Plant Molecular Biology

, Volume 44, Issue 6, pp 777–787 | Cite as

A gene expression screen identifies EARLI1 as a novel vernalization-responsive gene in Arabidopsis thaliana

  • Richard Wilkosz
  • Michael Schläppi


Vernalization promotes early flowering in late ecotypes of Arabidopsis thaliana. The mechanisms of vernalization are poorly understood. A subtractive hybridization approach was used to isolate vernalization-responsive genes from a late-flowering ecotype of Arabidopsis thaliana based on the premise that transcript levels of such genes would increase with cold treatment and remain high even after removal of the vernalization stimulus. EARLI1 is the first Arabidopsis gene shown to be stably activated by vernalization. The abundance of its RNA is progressively elevated by vernalization and remains high for at least 20 days at room temperature. The basal level of EARLI1 RNA is higher in early-flowering ecotypes, but is increased also after vernalization. Vernalization and subsequent growth in long-day photoperiods have an additive or synergistic effect on EARLI1 activation. EARLI1 RNA levels are also transiently induced by brief exposures to cold, but not to abscisic acid. EARLI1 is thus a novel vernalization-responsive gene in Arabidopsis thaliana that can be used to investigate vernalization-specific transcriptional regulation.

abscisic acid EARLI1 FLC photoperiod subtractive hybridization vernalization 


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  1. An, Y.-Q., McDowell, J.M., Hunag, S., McKinney, E.C., Chambliss, S. and Meagher, R.B. 1996. Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues. Plant J. 10: 107–121.PubMedGoogle Scholar
  2. Bagnall, D.J. 1993. Light quality and vernalization interact in controlling late flowering in Arabidopsis thaliana ecotypes and mutants. Ann. Bot. 71: 75–83.Google Scholar
  3. Baker, S.S., Wilhelm, K.S. and Tomashow, M.F. 1994. The 50-region of Arabidopsis thaliana COR15a has cis-acting elements that confer cold-, drought-and ABA-regulated gene expression. Plant Mol. Biol. 24: 701–713PubMedGoogle Scholar
  4. Burn, J.E., Bagnall, D.J., Metzger, J.D., Dennis, E.S. and Peacock, W.J. 1993. DNA methylation, vernalization, and the initiation of flowering. Proc. Natl. Acad. Sci. USA 90: 287–291.PubMedGoogle Scholar
  5. Chandler, J., Wilson, A. and Dean, C. 1996. Arabidopsis mutants showing an altered response to vernalization. Plant J. 10: 637–644.PubMedGoogle Scholar
  6. Chong, K., Wang, L.-P., Tan, K.-H., Huang, H.-L., and Liang, H.-G. 1994. Molecular cloning and characterization of vernalizationrelated (ver) genes in winter wheat. Physiol. Plant. 92: 511–515.Google Scholar
  7. Chong, K., Bao, S.-l., Xu, T., Tan, K.-h. Liang, T.-b., Zeng, J.-z., Huang, H.-l., Xu, J. and Xu, Z.-h. 1998. Functional analysis of the ver gene using antisense transgenic wheat. Physiol. Plant. 102: 87–92.Google Scholar
  8. Clark, J.H. and Dean, C. 1994. Mapping FRI, a locus controlling flowering time and vernalization response in Arabidopsis thaliana. Mol. Gen. Genet. 242: 81–89.PubMedGoogle Scholar
  9. Cook, D., Dreyer, D., Bonnet, D., Howell, M., Nony, E. and VandenBosch, K. 1995. Transient induction of a peroxidase gene in Medicago truncatula precedes infection by Rhizobium meliloti. Plant Cell 7: 43–55.PubMedGoogle Scholar
  10. Dennis, E.S., Finnegan, E.J., Bilodeau, P. Chaudhury, A., Genger, R., Helliwell, C.A., Sheldon, C.C., Bagnall, D.J. and Peacock, W.J. 1996. Vernalization and the initiation of flowering. Semin. Cell Dev. Biol. 7: 441–448.Google Scholar
  11. Dennis, E.S., Bilodeau, P., Burn, J., Finnegan, J.E., Genger, R., Helliwell, C., Kang, B.J., Sheldon, C.C. and Peacock, W.J. 1997. Methylation controls the low temperature induction of flowering in Arabidopsis. In: A.J. Greenland, E.M. Meyerowitz and M. Steer (Eds.) Control of Plant Development: Genes and Signals, Symposia of the Society for Experimental Biology Vol. 51, Company of Biologists, Cambridge, MD, pp. 97–103.Google Scholar
  12. Finnegan, E.J., Genger, R.K., Kovac, K., Peacock, W.J. and Dennis, E.S. 1998. DNA methylation and the promotion of flowering by vernalization. Proc. Natl. Acad. Sci. USA 95: 5824–5829.PubMedGoogle Scholar
  13. Hajela, R.K., Horvath, D.P., Gilmour, S.J. and Thomashow, M.F. 1990. Molecular expression of cor (cold-regulated) genes in Arabidopsis thaliana. Plant Physiol. 93: 1246–1252.Google Scholar
  14. Koornneef, M., Blankestijn-de Vries, H., Hanhart, C., Soppe, W. and Peeters, T. 1994. The phenotype of some late flowering mutants is enhanced by a locus on chromosome 5 that is not effective in the Landsberg erecta wild-type. Plant J. 6: 911–919.Google Scholar
  15. Koornneef, M., Alonso-Blanco, C., Peeters, A.J.M. and Soppe W. 1998. Genetic control of flowering time in Arabidopsis. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49: 345–370.PubMedGoogle Scholar
  16. Kurkela, S.M. and Franck, M. 1990. Cloning and characterization of a cold-and ABA-inducible Arabidopsis gene. Plant Mol. Biol. 15: 137–144.PubMedGoogle Scholar
  17. Lee, I., Bleecker, A. and Amasino, R. 1993. Analysis of naturally occurring late flowering in Arabidopsis thaliana. Mol.Gen. Genet. 237: 171–176.PubMedGoogle Scholar
  18. Lee, I., Michaels, S.D., Masshardt, A.S. and Amasino, R.M. 1994. The late-flowering phenotype and mutations in LUMINIDEPENDENS is suppressed in the Landsberg erecta strain of Arabidopsis. Plant J. 6: 903–909.Google Scholar
  19. Lee, I. and Amasino, R. 1995. Effect of vernalization, photoperiod, and light quality on the flowering phenotype of Arabidopsis plants containing the FRIGIDA gene. Plant Physiol. 108: 157–162.PubMedGoogle Scholar
  20. Levy, Y.Y. and Dean, C. 1998. The transition to flowering. Plant Cell 10: 1973–1989.PubMedGoogle Scholar
  21. Lin, C. and Thomashow, M.F. 1992. DNA sequence analysis of a complementary DNA for cold-regulated Arabidopsis gene cor15 and characterization of the COR15 polypeptide. Plant Physiol. 99: 519–525.Google Scholar
  22. Martínez-Zapater, J.M. and Somerville, C.R. 1990. Effect of light quality and vernalization on late-flowering mutants of Arabidopsis thaliana. Plant Physiol. 92: 770–776.Google Scholar
  23. Martínez-Zapater, J.M., Coupland, G., Dean, C. and Koornneef, M. 1994. The transition to flowering in Arabidopsis. In: E.M. Meyerowitz and C.R. Somerville (Eds.) Arabidopsis, Cold Spring Harbor Laboratory Press, Plainview, NY, pp. 403–433.Google Scholar
  24. Michaels, S.D. and Amasino, R.M. 1999. FLOWERING LOCUS C encodes a novel MADS domain protein that acts as a repressor of flowering. Plant Cell 11: 949–956.PubMedGoogle Scholar
  25. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15: 473–497.Google Scholar
  26. Piñ eiro, M. and Coupland, G. 1998. The control of flowering time and floral identity in Arabidopsis. Plant Physiol. 117: 1–8.PubMedGoogle Scholar
  27. Richards, K.D. and Gardner, R.C. 1995. pEARLI1 (Accession No. L43080): an Arabidopsis member of a conserved gene family (PGR95–099). Plant Physiol. 109: 1497.PubMedGoogle Scholar
  28. Riechmann, J.L. and Meyerowitz, E.M. 1997. MADS domain proteins in plant development. Biol. Chem. 378: 1079–1101.PubMedGoogle Scholar
  29. Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Plainview, NY.Google Scholar
  30. Sanda, S.L. and Amasino, R. 1996. Interaction of FLC and late-flowering mutations in Arabidopsis thaliana. Mol. Gen. Genet. 251: 69–74.PubMedGoogle Scholar
  31. Sanda, S., John, M. and Amasino, R. 1997. Analysis of flowering time in ecotypes of Arabidopsis thaliana. J. Hered. 88: 69–72.PubMedGoogle Scholar
  32. Schaffer, R., Ramsay, N. Samach, A., Corden, S., Putterill, J., Carré, I.A. and Coupland, G. 1999. LATE ELONGATED HYPOCOTYL, an Arabidopsis gene encoding a MYB transcription factor, regulates circadian rhythmicity and photoperiodic responses. Cell 93: 1219–1229.Google Scholar
  33. Sheldon, C.C., Burn, J.E., Perez, P.P., Metzger, J., Edwards, J.A., Peacock, W.J. and Dennis, E.S. 1999. The FLF MADS box gene: a repressor of flowering in Arabidopsis regulated by vernalization and methylation. Plant Cell 11: 445–458.PubMedGoogle Scholar
  34. Sheldon, C.C., Rouse, D.T., Finnegan, E.J., Peacock, W.J. and Dennis, E.S. 2000. The molecular basis of vernalization: the central role of FLOWERING LOCUS C (FLC). Proc. Natl. Acad. Sci. USA 97: 3753–3758.PubMedGoogle Scholar
  35. Simpson, G.G., Gendall, A.R. and Dean, C. 1999. When to switch to flowering. Annu. Rev. Cell Dev. Biol. 99: 519–550.Google Scholar
  36. Terzaghi, W.B. and Cashmore, A.R. 1995. Light-regulated transcription. Annu. Rev. Plant Physiol. Plant Mol. Biol. 46: 445–474.Google Scholar
  37. Thomashow, M.F. 1994. Arabidopsis thaliana as a model for studying mechanisms of plant cold tolerance. In: E.M. Meyerowitz and C.R. Somerville (Eds.) Arabidopsis, Cold Spring Harbor Laboratory Press, Plainview, NY, pp. 807–834.Google Scholar
  38. Wang, Z. and Brown, D.D. 1991. A gene expression screen. Proc. Natl. Acad. Sci. USA 88: 11505–11509.PubMedGoogle Scholar
  39. Wang, Z.-Y., Kenigsbuch, D., Sun, L., Harel, E., Ong, M.S. and Tobin, E.M. 1997. A Myb-related transcription factor is involved in the phytochrome regulation of an Arabidopsis Lhcb gene. Plant Cell 9: 491–507.PubMedGoogle Scholar
  40. Yeh, K.-W., Juang, R.-H., and Su, J.-C. 1990. A rapid and efficient method for RNA isolation from plants with high carbohydrate content. Focus 13: 102–103.Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Richard Wilkosz
    • 1
  • Michael Schläppi
    • 1
  1. 1.Department of BiologyMarquette UniversityMilwaukeeUSA

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