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Rapid induction of phenylalanine ammonia-lyase and chalcone synthase mRNAs during fungus infection of soybean (Glycine max L.) roots or elicitor treatment of soybean cell cultures at the onset of phytoalexin synthesis

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Abstract

The differential regulation of the activities and amounts of mRNAs for two enzymes involved in isoflavonoid phytoalexin biosynthesis in soybean was studied during the early stages after inoculation of primary roots with zoospores from either race 1 (incompatible, host resistant) or race 3 (compatible, host susceptible) of Phytophthora megasperma f.sp. glycinea, the causal fungus of root rot disease. In the incompatible interaction, cloned cDNAs were used to demonstrate that the amounts of phenylalanine ammonia-lyase and chalcone synthase mRNAs increased rapidly at the time of penetration of fungal germ tubes into epidermal cell layers (1–2 h after inoculation) concomitant with the onset of phytoalxxin accumulation; highest levels were reached after about 7 h. In the compatible interaction, only a slight early enhancement of mRNA levels was found and no further increase occurred until about 9 h after inoculation. The time course for changes in the activity of chalcone synthase mRNA also showed major differences between the incompatible and compatible interaction. The observed kinetics for the stimulation of mRNA expression related to phytoalexin synthesis in soybean roots lends further support to the hypothesis that phytoalexin production is an early defense response in the incompatible plant-fungus interaction. The kinetics for the enhancement of mRNA expression after treatment of soybean cell suspension cultures with a glucan elicitor derived from P. megasperma cell walls was similar to that measured during the early stages of the resistant response of soybean roots.

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Abbreviations

cDNA:

copy DNA

CHS:

chalcone synthase

PAL:

phenylalanine ammonia-lyase

References

  • Ayers, A.R., Ebel, J., Finelli, F., Berger, N., Albersheim, P. (1976) Host-pathogen interactions. IX. Quantitative assays of elicitor activity and characterization of the elicitor present in the extracellular medium of cultures of Phytophthora megasperma var. sojae. Plant Physiol. 57, 751–759

    Google Scholar 

  • Bailey, J.A., Mansfield, J.W. (1982) Phytoalexins. Blackie, Glasgow

    Google Scholar 

  • Beagle-Ristaino, J.E., Rissler, J.F. (1983) Histopathology of susceptible and resistant soybean roots inoculated with zoospores of Phytophthora megasperma f.sp. glycinea. Phytopathology 73, 590–595

    Google Scholar 

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

    Google Scholar 

  • Birnboim, H.C. (1983) A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol. 100, 243–255

    Google Scholar 

  • Börner, H., Grisebach, H. (1982) Enzyme induction in soybean infected by Phytophthora megasperma f.sp. glycinea. Arch. Biochem. Biophys. 217, 65–71

    Google Scholar 

  • Bonhoff, A., Loyal, R., Ebel, J., Grisebach, H. (1986a) Race: cultivar-specific induction of enzymes related to phytoalexin biosynthesis in soybean roots following infection with Phytophthora megasperma f.sp. glycinea. Arch. Biochem. Biophys. 246, 149–154

    Google Scholar 

  • Bonhoff, A., Loyal, R., Feller, K., Ebel, J., Grisebach, H. (1986b) Further investigations of race: cultivar-specific induction of enzymes related to phytoalexin biosynthesis in soybean roots following infection with Phytophthora megasperma f.sp. glycinea. Biol. Chem. Hoppe-Seyler 367, 797–802

    Google Scholar 

  • Bonner, W.M., Laskey, R.A. (1974) A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88

    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 

  • Darvill, A.G., Albersheim, P. (1984) Phytoalexins and their elicitors — A defense against microbial infection in plants. Annu. Rev. Plant Physiol. 35, 243–275

    Google Scholar 

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

    Google Scholar 

  • Ebel, J., Schmidt, W.E., Loyal, R. (1984) Phytoalexin synthesis in soybean cells: elicitor induction of phenylalanine ammonia-lyase and chalcone synthase mRNAs and correlation with phytoalexin accumulation. Arch. Biochem. Biophys. 232, 240–248

    Google Scholar 

  • Edwards, K., Cramer, C.L., Bolwell, G.P., Dixon, R.A., Schuch, W., Lamb, C.J. (1985) Rapid transient induction of phenylalanine ammonia-lyase mRNA in elicitor-treated bean cells. Proc. Natl. Acad. Sci. USA 82, 6731–6735

    Google Scholar 

  • Esnault, R., Chibbar, R.N., Lee, D., Van Huystee, R.B., Ward, E.W.B. (1987) Early differences in production of mRNAs for phenylalanine ammonia-lyase and chalcone synthase in resistant and susceptible cultivars of soybean inoculated with Phytophthora megasperma f.sp. Glycinea. Physiol. Mol. Plant Pathol. 30, 293–297

    Google Scholar 

  • Eye, L.L., Sneh, B., Lockwood, J.L. (1978) Factors affecting zoospore production by Phytophthora megasperma var. sojae. Phytopathology 68, 1766–1768

    Google Scholar 

  • Feinberg, A.P., Vogelstein, B. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13

    Google Scholar 

  • Groner, B., Herrlich, P., Kennedy, N., Ponta, H., Rahmsdorf, U., Hynes, N.E. (1982) Delimitation of a DNA sequence which confers inducibility by glucocorticoid hormones. J. Cell. Biochem. 20, 349–357

    Google Scholar 

  • Hahn, M.G., Bonhoff, A., Grisebach, H. (1985) Quantitative localization of the phytoalexin glyceollin I in relation to fungal hyphae in soybean roots infected with Phytophthora megasperma f.sp. glycinea. Plant Physiol. 77, 591–601

    Google Scholar 

  • Hille, A., Purwin, C., Ebel, J. (1982) Induction of enzymes of phytoalexin synthesis in cultured soybean cells by an elecitor from Phytophthora megasperma f.sp. glycinea. Plant Cell Rep. 1, 123–127

    Google Scholar 

  • Kahn, M., Kolter, R., Thomas, C., Figurski, D., Meyer, R., Remaut, E., Helinski, D.R. (1979) Plasmid cloning vehicles derived from plasmids Col E1, F, R6K and RK2. Methods Enzymol. 68, 268–280

    Google Scholar 

  • Lamb, C.J., Corbin, D.R., Lawton, M.A., Sauer, N., Wingate, V.P.M. (1986) Recognition and response in plant: pathogen interactions. In: Recognition in microbe-plant symbiotic and pathogenic interactions, pp 333–345, Lugtenberg, B., ed. Springer, Berlin

    Google Scholar 

  • Langridge, P., Pintor-Toro, J.A., Feix, G. (1982) Zein precursor mRNAs from maize endosperm. Mol. Gen. Genet. 187, 432–438

    Google Scholar 

  • Maniatis, T., Fritsch, E.F., Sambrook, J. (1982) Molecular cloning: A laboratory manual, pp. 202–203. Cold Spring Harbor, N.Y., Cold Spring Harbor Laboratory

    Google Scholar 

  • Nakamura, K., Inouye, M. (1982) Construction of versatile expression cloning vehicles using the lipoprotein of E. coli. EMBO J. 1, 771–775

    Google Scholar 

  • Palmiter, R.D. (1974) Magnesium precipitation of ribonucleo-protein complexes. Expedient techniques for the isolation of undegraded polysomes and messenger ribonucleic acid. Biochemistry 13, 3606–3615

    Google Scholar 

  • Pelham, H.R.B., Jackson, R.J. (1976) An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem. 67, 247–256

    Google Scholar 

  • Ryder, T.B., Cramer, C.L., Bell, J.N., Robbins, M.P., Dixon, R.A., Lamb, C.J. (1984) Elicitor rapidly induces chalcone synthase mRNA in Phaseolus vulgaris cells at the onset of the phytoalexin defense response. Proc. Natl. Acad. Sci. USA 81, 5724–5728

    Google Scholar 

  • Scheel, D., Hauffe, K.D., Jahnen, W., Hahlbrock, K. (1986) Stimulation of phytoalexin formation in fungus-infected plants and elicitor-treated cell cultures of parsley. In: Recognition in microbe-plant symbiotic and pathogenic interactions, pp. 325–331, Lugtenberg, B., ed. Springer, Berlin

    Google Scholar 

  • Schmelzer, E., Börner, H., Grisebach, H., Ebel, J., Hahlbrock, K. (1984) Phytoalexin synthesis in soybean (Glycine max). Similar time courses of mRNA induction in hypocotyls infected with a fungal pathogen and in cell cultures treated with fungal elicitor. FEBS Lett. 172, 59–63

    Google Scholar 

  • Schmidt, W.E. (1986) Spezifische Bindung eines β-Glucanelicitors aus Phytophthora megasperma f.sp. glycinea an Plasma-membranen aus Sojabohne (Glycine max). Doctoral Thesis, University of Freiburg, FRG

  • Schröder, J., Heller, W., Hahlbrock, K. (1979a) Flavanone synthase: simple and rapid assay for the key enzyme of flavonoid biosynthesis. Plant Sci. Lett. 14, 281–286

    Google Scholar 

  • Schröder, J., Kreuzaler, F., Schäfer, E., Hahlbrock, K. (1979b) Concomitant induction of phenylalanine ammonia-lyase and flavanone synthase mRNAs in irradiated plant cells. J. Biol. Chem. 254, 57–65

    Google Scholar 

  • Sharp, J.K., Valent, B., Albersheim, P. (1984) Purification and partial characterization of a β-glucan fragment that elicits phytoalexin accumulation in soybean. J. Biol. Chem. 259, 11312–11320

    Google Scholar 

  • Suissa, M. (1983) Spectrophotometric quantitation of silver grains eluted from autoradiograms. Anal. Biochem. 133, 511–514

    Google Scholar 

  • Zimmermann, A., Hahlbrock, K. (1975) Light-induced changes in enzyme activities in parsley cell suspension cultures. Arch. Biochem. Biophys. 166, 54–62

    Google Scholar 

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Habereder, H., Schröder, G. & Ebel, J. Rapid induction of phenylalanine ammonia-lyase and chalcone synthase mRNAs during fungus infection of soybean (Glycine max L.) roots or elicitor treatment of soybean cell cultures at the onset of phytoalexin synthesis. Planta 177, 58–65 (1989). https://doi.org/10.1007/BF00392154

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  • DOI: https://doi.org/10.1007/BF00392154

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