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Organization and differential activation of a gene family encoding the plant defense enzyme chalcone synthase in Phaseolus vulgaris

Molecular and General Genetics MGG volume 210, pages 219–233 (1987)Cite this article

Summary

Chalcone synthase (CHS) catalyzes the first and key regulatory step in the branch pathway of phenylpropanoid biosynthesis specific for synthesis of ubiquitous flavonoid pigments and UV protectants. In bean (Phaseolus vulgaris L.) and other members of the Leguminoseae, chalcone synthase is also involved in the synthesis of the isoflavonoid-derived phytoalexin antibiotics characteristic of this family. We have demonstrated that the haploid genome of bean contains a family of about six to eight CHS genes, some of which are tightly clustered. Treatment of bean cells with fungal elicitor activates several of these genes leading to the accumulation of at least five and probably as many as nine distinct CHS transcripts encoding a set of CHS isopolypeptides of Mr 42–43 kDa but with differing pI in the range pH 6–7. In elicited cells specific transcripts and encoded polypeptides are differentially induced with respect to both the extent and kinetics of accumulation. Wounding or infection of hypocotyl tissue also activates several CHS genes with marked differences in the pattern of accumulation of specific transcripts and encoded polypeptides in wounded compared to infected tissue or elicited cells, indicating operation of more than one cue for defense gene activation. Illumination induces accumulation of a different set of CHS transcripts including only one of the set hitherto demonstrated to be induced by biological stress. The organization and differential regulation of the CHS gene family in bean are discussed in relation to the functions of this enzyme in adaptative and protective responses to diverse enviromental stresses.

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References

  • Anderson-Prouty AJ, Albersheim P (1975) Host-pathogen interactions. VII. Isolation of a pathogen-synthesized glucan that elicits a defense response in the pathogen's host. Plant Physiol 56:286–291

    Google Scholar 

  • Bailey JA, Deverall BJ (1971) Formation and activity of phaseollin in the interaction between bean (Phaseolus vulgaris) hypocotyls and physiological races of Colletotrichum lindemuthianum. Physiol Plant Pathol 1:435–449

    Google Scholar 

  • Bell EA (1981) The physiological role(s) of secondary (natural) products. In: Conn EE (ed) The biochemistry of plants: A comprehensive treatise. Vol 7, Secondary plant products. Academic Press, New York, pp 1–19

    Google Scholar 

  • Bell JN, Dixon RA, Bailey JA, Rowell PM, Lamb CJ (1984) Differential induction of chalcone synthase mRNA activity at the onset of phytoalexin accumulation in compatible and incompatible plant: pathogen interactions. Proc Natl Acad Sci USA 81:3384–3388

    Google Scholar 

  • Bell JN, Ryder TB, Wingate VPM, Bailey JA, Lamb CJ (1986) Differential accumulation of plant defense gene transcripts in a compatible and an incompatible plant: pathogen interaction. Mol Cell Biol 6:1615–1623

    Google Scholar 

  • Bolton GW, Nester EW, Gordon MP (1986) Plant phenolic compounds induce expression of the Agrobacterium tumefaciens loci needed for virulence. Science 232:983–985

    Google Scholar 

  • Bolwell GP, Bell JN, Cramer CL, Schuch W, Lamb CJ, Dixon RA (1985) l-Phenylalanine ammonia-lyase from Phaseolus vulgaris: Characterisation and differential induction of multiple forms from elicitor-treated cell suspension cultures. Eur J Biochem 149:411–419

    Google Scholar 

  • Bolwell GP, Sap J, Cramer CL, Lamb CJ, Schuch W, Dixon RA (1986) l-Phenylalanine ammonia-lyase from Phaseolus vulgaris: Partial degradation of enzyme subunits in vitro and in vivo. Biochim Biophys Acta 881:210–221

    Google Scholar 

  • Chappell J, Hahlbrock K (1984) Transcription of plant defense genes in response to UV light or fungal elicitor. Nature 311:76–78

    Google Scholar 

  • Coruzzi G, Broglie R, Edwards C, Chua N-H (1984) Tissue-specific and light-regulated expression of a pea nuclear gene encoding the small subunit of ribulose-1,5-bisphosphate carboxylase. EMBO J 3:1671–1679

    Google Scholar 

  • Cramer CL, Ryder TB, Bell JN, Lamb CJ (1985) Rapid switching of plant gene expression by fungal elicitor. Science 227:1240–1243

    Google Scholar 

  • Davis R, Botstein D, Roth JR (1981) Advanced bacterial genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 140–141

    Google Scholar 

  • Dean C, van den Elzen P, Tamaki S, Dunsmuir P, Bedbrook J (1985a) Linkage and homology analysis divides the eight genes for the small subunit of petunia ribulose 1,5-bisphosphate carboxylase into three gene families. Proc Natl Acad Sci USA 82:4964–4968

    Google Scholar 

  • Dean C, van den Elzen P, Tamaki S, Dunsmuir P, Bedbrook J (1985b) Differential expression of the eight genes of the petunia ribulose bisphosphate carboxylase small subunit multi-gene family. EMBO J 4:3055–3061

    Google Scholar 

  • Dixon RA, Bendall DS (1978) Changes in the levels of enzymes of phenylpropanoid and flavonoid synthesis during phaseollin production in cell suspension cultures of Phaseolus vulgaris. Physiol Plant Pathol 13:295–306

    Google Scholar 

  • Dixon RA, Dey PM, Lamb CJ (1983) Phytoalexins: Enzymology and molecular biology. Adv Enzymol 55:1–135

    Google Scholar 

  • Firmin JL, Wilson KE, Rossen L, Johnston AWB (1986) Flavonoid activation of nodulation genes in Rhizobium reversed by other compounds present in plants. Nature 324:90–92

    Google Scholar 

  • Fyrberg EA, Mahaffey JW, Bond BJ, Davidson N (1983) Transcripts of the six Drosophila actin genes accumulate in a stage- and tissue-specific manner. Cell 33:115–123

    Google Scholar 

  • Grab D, Loyal R, Ebel J (1985) Elicitor-induced phytoalexin synthesis in soybean cells: changes in the activity of chalcone synthase mRNA and the total population of translatable mRNA. Arch Biochem Biophys 243:523–529

    Google Scholar 

  • Haffner MH, Chin MB, Lane BG (1978) Wheat embryo ribonucleates. XII. Formal characterization of terminal and penultimate nucleoside residues at the 5′-ends of ‘capped’ RNA from imbibing wheat embryos. Can J Biochem 56:729–733

    Google Scholar 

  • Hahlbrock K, Grisebach H (1979) Enzymic controls in the biosynthesis of lignin and flavonoids. Annu Rev Plant Physiol 30:105–130

    Google Scholar 

  • Hanahan D (1983) Studies on the transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    Google Scholar 

  • Heller W, Hahlbrock K (1980) Highly purified “flavanone synthase” from parsley catalyzes the formation of naringenin chalcone. Arch Biochem Biophys 200:617–619

    Google Scholar 

  • Hightower RC, Meagher RB (1985) Divergence and differential expression of soybean actin genes. EMBO J 4:1–8

    Google Scholar 

  • Kreuzaler F, Ragg H, Fautz E, Kuhn DN, Hahlbrock K (1983) UV-induction of chalcone synthase mRNA in cell suspension cultures of Petroselinum hortense. Proc Natl Acad Sci USA 80:2591–2593

    Google Scholar 

  • Lawton MA, Lamb CJ (1987) Transcriptional activation of plant defense genes by fungal elicitor, wounding and infection. Mol Cell Biol 7:335–341

    Google Scholar 

  • Lawton MA, Dixon RA, Lamb CJ (1980) Elicitor modulation of the turnover of l-phenylalanine ammonia-lyase in French bean cell suspension cultures. Biochim Biophys Acta 633:162–175

    Google Scholar 

  • Lawton MA, Dixon RA, Hahlbrock K, Lamb CJ (1983a) Rapid induction of the synthesis of phenylalanine ammonia-lyase and chalcone synthase in elicitor-treated bean cells. Eur J Biochem 129:593–601

    Google Scholar 

  • Lawton MA, Dixon RA, Hahlbrock K, Lamb CJ (1983b) 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

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base specific chemical cleavages. Methods Enzymol 65:499–560

    Google Scholar 

  • McMaster GK, Carmichael GG (1977) Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci USA 74:4835–4839

    Google Scholar 

  • Mehdy MC, Lamb CJ (1987) Chalcone isomerase cDNA cloning and mRNA induction by fungal elicitor, wounding and infection. EMBO J 6:1527–1533

    Google Scholar 

  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12:7035–7056

    Google Scholar 

  • Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325

    Google Scholar 

  • Peters NK, Frost JW, Long SR (1986) A plant flavone, luteolin, induces expression of Rhizobium meliloti nodulation genes. Science 233:977–980

    Google Scholar 

  • Redmond JW, Batley M, Djordjevic MA, Innes RW, Kuempel PL, Rolfe BG (1986) Flavones induce expression of the nodulation genes in Rhizobium. Nature 323:632–635

    Google Scholar 

  • Reif HJ, Niesbach U, Deumling B, Saedler H (1985) Cloning and analysis of two genes for chalcone synthase from Petunia hybrida. Mol Gen Genet 199:208–215

    Google Scholar 

  • Reimold U, Kröger M, Kreuzaler F, Hahlbrock K (1983) Coding and 3′ non-coding nucleotide sequence of chalcone synthase mRNA and assignment of amino acid sequence of the enzyme. EMBO J 2:1801–1805

    Google Scholar 

  • Ryder TB, Cramer CL, Bell JN, Robbins MP, Dixon RA, Lamb CJ (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 

  • Schröder J, Betz B, Hahlbrock K (1976) Light-induced enzyme synthesis in cell suspension cultures of Petroselinum hortense. Demonstration in a heterologous cell-free system of rapid changes in the rate of phenylalanine ammonia-lyase synthesis. Eur J Biochem 67:527–541

    Google Scholar 

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

    Google Scholar 

  • Sommer H, Saedler H (1986) Structure of the chalcone synthase gene of Antirrhinum majus. Mol Gen Genet 202:429–434

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Stachel SE, Messens E, Van Montagu M, Zambryski P (1985) Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318:624–629

    Google Scholar 

  • Talbot DR, Adang MJ, Slightom JL, Hall TC (1984) Size and organization of a multigene family encoding phaseolin, the major seed storage protein of Phaseolus vulgaris L. Mol Gen Genet 198:42–49

    Google Scholar 

  • Thomas PA (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 82:6551–6555

    Google Scholar 

  • Wienand U, Weydemann U, Niesbach-Klösgen U, Peterson PA, Saedler H (1986) Molecular cloning of the c2 locus of Zea mays, the gene coding for chalcone synthese. Mol Gen Genet 203:202–207

    Google Scholar 

  • Wiermann R (1981) Secondary plant products and cell and tissue differentiation. In: Conn EE (ed) The biochemistry of plants: A comprehensive treatise. Vol 7, Secondary plant products. Academic Press, New York, pp 86–117

    Google Scholar 

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Author notes

  1. Thomas B. Ryder

    Present address: Genprobe, 9880 Campus Point Drive, 92121, San Diego, CA, USA

  2. John N. Bell

    Present address: Dept. of Embryology, Carnegie Institute of Washington, 115 W. University Parkway, 21210, Baltimore, MD, USA

Affiliations

  1. Plant Biology Laboratory, Salk Institute for Biological Studies, P.O. Box 85800, 92138, San Diego, CA, USA

    Thomas B. Ryder, Susan A. Hedrick, John N. Bell, Xaiowu Liang, Steven D. Clouse & Christopher J. Lamb

  2. Graduate Program in Biology, University of California San Diego, 92093, La Jolla, CA, USA

    Xaiowu Liang

Authors
  1. Thomas B. Ryder
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  2. Susan A. Hedrick
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  3. John N. Bell
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  4. Xaiowu Liang
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  5. Steven D. Clouse
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  6. Christopher J. Lamb
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Communicated by H. Saedler

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Ryder, T.B., Hedrick, S.A., Bell, J.N. et al. Organization and differential activation of a gene family encoding the plant defense enzyme chalcone synthase in Phaseolus vulgaris . Molec Gen Genet 210, 219–233 (1987). https://doi.org/10.1007/BF00325687

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  • DOI: https://doi.org/10.1007/BF00325687

Key words

  • Chalcone synthase
  • Multigene family
  • cDNA sequence
  • Environmental regulation
  • Phaseolus vulgaris