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. 1997 Oct;115(2):437–451. doi: 10.1104/pp.115.2.437

Regulation of sesquiterpene cyclase gene expression. Characterization of an elicitor- and pathogen-inducible promoter.

S Yin 1, L Mei 1, J Newman 1, K Back 1, J Chappell 1
PMCID: PMC158501  PMID: 9342864

Abstract

The promoter for a tobacco (Nicotiana tabacum) sesquiterpene cyclase gene, a key regulatory step in sesquiterpene phytoalexin biosynthesis, has been analyzed. The EAS4 promoter was fused to the beta-glucuronidase (GUS) reporter gene, and the temporal and spatial expression patterns of GUS activity were examined in stably transformed plants and in transient expression assays using electroporated protoplasts of tobacco. No GUS activity was observed in any tissues under normal growth conditions. A low level of GUS activity was detected in wounded leaf, root, and stem tissues, whereas a much higher level was observed when these tissues were challenged with elicitors or microbial pathogens. The GUS expression pattern directed by the EAS4 promoter was identical to the induction patterns observed for the endogenous sesquiterpene cyclase genes. Neither exogenous salicylic acid nor methyl jasmonate induced GUS expression; and H2O2 induced GUS expression to only a limited extent. Although the EAS4 promoter contains cis-sequences resembling previously identified transcriptional control motifs, other cis-sequences important for quantitative and qualitative gene expression were identified by deletion and gain-of-function analyses. The EAS4 promoter differs from previously described pathogen-/elicitor-inducible promoters because it only supports inducible gene expression and directs unique spatial expression patterns.

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Selected References

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  1. A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
  2. Baillieul F., Genetet I., Kopp M., Saindrenan P., Fritig B., Kauffmann S. A new elicitor of the hypersensitive response in tobacco: a fungal glycoprotein elicits cell death, expression of defence genes, production of salicylic acid, and induction of systemic acquired resistance. Plant J. 1995 Oct;8(4):551–560. doi: 10.1046/j.1365-313x.1995.8040551.x. [DOI] [PubMed] [Google Scholar]
  3. Baranowskij N., Frohberg C., Prat S., Willmitzer L. A novel DNA binding protein with homology to Myb oncoproteins containing only one repeat can function as a transcriptional activator. EMBO J. 1994 Nov 15;13(22):5383–5392. doi: 10.1002/j.1460-2075.1994.tb06873.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biedenkapp H., Borgmeyer U., Sippel A. E., Klempnauer K. H. Viral myb oncogene encodes a sequence-specific DNA-binding activity. Nature. 1988 Oct 27;335(6193):835–837. doi: 10.1038/335835a0. [DOI] [PubMed] [Google Scholar]
  5. Blein J. P., Milat M. L., Ricci P. Responses of Cultured Tobacco Cells to Cryptogein, a Proteinaceous Elicitor from Phytophthora cryptogea: Possible Plasmalemma Involvement. Plant Physiol. 1991 Feb;95(2):486–491. doi: 10.1104/pp.95.2.486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chappell J., Nable R. Induction of sesquiterpenoid biosynthesis in tobacco cell suspension cultures by fungal elicitor. Plant Physiol. 1987 Oct;85(2):469–473. doi: 10.1104/pp.85.2.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen Z., Silva H., Klessig D. F. Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid. Science. 1993 Dec 17;262(5141):1883–1886. doi: 10.1126/science.8266079. [DOI] [PubMed] [Google Scholar]
  8. Delaney T. P., Uknes S., Vernooij B., Friedrich L., Weymann K., Negrotto D., Gaffney T., Gut-Rella M., Kessmann H., Ward E., Ryals J. A central role of salicylic Acid in plant disease resistance. Science. 1994 Nov 18;266(5188):1247–1250. doi: 10.1126/science.266.5188.1247. [DOI] [PubMed] [Google Scholar]
  9. Despres C., Subramaniam R., Matton D. P., Brisson N. The Activation of the Potato PR-10a Gene Requires the Phosphorylation of the Nuclear Factor PBF-1. Plant Cell. 1995 May;7(5):589–598. doi: 10.1105/tpc.7.5.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Devergne J. C., Bonnet P., Panabières F., Blein J. P., Ricci P. Migration of the Fungal Protein Cryptogein within Tobacco Plants. Plant Physiol. 1992 Jul;99(3):843–847. doi: 10.1104/pp.99.3.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dixon R. A., Dey P. M., Lamb C. J. Phytoalexins: enzymology and molecular biology. Adv Enzymol Relat Areas Mol Biol. 1983;55:1–136. doi: 10.1002/9780470123010.ch1. [DOI] [PubMed] [Google Scholar]
  12. Dixon R. A., Harrison M. J. Activation, structure, and organization of genes involved in microbial defense in plants. Adv Genet. 1990;28:165–234. doi: 10.1016/s0065-2660(08)60527-1. [DOI] [PubMed] [Google Scholar]
  13. Dixon R. A., Paiva N. L. Stress-Induced Phenylpropanoid Metabolism. Plant Cell. 1995 Jul;7(7):1085–1097. doi: 10.1105/tpc.7.7.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dron M., Clouse S. D., Dixon R. A., Lawton M. A., Lamb C. J. Glutathione and fungal elicitor regulation of a plant defense gene promoter in electroporated protoplasts. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6738–6742. doi: 10.1073/pnas.85.18.6738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ellard-Ivey M., Douglas C. J. Role of Jasmonates in the Elicitor- and Wound-Inducible Expression of Defense Genes in Parsley and Transgenic Tobacco. Plant Physiol. 1996 Sep;112(1):183–192. doi: 10.1104/pp.112.1.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Eyal Y., Meller Y., Lev-Yadun S., Fluhr R. A basic-type PR-1 promoter directs ethylene responsiveness, vascular and abscission zone-specific expression. Plant J. 1993 Aug;4(2):225–234. doi: 10.1046/j.1365-313x.1993.04020225.x. [DOI] [PubMed] [Google Scholar]
  17. Fromm M., Taylor L. P., Walbot V. Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824–5828. doi: 10.1073/pnas.82.17.5824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gaffney T., Friedrich L., Vernooij B., Negrotto D., Nye G., Uknes S., Ward E., Kessmann H., Ryals J. Requirement of salicylic Acid for the induction of systemic acquired resistance. Science. 1993 Aug 6;261(5122):754–756. doi: 10.1126/science.261.5122.754. [DOI] [PubMed] [Google Scholar]
  19. Gough C., Hemon P., Tronchet M., Lacomme C., Marco Y., Roby D. Developmental and pathogen-induced activation of an msr gene, str 246C, from tobacco involves multiple regulatory elements. Mol Gen Genet. 1995 May 10;247(3):323–337. doi: 10.1007/BF00293200. [DOI] [PubMed] [Google Scholar]
  20. Hahlbrock K., Lamb C. J., Purwin C., Ebel J., Fautz E., Schäfer E. Rapid Response of Suspension-cultured Parsley Cells to the Elicitor from Phytophthora megasperma var. sojae: INDUCTION OF THE ENZYMES OF GENERAL PHENYLPROPANOID METABOLISM. Plant Physiol. 1981 Apr;67(4):768–773. doi: 10.1104/pp.67.4.768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harrison M. J., Choudhary A. D., Dubery I., Lamb C. J., Dixon R. A. Stress responses in alfalfa (Medicago sativa L.). 8. Cis-elements and trans-acting factors for the quantitative expression of a bean chalcone synthase gene promoter in electroporated alfalfa protoplasts. Plant Mol Biol. 1991 May;16(5):877–890. doi: 10.1007/BF00015079. [DOI] [PubMed] [Google Scholar]
  22. Hart C. M., Nagy F., Meins F., Jr A 61 bp enhancer element of the tobacco beta-1,3-glucanase B gene interacts with one or more regulated nuclear proteins. Plant Mol Biol. 1993 Jan;21(1):121–131. doi: 10.1007/BF00039623. [DOI] [PubMed] [Google Scholar]
  23. Hildmann T., Ebneth M., Peña-Cortés H., Sánchez-Serrano J. J., Willmitzer L., Prat S. General roles of abscisic and jasmonic acids in gene activation as a result of mechanical wounding. Plant Cell. 1992 Sep;4(9):1157–1170. doi: 10.1105/tpc.4.9.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Huang H. C., He S. Y., Bauer D. W., Collmer A. The Pseudomonas syringae pv. syringae 61 hrpH product, an envelope protein required for elicitation of the hypersensitive response in plants. J Bacteriol. 1992 Nov;174(21):6878–6885. doi: 10.1128/jb.174.21.6878-6885.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lam E., Benfey P. N., Gilmartin P. M., Fang R. X., Chua N. H. Site-specific mutations alter in vitro factor binding and change promoter expression pattern in transgenic plants. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7890–7894. doi: 10.1073/pnas.86.20.7890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lamb C. J., Lawton M. A., Dron M., Dixon R. A. Signals and transduction mechanisms for activation of plant defenses against microbial attack. Cell. 1989 Jan 27;56(2):215–224. doi: 10.1016/0092-8674(89)90894-5. [DOI] [PubMed] [Google Scholar]
  28. Lawton M. A., Lamb C. J. Transcriptional activation of plant defense genes by fungal elicitor, wounding, and infection. Mol Cell Biol. 1987 Jan;7(1):335–341. doi: 10.1128/mcb.7.1.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Levine A., Tenhaken R., Dixon R., Lamb C. H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell. 1994 Nov 18;79(4):583–593. doi: 10.1016/0092-8674(94)90544-4. [DOI] [PubMed] [Google Scholar]
  30. Liang X. W., Dron M., Schmid J., Dixon R. A., Lamb C. J. Developmental and environmental regulation of a phenylalanine ammonia-lyase-beta-glucuronidase gene fusion in transgenic tobacco plants. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9284–9288. doi: 10.1073/pnas.86.23.9284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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. 1989 Jun;8(6):1641–1648. doi: 10.1002/j.1460-2075.1989.tb03554.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Malamy J., Carr J. P., Klessig D. F., Raskin I. Salicylic Acid: a likely endogenous signal in the resistance response of tobacco to viral infection. Science. 1990 Nov 16;250(4983):1002–1004. doi: 10.1126/science.250.4983.1002. [DOI] [PubMed] [Google Scholar]
  33. Mehdy M. C. Active Oxygen Species in Plant Defense against Pathogens. Plant Physiol. 1994 Jun;105(2):467–472. doi: 10.1104/pp.105.2.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Métraux J. P., Signer H., Ryals J., Ward E., Wyss-Benz M., Gaudin J., Raschdorf K., Schmid E., Blum W., Inverardi B. Increase in salicylic Acid at the onset of systemic acquired resistance in cucumber. Science. 1990 Nov 16;250(4983):1004–1006. doi: 10.1126/science.250.4983.1004. [DOI] [PubMed] [Google Scholar]
  35. Ohme-Takagi M., Shinshi H. Structure and expression of a tobacco beta-1,3-glucanase gene. Plant Mol Biol. 1990 Dec;15(6):941–946. doi: 10.1007/BF00039434. [DOI] [PubMed] [Google Scholar]
  36. Oommen A., Dixon R. A., Paiva N. L. The elicitor-inducible alfalfa isoflavone reductase promoter confers different patterns of developmental expression in homologous and heterologous transgenic plants. Plant Cell. 1994 Dec;6(12):1789–1803. doi: 10.1105/tpc.6.12.1789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Peach C., Velten J. Transgene expression variability (position effect) of CAT and GUS reporter genes driven by linked divergent T-DNA promoters. Plant Mol Biol. 1991 Jul;17(1):49–60. doi: 10.1007/BF00036805. [DOI] [PubMed] [Google Scholar]
  38. Qin X. F., Holuigue L., Horvath D. M., Chua N. H. Immediate early transcription activation by salicylic acid via the cauliflower mosaic virus as-1 element. Plant Cell. 1994 Jun;6(6):863–874. doi: 10.1105/tpc.6.6.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Reinbothe S., Mollenhauer B., Reinbothe C. JIPs and RIPs: the regulation of plant gene expression by jasmonates in response to environmental cues and pathogens. Plant Cell. 1994 Sep;6(9):1197–1209. doi: 10.1105/tpc.6.9.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ricci P., Bonnet P., Huet J. C., Sallantin M., Beauvais-Cante F., Bruneteau M., Billard V., Michel G., Pernollet J. C. Structure and activity of proteins from pathogenic fungi Phytophthora eliciting necrosis and acquired resistance in tobacco. Eur J Biochem. 1989 Aug 15;183(3):555–563. doi: 10.1111/j.1432-1033.1989.tb21084.x. [DOI] [PubMed] [Google Scholar]
  41. Roby D., Broglie K., Gaynor J., Broglie R. Regulation of a chitinase gene promoter by ethylene and elicitors in bean protoplasts. Plant Physiol. 1991 Sep;97(1):433–439. doi: 10.1104/pp.97.1.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ryals J., Lawton K. A., Delaney T. P., Friedrich L., Kessmann H., Neuenschwander U., Uknes S., Vernooij B., Weymann K. Signal transduction in systemic acquired resistance. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4202–4205. doi: 10.1073/pnas.92.10.4202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ryals J., Uknes S., Ward E. Systemic Acquired Resistance. Plant Physiol. 1994 Apr;104(4):1109–1112. doi: 10.1104/pp.104.4.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Schardl C. L., Byrd A. D., Benzion G., Altschuler M. A., Hildebrand D. F., Hunt A. G. Design and construction of a versatile system for the expression of foreign genes in plants. Gene. 1987;61(1):1–11. doi: 10.1016/0378-1119(87)90359-3. [DOI] [PubMed] [Google Scholar]
  45. Shen Q., Zhang P., Ho T. H. Modular nature of abscisic acid (ABA) response complexes: composite promoter units that are necessary and sufficient for ABA induction of gene expression in barley. Plant Cell. 1996 Jul;8(7):1107–1119. doi: 10.1105/tpc.8.7.1107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Shinshi H., Usami S., Ohme-Takagi M. Identification of an ethylene-responsive region in the promoter of a tobacco class I chitinase gene. Plant Mol Biol. 1995 Mar;27(5):923–932. doi: 10.1007/BF00037020. [DOI] [PubMed] [Google Scholar]
  47. Tenhaken R., Levine A., Brisson L. F., Dixon R. A., Lamb C. Function of the oxidative burst in hypersensitive disease resistance. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4158–4163. doi: 10.1073/pnas.92.10.4158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Uknes S., Dincher S., Friedrich L., Negrotto D., Williams S., Thompson-Taylor H., Potter S., Ward E., Ryals J. Regulation of pathogenesis-related protein-1a gene expression in tobacco. Plant Cell. 1993 Feb;5(2):159–169. doi: 10.1105/tpc.5.2.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Vögeli U., Chappell J. Induction of sesquiterpene cyclase and suppression of squalene synthetase activities in plant cell cultures treated with fungal elicitor. Plant Physiol. 1988 Dec;88(4):1291–1296. doi: 10.1104/pp.88.4.1291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Vögeli U., Chappell J. Regulation of a sesquiterpene cyclase in cellulase-treated tobacco cell suspension cultures. Plant Physiol. 1990 Dec;94(4):1860–1866. doi: 10.1104/pp.94.4.1860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Vögeli U., Freeman J. W., Chappell J. Purification and characterization of an inducible sesquiterpene cyclase from elicitor-treated tobacco cell suspension cultures. Plant Physiol. 1990 May;93(1):182–187. doi: 10.1104/pp.93.1.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Ward E. R., Uknes S. J., Williams S. C., Dincher S. S., Wiederhold D. L., Alexander D. C., Ahl-Goy P., Metraux J. P., Ryals J. A. Coordinate Gene Activity in Response to Agents That Induce Systemic Acquired Resistance. Plant Cell. 1991 Oct;3(10):1085–1094. doi: 10.1105/tpc.3.10.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Weisshaar B., Armstrong G. A., Block A., da Costa e Silva O., Hahlbrock K. Light-inducible and constitutively expressed DNA-binding proteins recognizing a plant promoter element with functional relevance in light responsiveness. EMBO J. 1991 Jul;10(7):1777–1786. doi: 10.1002/j.1460-2075.1991.tb07702.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Williams M. E., Foster R., Chua N. H. Sequences flanking the hexameric G-box core CACGTG affect the specificity of protein binding. Plant Cell. 1992 Apr;4(4):485–496. doi: 10.1105/tpc.4.4.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Yu L. M., Lamb C. J., Dixon R. A. Purification and biochemical characterization of proteins which bind to the H-box cis-element implicated in transcriptional activation of plant defense genes. Plant J. 1993 Jun;3(6):805–816. doi: 10.1111/j.1365-313x.1993.00805.x. [DOI] [PubMed] [Google Scholar]
  56. Zhu Q., Chappell J., Hedrick S. A., Lamb C. Accurate in vitro transcription from circularized plasmid templates by plant whole cell extracts. Plant J. 1995 Jun;7(6):1021–1030. doi: 10.1046/j.1365-313x.1995.07061021.x. [DOI] [PubMed] [Google Scholar]
  57. da Costa e Silva O., Klein L., Schmelzer E., Trezzini G. F., Hahlbrock K. BPF-1, a pathogen-induced DNA-binding protein involved in the plant defense response. Plant J. 1993 Jul;4(1):125–135. doi: 10.1046/j.1365-313x.1993.04010125.x. [DOI] [PubMed] [Google Scholar]
  58. van de Löcht U., Meier I., Hahlbrock K., Somssich I. E. A 125 bp promoter fragment is sufficient for strong elicitor-mediated gene activation in parsley. EMBO J. 1990 Sep;9(9):2945–2950. doi: 10.1002/j.1460-2075.1990.tb07486.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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