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Phytohormone control of the tobacco anionic peroxidase promoter

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Abstract

The tobacco anionic peroxidase gene encodes the predominant peroxidase isoenzyme in the aerial portions of tobacco. Three kb of the peroxidase promoter was joined to the coding region of theEscherichia coli β-glucuronidase gene (GUS), and transiently expressed in tobacco mesophyll protoplasts in the presence or absence of plant growth regulators. Benzyladenine, ethylene, and gibberellic acid did not affect peroxidase gene expression. Abscisic acid slightly inhibited expression at high concentrations. The auxins indole-3-acetic acid (IAA) and naphthaleneacetic acid strongly suppressed peroxidase expression. We observed half maximal suppression at 30 μM IAA. An antiauxin,p-chlorophenoxyisobutyric acid (PCIB), enhanced expression from the peroxidase promoter above that of untreated controls or restored activity when used in combination with IAA. Sequencing 3 kb of the peroxidase promoter revealed many potential regulatory elements based on sequence homology to previously characterized genes. This includes several consensus transcription factor binding sites found in auxin-regulated promoters. 5′ deletions of the peroxidase promoter/GUS fusion revealed several positive and negative regulatory elements. An upstream enhancer element was found between −3146 and −638 from the start of transcription. A strong silencer element was observed between −638 and −220. Removal of this silencer resulted in a truncated promoter (−220) with 100% activity of the full-length promoter (−3146). Inhibition by auxin was observed with all 5′ deletions.

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References

  1. Ainley W, Walker J, Nagao R, Key J: Sequence and characterization of two auxin-regulated genes from soybean. J Biol Chem 263: 10658–10666 (1988).

    PubMed  Google Scholar 

  2. Ballas N, Wong L-M, Theologis A: Identification of the auxinresponsive element,AuxRE, in the primary indoleacetic acid-inducible gene,PS-IAA4/5, of pea (Pisum sativum). J Mol Biol 233: 580–596 (1993).

    Article  PubMed  Google Scholar 

  3. Boulikas T: Nature of DNA sequences at the attachment regions of genes to the nuclear matrix. J Cell Biochem52: 14–22 (1993).

    PubMed  Google Scholar 

  4. Bradford M M: A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).

    Article  PubMed  Google Scholar 

  5. Bustos M, Guiltinan M, Jordano J, Begum D, Kalkan F, Hall T: Regulation of β-glucuronidase expression in transgenic tobacco plants by an A/T-rich,cis-acting sequence found upstream of a french bean β-phaseolin gene. Plant Cell 1: 839–853 (1989).

    Article  PubMed  Google Scholar 

  6. Conner T, Goekjian V, LaFayette P, Key J: Structure and expression of two auxin-inducible genes fromArabidopsis. Plant Mol Biol 15: 623–632 (1990).

    PubMed  Google Scholar 

  7. Czarnecka E, Nagao R, Key J, Gurley W: Characterization ofGmhsp26-A, a stress gene encoding a divergent heat shock protein of soybean: heavy-metal induced inhibition of intron processing. Mol Cell Biol 8: 1113–1122 (1988).

    PubMed  Google Scholar 

  8. Dunsmuir P, Bond D, Lee K, Gidoni D, Townsend J: Stability of introduced genes and stability in expression. In: Gelvin SB, Schilperoort RA, Verma DPS (eds) Plant Molecular Biology Manual, pp. C1: 5–7. Kluwer Academic Publishers. Dordrecht, Netherlands (1988).

    Google Scholar 

  9. Fry SC. The Growing Plant Cell Wall: Chemical and Metabolic Analysis. Longman Scientific and Technical, Essex, UK (1988).

    Google Scholar 

  10. Goldsbrough A, Albrecht H, Stratford R: Salicylic acid-inducible binding of a tobacco nuclear protein to a 10 bp sequence which is highly conserved amongst stress-inducible genes. Plant J 3: 563–571 (1993).

    PubMed  Google Scholar 

  11. Hagen G, Martin G, Li Y, Guilfoyle T: Auxin-induced expression of the soybean GH3 promoter in transgenic tobacco plants. Plant Mol Biol 17: 567–579 (1991).

    PubMed  Google Scholar 

  12. Hinnman RL, Lang J: Peroxidase catalyzed oxidation of indole-3-acetic acid. Biochemistry 4: 144–158 (1965).

    Google Scholar 

  13. Intapruk C, Higashimura N, Yamamoto K, Okada N, Shinmyo A, Takano M: Nucleotide sequences of two genomic DNAs encoding peroxidase ofArabidopsis thaliana. Gene 98: 237–241 (1991).

    Article  PubMed  Google Scholar 

  14. Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).

    PubMed  Google Scholar 

  15. Jourdan PS, Earle ED: Genotypic variability in the frequency of plant regeneration from leaf protoplasts of fourBrassica spp. and ofRaphanus sativus. J Am Soc Hort Sci 114: 343–349 (1989).

    Google Scholar 

  16. Kawagoe Y, Murai N: Four distinc nuclear proteins recognizein vitro the proximal promoter of the bean seed storage protein β-phaseolin gene conferring spatial and temporal control. Plant J 2: 927–936 (1992).

    Article  PubMed  Google Scholar 

  17. Kawagoe Y, Campbell B, Murai N: Synergism between CACGTG (G-box) and CACCTGcis elements is required for activation of the bean seed storage protein β-phaseolin gene. Plant J 5: 885–890 (1994).

    Article  PubMed  Google Scholar 

  18. Kim S, Choi J, Costa M, An G: Identification of G-box sequence as an essential element for methyl jasmonate response of potato proteinase inhibitor II promoter. Plant Physiol 99: 627–631 (1992).

    Google Scholar 

  19. Knee M: Role of ethylene in chlorophyll degradation in radish cotyledons. J Plant Growth Regul 10: 157–162 (1991).

    Google Scholar 

  20. Kraft R, Tardiff J, Krauter KS, Leinwand LA: Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques 6: 544–549 (1988).

    PubMed  Google Scholar 

  21. Lagrimini LM: Analysis of peroxidase function in transgenic plants. In: Welinder KG, Rasmussen SK, Penel C, Greppin H (eds) Plant Peroxidases: Biochemistry and Physiology, pp. 301–306. University of Geneva, Switzerland (1993).

    Google Scholar 

  22. Lagrimini LM, Burkhart W, Moyer M, Rothstein S: Molecular cloning of complementary DNA encoding the lignin-forming peroxidase from tobacco: molecular analysis and tissue-specific expression. Proc Natl Acad Sci USA 84: 7542–7546 (1987).

    Google Scholar 

  23. Lagrimini LM, Rothstein S: Tissue specificity of tobacco peroxidase isozymes and their induction by wounding and tobacco mosaic virus infection. Plant Physiol 84: 438–442 (1987).

    Google Scholar 

  24. Lam E, Chua N-H: ASF-2: a factor that binds to the cauliflower mosaic virus 35S promoter and a conserved GATA motif inCab promoters. Plant Cell 1: 1147–1156 (1989).

    Article  PubMed  Google Scholar 

  25. Liu Z-B, Ulmasov T, Shi X, Hagen G, Guilfoyle T: SoybeanGH3 promoter contains multiple auxin-inducible elements. Plant Cell 6: 645–657 (1994).

    PubMed  Google Scholar 

  26. Mader M, Meyer Y, Bopp M: Localization of peroxidase isozymes in protoplasts and cell walls ofNicotiana tabacum L. Planta 122: 259–268 (1975).

    Google Scholar 

  27. Mader M, Nessel A, Bopp M: On the physiological significance of the isozyme groups of peroxidase from tobacco demonstrated by biochemical properties. II. pH optima, Michaelis constants, maximal oxidation rates. Z Pflanzenphysiol 82: 247–260 (1977).

    Google Scholar 

  28. McClure B, Hagen G, Brown C, Gee M, Guilfoyle T: Transcription, organization, and sequence of an auxin-regulated gene cluster in soybean. Plant Cell 1: 229–239 (1989).

    Article  PubMed  Google Scholar 

  29. McRae DH, Bonner J: Chemical structure and antiauxin activity. Physiol Plant 6: 485–510 (1953).

    Google Scholar 

  30. Nagao R, Goekjian V, Hong J, Key J: Identification of protein-binding DNA sequences in an auxin-regulated gene of soybean. Plant Mol Biol 21: 1147–1162 (1993).

    Article  PubMed  Google Scholar 

  31. Negrutiu I, Shillito RD, Potrykus I, Biasini G, Sala F: Hybrid genes in the analysis of transformation conditions. I. Setting up a simple method for direct gene transfer in plant protoplasts. Plant Mol Biol 8: 363–373 (1987).

    Google Scholar 

  32. Ockerse R, Siegel BZ, Galston AW: Hormone-induced repression of a peroxidase isozyme in plant tissue. Science 18: 452–453 (1966).

    Google Scholar 

  33. Oeller P, Keller J, Parks J, Silbert J, Theologis A: Structural characterization of the early indoleacetic acid-inducible genes,PS-IAA4/5 andPS-IAA6, of pea (Pisum sativum L.). J Mol Biol 233: 789–798 (1993).

    Article  PubMed  Google Scholar 

  34. Rieping M, Schoffl F: Synergistic effect of upstream sequences, CCAAT box elements, and HSE sequences for enhanced expression of chimaeric heat shock genes in transgenic tobacco. Mol Gen Genet 231: 226–232 (1992).

    PubMed  Google Scholar 

  35. Roberts E, Kutchan T, Kolattukudy PE: Cloning and sequencing of cDNA for a highly anionic peroxidase from potato and the induction of its mRNA in suberizing potato tubers and tomato fruits. Plant Mol Biol 11: 15–26 (1988).

    Google Scholar 

  36. Takahashi Y, Niwa Y, Machida Y, Nagata T: Location of thecis-acting auxin-responsive region in the promoter of thePar gene from tobacco mesophyll protoplasts. Proc Natl Acad Sci USA 87: 8013–8016 (1990).

    PubMed  Google Scholar 

  37. Theologis A: Rapid gene regulation by auxin. Annu Rev Plant Physiol 37: 407–438 (1986).

    Google Scholar 

  38. Thorpe TA, Van MTT, Gaspar T: Isoperoxidases in epidermal layers of tobacco and changes during organ formationin vitro. Physiol Plant 44: 388–394 (1978).

    Google Scholar 

  39. Williams M, Foster R, Chua N-H: Sequences flanking the hexameric G-box core CACGTG affect the specificity of protein binding. Plant Cell 4: 485–496 (1992).

    Article  PubMed  Google Scholar 

  40. Ye XS, Pan SQ, Kuc J: Activity, isozyme pattern, and cellular localization of peroxidase as related to systemic resistance of tobacco to blue mold (Peronospora tabacina) and to tobacco mosaic virus. Phytopathology 80: 1295–1298 (1990).

    Google Scholar 

  41. Zarembinski T, Theologis A: Anaerobiosis and plant growth hormones induce two genes encoding 1-aminocyclopropane-1-carboxylate synthase in rice (Oryza sativa L.). Mol Biol Cell 4: 363–373 (1993).

    PubMed  Google Scholar 

  42. Zhou C, Abaigar L, Jong AY: A protocol for using T7 DNA polymerase in oligonucleotide site-directed mutagenesis. Biotechniques 8: 503 (1990).

    PubMed  Google Scholar 

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  1. Department of Horticulture and Crop Sciences, Ohio State University, 43210-1096, Columbus, OH, USA

    Karen L. Klotz & L. Mark Lagrimini

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  1. Karen L. Klotz
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  2. L. Mark Lagrimini
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Klotz, K.L., Lagrimini, L.M. Phytohormone control of the tobacco anionic peroxidase promoter. Plant Mol Biol 31, 565–573 (1996). https://doi.org/10.1007/BF00042229

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  • DOI: https://doi.org/10.1007/BF00042229

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