Plant Molecular Biology

, Volume 24, Issue 5, pp 743–755

Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.)

  • Francesca Sparvoli
  • Cathie Martin
  • Attilio Scienza
  • Giuseppe Gavazzi
  • Chiara Tonelli
Research Articles

Abstract

Genes involved in flavonoid and stilbene biosynthesis were isolated from grape (Vitis vinifera L.). Clones coding for phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydoxylase (F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX) and UDP glucose:flavonoid 3-O-glucosyl transferase (UFGT), were isolated by screening a cDNA library, obtained from mRNA from seedlings grown in light for 48 h using snapdragon (Antirrhinum majus) and maize heterologous probes. A cDNA clone coding for stilbene synthase (StSy) was isolated by probing the library with a specific oligonucleotide. These clones were sequenced and when the putative products were compared to the published amino acid sequence for corresponding enzymes, the percentages of similarity ranged from 65% (UFGT) to 90% (CHS and PAL). The analysis of the genomic organization and expression of these genes in response to light shows that PAL and StSy genes belong to large multigene families, while the others are present in one to four copies per haploid genome. The steady-state level of mRNAs encoded by the flavonoid biosynthetic genes as determined in young seedlings is coordinately induced by light, except for PAL and StSy, which appear to be constitutively expressed.

Key words

anthocyanins cDNA cloning flavonoids gene expression genomic organization stilbenes Vitis vinifera

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References

  1. 1.
    Akada S, Kung SD, Dube SK: The nucleotide sequence of gene 1 of soybean chalcone synthase multigene family. Plant Mol Biol 16: 751–752 (1991).Google Scholar
  2. 2.
    Beld M, Martin C, Huits H, Stuitje AR, Gerats AGM: Flavonoid synthesis in Petunia hybrida: partial characterisation of dihydroflavonol-4-reductase genes. Plant Mol Biol 13: 491–502 (1989).Google Scholar
  3. 3.
    Blyden ER, Doerner PW, Lamb CJ, Dixon RA: Sequence analysis of a chalcone isomerase cDNA of Phaseolus vulgaris L.. Plant Mol Biol 16: 167–169 (1991).Google Scholar
  4. 4.
    Britsch L, Ruhnau-Brich B, Forkmann G: Molecular cloning, sequence analysis, and in vitro expression of flavanone 3β-hydroxilase from Petunia hybrida. J Biol Chem 267: 5380–5387 (1992).Google Scholar
  5. 5.
    Chandler VL, Radicella PJ, Robbins TP, Chen J, Turks D: Two regulatory genes of the maize anthocyanin pathway are homologous: isolation of B utilizing R genomic sequences. Plant Cell 1: 1175–1183 (1989).Google Scholar
  6. 6.
    Cone KC, Burr FA, Burr B: Molecular analysis of the maize anthocyanin regulatory locus C1. Proc Natl Acad Sci USA 83: 9631–9635 (1986).Google Scholar
  7. 7.
    Consonni G, Geuna F, Gavazzi G, Tonelli C: Molecular homology among members of the R gene family in maize. Plant J 3: 335–346 (1993).Google Scholar
  8. 8.
    Dalkin K, Edwards R, Edington B, Dixon RA: Stress responses in alfalfa (Medicago sativa L.). 1. induction of phenylpropanoid biosynthesis and hydrolytic enzymes in elicitor treated cell suspension cultures. Plant Physiol 92: 440–446 (1989).Google Scholar
  9. 9.
    Dangl JL: Regulatory elements controlling developmental and stress induced expression of phenylpropanoid genes. In: Boller T, Meins F (eds) Plant Gene Research, vol 8: Genes Involved in Plant Defense, pp. 225–286. Springer-Verlag. New York (1991).Google Scholar
  10. 10.
    Davies KM: A Malus cDNA with homology to the Antirrhinum Candica and Zea A2 genes. Plant Physiol 103: 1015 (1993).Google Scholar
  11. 11.
    Dellaporta SL, Greenblatt I, Kermicle JL, Hicks JB, Wessler SR: Molecular cloning of the maize R-nj allele by transposon tagging with Ac. In: Gustafson JP, Appels R (eds) Chromosome Structure and Function; 18th Stadler Genetics Symposium, pp. 263–282. Plenum, New York (1988).Google Scholar
  12. 12.
    Derks W, Creasy LL: The significance of stilbene phytoalexins in the Plasmopara viticola-grapevine interaction. Physiol Mol Plant Pathol 34: 189–202 (1989).Google Scholar
  13. 13.
    Devereaux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).Google Scholar
  14. 14.
    Dixon RA: The phytoalexins response: elicitation, signalling and control of host gene expression. Biol Rev 61: 239–291 (1986).Google Scholar
  15. 15.
    Dooner HK, Robbins TP, Jorgensen RA: Genetic and developmental control of anthocyanin biosynthesis. Ann Rev Genet 25: 173–199 (1991).Google Scholar
  16. 16.
    Ebel J, Hahlbrock K: Biosynthesis. In: Harborne JB, Marby TJ (eds) The Flavonoids, pp 641–680. Chapman and Hall, London (1982).Google Scholar
  17. 17.
    Edwards K, Cramer CL, Bolwell GP, Dixon RA, Schulch W, Lamb CJ: Rapid transient induction of phenylalanine ammonia-lyase mRNA in elicitor-treated bean cells. Proc Natl Acad Sci USA 82: 6731–6735 (1985).Google Scholar
  18. 18.
    Ehmann B, Schaefer E: Nucleotide sequences encoding two different chalcone synthases expressed in cotyledons of SAN 9789 treated mustard (Sinapsis alba L.). Plant Mol Biol 11: 869–870 (1988).Google Scholar
  19. 19.
    Fliegmann J, Schroeder G, Schanz S, Britsch L, Schroeder J: Molecular analysis of chalcone and dihydropinosylvin synthase from Scots pine (Pinus sylvestris), and differential regulation of these and related enzyme activities in stressed plants. Plant Mol Biol 18: 489–503 (1992).Google Scholar
  20. 20.
    Frank RL, Vodkin LO: Sequence and structure of a phenylalanine ammonia-lyase gene from Glycine max. DNA Seq 1: 335–346 (1991).Google Scholar
  21. 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. 22.
    Fritzmeier KH, Rolfs CH, Pfau J, Kindl H: Action of ultraviolet-C on stilbene formation in callus of Arachis hypogaea. Planta 159: 25–29 (1983).Google Scholar
  23. 23.
    Fritzmeier KH, Kindl H: Coordinate induction by UV light of stilbene synthase, phenylalanine ammonia-lyase and cinnamate 4-hydroxylase in leaves of Vitaceae. Planta 151: 48–52 (1981).Google Scholar
  24. 24.
    Furtek D, Schiefelbein JW, Johnston F, Nelson OEJr: Sequence comparison of three wild-type Bronze-1 allele from Zea mays. Plant Mol Biol 11: 473–482 (1988).Google Scholar
  25. 25.
    Goodrich J, Carpenter R, Coen ES: A common gene regulates pigmentation pattern in diverse plant species. Cell 68: 955–964 (1992).Google Scholar
  26. 26.
    Gowri G, Paiva NL, Dixon RA: Stress responses in alfalfa (Medicago sativa L.) sequence analysis of phenylalanine ammonia-lyase (PAL) cDNA clones and appearance of PAL transcripts in elicitor-treated cell cultures and developing plants. Plant Mol Biol 17: 415–429 (1991).Google Scholar
  27. 27.
    Hahlbrock K, Scheele D: Physiology and molecular biology of phenylpropanoid metabolism. Ann Rev Plant Physiol Plant Mol Biol 40: 347–364 (1989).Google Scholar
  28. 28.
    Hain R, Reif HJ, Krause E, Langebartels R, Kindl H, Vornam B, Wiese W, Schmelzer E, Schreier PH, Stöcker RH, Stenzel K: Disease resistance results from foreign phytoalexin expression in a novel plant. Nature 361: 153–156 (1993).Google Scholar
  29. 29.
    Jackson D, Roberts K, Martin CR: Temporal and spatial control of expression of anthocyanin biosynthetic genes in developing flowers of Antirrhinum majus. Plant J 2: 425–434 (1992).Google Scholar
  30. 30.
    Joos HJ, Hahlbrock H: Phenylalanine ammonia-lyase in potato (Solanum tuberosum L.): genomic complexity, structural comparison of two selected genes and modes of expression. Eur J Biochem 204: 621–629 (1992).Google Scholar
  31. 31.
    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: 159–169 (1987).Google Scholar
  32. 32.
    Koes RE, Spelt CE, van den Elzen PJM, Mol JNM: Cloning and molecular characterization of the chalcone synthase multigene family of Petunia. Gene 81: 245–257 (1989).Google Scholar
  33. 33.
    Kristiansen KN, Rohde W: Structure of the Hordeum vulgare gene encoding dihydroflavonol-4-reductase and molecular analysis of ant 18 mutants blocked in flavonoid synthesis. Mol Gen Genet 230: 49–59 (1991).Google Scholar
  34. 34.
    Kubasek WL, Shirley BW, McKillop A, Goodman HM, Briggs W, Ausubel FM: Regulation of flavonoid biosynthetic genes in germinating Arabidopsis seedlings. Plant Cell 4: 1229–1236 (1992).Google Scholar
  35. 35.
    Lamb CJ, Lawton MA, Dron M, Dixon RA: Signals and tansduction mechanisms for activation of plant defenses against microbial attack. Cell 56: 215–225 (1989).Google Scholar
  36. 36.
    Langcake P: Disease resistance of Vitis spp. and the production of the stress metabolites resveratrol, ε-viniferin, α-viniferin and pterostilbene. Physiol Plant Pathol 18: 213–226 (1981).Google Scholar
  37. 37.
    Lawton MA, Lamb CJ: Transcriptional activation of plant defense genes by fungal elicitor, wounding and infection. Mol Cell Biol 7: 335–341 (1981).Google Scholar
  38. 38.
    Liswidowati MF, Hohomann F, Burkhardt S, Kindl H: Induction of stilbene synthase by Botrytis cinerea in cultured grapevine cells. Planta 18: 307–314 (1991).Google Scholar
  39. 39.
    Lois R, Dietrich A, Hahlbrock K, Schulz W: A phenylalanine ammonia-lyase gene from parsley: structure, regulation and identification of elicitor and light responsive cis-acting elements. EMBO J 8: 1641–1648 (1989).Google Scholar
  40. 40.
    Long S: Rhizobium-legume nodulation: life together in the underground. Cell 3: 829–840. (1989).Google Scholar
  41. 41.
    Ludwig SR, Habera LF, Dellaporta SL Wessler SR: Lc, a member of the maize R gene family responsible for tissue-specific anthocyanin production encodes a protein similar to transcriptional activators and contains the myc-homology region. Proc Natl Acad Sci USA 86: 7092–7096 (1989).Google Scholar
  42. 42.
    Martin CR, Gerats AGM: The control of flower colouration. In: Jordan B (ed) The Molecular Biology of Flowering, pp. 219–255. CAB Int., Wallingford, Oxford (1991).Google Scholar
  43. 43.
    Martin CR, Prescott A, Mackay S, Bartlett J, Vrijlandt E: Control of anthocyanin biosynthesis in flowers of Antirrhinum majus. Plant J 1: 37–49 (1991).Google Scholar
  44. 44.
    Mattivi F, Scienza A, Failla O, Villa PL, Anzani R, Tedesco G, Gianazza E, Righetti PG: Vitis vinifera: a chemotaxonomic approach-antocianyns in the skin. 5th International Symposium on Grape Breeding, Geilweilerhof, Vitis special issue (1989).Google Scholar
  45. 45.
    Melchior F, Kindl H: Grapevine stilbene synthase cDNA only slightly differing from chalcone synthase cDNA is expressed in Escherichia coli into catalytically active enzyme. FEBS Lett 268: 17–20 (1990).Google Scholar
  46. 46.
    Meldgaard M: Expression of chalcone synthase, dihydroflavonol reductase and flavanone-3-hydroxylase in mutants of barley deficient in anthocyanin and proantocyanidin biosynthesis. Theor Appl Genet 83: 695–706 (1992).Google Scholar
  47. 47.
    Menssen A, Hohmann WM, Schnable PS, Peterson PA, Saedler H: The En/Spm transposable element of Zea mays contains splice sites at the termini generating a novel intron from dSpm element in the A2 gene. EMBO J 9: 3051–3057 (1990).Google Scholar
  48. 48.
    Mereghetti M, Tonelli C, Gavazzi G: Developmental expression of light inducible genes of the R family in immature seeds of maize. Maydica 36: 337–342 (1991).Google Scholar
  49. 49.
    Minami E, Ozeki Y, Matsuoka M, Koizuka N, Tanaka Y: Structure and some characterization of the gene for phenylalanine ammonia-lyase from rice plants. Eur J Biochem 185: 19–25 (1989).Google Scholar
  50. 50.
    Paz-Ares J, Ghosal D, Wienand U, Peterson PA, Saedler H: The regulatory C1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. EMBO J 6: 3553–3558 (1987).Google Scholar
  51. 51.
    Prescott AG: A dilemma of dioxygenase (or where biochemistry and molecular biology fail to meet). J Exp Bot 44: 849–861 (1993).Google Scholar
  52. 52.
    Quattrocchio F, Wing JF, Leppen HTC, Mol JNM, Koes RE: Regulatory genes controlling anthocyanin pigmentation are functionally conserved among plant species and have distinct sets of target genes. Plant Cell 5: 1497–1512 (1993).Google Scholar
  53. 53.
    Ralston EJ, English JJ, Dooner HK: Sequence of three bronze alleles of maize and correlation with the genetic fine structure. Genetics 119: 185–197 (1988).Google Scholar
  54. 54.
    Robbins MP, Bollwell GP, Dixon RA: Metabolic changes in elicitor treated bean cells: selectivity of enzyme induction in relation to phytolalexins accumulation. Eur J Biochem 148: 563–569 (1985).Google Scholar
  55. 55.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).Google Scholar
  56. 56.
    Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).Google Scholar
  57. 57.
    Schröeder G, Brown JWS, Schröeder J: Molecular analysis of resveratrol synthase. cDNA, genomic clones and relationship with chalcone synthase. Eur J Biochem 172: 161–169 (1988).Google Scholar
  58. 58.
    Schulz W, Eiben H-G, Hahlbrock K: Expression in Escherichia coli of catalytically active phenylalanine ammonia-lyase from parsley. FEBS Lett 258: 335–338 (1989).Google Scholar
  59. 59.
    Schwarz-Sommer Z, Shepherd N, Tacke E, Gierl A, Rohde W, Leclercq L, Mattes M, Berndtgen R, Peterson PA, Saedler H: Influence of transposable elements on the structure and function of A1 gene of Zea mays. EMBO J 6: 287–294 (1987).Google Scholar
  60. 60.
    Sommer H, Saedler H: Structure of the chalcone synthase gene of Antirrhinum majus. Mol Gen Genet 202: 429–434 (1986).Google Scholar
  61. 61.
    Styles DE, Ceska O: The genetic control of flavonoid synthesis in maize. Can J Genet Cytol 19: 289–302 (1977).Google Scholar
  62. 62.
    Taylor LP, Briggs WR: Genetic regulation and photocontrol of anthocyanin accumulation in maize seedlings. Plant Cell 2: 115–127 (1990).Google Scholar
  63. 63.
    Tonelli C, Consonni G, Faccio Dolfini S, Dellaporta SL, Viotti A, Gavazzi G: Genetic and molecular analysis of Sn, a light-inducible, tissue specific regulatory gene in maize. Mol Gen Genet 225: 401–410 (1991).Google Scholar
  64. 64.
    van Tunen AJ, Koes RE, Spelt CE, van der Krol AR, Stuitje AR, Mol JNM: Cloning of the two chalcone flavanone isomerase genes from Petunia hybrida: coordinate, light-regulated and differential expression of flavonoid genes. EMBO J 7: 1257–1263 (1988).Google Scholar
  65. 65.
    van Tunen AJ, Hartman SA, Mur LA, Mol JNM: Regulation of chalone flavanone isomerase (CHI) gene expression in Petunia hybrida: the use of alternative promoters in corolla anthers and pollen. Plant Mol Biol 12: 539–551 (1989).Google Scholar
  66. 66.
    Wienand U, Weydemann U, Niesbach-Kloesgen U, Peterson PA, Saedler H: Molecular cloning of C2 locus of Zea mays, the gene coding for chalcone synthase. Mol Gen Genet 203: 202–207 (1986).Google Scholar
  67. 67.
    Weiss D, van der Huit AH, Kroon JTM, Mol JNM, Kooter JM. The Petunia homologue of the Antirrhinum candi and Zea mays A2 flavonoid genes; homology to flavanone 3-hydroxylase and ethylene forming enzyme. Plant Mol Biol 22: 893–897 (1993).Google Scholar
  68. 68.
    Wise RP, Rohde W, Salamini F: Nucleotide sequence of the Bronze-1 homologous gene from Hordeum vulgare. Plant Mol Biol 14: 277–279 (1990).Google Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Francesca Sparvoli
    • 1
  • Cathie Martin
    • 3
  • Attilio Scienza
    • 2
  • Giuseppe Gavazzi
    • 1
  • Chiara Tonelli
    • 1
  1. 1.Dipartmento di Genetica e di Biologia dei MicroorganismiUniversità degli Studi di MilanoMilanoItalia
  2. 2.Istituto di Coltivazioni ArboreeUniversità degli Studi di MilanoMilanoItalia
  3. 3.Department of GeneticsJohn Innes InstituteNorwichUK

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