Skip to main content
NCBI home page
The Plant Cell logoLink to The Plant Cell
. 1994 Dec;6(12):1789–1803. doi: 10.1105/tpc.6.12.1789

The elicitor-inducible alfalfa isoflavone reductase promoter confers different patterns of developmental expression in homologous and heterologous transgenic plants.

A Oommen 1, R A Dixon 1, N L Paiva 1
PMCID: PMC160562  PMID: 7866024

Abstract

In legumes, the synthesis of infection- and elicitor-inducible antimicrobial phytoalexins occurs via the isoflavonoid branch of the phenylpropanoid pathway. To study transcriptional regulation of isoflavonoid pathway-specific genes, we have isolated the gene encoding isoflavone reductase (IFR), which is the enzyme that catalyzes the penultimate step in the synthesis of the phytoalexin medicarpin in alfalfa. Chimeric gene fusions were constructed between 765- and 436-bp promoter fragments of the IFR gene and the beta-glucuronidase reporter gene and transferred to alfalfa and tobacco by Agrobacterium-mediated transformation. Both promoter fragments conferred elicitor-mediated expression in cell suspension cultures derived from transgenic plants of both species and fungal infection-mediated expression in leaves of transgenic alfalfa. Developmental expression directed by both promoter fragments in transgenic alfalfa was observed only in the root meristem, cortex, and nodules, which is consistent with the accumulation of endogenous IFR transcripts. However, in transgenic tobacco, expression from the 765-bp promoter was observed in vegetative tissues (root meristem and cortex, inner vascular tissue of stems and petioles, leaf tips, and stem peripheries adjacent to petioles) and in reproductive tissues (stigma, placenta, base of the ovary, receptacle, seed, tapetal layer, and pollen grains), whereas the 436-bp promoter was expressed only in fruits, seed, and pollen. These data indicate that infection/elicitor inducibility of the IFR promoter in both species and developmental expression in alfalfa are determined by sequences downstream of position -436, whereas sequences between -436 and -765 confer a complex pattern of strong ectopic developmental expression in the heterologous species that lacks the isoflavonoid pathway.

Full Text

The Full Text of this article is available as a PDF (3.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Benfey P. N., Chua N. H. Regulated genes in transgenic plants. Science. 1989 Apr 14;244(4901):174–181. doi: 10.1126/science.244.4901.174. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Drews G. N., Beals T. P., Bui A. Q., Goldberg R. B. Regional and cell-specific gene expression patterns during petal development. Plant Cell. 1992 Nov;4(11):1383–1404. doi: 10.1105/tpc.4.11.1383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  7. Fritze K., Staiger D., Czaja I., Walden R., Schell J., Wing D. Developmental and UV Light Regulation of the Snapdragon Chalcone Synthase Promoter. Plant Cell. 1991 Sep;3(9):893–905. doi: 10.1105/tpc.3.9.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Harrison M. J., Lawton M. A., Lamb C. J., Dixon R. A. Characterization of a nuclear protein that binds to three elements within the silencer region of a bean chalcone synthase gene promoter. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2515–2519. doi: 10.1073/pnas.88.6.2515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jacobs M., Rubery P. H. Naturally occurring auxin transport regulators. Science. 1988 Jul 15;241(4863):346–349. doi: 10.1126/science.241.4863.346. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Junghans H., Metzlaff M. A simple and rapid method for the preparation of total plant DNA. Biotechniques. 1990 Feb;8(2):176–176. [PubMed] [Google Scholar]
  12. Kessmann H., Edwards R., Geno P. W., Dixon R. A. Stress Responses in Alfalfa (Medicago sativa L.) : V. Constitutive and Elicitor-Induced Accumulation of Isoflavonoid Conjugates in Cell Suspension Cultures. Plant Physiol. 1990 Sep;94(1):227–232. doi: 10.1104/pp.94.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Loake G. J., Faktor O., Lamb C. J., Dixon R. A. Combination of H-box [CCTACC(N)7CT] and G-box (CACGTG) cis elements is necessary for feed-forward stimulation of a chalcone synthase promoter by the phenylpropanoid-pathway intermediate p-coumaric acid. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9230–9234. doi: 10.1073/pnas.89.19.9230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lynn D. G., Chen R. H., Manning K. S., Wood H. N. The structural characterization of endogenous factors from Vinca rosea crown gall tumors that promote cell division of tobacco cells. Proc Natl Acad Sci U S A. 1987 Feb;84(3):615–619. doi: 10.1073/pnas.84.3.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mason H. S., DeWald D. B., Mullet J. E. Identification of a methyl jasmonate-responsive domain in the soybean vspB promoter. Plant Cell. 1993 Mar;5(3):241–251. doi: 10.1105/tpc.5.3.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ni W., Trelease R. N. Post-Transcriptional Regulation of Catalase Isozyme Expression in Cotton Seeds. Plant Cell. 1991 Jul;3(7):737–744. doi: 10.1105/tpc.3.7.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ohl S., Hedrick S. A., Chory J., Lamb C. J. Functional properties of a phenylalanine ammonia-lyase promoter from Arabidopsis. Plant Cell. 1990 Sep;2(9):837–848. doi: 10.1105/tpc.2.9.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Paiva N. L., Sun Y., Dixon R. A., VanEtten H. D., Hrazdina G. Molecular cloning of isoflavone reductase from pea (Pisum sativum L.): evidence for a 3R-isoflavanone intermediate in (+)-pisatin biosynthesis. Arch Biochem Biophys. 1994 Aug 1;312(2):501–510. doi: 10.1006/abbi.1994.1338. [DOI] [PubMed] [Google Scholar]
  20. Pankhurst C. E., Biggs D. R. Sensitivity of Rhizobium to selected isoflavonoids. Can J Microbiol. 1980 Apr;26(4):542–545. doi: 10.1139/m80-092. [DOI] [PubMed] [Google Scholar]
  21. Traut T. W. Do exons code for structural or functional units in proteins? Proc Natl Acad Sci U S A. 1988 May;85(9):2944–2948. doi: 10.1073/pnas.85.9.2944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES