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Plant Molecular Biology

, Volume 17, Issue 3, pp 567–579 | Cite as

Auxin-induced expression of the soybean GH3 promoter in transgenic tobacco plants

  • Gretchen Hagen
  • Gary Martin
  • Yi Li
  • Tom J. Guilfoyle
Full Paper

Abstract

The gene encoding the auxin-responsive GH3 mRNA (G. Hagen, A. Kleinschmidt, TJ. Guilfoyle, Planta 162: 147–153 (1984)) from soybean was cloned, and its sequence and transcription initiation site were determined. The promoter of the GH3 gene has been fused to the open reading frame of theEscherichia coli uidA gene which encodes β-glucuronidase (GUS). This fusion gene was introduced into tobacco viaAgrobacterium tumefaciens-mediated transformation, and the expression of the gene was examined by fluorometric assay and histochemical staining of young R1 tobacco seedlings and mature plants. In transgenic tobacco plants that have not been exposed to exogenous auxin, expression of the fusion gene is largely restricted to roots of young green plants and developing floral organs, including ovules, developing seeds, and pollen, of mature plants. Application of exogenous auxin to tobacco seedlings or plant organs results in a greater than 50-fold increase in expression of GUS. Auxin-induced GUS expression is greates in vascular tissue, but not restricted to this tissue. The auxin-deduced GUS expression was characterized for kinetics, auxin specificity and dose response.

Key words

auxin GH3 gene transgenic tobacco auxin-regulated gene expression 

Abbreviations

NAA

α-naphthaleneacetic acid

IAA

indole-3-acetic acid

IBA

indole-3-butyric acid

2,4-D

2,4-dichlorophenoxyacetic acid

GUS

β-glucuronidase

NOS

nopaline synthase

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References

  1. 1.
    Ainley WM, Walker JC, Nagao RT, Key JL: Sequence and characterization of two auxin-regulated genes from soybean. J Biol Chem 263: 10658–10666 (1988).Google Scholar
  2. 2.
    An G, Costa MA, Ha S-B: Nopaline synthase promoter is wound inducible and auxin inducible. Plant Cell 2: 225–233 (1990).Google Scholar
  3. 3.
    Arndt KT, Styles CA, Fink GR: A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases. Cell 56: 527–537 (1989).Google Scholar
  4. 4.
    Dale RMK, McClure BA, Houchins JP: A rapid singlestranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: Application to sequencing corn mitochondrial 18S rDNA. Plasmid 13: 31–40 (1985).Google Scholar
  5. 5.
    Evans ML: The action of auxin on plant cell elongation. CRC Crit Rev Plant Sci 2: 317–365 (1985).Google Scholar
  6. 6.
    Gee MA, Hagen G, Guilfoyle TJ: Tissue-specific and organ-specific expression of soybean auxin-responsive transcripts GH3 and SAURs. Plant Cell 3, in press (1991).Google Scholar
  7. 7.
    Gielen J, De Beuckeleer M, Seurinck J, Deboeck F, De Greve H, Lemmers M, Van Montagu M, Schell J: The complete nucleotide sequence of the TL-DNA of theAgrobacterium tumefaciens plasmid pTiAch5. EMBO J 3: 835–846 (1984).Google Scholar
  8. 8.
    Guilfoyle TJ: Auxin-regulated gene expression in higher plants. CRC Crit Rev Plant Sci 4: 247–276 (1986).Google Scholar
  9. 9.
    Hagen G, Kleinschmidt A, Guilfoyle T: Auxin regulated gene expression in intact soybean hypocotyl and excised hypocotyl sections. Planta 162: 147–153 (1984).Google Scholar
  10. 10.
    Hagen G, Guilfoyle T: Rapid induction of selective transcription by auxins. Mol Cell Biol 5: 1197–1203 (1985).Google Scholar
  11. 11.
    Hagen G, Uhrhammer N, Guilfoyle TJ: Regulation of expression of an auxin-induced soybean sequence by cadmium. J Biol Chem 263: 6442–6226 (1988).Google Scholar
  12. 12.
    Hagen G: Molecular approaches to understanding auxin action. New Biol 1: 19–23 (1989).Google Scholar
  13. 13.
    Horsch RB, Fry JE, Hoffmann NL, Wallroth M, Eichholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).Google Scholar
  14. 14.
    Horsch RB, Klee HJ: Rapid assay of foreign gene expression in leaf discs transformed byAgrobacterium tumefaciens: Role of T-DNA borders in the transfer process. Proc Natl Acad Sci USA 83: 4428–4432 (1986).Google Scholar
  15. 15.
    Huynh TV, Young RA, Davis RW: Constructing and screening cDNA libraries in lambda gt10. In: Glover DM (ed) DNA Cloning Techniques: A Practical Approach, vol 1, pp. 49–78. IRL Press, London (1985).Google Scholar
  16. 16.
    Jefferson RA: Assay for chimeric genes in plants: the GUS fusion system. Plant Mol Biol Rep 5: 387–405 (1987).Google Scholar
  17. 17.
    Katagiri F, Lam E, Chua N-H: Two tobacco DNA-binding proteins with homology to the nuclear factor CREB. Nature 340: 225–233 (1989).Google Scholar
  18. 18.
    Key JL: Modulation of gene expression by auxin. BioEssays 11: 52–58 (1989).Google Scholar
  19. 19.
    McClure BA, Hagen G, Brown CS, Gee MA, Guilfoyle TJ: Transcription, organization and sequence of an auxin-regulated gene cluster in soybean. Plant Cell 1: 229–239 (1989).Google Scholar
  20. 20.
    Mitchell PJ, Tjian R: Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science 245: 371–378 (1989).Google Scholar
  21. 21.
    Rogers SG, Klee HJ, Horsch RB, Fraley RT: Improved vectors for plant tranformation: Expression cassette vectors and new selectable markers. Meth Enzymol 153: 253–277 (1987).Google Scholar
  22. 22.
    Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).Google Scholar
  23. 23.
    Slightom JL, Durand-Tardif M, Jouanin L, Tepfer D: Nucleotide sequence analysis of TL-DNA ofAgrobacterium rhizogenes agropine type plasmid. J Biol Chem 261: 108–121 (1986).Google Scholar
  24. 24.
    Skuzeski JM, Nichols LM, Gesteland RF: Analysis of leaky viral translation termination codonsin vivo by transient expression of improved β-glucuronidase vectors. Plant Mol Biol 15: 65–79 (1990).Google Scholar
  25. 25.
    Tabata T, Takase H, Takayama S, Mikami K, Nakatsuka A, Kawata T, Nakayama T, Iwabuchi M. A protein that binds to a cis-acting element of wheat histone genes has a leucine zipper motif. Science 245: 965–967 (1989).Google Scholar
  26. 26.
    Wright RM, Hagen G, Guilfoyle T: An auxin-induced polypeptide in dicotyledonous plants. Plant Mol Biol 9: 625–643 (1987).Google Scholar

Copyright information

© Kluwer Academic Publishers 1991

Authors and Affiliations

  • Gretchen Hagen
    • 1
  • Gary Martin
    • 1
  • Yi Li
    • 1
  • Tom J. Guilfoyle
    • 1
  1. 1.Department of BiochemistryUniversity of MissouriColumbiaUSA

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