Skip to main content
Log in

Characterization and expression of chalcone synthase in different genotypes of Matthiola incana R.Br. during flower development

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The expression of the key enzyme of flavonoid biosynthesis, chalcone synthase (CHS), has been followed in different genotypes of Matthiola incana R.Br. (Brassicaceae) which are genetically defined with respect to anthocyanin production. Enzyme activity was determined by a radioactive assay in crude flower extracts. The amount of enzyme protein in the developing flower was determined by use of SDS-PAGE, protein blotting, reaction with an antiserum against CHS of parsley (Petroselinum hortense), and PAP staining. The molecular weight of about 41 500 of the CHS subunits corresponds with that obtained from other higher plants. Steps of flower development were subdivided into stages-1,0, I–IV. During flower development of a Matthiola line with coloured petals (line 07) a defined pattern of CHS enzyme production can be observed: At the stage of bud opening (stage 0–I) a dramatic increase of the amount of CHS enzyme prodein in the petals occurs. This is quite different from results obtained with petals of the white flowering mutant line 18 bearing a genetic defect in the gene f coding for CHS. Here a reduced and nearly constant level of CHS enzyme protein can be observed during flower development. This line is most attractive for our studies of the regulation of enzyme synthesis because under stress conditions a slight colouring of the flower petals occurs, which is uniformly distributed and line-specific. This suggests that we are dealing with a CHS mutant producing a rather inactive enzyme protein at a low level. This protein may regain enzyme activity under certain environmental conditions. Preliminary investigations suggest a rather high level of CHS mRNA transcription at the bud opening stage of the flowers. Other white flowering mutant lines, line 17 (genotype ee) and line 19 (gg) with a late block in the flavonoid biosynthesis pathway, exhibit a concomitant reduction of CHS enzyme activity and protein content in comparison to anthocyanin-producing lines with the f+f+e+e+g+g+-genotype.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Dangelmayr B, Stotz G, Spribille R, Forkmann G: Relationship between flower development, anthocyanin accumulation and activity of enzymes involved in flavonoid biosynthesis in Matthiola incana R. Br. Z. Naturforsch. 38c:551–555, 1983.

    Google Scholar 

  2. Forkmann G: Precursors and genetic control of anthocyanin synthesis in Matthiola incana R. Br. Planta 137:159–163, 1977.

    Google Scholar 

  3. Grisebach H, Barz W: Biochemie der Flavonoide. Naturwiss 56:538–544, 1969.

    Google Scholar 

  4. Hahlbrock K: Flavonoids. In: Stumpf PK, Conn EE (eds), The biochemistry of plants. Vol. 7, Acad. Press, New York, 1981, pp 425–456.

    Google Scholar 

  5. Hahlbrock K, Grisebach H: Enzymatic controls in the biosynthesis of lignin and flavonoids. Ann Rev Plant Physiol 30:105–130, 1979.

    Google Scholar 

  6. Heller W, Hahlbrock K: Highly purified ‘flavanone synthase’ from parsley catalyzes the formation of naringenin chalcone. Arch Biochem Biophys 200:617–619, 1980.

    Google Scholar 

  7. Hemleben V, Frey M, Rall S, Koch M, Kittel M, Kreuzaler F, Ragg H, Fautz E, Hahlbrock K: Studies on the chalcone synthase gene of two higher plants: Petroselinum hortense and Matthiola incana. In: Burger MM, Weber R (eds), Embryonic development B: Cellular aspects: Alan R. Liss, New York, 1982, pp 555–566.

    Google Scholar 

  8. Hille A, Purwin C, Ebel J: Induction of enzymes of phytoalexin synthesis in cultured soybean cells by an elicitor from Phytophtora megasperma f. sp. glycinea. Plant Cell Reports 1:123–127, 1982.

    Google Scholar 

  9. Kreuzaler F, Hahlbrock K: Enzymatic synthesis of aromatic compounds in higher plants: Formation of naringenin (5,7,4′-trihydroxyflavanone) from p-coumaroyl coenzyme A and malonyl coenzyme A. FEBS Letters 28:69–72, 1972.

    Google Scholar 

  10. Kreuzaler F, Hahlbrock K: Enzymatic synthesis of an aromatic ring from acetate units. Partial purification and some properties of flavanone synthase from cell-suspension cultures of petroselinum hortense. Eur J Biochem 56:205–213, 1975.

    Google Scholar 

  11. Kreuzaler F, Ragg H, Heller W, Tesch R, Witt I, Hammer D, Hahlbrock K: Flavanone synthase from Petroselinum hortense. Molecular weight, subunit composition, size of messenger RNA, and absence of pantetheinyl residue. Eur J Biochem 99:89–96, 1979.

    Google Scholar 

  12. Kreuzaler F, Ragg H, Fautz E, Kuhn DN, Hahlbrock K: UV-induction of chalcone synthase mRNA in cell suspension cultures of Petroselinum hortense. Proc Natl Acad Sci USA 81:2591–2593, 1983.

    Google Scholar 

  13. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970.

    Google Scholar 

  14. Lawton MA, Dixon RA, Hahlbrock K, Lamb C: Elicitor induction of mRNA activity. Rapid effects of elicitor on phenylalanine ammonia-lyase and chalcone synthase mRNA activities in bean cells. Eur J Biochem 130:131–139, 1983.

    Google Scholar 

  15. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275, 1951.

    Google Scholar 

  16. Mol JNM, Schram AW, deVlaming P, Gerats AGM, Kreuzaler F, Hahlbrock K, Reif HJ, Veltkamp E: Regulation of flavonoid gene expression in Petunia hybrida: Description and partial characterization of a conditional mutant in chalcone synthase gene expression. Mol Gen Genet 192:424–429, 1983.

    Google Scholar 

  17. Rall S: Charakterisierung von Anthocyanen und den Enzymen Phenylalanin-Ammonium-Lyase und Chalkonsynthase in verschiedenen Entwicklungsstadien definierter Genotypen von Matthiola incana R Br Dissertation Tübingen, 1983.

  18. Reimold U, Krüger M, Kreuzaler F, Hahlbrock K: Coding and 3′ non-coding nucleotide sequence of chalcone synthase mRNA and assignment of amino acid sequence of the enzyme. The EMBO Journal 2:1801–1805, 1983.

    Google Scholar 

  19. Ritter-Thomas U: Beitrag zur Kenntnis der Inhaltsstoffe von Matthiola incana. Planta 114:259–268, 1973.

    Google Scholar 

  20. Schleif RJ, Wensink PC: Working with proteins. In: Practical methods in molecular biology. Springer-Verlag, New York, 1981, pp 61–88.

    Google Scholar 

  21. Schröder J, Schäfer E: Radioiodinated antibodies, a tool in studies on the presence and role of inactive enzyme forms: Regulation of chalcone synthase in parsley cell suspension cultures. Arch Biochem Biophys 203:800–808, 1980.

    Google Scholar 

  22. Seyffert W: Simulation of quantitative characters by genes with biochemically definable action. II. The material. Theor Appl Genet 41:285–291, 1971.

    Google Scholar 

  23. Seyffert W: Beiträge zur Genetik und Enzymologie der Flavonoide. Biol Zbl 101:465–483, 1982.

    Google Scholar 

  24. Spribille R: Nachweis und Charakterisierung der Chalkonsynthase unter Verwendung definierter Genotypen höherer Pflanzen. Dissertation Tübingen, 1982.

  25. Spribille R, Forkmann G: Genetic control of chalcone synthase activity in flowers of Matthiola incana R Br Z Naturforsch 36c:619–624, 1981.

    Google Scholar 

  26. Sternberger LA: The unlabeled antibody peroxidase-antiperoxidase (PAP) method. In: Immunocytochemistry. 2nd edn. John Wiley & Sons, New York, 1979, pp 104–169.

    Google Scholar 

  27. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354, 1979.

    Google Scholar 

  28. Wienand U, Sommer H, Schwarz Z, Shepherd N, Saedler H, Kreuzaler F, Ragg H, Fautz E, Hahlbrock K, Harrison B, Peterson PA: A general method to identify plant structural genes among genomic DNA clones using transposable element induced mutations. Mol Gen Genet 187:195–202, 1982.

    Google Scholar 

  29. Wiermann R: Secondary plant products and cell and tissue differentiation. In: Stumpf PK, Conn EE (eds) The biochemistry of plants. Vol. 7, Acad Press, New York, 1981, pp 85–116.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rall, S., Hemleben, V. Characterization and expression of chalcone synthase in different genotypes of Matthiola incana R.Br. during flower development. Plant Mol Biol 3, 137–145 (1984). https://doi.org/10.1007/BF00016061

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00016061

Keywords

Navigation