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Cloning of cDNA coding for dihydroflavonol-4-reductase (DFR) and characterization of dfr expression in the corollas of Gerbera hybrida var. Regina (Compositae)

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Abstract

We are approaching corolla differentiation in Compositae by studying the regulation of flavonoid pathway genes during inflorescence development in gerbera. We have cloned a dfr cDNA from a ray floret corolla cDNA library of Gerbera hybrida var. Regina by a PCR technique based on homologies found in genes isolated from other plant species. The functionality of the clone was tested in vivo by complementing the dihydrokaempferol accumulating petunia mutant line RL01. By Southern blot analysis, G. hybrida var. Regina was shown to harbour a small family of dfr genes, one member of which was deduced to be mainly responsible for the DFR activity in corolla. Dfr expression in corolla correlates with the anthocyanin accumulation pattern: it is basipetally induced, epidermally specific and restricted to the ligular part of corolla. By comparing the dfr expression in different floret types during inflorescence development, we could see that dfr expression reflects developmental schemes of the outermost ray and trans florets, contrasted with that of the disc florets.

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References

  1. Almeida J, Carpenter R, Robbins TP, Martin C, Coen ES: Genetic interactions underlying flower color patterns in Antirrhinum majus. Genes Devel. 3: 1758–1767 (1989).

    Google Scholar 

  2. Asen S: High pressure liquid chromatographic analysis of flavonoid chemical markers in petals from Gerbera flowers as an adjunct for cultivar and germplasm identification. Phytochemistry 23: 2523–2526 (1984).

    Google Scholar 

  3. Beld M, Martin C, Huits H, Stuitje AR, Gerats AGM: Flavonoid synthesis in Petunia hybrida: partial characterization of dihydroflavonol-4-reductase genes. Plant Mol Biol 13: 491–502 (1989).

    Google Scholar 

  4. Burtt BL: Aspects of diversification in the capitulum. In: Heywood VH, Harborne JB, Turner BL (eds) The Biology and Chemistry of the Compositae, pp. 41–59. Academic Press, London (1977).

    Google Scholar 

  5. Coen ES, Almeida J, Robbins TP, Hudson A, Carpenter R: Molecular analysis of genes determining spatial patterns in Antirrhinum majus. In Verma DPS, Goldberg RB (eds) Temporal and Spatial Regulation of Plant Genes, pp. 63–82. Springer-Verlag, Wien (1988).

    Google Scholar 

  6. Cox KH, Goldberg RB: Analysis of plant gene expression. In: Shaw CH (ed) Plant Molecular Biology: A Practical Approach, pp. 1–34. IRL Press, Oxford (1988).

    Google Scholar 

  7. Deblaere R, Bytebier B, De Greve H, Deboeck F, Schell J, Van Montagu M, Leemans J: Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucl Acids Res 13: 4777–4788 (1985).

    Google Scholar 

  8. Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: Version II. Plant Mol Biol Rep 1: 19–21 (1983).

    Google Scholar 

  9. Drennan D, Harding J, Byrne TG: Heritability of inflorescence and floret traits in Gerbera. Euphytica 35: 319–330 (1986).

    Google Scholar 

  10. Elomaa P, Honkanen J, Puska R, Seppänen P, Helariutta Y, Mehto M, Kotilainen M, Nevalainen L, Teeri TH: Agrobacterium-mediated transfer of antisense chalcone synthase cDNA to Gerbera hybrida inhibits flower pigmentation. Bio/technology (in press).

  11. Forkmann G: Flavonoids as flower pigments: The formation of the natural spectrum and its extension by genetic engineering. Plant Breed 106: 1–26 (1991).

    Google Scholar 

  12. Hain R, Stabel P, Czernilofsky AP, Steinbiss HH, Herrera-Estrulla L, Schell J: Uptake, integration, expression and genetic transmission of a selectable chimeric gene by plant protoplasts. Mol Gen Genet 199: 161–168 (1985).

    Google Scholar 

  13. Hattori M, Yoshiyuki S: Dideoxy sequencing method using denatured plasmid templates. Anal Biochem 152: 232–238 (1986).

    Google Scholar 

  14. Henikoff S: Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28: 351–359 (1984).

    Google Scholar 

  15. Horsch RB, Fry J, Hoffmann N, Neidermeyer J, Rogers SG, Fraley RT: Leaf disc transformation. In: Gelvin SB, Schilperoort RA, Verma DPS (eds) Plant Molecular Biology Manual, pp. A5/1-A5/9. Kluwer Academic Publishers, Dordrecht (1988).

    Google Scholar 

  16. Jackson D, Roberts K, Martin C: Temporal and spatial control of expression of anthocyanin biosynthetic genes in developing flowers of Antirrhinum majus. Plant J 2: 425–434 (1992).

    Google Scholar 

  17. Jones JDG, Duinsmuir P, Bedbrook J: High level expression of introduced chimeric genes in regenerated transformed plants. EMBO J 4: 2411–2418 (1985).

    Google Scholar 

  18. Koes RE, Spelt CE, Mol JNM: The chalcone synthase multigene family of Petunia hybrida (V30): differential, light-regulated expression during flower development and UV light induction. Plant Mol Biol 12: 213–225 (1989).

    Google Scholar 

  19. Koes RE, van Blokland R, Quattrocchio F, van Tunen AJ, Mol JNM: Chalcone synthase promoters in petunia are active in pigmented and unpigmented cell types. Plant Cell 2: 379–392 (1990).

    Google Scholar 

  20. Kristiansen KN, Rohde W: Structure of the Hordeum vulgare gene encoding dihydroflavonol-4-reductase and molecular analysis of ant18 mutants blocked in flavonoid synthesis. Mol Gen Genet 230: 49–59 (1991).

    Google Scholar 

  21. Leppik EE: The evolution of capitulum types of the Compositae in the light of insect-flower interaction. In: Heywood VH, Harborne JB, Turner BL (eds) The Biology and Chemistry of the Compositae, pp. 61–89. Academic Press, London (1977).

    Google Scholar 

  22. Lutcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA: Selection of AUG initiation codon differs in plants and animals. EMBO J 6: 43–48 (1987).

    Google Scholar 

  23. Martin C, Carpenter R, Sommer H, Saedler H, Coen ES: Molecular analysis of instability in flower pigmentation of A. majus, following isolation of the pallida locus by transposition tagging. EMBO J 4: 1625–1630 (1985).

    Google Scholar 

  24. Martin C, Prescott A, Mackay S, Bartlett J, Vrijlandt E: Control of anthocyanin biosynthesis in flowers of Antirrhinum majus. Plant J 1: 37–49 (1991).

    Google Scholar 

  25. Meyer P, Heidmann I, Forkmann G, Saedler H: A new petunia flower colour generated by transformation of a mutant with a maize gene. Nature 330: 677–678 (1987).

    Google Scholar 

  26. O'Reilly C, Shepherd NS, Pereira A, Schwarz-Sommer Z, Bertram I, Robertson DS, Peterson PA, Saedler H: Molecular cloning of the a1 locus of Z. mays using the transposable elements En and Mu1. EMBO J 4: 877–882 (1985).

    Google Scholar 

  27. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press. Cold Spring Harbor (NY) (1989).

    Google Scholar 

  28. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  29. Schwarz-Sommer Z, Shepherd N, Tacke E, Gierl A, Rohde W, Leclercq L, Mattes M, Berndtgen R, Peterson PA, Saedler H: Influence of transposable elements on the structure and function of the A1 gene of Zea mays. EMBO J 6: 287–294 (1987).

    Google Scholar 

  30. van Tunen AJ, Hartman SA, Mur LA, Mol JNM: Regulation of chalcone flavanone isomerase (CHI) gene expression in Petunia hybrida: the use of alternative promoters in corolla anthers and pollen. Plant Mol Biol 12: 539–551 (1988).

    Google Scholar 

  31. van Tunen AJ, Mur LA, Brouns GS, Rienstra J-D, Koes RE, Mol JNM: Pollen- and anther-specific chi promoters from petunia: tandem promoter regulation of the chiA gene. Plant Cell 2: 393–401 (1990).

    Google Scholar 

  32. van Tunen AJ, Mol JNM: Control of flavonoid biosynthesis and manipulation of flower colour. In: Grierson D (ed) Plant Biotechnology series, pp. 94–130. Blackie and Son, Limited, Glasgow (1991).

    Google Scholar 

  33. Weiss D, Halevy AH: Stamens and gibberellin in the regulation of corolla pigmentation and growth in Petunia hybrida. Planta 179: 89–96 (1989).

    Google Scholar 

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Helariutta, Y., Elomaa, P., Kotilainen, M. et al. Cloning of cDNA coding for dihydroflavonol-4-reductase (DFR) and characterization of dfr expression in the corollas of Gerbera hybrida var. Regina (Compositae). Plant Mol Biol 22, 183–193 (1993). https://doi.org/10.1007/BF00014927

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