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The Arabidopsis thaliana 4-coumarate:CoA ligase (4CL) gene: stress and developmentally regulated expression and nucleotide sequence of its cDNA

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Abstract

An Arabidopsis cDNA clone encoding 4-coumarate:CoA ligase (4CL), a key enzyme of phenylpropanoid metabolism, was identified and sequenced. The predicted amino acid sequence is similar to those of other cloned 4CL genes. Southern blot analysis indicated that 4CL is single-copy gene in Arabidopsis. Northern blots showed that 4CL expression was activated early during seedling development. The onset of 4CL expression was correlated with the onset of lignin deposition in cotyledons and roots 2–3 days after germination. The timing of the expression of a parsley 4CL1-GUS fusion in transgenic Arabidopsis seedlings was examined in parallel and was very similar to that of endogenous 4CL. In mature plants, highest 4CL expression was observed in bolting stems, where relatively large amounts of lignin accumulate. Both 4CL and 4CL1-GUS mRNA accumulation was strongly and transiently activated by wounding of mature Arabidopsis leaves. 4CL expression was specifically activated within 6 h after infiltration of Arabidopsis ecotype Columbia leaves with a Pseudomonas syringae pv. maculicola strain harboring the bacterial avirulence gene avrB, which causes in incompatible interaction. The timing of 4CL activation was identical to the previously observed activation of PAL gene expression in this interaction. No activation of 4CL expression was observed in a compatible interaction caused by a Pseudomonas syringae pv. maculicola strain without avrB.

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References

  1. Allina SM, Douglas CJ: Isolation and characterization of the 4-coumarate:CoA ligase gene family in a poplar hybrid. Plant Physiol 105 (Suppl): 154 (1994).

    Google Scholar 

  2. Becker-André M, Schultze-Lefert P, Hahlbrock K: Structural comparison, modes of expression, and putative cis-acting elements of the two 4-coumarate:CoA ligase genes in potato. J Biol Chem 266: 8551–8559 (1991).

    Google Scholar 

  3. Bevan M, Schufflebottom D, Edwards K, Jefferson R, Schuch W: Tissue- and cell-specific activity of a phenylalanine ammonia-lyase promoter in transgenic plants. EMBO J 8: 1899–1906 (1989).

    Google Scholar 

  4. Bisgrove SR, Simonich MT, Smith NM, Sattler A, Innes RW: A disease resistance gene in Arabidopsis with specificity for two different pathogen avirulence genes. Plant Cell 6: 927–933 (1994).

    Google Scholar 

  5. daCosta e Silva O, Klein L, Schmelzer E, Trezzini GF, Hahlbrock K: BPF-1, a pathogen-induced DNA-binding protein involved in the plant defense response. Plant J 4: 125–135 (1993).

    Google Scholar 

  6. Debener T, Lehnackers H, Arnold M, Dangl JL: Identification and molecular mapping of a single Arabidopsis thaliana locos determining resistance to a phytopathogenic Pseudomonas syringae isolate. Plant J 1: 289–302 (1991).

    Google Scholar 

  7. Dharmawardhana DP, Ellis BE, Carlson JE: Characterization of vascular lignification in Arabidopsis thaliana. Can J Bot 70: 2238–2244 (1992).

    Google Scholar 

  8. Dong X, Mindrinos M, Davis K, Ausubel F: Induction of Arabidopsis defense genes by virulent and avirulent Pseudomonas syringae strains and by a cloned avirulence gene. Plant Cell 3: 61–72 (1991).

    Google Scholar 

  9. Douglas CJ, Hauffe KD, Ites-Morales ME, Ellard M, Paszkowski U, Hahlbrock K, Dangl JL. Exonic sequences are required for elicitor and light activation of a plant defense gene, but promoter sequences are sufficient for tissue specific expression. EMBO J 10: 1767–1775 (1991).

    Google Scholar 

  10. Doyle JJ, Doyle JL: Isolation of plant DNA from fresh tissue. Focus 12: 13–15 (1990).

    Google Scholar 

  11. Feinbaum RL, Ausubel FM: Transcriptional regulation of the Arabidopsis thaliana chalcone synthase gene. Mol Cell Biol 8: 1885–1892 (1988).

    Google Scholar 

  12. Fritzemeier K-H, Cretin C, Kombrink E, Rohwer F, Taylor J, Scheel D, Hahlbrock K: Transient induction of phenylalanine ammonia-lyase and 4-coumarate:CoA ligase mRNAs in potato leaves infected with virulent or avirulent races of Phytophthora infestans. Plant Physiol 85: 34–41 (1987).

    Google Scholar 

  13. Grand C, Boudet A, Boudet AM: Isoenzymes of hydroxycinnamate: CoA ligase prom poplar stems properties and tissue distribution. Planta 158: 225–229 (1983).

    Google Scholar 

  14. Hahlbrock K, Scheel D: Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Plant Mol Biol 40: 347–469 (1989).

    Google Scholar 

  15. Hauffe KD, Paszkowski U, Schulzelefert P, Hahlbrock K, Dangl JL, Douglas CJ: A parsley 4CL-1 promoter fragment specifies complex expression patterns in transgenic tobacco. Plant Cell 3: 435–443 (1991).

    Google Scholar 

  16. Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).

    Google Scholar 

  17. Jorgensen A, Cuellar E, Thompson F: Modes and tempos in the evolution of nuclear encoded ribosomal RNA genes in legumes. Carnegie Inst of Washington Year Book 81: 98–101 (1982).

    Google Scholar 

  18. Joshi CP: An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucl Acids Res 15: 6643–6653 (1987).

    Google Scholar 

  19. Keen NT, Buzzell R: New disease resistance genes in soybean against Pseudomonas syringae pv. glycinea: evidence that one of them interacts with a bacterial elicitor. Theor Appl Genet 81: 133–138 (1991).

    Google Scholar 

  20. Keen NT, Tamaki S, Kobayashi D, Trollinger DJ: Improved broad host range plasmids for DNA cloning in gram-negative bacteria. Gene 70: 191–197 (1988).

    Google Scholar 

  21. Kiedrowski S, Kawalleck P, Hahlbrock K, Somssich IE, Dangl JL: Rapid activation of a novel plant defense gene is strictly dependent on the Arabidopsis RPM1 disease resistance locus. EMBO J 11: 4677–4684 (1992).

    Google Scholar 

  22. Knobloch K-H, Hahlbrock K: Isoenzymes of p-coumarate: CoA ligase from suspension cultures of Glycine max. Eur J Biochem 52: 311–320 (1975).

    Google Scholar 

  23. Kozak M: Point mutations define a sequence flanking the AUG initiatior codon that modulates translation by eukaryotic ribosomes. Cell 44: 283–292 (1986).

    Google Scholar 

  24. Kubasek WL, Shirley BW, McKillop A, Goodman HM, Briggs W, Ausubel FM: Regulation of flavonoid biosynthesis genes in germinating Arabidopsis seedlings. Plant Cell 4: 1229–1236 (1992).

    Google Scholar 

  25. Lamb CJ, Lawton MA, Dron M, Dixon RA: Signals and tranduction mechanisms for activation of plant defenses against microbial attack. Cell 56: 215–224 (1989).

    Google Scholar 

  26. Liang X, Dron M, Schmid J, Dixon RA, Lamb CJ: Developmental and environmental regulation of a phenylalanine ammonia-lyase beta-glucuronidase gene fusion in transgenic tobacco plants. Proc Natl Acad Sci USA 86: 9284–9288 (1989).

    Google Scholar 

  27. Logemann J, Schell J, Willmitzer L: Improved method for the isolation of RNA from plant tissues. Anal Biochem 163: 16–20 (1987).

    Google Scholar 

  28. 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 

  29. 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 C 47: 90–94 (1992).

    Google Scholar 

  30. Lozoya E, Hoffmann H, Douglas CJ, Schulz W, Scheel D, Hahlbrock K: Primary structure and catalytic properties of isoenzymes encoded by the two 4-coumarate:CoA ligase genes in parsley. Eur J Biochem 176: 661–667 (1989).

    Google Scholar 

  31. Mindrinos M, Katagiri F, Yu G-L, Ausubel FM: The A. thaliana disease resistance gene RPS2 encodes a nucleotide-binding site and leucine-rich repeats. Cell 78: 1089–1099 (1994).

    Google Scholar 

  32. Moniz de Sá M, Subramaniam R, Williams FE, Douglas CJ: Rapid activation of phenylpropanoid metabolism in elicitor-treated hybrid poplar (Populus trichocarpa Torr and Gray × Populus deltoides Marsh) suspension-cultured cells. Plant Physiol 98: 728–737 (1992).

    Google Scholar 

  33. Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).

    Google Scholar 

  34. Ohl S, Hedrick SA, Chory J, Lamb CJ: Functional properties of a phenylalanine ammonia-lyase promoter from Arabidopsis. Plant Cell 2: 237–248 (1990).

    Google Scholar 

  35. Orr JD, Lynn DG: Biosynthesis of dehydrodiconiferyl alcohol glucosides: implications for the control of tobacco cell growth. Plant Physiol 98: 343–352 (1992).

    Google Scholar 

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

    Google Scholar 

  37. Schmelzer E, Kruger-Lebus S, Hahlbrock K: Temporal and spatial patterns of gene expression around sites of attempted fungal infection in parsley leaves. Plant Cell 1: 993–1001 (1989).

    Google Scholar 

  38. Schmid J, Doerner PW, Clouse SD, Dixon RA, Lamb CJ: Developmental and environmental regulation of a bean chalcone synthase promoter in transgenic tobacco. Plant Cell 2: 619–631 (1990).

    Google Scholar 

  39. Sharma YK, Davbis KR: Ozone-induced expression of stress-related genes in Arabidopsis thaliana. Plant Physiol 105: 1089–1096 (1994).

    Google Scholar 

  40. Subramaniam R, Reinold S, Molitor EK, Douglas CJ: Structure, inheritance, and expression of hybrid poplar (Populus trichocarpa × Populus deltoides) phenylalanine ammonia-lyase genes. Plant Physiol 102: 71–83 (1993).

    Google Scholar 

  41. Trezzini GF, Horrichs A, Somssich IE: Isolation of putative defense-related genes from Arabidopsis thaliana and expression in fungal elicitor-treated cells. Plant Mol Biol 21: 385–389 (1993).

    Google Scholar 

  42. Uhlmann A, Ebel J: Molecular cloning and expression of 4-coumarate:coenzyme A ligase, an enzyme involved in the resistance of soybean (Glycine max L.) against pathogen infection. Plant Physiol 102: 1147–1156 (1993).

    Google Scholar 

  43. Valvekens D, VanMontagu M, VanLijsebettens M: Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85: 5536–5540 (1988).

    Google Scholar 

  44. Wanner LA, Mittal S, Davis KR: Recognition of the avirulence gene avrB from Pseudomonas syringae pv. glycinea by Arabidopsis thaliana. Mol Plant-Microbe Interact 6: 582–591 (1993).

    Google Scholar 

  45. Wanner LE, Li G, Ware DH, Somssich I, Davis KR: The phenylalanine ammonia-lyase gene family in Arabidopsis thaliana. Plant Mol Biol 27: 327–338 (1995).

    Google Scholar 

  46. Wu SC, Hahlbrock K: In situ localization of phenylpropanoid-related gene expression in different tissues of light-grown and dark-grown parsley seedlings. Z Naturforsch C 47: 591–600 (1992).

    Google Scholar 

  47. Zhao Y, Kung SD, Dube SK: Nucleotide sequence of rice 4-coumarate:CoA ligase gene, 4-CL.1. Nucl Acids Res 18: 6144 (1990).

    Google Scholar 

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Author notes

  1. Mary Ellard

    Present address: Department of Botany and Plant Pathology, Oregon State University, 97331-2902, Corvallis, OR, USA

  2. Leslie A. Wanner

    Present address: Institute for Biology and Geology, Drammsveien 201, N-9037, Tromsø, Norway

Authors and Affiliations

  1. Department of Botany, University of British Columbia, V6T 1Z4, Vancouver, Canada

    Diana Lee, Mary Ellard & Carl J. Douglas

  2. Ohio State Plant Biotechnology Center, The Ohio State University, 43210, Columbus, OH, USA

    Leslie A. Wanner & Keith R. Davis

  3. Department of Plant Biology, The Ohio State University, 43210, Columbus, OH, USA

    Leslie A. Wanner & Keith R. Davis

Authors
  1. Diana Lee
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  3. Leslie A. Wanner
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  4. Keith R. Davis
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  5. Carl J. Douglas
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Lee, D., Ellard, M., Wanner, L.A. et al. The Arabidopsis thaliana 4-coumarate:CoA ligase (4CL) gene: stress and developmentally regulated expression and nucleotide sequence of its cDNA. Plant Mol Biol 28, 871–884 (1995). https://doi.org/10.1007/BF00042072

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  • DOI: https://doi.org/10.1007/BF00042072

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