dgl,dgl] mutant, the brz mutant, and the chloronerva mutant. We also thank the Arabidopsis Biological Resource Center (Ohio State University, USA) for providing seeds of the frd3-3 mutant. This work was supported by the European Regional Development Fund from the European Union, the “Ministerio de Educación y Ciencia” (Projects AGL2007-64372 and AGL2010-17121), and the “Junta de Andalucía” (Research Groups AGR115 and BIO159, and Project AGR-3849). Funding to MGS and GS was provided by a Grant from the USA National Science Foundation Plant Genome Program (Award #0820769).


  1. Bacaicoa E, Mora V, Zamarreño AM, Fuentes M, Casanova E, García-Mina JM (2011) Auxin: a major player in the shoot-to-root regulation of root Fe-stress physiological responses to Fe deficiency in cucumber plants. Plant Physiol Biochem 49:545–556PubMedCrossRefGoogle Scholar
  2. Bethke G, Unthan T, Uhrig JF, Pöschl Y, Gust AA, Scheel D, Lee J (2009) Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in Arabidopsis thaliana via ethylene signaling. Proc Natl Acad Sci USA 106:8067–8072PubMedCrossRefGoogle Scholar
  3. Bienfait HF, De Weger LA, Kramer D (1987) Control of development of iron-efficiency reactions in potato as a response to iron deficiency is located in the roots. Plant Physiol 83:244–247PubMedCrossRefGoogle Scholar
  4. Brumbarova T, Bauer P (2005) Iron-mediated control of the basic helix-loop-helix protein FER, a regulator of iron uptake in tomato. Plant Physiol 137:1018–1026PubMedCrossRefGoogle Scholar
  5. Buhtz A, Pieritz J, Springer F, Kehr J (2010) Phloem small RNAs, nutrient stress responses, and systemic mobility. BMC Plant Biol 10:64PubMedCrossRefGoogle Scholar
  6. Chen WW, Yang JL, Qin C, Jin CW, Mo JH, Ye T, Zheng SJ (2010) Nitric oxide acts downstream of auxin to trigger root ferric-chelate reductase activity in response to iron deficiency in Arabidopsis thaliana. Plant Physiol 154:810–819PubMedCrossRefGoogle Scholar
  7. Clark RJ, Tan CC, Preza GC, Nemeth E, Ganz T, Craik DJ (2011) Understanding the structure/activity relationships of the iron regulatory peptide hepcidin. Chem Biol 18:336–343PubMedCrossRefGoogle Scholar
  8. Durrett TP, Gassmann W, Rogers EE (2007) The FRD3-mediated efflux of citrate into the root vasculature is necessary for efficient iron translocation. Plant Physiol 144:197–205PubMedCrossRefGoogle Scholar
  9. Enomoto Y, Hodoshima H, Shimada H, Shoji K, Yoshihara T, Goto F (2007) Long-distance signals positively regulate the expression of iron uptake genes in tobacco roots. Planta 227:81–89PubMedCrossRefGoogle Scholar
  10. García MJ, Lucena C, Romera FJ, Alcántara E, Pérez-Vicente R (2010) Ethylene and nitric oxide involvement in the up-regulation of key genes related to iron acquisition and homeostasis in Arabidopsis. J Exp Bot 61:3885–3899PubMedCrossRefGoogle Scholar
  11. García MJ, Suárez V, Romera FJ, Alcántara E, Pérez-Vicente R (2011) A new model involving ethylene, nitric oxide and Fe to explain the regulation of Fe-acquisition genes in Strategy I plants. Plant Physiol Biochem 49:537–544PubMedCrossRefGoogle Scholar
  12. Germain H, Chevalier E, Matton DP (2006) Plant bioactive peptides: an expanding class of signaling molecules. Can J Bot 84:1–19CrossRefGoogle Scholar
  13. Graziano M, Lamattina L (2007) Nitric oxide accumulation is required for molecular and physiological responses to iron deficiency in tomato roots. Plant J 52:949–960PubMedCrossRefGoogle Scholar
  14. Grusak MA, Pezeshgi S (1996) Shoot-to-root signal transmission regulates root Fe(II) reductase activity in the dgl mutant of pea. Plant Physiol 110:329–334PubMedGoogle Scholar
  15. Grusak MA, Welch RM, Kochian LV (1990) Physiological characterization of a single-gene mutant of Pisum sativum exhibiting excess iron accumulation. I. Root iron reduction and iron uptake. Plant Physiol 93:976–981PubMedCrossRefGoogle Scholar
  16. Han ZH, Han CQ, Xu XF, Wang Q (2005) Relationship between iron deficiency stress and endogenous hormones in iron-efficient versus inefficient apple genotypes. J Plant Nutr 28:1887–1895CrossRefGoogle Scholar
  17. Ivanov R, Brumbarova T, Bauer P (2011) Fitting into the harsh reality: regulation of iron-deficiency responses in dicotyledonous plants. Mol Plant 5:27–42PubMedCrossRefGoogle Scholar
  18. Jung JY, Shin R, Schachtman DP (2009) Ethylene mediates response and tolerance to potassium deprivation in Arabidopsis. Plant Cell 21:607–621PubMedCrossRefGoogle Scholar
  19. Klatte M, Schuler M, Wirtz M, Fink-Straube C, Hell R, Bauer P (2009) The analysis of Arabidopsis nicotianamine synthase mutants reveals functions for nicotianamine in seed iron loading and iron deficiency responses. Plant Physiol 150:257–271PubMedCrossRefGoogle Scholar
  20. Kneen BE, LaRue T, Welch RM, Weeden NF (1990) Pleiotropic effects of brz. A mutation in Pisum sativum (L.) cv ‘Sparkle’ conditioning decreased nodulation and increased iron uptake and leaf necrosis. Plant Physiol 93:717–722PubMedCrossRefGoogle Scholar
  21. Landsberg EC (1984) Regulation of iron-stress-response by whole plant activity. J Plant Nutr 7:609–621CrossRefGoogle Scholar
  22. Lei M, Zhu C, Liu Y, Karthikeyan AS, Bressan RA, Raghothama KG, Liu D (2011) Ethylene signalling is involved in regulation of phosphate starvation-induced gene expression and production of acid phosphatases and anthocyanin in Arabidopsis. New Phytol 189:1084–1095PubMedCrossRefGoogle Scholar
  23. Li C, Zhu X, Zhang F (2000) Role of shoot in regulation of iron deficiency responses in cucumber and bean plants. J Plant Nutr 23:1809–1818CrossRefGoogle Scholar
  24. Ling HQ, Koch G, Baumlein H, Ganal MW (1999) Map-based cloning of chloronerva, a gene involved in iron uptake of higher-plants encoding nicotianamine synthase. Proc Natl Acad Sci USA 96:7098–7103PubMedCrossRefGoogle Scholar
  25. Lingam S, Mohrbacher J, Brumbarova T, Potuschak T, Fink-Straube C, Blondet E, Genschik P, Bauer P (2011) Interaction between the bHLH transcription factor FIT and the ETHYLENE INSENSITIVE3/ETHYLENE INSENSITIVE3-LIKE1 reveals molecular linkage between the regulation of iron acquisition and ethylene signaling in Arabidopsis. Plant Cell 23:1815–1829PubMedCrossRefGoogle Scholar
  26. Liu TY, Chang CY, Chiou TJ (2009) The long-distance signaling of mineral macronutrients. Curr Opin Plant Biol 12:312–319PubMedCrossRefGoogle Scholar
  27. Lubkowitz M (2011) The oligopeptide transporters: a small gene family with a diverse group of substrates and functions? Mol Plant 4:407–415PubMedCrossRefGoogle Scholar
  28. Lucena C, Waters BM, Romera FJ, García MJ, Morales M, Alcántara E, Pérez-Vicente R (2006) Ethylene could influence ferric reductase, iron transporter, and H+-ATPase gene expression by affecting FER (or FER-like) gene activity. J Exp Bot 57:4145–4154PubMedCrossRefGoogle Scholar
  29. Lucena C, Romera FJ, Rojas CL, García MJ, Alcántara E, Pérez-Vicente R (2007) Bicarbonate blocks the expression of several genes involved in the physiological responses to Fe deficiency of Strategy I plants. Funct Plant Biol 34:1002–1009CrossRefGoogle Scholar
  30. Maas FM, van de Wetering DAM, van Beusichem Ml, Bienfait HF (1988) Characterization of phloem iron and its possible role in the regulation of Fe-efficiency reactions. Plant Physiol 87:167–171PubMedCrossRefGoogle Scholar
  31. Marentes E, Grusak MA (1998) Mass determination of low-molecular-weight proteins in phloem sap using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. J Exp Bot 49:903–911Google Scholar
  32. Marentes E, Stephens BW, Grusak MA (1997) Characterization of a phloem mobile chelator involved in the phloem transport of iron from vegetative tissues to developing seeds of pea. Abstract 9th International Symposium on Iron Nutrition and Interactions in Plants, Stuttgart/Germany, p 74Google Scholar
  33. Meiser J, Lingam S, Bauer P (2011) Post-translational regulation of the Fe deficiency bHLH transcription factor FIT is affected by iron and nitric oxide. Plant Physiol 157:2154–2166PubMedCrossRefGoogle Scholar
  34. Pich A, Manteuffel R, Hillmer S, Scholz G, Schmidt W (2001) Fe homeostasis in plant cells: Does nicotianamine play multiple roles in the regulation of cytoplasmic Fe concentration? Planta 213:967–976PubMedCrossRefGoogle Scholar
  35. Ramírez L, Simontacchi M, Murgia I, Zabaleta E, Lamattina L (2011) Nitric oxide, nitrosyl iron complexes, ferritin and frataxin: a well equipped team to preserve plant iron homeostasis. Plant Sci 181:582–592PubMedCrossRefGoogle Scholar
  36. Rodriguez-Castrillón JA, Moldovan M, García JI, Lucena JJ, García-Tomé ML, Hernández-Apaolaza L (2008) Isotope pattern deconvolution as a tool to study iron metabolism in plants. Anal Bioanal Chem 390:579–590PubMedCrossRefGoogle Scholar
  37. Rogers EE, Guerinot ML (2002) FRD3, a member of the multidrug and toxin efflux family, controls iron deficiency responses in Arabidopsis. Plant Cell 14:1787–1799PubMedCrossRefGoogle Scholar
  38. Romera FJ, Alcántara E, De la Guardia MD (1992) Role of roots and shoots in the regulation of the Fe efficiency responses in sunflower and cucumber. Physiol Plant 85:141–146CrossRefGoogle Scholar
  39. Romera FJ, García MJ, Alcántara E, Pérez-Vicente R (2011) Latest findings about the interplay of auxin, ethylene and nitric oxide in the regulation of Fe deficiency responses by Strategy I plants. Plant Signal Behav 6:167–170PubMedCrossRefGoogle Scholar
  40. Roschzttardtz H, Séguéla-Arnaud M, Briat JF, Vert G, Curie C (2011) The FRD3 citrate effluxer promotes iron nutrition between symplastically disconnected tissues throughout Arabidopsis development. Plant Cell 23:2725–2737PubMedCrossRefGoogle Scholar
  41. Santi S, Schmidt W (2009) Dissecting iron deficiency-induced proton extrusion in Arabidopsis roots. New Phytol 183:1072–1084PubMedCrossRefGoogle Scholar
  42. Santi S, Cesco S, Varanini Z, Pinton R (2005) Two plasma membrane H+-ATPase genes are differentially expressed in iron-deficient cucumber plants. Plant Physiol Biochem 43:287–292PubMedCrossRefGoogle Scholar
  43. Schmidke I, Krüger C, Frömmichen R, Scholz G, Stephan UW (1999) Phloem loading and transport characteristics of iron in interaction with plant-endogenous ligands in castor bean seedlings. Physiol Plant 106:82–89CrossRefGoogle Scholar
  44. Scholz G, Schlesier G, Seifert K (1985) Effect of nicotianamine on iron uptake by the tomato mutant ‘chloronerva’. Physiol Plant 63:99–104CrossRefGoogle Scholar
  45. Schuler M, Rellán-Álvarez R, Fink-Straube C, Abadía J, Bauer P (2012) Nicotianamine functions in the phloem-based transport of iron to sink organs, in pollen development and pollen tube growth in Arabidopsis. Plant Cell 24:2380–2400PubMedCrossRefGoogle Scholar
  46. Stacey MG, Osawa H, Patel A, Gassmann W, Stacey G (2006) Expression analyses of Arabidopsis oligopeptide transporters during seed germination, vegetative growth and reproduction. Planta 223:291–305PubMedCrossRefGoogle Scholar
  47. Stacey MG, Patel A, McClain WE, Mathieu M, Remley M, Rogers EE, Gassmann W, Blevins DG, Stacey G (2008) The Arabidopsis AtOPT3 protein functions in metal homeostasis and movement of iron to developing seeds. Plant Physiol 146:589–601PubMedCrossRefGoogle Scholar
  48. Stephan UW, Scholz G (1993) Nicotianamine: mediator of transport of iron and heavy metals in the phloem? Physiol Plant 88:522–529CrossRefGoogle Scholar
  49. Venkatraju K, Marschner H (1981) Inhibition of iron-stress reactions in sunflower by bicarbonate. Z Pflanzenernähr Bodenkd 144:339–355CrossRefGoogle Scholar
  50. Walker EL, Connolly EL (2008) Time to pump iron: iron-deficiency-signaling mechanisms of higher plants. Curr Opin Plant Biol 11:530–535PubMedCrossRefGoogle Scholar
  51. Waters BM, Lucena C, Romera FJ, Jester GG, Wynn AN, Rojas CL, Alcántara E, Pérez-Vicente R (2007) Ethylene involvement in the regulation of the H+-ATPase CsHA1 gene and of the new isolated ferric reductase CsFRO1 and iron transporter CsIRT1 genes in cucumber plants. Plant Physiol Biochem 45:293–301PubMedCrossRefGoogle Scholar
  52. Wu T, Zhang HT, Wang Y, Jia WS, Xu XF, Zhang XZ, Han ZH (2012) Induction of root Fe(III) reductase activity and proton extrusion by iron deficiency is mediated by auxin-based systemic signalling in Malus xiaojinensis. J Exp Bot 63:859–870PubMedCrossRefGoogle Scholar
  53. Yuan YX, Wu HL, Wang N, Li J, Zhao WN, Du J, Wang DW, Ling HQ (2008) FIT interacts with AtbHLH038 and AtbHLH039 in regulating iron uptake gene expression for iron homeostasis in Arabidopsis. Cell Res 18:385–397PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • María J. García
    • 3
  • Francisco J. Romera
    • 1
    Email author
  • Minviluz G. Stacey
    • 2
  • Gary Stacey
    • 2
  • Eduardo Villar
    • 1
  • Esteban Alcántara
    • 1
  • Rafael Pérez-Vicente
    • 3
  1. 1.Department of Agronomy, Edificio Celestino Mutis (C-4), Campus de Excelencia Internacional Agroalimentario de Rabanales (ceiA3)University of CórdobaCórdobaSpain
  2. 2.Divisions of Plant Science and BiochemistryUniversity of MissouriColumbiaUSA
  3. 3.Department of Botany, Ecology and Plant Physiology, Edificio Celestino Mutis (C-4), Campus de RabanalesUniversity of CórdobaCórdobaSpain

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