Plant Molecular Biology

, Volume 52, Issue 5, pp 1025–1036 | Cite as

Molecular characterization and spatial expression of the sunflower ABP1 gene

  • Clément Thomas
  • Denise Meyer
  • Michel Wolff
  • Christophe Himber
  • Malek Alioua
  • André Steinmetz


We have used RT-PCR and low-stringency cDNA library screening to isolate the coding sequence of the sunflower auxin-binding protein (ABP1). All the clones analysed contained the same nucleotide sequence, suggesting that ABP1 is encoded by a single-copy gene in sunflower. The deduced amino acid sequence shows a high degree of similarity with ABP1 proteins from other plant species. Most remarkably, the sunflower protein lacks two cysteine residues present in all other plant ABPs known to date and shown to be involved in a disulfide bridge in the maize protein. Genomic Southern hybridization data support the existence of a single copy of the ABP1 gene in the sunflower genome. Northern hybridization corroborated earlier observations indicating that the steady-state level of ABP1 transcript is higher in actively dividing and growing organs than in the rest of the plant: it is more abundant in the shoot apex, floral buds and immature embryos than in mature leaves, stem, roots and ray flowers. To characterize the tissular ABP1 transcript distribution in sunflower, various organ sections were analysed upon in situ hybridization. Localized accumulation of the ABP1 transcript suggests that its spatial expression is highly regulated at the tissue level. In addition, the transcript preferentially accumulates in tissues having a high rate of cellular division, such as shoot and root apical meristems, leaf primordia and pro-vascular tissues. The ABP1 expression pattern was also studied at a temporal scale during lateral root formation. Real time PCR showed an elevation of the steady state level of the ABP1 transcript in root axes after 36 h of seed germination. In situ hybridization revealed that this global increase is the result of local accumulation of the ABP1 transcript in lateral root primordia, which are known to develop under auxin action. The possibility that a high ABP1 expression level correlates with a high cellular sensitivity to auxin is discussed.

Abbreviations: ABP1, auxin-binding protein 1; cDNA, complementary DNA; cds, coding DNA sequence

auxin auxin-binding protein in situ hybridization lateral roots real-time PCR sunflower 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anai, T., Miyata, M., Kosemura, S., Yamamura, S., Tsuge, T., Matsui, M., Uchida, H. and Hasegawa, K. 1997. Comparison of abp1 primary sequences from monocotyledonous and dicotyledonous species. J. Plant Physiol. 151: 446–449.Google Scholar
  2. Bauly, J.M., Sealy, I.M., Macdonald, H., Brearley, J., Dröge, S., Hillmer, S., Robinson, D.G., Venis, M.A., Blatt, M.R., Lazarus, C.M. and Napier, R.M. 2000. Overexpression of auxin-binding protein enhances the sensitivity of guard cells to auxin. Plant Physiol. 124: 1229–1238.Google Scholar
  3. Bhalerao, R.P., Eklöf, J., Ljung, K., Marchant, A., Bennett, M. and Sandberg, G. 2002. Shoot derived auxin is essential for early lateral root emergence in Arabidopsis seedlings. Plant J. 29: 325–332.Google Scholar
  4. Blakely, L.M., Blakely, R.M., Colowit, P.M. and Elliot, D.S. 1988. Experimental studies on lateral root formation in radish seedling roots. II. Analysis of the dose-response to exogenous auxin. Plant Physiol. 87: 414–419.Google Scholar
  5. Bronner, R., Jeannin, G. and Hahne, G. 1994. Early events during organogenesis and somatic embryogenesis induced on immature zygotic embryos of sunflower (Helianthus annuus L.). Can. J. Bot. 72: 239–248.Google Scholar
  6. Bronsema, F.B.F., van Oostveen, W.J.F. and Lammeren, A.A.M. 1998. Immunocytochemical localization of auxin-binding proteins in coleoptiles and embryos of Zea mays L. Protoplasma 202: 65–75.Google Scholar
  7. Celenza, J.L., Grisafi, P.L. and Fink, G.R. 1995. A pathway for lateral root formation in Arabidopsis thaliana. Genes Dev. 9: 2131–2142.Google Scholar
  8. Chen, J.G., Ullah, H., Young, J.C., Sussmann, M.R. and Jones, A.M. 2001. ABP1 is required for organized cell elongation and division in Arabidopsis embryogenesis. Genes Dev. 15: 902–911.Google Scholar
  9. Cox, K.H., DeLeon, D.V., Angerer, L.M. and Angerer, R.C. 1984. Detection of mRNAs in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev. Biol. 101: 485–503.Google Scholar
  10. David, K., Carnero-Diaz, E., Leblanc, N., Monestiez, M., Grosclaude, J. and Perrot-Rechenmann, C. 2001. Conformational dynamics underlie the activity of the auxin-binding protein, Nt-abp1. J. Biol. Chem. 276: 34517–34523.Google Scholar
  11. Davies, P.J. 1995. The plant hormones: their nature, occurence and functions. In: P.J. Davies (Ed.) Plant Hormones: Physiology, Biochemistry and Molecular Biology, 5th ed., Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 1–12.Google Scholar
  12. Dellaporta, S.L., Wood, J. and Hicks, J.B. 1983. A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1: 19–21.Google Scholar
  13. Diekmann, W., Venis, M.A. and Robinson, D.G. 1995. Auxin induces clustering of the auxin-binding protein at the surface of maize coleoptile protoplasts. Proc. Natl. Acad. Sci. USA 92: 3425–3429.Google Scholar
  14. Feckler, C., Muster, G., Römer, W. and Palme, K. 2001. Mass spectrometric analysis reveals a cysteine bridge between residues 2 and 61 of the auxin-binding protein 1 from Zea mays L. FEBS Lett. 509: 446–450.Google Scholar
  15. Hesse, T., Feldwisch, J., Balshüsemann, D., Bauw, G., Puype, M., Vanderkerckhove, J., Löbler, M., Klämbt, D., Schell, J. and Palme, K. 1989. Molecular cloning and structural analysis of a gene from Zea mays (L.) coding for a putative receptor for the plant hormone auxin. EMBO J. 8: 2453–2461.Google Scholar
  16. Hesse, T., Garbers, C., Brzobohaty, B., Kreimer, G., Söll, D., Melkonian, M., Schell, J. and Palme, K. 1993. Two members of the ERabp gene family are expressed differentially in reproductive organs but to similar leels in the coleoptile of maize. Plant Mol. Biol. 23: 57–66.Google Scholar
  17. Jones, A.M. and Hermann, E.M. 1993. KDEL-containing auxinbinding protein is secreted to the plasma membrane and cell wall. Plant Physiol. 101: 595–606.Google Scholar
  18. Jones, A.M., Im, K.H., Savka, M.A., Wu, M.J., DeWitt, N.G., Shillito, R. and Binns, A.N. 1998. Auxin-dependent cell expansion mediated by overexpressed auxin-binding protein 1. Science 282: 1114–1117.Google Scholar
  19. Laskowski, M.J., Williams, M.E., Nusbaum, C. and Sussex, I.M. 1995. Formation of lateral root meristems is a two-stage process. Development 121: 3303–3310.Google Scholar
  20. Leblanc, N., Roux, C., Pradier, J.M. and Perrot-Rechenmann, C. 1997. Characterization of two cDNAs encoding auxin-binding proteins in Nicotiana tabacum. Plant Mol. Biol. 33: 679–689.Google Scholar
  21. Leblanc, N., Perrot-Rechenmann, C. and Barbier-Brygoo, H. 1999a. The auxin-binding protein Nt-ERabp1 alone activates an auxinlike transduction pathway. FEBS Lett. 449: 57–60.Google Scholar
  22. Leblanc, N., David, K., Grosclaude, J., Pradier, J.M., Barbier-Brygoo, H., Labiau, S. and Perrot-Rechenmann, C. 1999b. A novel immunological approach establishes that the auxin-binding protein, Nt-abp1, is an element involved in auxin signaling at the plasma membrane. J. Biol. Chem. 274: 28314–28320.Google Scholar
  23. Löbler, M. and Klämbt, D. 1985. Auxin-binding protein from coleoptile membranes of corn (Zea mays L.). I. Purification by immunological methods and characterization. J. Biol. Chem. 260: 9848–9853.Google Scholar
  24. Marchant, A., Bhalerao, R., Casimiro, I., Eklöf, J., Casero, P.J., Bennett, M. and Sandberg, G. 2002. AUX1 promotes lateral root formation by facilitating indole-3-acetic acid distribution between sink and source tissues in the Arabidopsis seedlings. Plant Cell 14: 589–597.Google Scholar
  25. Massotte, D., Fleig, U. and Palme, K. 1995. Purification and characterization of an auxin-binding from Arabidopsis thaliana expressed in baculovirus-infected insect cells. Protein Expr. Purif. 6: 220–227.Google Scholar
  26. Napier, R.M., David, K.M. and Perrot-Rechenmann, C. 2002. A short history of auxin-binding proteins. Plant Mol. Biol. 49: 339–348.Google Scholar
  27. Palme, K., Hesse, T., Campos, N., Garbers, C., Yanofsky, M.F. and Schell, J. 1992. Molecular analysis of an auxin binding protein gene located on chromosome 4 of Arabidopsis. Plant Cell 4: 193–201.Google Scholar
  28. Sali, A. and Blundell, T.L. 1993. Comparative modelling by satisfaction of spatial restraints. J. Mol. Biol. 234: 779–815.Google Scholar
  29. Steffens, B., Feckler, C., Palme, K., Christian, M., Böttger, M. and Lüthen, H. 2001. The auxin signal for protoplast swelling is perceived by extracellular ABP1. Plant J. 27: 591–999.Google Scholar
  30. Schwob, E., Choi, S.Y., Simmons, C., Migliaccio, F., Ilag, L., Hesse, T., Palme, K. and Söll, D. 1993. Molecular analysis of three maize 22 kDa auxin-binding protein genes: transient expression and regulatory regions. Plant J. 4: 423–432.Google Scholar
  31. Thomas, C., Bronner, R., Molinier, J., Prinsen, E., van Onckelen, H. and Hahne, G. 2002. Immuno-cytochemical localization of indole-3-acetic acid during induction of somatic embryogenesis in cultured sunflower embryos. Planta 215: 577–583.Google Scholar
  32. Timpte, C. 2001. Auxin binding protein: curiouser and curiouser. Trends Plant Sci. 6: 586–590.Google Scholar
  33. Woo, E.J., Marshall, J., Bauly, J., Chen, J.G., Venis, M., Napier, R. and Pickersgill, R. 2002. Crystal structure of auxin-binding protein 1 in complex with auxin. EMBO J. 21: 2877–2885.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Clément Thomas
    • 1
  • Denise Meyer
    • 1
  • Michel Wolff
    • 1
  • Christophe Himber
    • 1
  • Malek Alioua
    • 1
  • André Steinmetz
    • 1
  1. 1.Institut de Biologie Moléculaire des Plantes CNRS, and Université Louis PasteurStrasbourg CedexFrance

Personalised recommendations