Skip to main content
SpringerLink
Log in

Stress responses in alfalfa (Medicago sativa L.). 15. Characterization and expression patterns of members of a subset of the chalcone synthase multigene family

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

We have identified five different full length chalcone synthase (CHS) cDNA clones from a cDNA library produced from transcripts isolated from an elicitor-treated alfalfa cell suspension culture. Nucleotide sequence similarity between the clones varied from 88–93%. Oligonucleotides based on divergent sequences in the 5′-untranslated regions of the clones could distinguish individual genes, or groups of genes, and their corresponding transcripts. Developmentally regulated expression of the CHS transcripts was predominantly in roots and root nodules; other unidentified members of the CHS gene family are expressed in stems, leaves and nodules. One of the CHS transcripts was strongly expressed in floral tissue. All the CHS transcripts studied were induced in elicitor-treated cell suspension cultures. Transcripts were also induced in roots in response to wounding or spraying with various elicitors, and in leaves infected with Phoma medicaginis (but not in wounded leaves). The induction kinetics of CHS2 transcripts were more rapid and/or transient than those of other members of the CHS family in CuCl2-treated roots and Phoma-infected leaves. The results are discussed in terms of the evolution and functions of the CHS gene family in legumes.

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

References

  1. Akada S, Kung SD, Dube SK: The nucleotide sequence of gene 1 of the soybean chalcone synthase multigene family. Plant Mol Biol 16: 751–752 (1991).

    Google Scholar 

  2. Akada S, Kung SD, Dube SK: The nucleotide sequence of gene 3 of the soybean chalcone synthase multigene family. Nucleic Acids Res 18: 19 (1990).

    Google Scholar 

  3. Barker DG, Bianchi S, Blondon F, Dattée Y, Duc G, Essad S, Flament P, Gallusci P, Génier G, Guy P, Muel X, Tourneur J, Dénarié J, Huguet T: Medicago truncatula, a model plant for studying the molecular genetics of the Rhizobium-legume symbiosis. Plant Mol Biol Rep 8: 40–49 (1990).

    Google Scholar 

  4. Batschauer A, Ehmann B, Schäfer E: Cloning and characterization of a chalcone synthase gene from mustard and its light-dependent expression. Plant Mol Biol 16: 175–185 (1991).

    Google Scholar 

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

    Google Scholar 

  6. Brummer EC, Kochert G, Bouton JH: RFLP variation in diploid and tetraploid alfalfa. Theor Appl Genet 832: 89–96 (1991).

    Google Scholar 

  7. Chomczynski P, Sacchi N: Single step method of RNA isolation by guanidinium thiocyanate phenol chloroform extraction. Anal Biochem 162: 156–159 (1987).

    Google Scholar 

  8. Cramer CL, Edwards K, Dron M, Liang X, Dildine SL, Bolwell GP, Dixon RA, Lamb CJ, Schuch W: Phenylalanine ammonia-lyase gene organization and structure. Plant Mol Biol 12: 367–383 (1989).

    Google Scholar 

  9. Dalkin K, Jorrin J, Dixon RA: Stress responses in alfalfa (Medicago sativa L.) VII. Induction of defense related mRNAs in elicitor-treated cell suspension cultures. Physiol Mol Plant Path 37: 293–307 (1990).

    Google Scholar 

  10. Daniel S, Barz W: Elicitor-induced metabolic changes in cell cultures of chickpea (Cicer arietinum L.) cultivars resistant and susceptible to Ascochyta rabiei. II. Differential induction of chalcone-synthase-mRNA activity and analysis of in vitro-translated protein patterns. Planta 182: 279–286 (1990).

    Google Scholar 

  11. Danso SKA, Alexander M: Survival of two strains of Rhizobium in soil. Soil Sci Soc Am Proc 38: 86–89 Z (1974).

    Google Scholar 

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

    Google Scholar 

  13. Dhawale S, Souciet G, Kuhn DN: Increase of chalcone synthase mRNA in pathogen-inoculated soybeans with race-specific resistance is different in leaves and roots. Plant Physiol 91: 911–916 (1989).

    Google Scholar 

  14. Dixon RA, Choudhary AD, Dalkin K, Edwards R, Fahrendorf T, Gowri G, Harrison MJ, Lamb CJ, Loake GJ, Maxwell CA, Orr JO, Paiva NL: Molecular biology of stress-induced phenylpropanoid and isoflavonoid biosynthesis in alfalfa. In: Stafford HA, Ibrahim RK (eds) Phenolic Metabolism in Plants, pp. 91–138, Plenum Press, New York (1992).

    Google Scholar 

  15. Dixon RA, Dey PM, Lamb CJ: Phytoalexins: enzymology and molecular biology. Adv Enzymol Rel Areas Mol Biol 55: 1–136 (1983).

    Google Scholar 

  16. Epping B, Kittel M, Ruhnau B, Hemleben V: Isolation and sequence analysis of a chalcone synthase cDNA of Matthiola incana R. Br. (Brassicaceae). Plant Mol Biol 14: 1061–1063 (1990).

    Google Scholar 

  17. Estabrook EM, Sengupta-Gopalan: Differential expression of phenylalanine ammonia-lyase and chalcone synthase during soybean nodule development. Plant Cell 3: 299–308 (1991).

    Google Scholar 

  18. Feinbaum RL, Ausubel FM: Transcriptional regulation of the Arabidopsis thaliana chalcone synthase gene. Mol Cell Biol 8: 1985–1992 (1988).

    Google Scholar 

  19. Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to a high specific activity. Anal Biochem 137: 266–267 (1984).

    Google Scholar 

  20. Flood J, Khan FZ, Milton JM: The role of phytoalexins in Verticillium wilt of lucerne (Medicago sativa). Ann Appl Biol 89: 329–332 (1978).

    Google Scholar 

  21. Franken P, Niesbach-Klösgen U, Weydemann U, Maréchal-Drouard L, Saedler H, Wienand U: The duplicated chalcone synthase genes C2 and Whp (white pollen) of Zea mays are independently regulated; evidence for translational control of Whp expression by the anthocyanin intensifying gene in. EMBO J 10: 2605–2612 (1991).

    Google Scholar 

  22. Hahlbrock K, Scheel D: Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Plant Mol Biol 40: 347–369 (1989).

    Google Scholar 

  23. Harker CL, Ellis THN, Coen ES: Identification and genetic regulation of the chalcone synthase multigene family in pea. Plant Cell 2: 185–194 (1990).

    Google Scholar 

  24. Hartwig UA, Joseph CM, Phillips DA: Flavonoids released naturally from alfalfa seeds enhance growth rate of Rhizobium meliloti. Plant Physiol 95: 797–803 (1991).

    Google Scholar 

  25. Hirsch AM, Bhuvaneswari TV, Torrey JG, Bisseling T: Early nodulin genes are induced in alfalfa root outgrowths elicited by auxin transport inhibitors. Proc Natl Acad Sci USA 86: 1244–1249 (1989).

    Google Scholar 

  26. Jacobs M, Rubery PH: Naturally occurring auxin transport regulators. Science 241: 1374–1377 (1988).

    Google Scholar 

  27. Junghans H, Metzlaff M: A simple and rapid method for the preparation of total plant DNA. Bio Techniques 2: 176 (1990).

    Google Scholar 

  28. Kessman H, Choudhary AD, Dixon RA: Stress responses in alfalfa (Medicago sativa L.) V. Constitutive and elicitor induced accumulation of isoflavonoid conjugates in cell suspension cultures and protoplasts. Plant Cell Rep 9: 38–41 (1990).

    Google Scholar 

  29. Kessman H, Edwards R, Geno PW, Dixon RA: Stress responses in alfalfa (Medicago sativa L.). V. Constitutive and elicitor-induced accumulation of isoflavonoid conjugates in cell suspension cultures. Plant Physiol 94: 227–232 (1990).

    Google Scholar 

  30. 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 

  31. Koes RE, Spelt CE, Reif HJ, van den Elzen PJM, Veltkamp E, Mol JNM: Floral tissue of Petunia hybrida (V30) expresses only one member of the chalcone synthase multigene family. Nucl Acids Res 14: 5229–5239 (1986).

    Google Scholar 

  32. Koes RE, Spelt CE, Mol JNM, Gerats AGM: The chalcone synthase multigene family of Petunia hybrida (V30): sequence homology, chromosomal localization and evolutionary aspects. Plant Mol Biol 10: 375–385 (1987).

    Google Scholar 

  33. Latunde-Dada AO, Lucas JA: Involvement of the phytoalexin medicarpin in the differential response of callus lines of lucerne (Medicago sativa) to infection by Verticillium albo-atrum. Physiol Plant Path 26: 31–42 (1985).

    Google Scholar 

  34. Lois R, Hahlbrock K: Differential wound activation of members of the phenylalanine ammonia-lyase and 4-coumarate: CoA ligase gene families in various organs of parsley plants. Z Naturforsch 47c: 90–94 (1992).

    Google Scholar 

  35. Maxwell CA, Hartwig UA, Joseph CM, Phillips DA: A chalcone and two related flavonoids released from alfalfa roots induce nod genes of Rhizobium meliloti. Plant Physiol 91: 842–847 (1989).

    Google Scholar 

  36. Niesbach-Klösgen U, Barzen E, Bernhardt J, Rohde W, Schwarz-Sommer Z, Reif HJ, Wienand U, Saedler H: Chalcone synthase genes in plants: A tool to study evolutionary relationships. J Mol Evol 26: 213–225 (1987).

    Google Scholar 

  37. O'Neill SD, Tong Y, Spörlein B, Forkmann G, Yoder JI: Molecular genetic analysis of chalcone synthase in Lycopersicon esculentum and an anthocyanin-deficient mutant. Mol Gen Genet 224: 279–288 (1990).

    Google Scholar 

  38. Pierce L, McCain AH: Alfalfa sprout rot caused by Erwinia chrysanthemi. Plant Dis 71: 786–788 (1987).

    Google Scholar 

  39. Rohde W, Dörr S, Salamini F, Becker D: Structure of a chalcone synthase gene from Hordeum vulgare. Plant Mol Biol 16: 1103–1106 (1991).

    Google Scholar 

  40. Ryder TB, Hedrick SA, Bell JN, Liang X, Clouse SD, Lamb CJ: Organization and differential activation of a gene family encoding the plant defense enzyme chalcone synthase in Phaseolus vulgaris. Mol Gen Genet 210: 219–233 (1987).

    Google Scholar 

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

    Google Scholar 

  42. Schumacher H-M, Grundlach H, Fiedler F, Zenk MH: Elicitation of benzophenanthridine alkaloid synthesis in Eschscholtzia cell cultures. Plant Cell Rep 6: 410–413 (1987).

    Google Scholar 

  43. Timberlake CF, Bridle P: The anthocyanins. In: Harborne JB, Mabry TJ, Mabry H (eds) The Flavonoids, Part 1, pp 214–266. Academic Press, New York/San Francisco (1975).

    Google Scholar 

  44. Yang WC, Cremers HCJC, Hogendijk P, Katinakis P, Wijffelman CA, Franssen H, Van Kammen A, Bisseling T: In-situ localization of chalcone synthase mRNA in pea root nodule development. Plant J 2: 143–151 (1992).

    Google Scholar 

Download references

Author information

Author notes

  1. Karen Dalkin

    Present address: Department of Biotechnology Research, Pioneer Hi-Bred International, Inc., P.O. Box 38, 50131-0038, Johnston, IA, USA

Authors and Affiliations

  1. Plant Biology Division, The Samuel Roberts Noble Foundation, P.O. Box 2180, 73402, Ardmore, OK, USA

    Holger Junghans, Karen Dalkin & Richard A. Dixon

Authors
  1. Holger Junghans
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Karen Dalkin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard A. Dixon
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Junghans, H., Dalkin, K. & Dixon, R.A. Stress responses in alfalfa (Medicago sativa L.). 15. Characterization and expression patterns of members of a subset of the chalcone synthase multigene family. Plant Mol Biol 22, 239–253 (1993). https://doi.org/10.1007/BF00014932

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00014932

Key words

Search

Navigation