Skip to main content
SpringerLink
Log in

Phenylalanine ammonia-lyase and chalcone synthase in glands ofPrimula kewensis (W. Wats): immunofluorescence and immunogold localization

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS) were localized by indirect immunofluorescence and immunogold labeling in glands ofPrimula kewensis. Both enzymes were exclusively present in the head cells of the glands. Phenylalanine ammonialyase was located in the regions of the dense tubular endoplasmic reticulum and occasionally found in more or less spherical organelles that have not yet been identified. Furthermore, an appreciable proportion of the enzyme protein was associated with the plasmalemma and the cell wall of the head cell. In contrast, the occurrence of CHS was restricted to the spherical, unidentified cell compartments. Our findings indicate that the gland cells have the potential for flavonoid biosynthesis. When a mutant ofP. kewensis forming structurally intact glands but incapable of farina excretion was studied, neither PAL nor CHS were found in the head cells.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

CHS:

chalcone synthase

IgG:

immunoglobulin G

PAL:

phenylalanine ammonia-lyase

References

  • Amrhein, N., Zenk, M.M. (1971) Untersuchungen zur Rolle der Phenylalanin-Ammonium-Lyase (PAL) bei der Regulation der Flavonoidsynthese in Buchweizen (Fagopyrum esculentum Moench). Z. Pflanzenphysiol.64, 145–168

    Google Scholar 

  • Asai, F., Iinuma, M., Tanaka, T., Takenaka, M., Mizuno, M. (1992) Five complex flavonoids in the farinose exudate ofPityrogramma calomelanos. Phytochemistry31, 2487–2490

    Google Scholar 

  • Bäumker, P.A., Arendt, S., Wiermann, R. (1988) Metabolism of ferulic acid sucrose esters in anthers ofTulipa cv. Apeldoorn. II. Highly specific degradation of the esters by different esterase activities. Z. Naturforsch.43c, 647–655

    Google Scholar 

  • Beerhues, L., Wiermann, R. (1988) Chalcone synthases from spinach (Spinacia oleracea L.). I. Purification, peptide patterns, and immunological properties of different forms. Planta173, 532–543

    Google Scholar 

  • Beerhues, L., Robenek, H., Wiermann, R. (1988) Chalcone synthases from spinach (Spinacia oleracea L.). II. Immunofluorescence and immunogold localization. Planta173, 544–553

    Google Scholar 

  • Bell, A.A. (1981) Biochemical mechanisms of disease resistance. Annu. Rev. Plant Physiol.32, 21–81

    Google Scholar 

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

    Google Scholar 

  • Brouillard, R. (1988) Flavonoids and flower colour. In: The flavonoids, pp. 525–538, Harborne, J.B., ed. Chapman and Hall, London

    Google Scholar 

  • Caldwell, M.M., Robberecht, R., Flint, S.D. (1983) Internal filters: Prospects for UV-acclimation in higher plants. Physiol. Plant58, 445–450

    Google Scholar 

  • Cosio, E.G., Weissenböck, G., McClure, J.W. (1985) Acifluorfeninduced isoflavonoids and enzymes of their biosynthesis in mature soybean leaves. Plant Physiol.78, 14–19

    Google Scholar 

  • Ebel, J. (1986) Phytoalexin synthesis: The biochemical analysis of the induction process. Annu. Rev. Phytopathol.24, 235–264

    Google Scholar 

  • Fahn, A. (1988) Secretory tissues in vascular plants. New Phytol.108, 229–257

    Google Scholar 

  • Frens, G. (1973) Controlled nucleation for the regulation of the particle size in mono disperse gold suspensions. Nature (London) Phys. Sci.241, 20–22

    Google Scholar 

  • Gershenzon, J., Croteau, R. (1990) Regulation of monoterpene biosynthesis in higher plants. Rec. Adv. Phytochem.24, 99–160

    Google Scholar 

  • Gershenzon, J., McCaskill, D., Rajaonarivony, J.I.M., Mihaliak, C., Karp, F., Croteau, R. (1992) Isolation of secretory cells from plant glandular trichomes and their use in biosynthetic studies of monoterpenes and other gland products. Anal. Biochem.200, 130–138

    Google Scholar 

  • Gleitz, J., Scitz, H.U. (1989) Induction of chalcone synthase in cell suspension cultures of carrot (Daucus carota L. ssp.sativus) by ultraviolet light: evidence for two different forms of chalcone synthase. Planta179, 323–330

    Google Scholar 

  • Greenaway, W., English, S., Whatley, F.R. (1990) Phenolic composition of bud exudates ofPopulus deltoides. Z. Naturforsch.45c, 587–593

    Google Scholar 

  • Greenaway, W., English, S., May, J., Whatley, F.R. (1991) Analysis of phenolics of bud exudate ofPopulus sieboldii by GC-MS. Phytochemistry30, 3005–3008

    Google Scholar 

  • Hansel, R. (1962) Flavonoide-Endausgestaltung und Verteilung über das Pflanzensystem. Planta Med.10, 361–384

    Google Scholar 

  • Hanson, K.R., Havir, E.A. (1981) Phenylalanine ammonia-lyase. In: The biochemistry of plants, vol. 7: Secondary plant products, pp. 577–625, Stumpf, P.K., Conn, E.E., eds. Academic Press, New York

    Google Scholar 

  • Harborne, J.B. (1968) Comparative biochemistry of the flavonoids — VII: Correlations between flavonoid pigmentation and systematics in the family Primulaceae. Phytochemistry7, 1215–1230

    Google Scholar 

  • Harborne, J.B. (1981) Variation in and functional significance of phenolic conjugation in plants. Rec. Adv. Phytochem.12, 457–474

    Google Scholar 

  • Harborne, J.B. (1986) Nature, distribution and function of plant flavonoids. Prog. Clin. Biol. Res.213, 14–24

    Google Scholar 

  • Hedin, P.A., Waage, S.K. (1986) Roles of flavonoids in plant resistance to insects. Prog. Clin. Biol. Res.213, 87–100

    Google Scholar 

  • Hirsch, A.M. (1992) Developmental biology of legume nodulation. New Phytol.122, 211–237

    Google Scholar 

  • Hrazdina, G., Wagner, G.J. (1985) Metabolic pathways as enzyme complexes: Evidence for the synthesis of phenylpropanoids on membrane associated enzyme complexes. Arch. Biochem. Biophys.237, 88–100

    Google Scholar 

  • Hrazdina, G., Marx, G.A., Hoch, M.C. (1982) Distribution of secondary plant metabolites and their biosynthetic enzymes in pea leaves. Anthocyanins and flavonol glycosides. Plant Physiol.70, 745–748

    Google Scholar 

  • Jahnen, W., Hahlbrock, K. (1988) Differential regulation and tissuespecific distribution of enzymes of phenylpropanoid pathways in developing parsley seedlings. Planta173, 453–458

    Google Scholar 

  • Kandra, L., Wagner, G.J. (1988) Studies of the site and mode of biosynthesis of tobacco trichome exudate components. Arch. Biochem. Biophys.265, 425–432

    Google Scholar 

  • Karwatzki, B., Herget, A., Beerhues, L., Wiermann, R. (1993) In situ localization of chalcone synthase in tannin-containing plants. Phytochemistry32, 585–590

    Google Scholar 

  • Keene, C.K., Wagner, G.J. (1985) Direct demonstration of duvatrienediol biosynthesis in glandular heads of tobacco trichomes. Plant Physiol.79, 1026–1032

    Google Scholar 

  • Kelsley, R.G., Reynolds, G.W., Rodriguez, E. (1984) The chemistry of biologically active constituents secreted and stored in plant glandular trichomes. In: Biology and chemistry of plant trichomes, pp. 187–241. Rodriguez, E., Healey, O.L., Metha, J. eds. Plenum Press, New York

    Google Scholar 

  • Latchinian-Sadek, L., Ibrahim, R.K. (1991) Flavonol ring B-specific O-glucosyltransferases: Purification, production of polyclonal antibodies, and immunolocalization. Arch. Biochem. Biophys.289, 230–236

    Google Scholar 

  • Lembach, A., Beerhues, L., Wiermann, R. (1989) In situ localization of chalcone synthase inLarix needles by indirect immunofluorescence. Protoplasma153, 58–61

    Google Scholar 

  • McCaskill, D., Gershenzon, J., Croteau, R. (1992) Morphology and monoterpene biosynthetic capabilities of secretory cell clusters isolated from glandular trichomes of peppermint (Mentha piperita L.). Planta187, 445–454

    Google Scholar 

  • Read, S.M., Northcote, D.H. (1981) Minimization of variation in the response for different proteins of the Coomassie Blue G dye binding assay for protein. Anal. Biochem.116, 53–64

    Google Scholar 

  • Rommeswinkel, M., Karwatzki, B., Beerhues, L., Wiermann, R. (1992) Immunofluorescence localization of chalcone synthase in roots ofPisum sativum L. andPhaseolus vulgaris L. and comparable immunochemical analysis of chalcone synthase from pea leaves. Protoplasma166, 115–121

    Google Scholar 

  • Roth, J., Bendayan, J., Orci, L. (1982) The protein A-gold (pAg) technique-a qualitative and quantitative approach for antigen localization on thin sections. In: Techniques in immunochemistry, pp. 107–133, Bu'Lock, G.R., Petrisz, P., eds. Academic Press, New York London Toronto Sydney San Francisco

    Google Scholar 

  • Schachtele, C., Steup, M. (1986) α-1,4-glucan phosphorylase forms from leaves of spinach (Spinacia oleracea L.). I. In situ localization by indirect immunofluorescence. Planta167, 444–451

    Google Scholar 

  • Schnepf, E. (1972) Tubuläres endoplasmatisches Reticulum in Drüsen mit lipophilen Ausscheidungen vonFicus, Ledum undSalvia. Biochem. Physiol. Pflanzen163, 113–125

    Google Scholar 

  • Schnepf, E., Klasová, A. (1972) Zur Feinstruktur von Öl- und Flavon-Drüsen. Ber. Dtsch. Bot. Ges.85, 249–258

    Google Scholar 

  • Spring, O., Bienert, U. (1987) Capitate glandular hairs from sunflower leaves: Development, distribution and sesquiterpene lactone content. J. Plant Physiol.130, 441–448

    Google Scholar 

  • Verma, D.P.S. (1992) Signals in root nodule organogenesis and endocytosis ofRhizobium. The Plant Cell4, 373–382

    Google Scholar 

  • Völker, W., Frick, B., Robenek, H. (1985) A simple device for low temperature polymerization of Lowicryl K4M. J. Microsc.138, 91–93

    Google Scholar 

  • Voirin, B., Bayet, Ch., Colson, M. (1993) Demonstration that flavone aglycones accumulate in the peltate glands ofMentha xpiperita leaves. Phytochemistry34, 85–87

    Google Scholar 

  • Wagner, G.H. (1991) Secreting glandular trichomes: More than just hairs. Plant Physiol.96, 675–679

    Google Scholar 

  • Wagner, G.J., Hrazdina, G. (1984) Endoplasmic reticulum as a site of phenylpropanoid and flavonoid metabolism inHippeastrum. Plant Physiol.74, 901–906

    Google Scholar 

  • Wollenweber, E. (1974) Die Verbreitung spezifischer Flavone in der GattungPrimula. Biochem. Physiol. Pflanzen166, 419–424

    Google Scholar 

  • Wollenweber, E. (1989) Exkret-Flavonoide bei Blütenpflanzen und Farnen. Naturwissenschaften76, 458–463

    Google Scholar 

  • Wollenweber, E. (1990) On the distribution of exudate flavonoids among angiosperms. Rev. Latinoama. Quim.21, 115–121

    Google Scholar 

  • Wollenweber, E., Dietz, V.H. (1981) Occurrence and distribution of free flavonoid aglycones in plants. Phytochemistry20, 869–932

    Google Scholar 

  • Wollenweber, E., Jay, M. (1988) Flavones and flavonols. In: The flavonoids, pp 233–302. Harborne, J.B., ed. Chapman and Hall, London

    Google Scholar 

  • Wollenweber, E., Schnepf, E. (1970) Vergleichende Untersuchungen über die flavonoiden Exkrete von “Mehl”- und “Öl” -Drüsen bei Primeln und die Feinstruktur der Drüsenzellen, Z. Pflanzenphysiol.62, 216–227

    Google Scholar 

  • Wollen weber, E., Egger, K., Schnepf, E. (1971) Flavonoid-Aglykone in Alnus-Knospen und die Feinstruktur der Drüsenzellen. Biochem. Physiol. Pflanzen162, 193–202

    Google Scholar 

  • Wollenweber, E., Iinuma, M., Tanaka, T., Mizuno, M. (1990) 5- Hydroxy-6,2′-dimethoxyflavone fromPrimula denticulata. Phytochemistry29, 633–637

    Google Scholar 

  • Wollenweber, E., Dörr, M., Stelzer, R., Arriaga-Giner, F.J. (1992) Lipophilic phenolics from the leaves ofEmpetrum nigrum - Chemical structures and exudate localization. Bot. Acta.105, 300–305

    Google Scholar 

  • Yamaura, T., Tanaka, S., Tabata, M. (1992) Localization of the biosynthesis and accumulation of monoterpenoids in glandular trichomes of thyme. Planta Med.58, 153–158

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Botanik, Universität Münster, Schlossgarten 3, D-48149, Münster, Germany

    H. Schöpker, M. Kneisel & R. Wiermann

  2. Institut für Pharmazeutische Biologie, Universität Bonn, Nussallee 6, D-53115, Bonn, Germany

    L. Beerhues

  3. Institut für Arterioskleroseforschung, Abteilung für Zellbiologie und Ultrastrukturforschung, Universität Münster, Domagkstrasse 3, D-48149, Münster, Germany

    H. Robenek

Authors
  1. H. Schöpker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Kneisel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L. Beerhues
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H. Robenek
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R. Wiermann
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Financial support from the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie is gratefully acknowledged. We are grateful to Mrs. Karin Schlattmann and Mrs. Susanne Otter for preparing the ultrathin sections and to Mrs. Marianne Opalka for taking the photographs.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schöpker, H., Kneisel, M., Beerhues, L. et al. Phenylalanine ammonia-lyase and chalcone synthase in glands ofPrimula kewensis (W. Wats): immunofluorescence and immunogold localization. Planta 196, 712–719 (1995). https://doi.org/10.1007/BF01106765

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01106765

Key words

Search

Navigation