Plant Molecular Biology

, Volume 12, Issue 5, pp 539–551 | Cite as

Regulation of chalcone flavanone isomerase (CHI) gene expression inPetunia hybrida: the use of alternative promoters in corolla, anthers and pollen

  • Arjen J. van Tunen
  • Susan A. Hartman
  • Leon A. Mur
  • Joseph N. M. Mol


In this paper we report on the organization and expression of the two chalcone flavanone isomerase (CHI) genes A and B from thePetunia hybrida inbred line V30. From a combination of sequence data, primer extension and RNAse protection experiments we infer the presence of two promoters PA1 and PA2 upstream of the CHI gene A coding region. It is shown that both promoters are used differentially in various flower tissues: the PA1 promoter is active in corolla and tube tissue whereas the PA2 promoter, which gives rise to a 437 bp longer transcript, is only active in late stages of anther development and more specifically in pollen grains. The CHI-B gene, on the other hand, has only one promoter (PB) which is active only in immature anther tissue. Thus, in addition to the use of two alternative promoters in front of the same CHI coding region (CHI-A), the promoters in front of the two distinct CHI gene copies are also used differentially as a mechanism to regulate their expression. Comparison of PB with other flavonoid gene promoters active in immature anther tissue revealed a highly conserved region which was designated as ‘anther box’. We hypothesize that it plays a regulatory role in anther-specific gene expression. Finally, a model describing the evolutionary relationship between both CHI genes is presented.

Key words

alternative promoters anther box chalcone flavanone isomerase genes flower development Petunia hybrida 


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  1. 1.
    Brown JWS: A catalogue of splice junction and putative branch point sequences from plant introns. Nucl Acids Res 14: 9549–9559 (1986).Google Scholar
  2. 2.
    Chappell J, Hahlbrock K: Transcription of plant defence genes in response to UV light or fungal elicitor. Nature 311: 76–78 (1984).Google Scholar
  3. 3.
    Chen J, Varner JE: An extracellular matrix protein in plants: characterization of a genomic clone for carrot extensin. EMBO J 4: 2145–2151 (1985).Google Scholar
  4. 4.
    Dean C, Tamaki S, Dunsmuir P, Favreau M, Katayama C, Dooner H, Bedbrook J: mRNA transcripts of several plant genes are polyadenylated at multiple sitesin vivo. Nucl Acids Res 14: 2229–2240 (1986).Google Scholar
  5. 5.
    Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).Google Scholar
  6. 6.
    Dixon RA: The phytoalexin response: elicitation, signalling and control of host gene expression. Biol Rev 61: 239–291 (1986).Google Scholar
  7. 7.
    Drouin G, Dover GA: A plant processed pseudo gene. Nature 328: 557–558 (1987).Google Scholar
  8. 8.
    Fedoroff NV, Furtek DB, Nelson OE: Cloning of the bronze locus in maize by a simple and generalisable procedure using the transposable controlling element Activator (Ac). Proc Natl Acad Sci USA 81: 3825–3829 (1984).Google Scholar
  9. 9.
    Firmin JL, Wilson KE, Rossen L, Johnston AWB: Flavonoid activation of nodulation genes inRhizobium reversed by other compounds in plants. Nature 324: 90–92 (1986).Google Scholar
  10. 10.
    Guilley H, Richards KE, Jonard G: Observations concerning the discontinous DNA of cauliflower mosaic virus. EMBO J 2: 277–282 (1983).Google Scholar
  11. 11.
    Henikoff S: Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28: 351–359 (1984).Google Scholar
  12. 12.
    Hull R, Covey SN: Does cauliflower mosaic virus replicate by reverse transcription? Trends Biochem Sci 8: 119–121 (1983).Google Scholar
  13. 13.
    Johns MA, Mottinger V, Freeling M: A low copy number copia-like transposon in maize. EMBO J 4: 1093–1101 (1984).Google Scholar
  14. 14.
    Joshi CP: Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucl Acids Res 15: 9627–9640 (1987).Google Scholar
  15. 15.
    Kehrel B, Wiermann R: Immunochemical localization of phenylalanine ammonia-lyase and chalcone synthase in anthers. Planta 163: 183–190 (1985).Google Scholar
  16. 16.
    Koes RE, Spelt CE, Reif HJ, van denElzen PJM, Veltkamp E, Mol JNM: Floral tissue ofPetunia hybrida (V30) expresses only one member of the chalcone synthase multigene family. Nucl Acids Res 14: 5229–5239 (1986).Google Scholar
  17. 17.
    Koes RE, Spelt CE, Mol JNM, Gerats AGM: The chalcone synthase multigene family ofPetunia hybrida: sequence homology, chromosomal localization and evolutionary aspects. Plant Mol Biol 10: 159–169 (1987).Google Scholar
  18. 18.
    Koes RE, Spelt CE, Mol JNM: The chalcone synthase multigene family ofPetunia hybrida (V30): differential, light-regulated expression during flower development and UV-light induction. Plant Mol Biol 12: 213–225.Google Scholar
  19. 19.
    Langridge P, Feix G: A zein gene of maize is transcribed from two widely separated promoter regions Cell 34: 1015–1022 (1983).Google Scholar
  20. 20.
    Maniatis T, Fritsch EF, Sambrook J: Molecular cloning; a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).Google Scholar
  21. 21.
    Martin C, Carpenter R, Sommer H, Saedler H, Coen E: Molecular analysis of instability in flower pigmentation ofAnitirrhinum majus, following isolation of the pallida locus by transposon tagging. EMBO J 4: 1625–1630 (1985).Google Scholar
  22. 22.
    Mehdy MC, Lamb CJ: Chalcone isomerase cDNA cloning and mRNA induction by fungal elicitor, wounding and infection. EMBO J 6: 1527–1533 (1987).Google Scholar
  23. 23.
    Meyer P, Heidman I, Forkman G, Saedler H: A newPetunia flower colour generated by transformation of a mutant with a maize gene. Nature 330: 677–678 (1987).Google Scholar
  24. 24.
    Mol JNM, Schram AW, deVlaming P, Gerats AGM, Kreuzaler F, Hahlbrock K, Reif HJ, Veltkamp E: Regulation of flavonoid gene expression inPetunia hybrida: description and partial characterization of a conditional mutant in chalcone synthase gene expression. Mol Gen Genet 192: 424–429 (1983).Google Scholar
  25. 25.
    Paz-Ares J, Wienand U, Peterson PA, Saedler H: Molecular cloning of the c locus ofZea mays: a locus regulating the anthocyanin pathway. EMBO J 5: 829–833 (1986).Google Scholar
  26. 26.
    Peters NK, Frost JW, Long SR: A plant flavone, luteolin influences expression ofRhizobium meliloti nodulation genes. Science 233: 977–980 (1986).Google Scholar
  27. 27.
    Pfeiffer P, Hohn T: Involvement of reverse transcription in the replication of cauliflower mosaic virus: a detailed model and test of some aspects. Cell 33: 781–789 (1983).Google Scholar
  28. 28.
    Redmond JW, Baley M, Djordjevic MA, Innes RW, Kuempel PL, Rolfe BG: Flavones induce expression of the nodulation genes inRhizobium. Nature 323: 632–635 (1986).Google Scholar
  29. 29.
    Reif HJ, Niesbach U, Deumling B, Saedler H: Cloning and analysis of two genes for chalcone synthase fromPetunia hybrida. Mol Gen Genet 199: 208–215 (1985).Google Scholar
  30. 30.
    Rudall P: Anatomy of flowering plants: an introduction to structure and development Edward Arnold (Publishers), London (1987).Google Scholar
  31. 31.
    Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).Google Scholar
  32. 32.
    Schibler U, Sierra F: Alternative promoters in developmental gene expression Ann Rev Genet 21: 237–257 (1987).Google Scholar
  33. 33.
    Schwarz-Sommer Z, Shepherd N, Tacke E, Gierl A, Rohde W, Lecercq L, Mattes M, Berndtgen R, Peterson PA, Saedler H: Influence of transposable elements on the structure of the A1 gene ofZea mays. EMBO J 6: 287–294 (1987).Google Scholar
  34. 34.
    Shepherd NS, Schwarz-Sommer Z, Blumberg vel Spalve J, Gupta M, Wienand U and Saedler H: Similarity of theCinl repetitive family ofZea mays to eukaryotic transposable elements. Nature 307: 185–187 (1984).Google Scholar
  35. 35.
    Sommer H, Saedler H: Structure of the chalcone synthase gene ofAntirrhinum majus. Mol Gen Genet 202: 429–434 (1986).Google Scholar
  36. 36.
    Sommer H, Bonas U, Saedler H: Transposon-induced alterations in the promoter region affect transcription of the chalcone synthase gene ofAntirrhinum majus. Mol Gen Genet 211: 49–55 (1988).Google Scholar
  37. 37.
    van derKrol AR, Lenting PE, Veenstra J, van derMeer IM, Koes RE, Gerats AGM, Mol JNM, Stuitje AR: An anti-sense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333: 866–869 (1988).Google Scholar
  38. 38.
    vanTunen AJ, Mol JNM: A novel purification procedure for chalcone flavanone isomerase fromPetunia hybrida and the use of its antibodies to characterize the Po mutation. Arch Biochem Biophys 257: 85–91 (1987).Google Scholar
  39. 39.
    vanTunen AJ, Koes RE, Spelt CE, van derKrol AR, Stuitje AR, Mol JNM: Cloning of the two chalcone flavanone isomerase genes fromPetunia hybrida; coordinate, light regulated and differential expression of flavonoid genes. EMBO J 7: 1257–1263 (1988).Google Scholar
  40. 40.
    Wienand U, Weydemanu, Niesbach-Klösgen U, Peterson PA, Saedler H: Molecular cloning of the c2 locus ofZea mays, the gene coding for chalcone synthase. Mol Gen Genet 203: 202–207 (1986).Google Scholar

Copyright information

© Kluwer Academic Publishers 1989

Authors and Affiliations

  • Arjen J. van Tunen
    • 1
  • Susan A. Hartman
    • 1
  • Leon A. Mur
    • 1
  • Joseph N. M. Mol
    • 1
  1. 1.Department of Genetics, Section Biosynthesis of Secondary MetabolitesVrije UniversiteitAmsterdamNetherlands

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