Skip to main content

A complex ensemble of cis-regulatory elements controls the expression of a Vicia faba non-storage seed protein gene


We have identified cis-regulatory elements within the 5′-upstream region of a Vicia faba non-storage seed protein gene, called usp, by studying the expression of usp-promoter deletion fragments fused to reporter genes in transgenic tobacco seeds. 0.4 kb of usp upstream sequence contain at least six, but probably more, distinct cis-regulatory elements which are responsible for seemingly all quantitative, spatial and temporal aspects of expression. Expression-increasing and-decreasing elements are interspersed and include an AT-rich sequence, a G-box element and a CATGCATG motif. The latter acts as a negative element in contrast to what has been found for the same motif in legumin-and vicilin-type seed storage protein gene promoters. Seed specificity of expression is mainly determined by the −68/+51 region which confers, however, only very low levels of expression. The data support the combinatiorial model of promoter function.

This is a preview of subscription content, access via your institution.


  1. 1.

    An G, Watson BD, Stachel S, Gordon MP, Nester EW: New cloning vehicles for transformation of higher plants. EMBO J 4: 277–284 (1985).

    Google Scholar 

  2. 2.

    Bassüner R, Manteuffel R, Müntz K, Püchel M, Schmidt P, Weber E: Analysis of in vivo and in vitro globulin formation during cotyledon development of field beans (Vicia faba L. var. minor). Biochem Physiol Pflanzen 178: 665–684 (1983).

    Google Scholar 

  3. 3.

    Bassüner R, Bäumlein H, Huth A, Jung R, Wobus U, Rapoport TA, Saalbach G, Müntz K: Abundant embryonic mRNA in field beans (Vicia faba L.) codes for a new class of seed protiens: cDNA cloning and characterization of the primary translation product. Plant Mol Biol 11: 321–334 (1988).

    Google Scholar 

  4. 4.

    Bäumlein H, Müller AJ, Schiemann J, Helbing D, Manteuffel R, Wobus U: A legumin B gene of Vicia faba is expressed in developing seeds of transgenic tobacco. Biol Zbl 106: 569–575 (1987).

    Google Scholar 

  5. 5.

    Bäumlein H, Boerjan W, Nagy I, Bassüner R, VanMontagu M, Inze D, Wobus U: A novel seed protein gene from Vicia faba is developmentally regulated in transgenic tobacco and Arabidopsis plants. Mol Gen Genet 225: 459–467 (1991).

    Google Scholar 

  6. 6.

    Bäumlein H, Boerjan W, Nagy I, Panitz R, Inze D, Wobus U: Upstream sequences regulating legumin gene expression in heterologous transgenic plants. Mol Gen Genet 225: 121–128 (1991).

    Google Scholar 

  7. 7.

    Bäumlein H, Nagy I, Villarroel R, Inze D, Wobus U: Cis-analysis of a seed protein gene promoter: the conservative RY repeat CATGCATG within the legumin box is essential for tissue-specific expression of a legumin gene. Plant J 2: 233–239 (1992).

    Google Scholar 

  8. 8.

    Benfey PN, Chua NH: The cauliflower mosaic virus 35 S promoter: combinatorial regulation of transcription in plants. Science 250: 959–966 (1990).

    Google Scholar 

  9. 9.

    Bradford MM: A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).

    Google Scholar 

  10. 10.

    Bustos MM, Guiltinan MJ, Jordan J, Begum D, Kalkan FA, Hall TC. Regulation of β-glucuronidase expression in transgenic tobacco plants by an A/T rich cis acting sequence found upstream of french bean β-phaseolin gene. Plant Cell 1: 839–853 (1989).

    Google Scholar 

  11. 11.

    Bustos MM, Begum D, Kalkan FA, Battrow MJ, Hall TC: Positive and negative cis-acting DNA domains are required for spatial and temporal regulation of gene expression by a seed storage promoter. EMBO J 10: 1469–1479 (1991).

    Google Scholar 

  12. 12.

    Burrow MD, Sen P, Chlan CA, Murai N: Developmental control of the β-phaseolin gene requires positive, negative and temporal seed-specific transcriptional regulatory elements and a negative element for stem and root expression. Plant J 2: 537–548 (1992).

    Google Scholar 

  13. 13.

    Chamberland S, Daigle N, Bernier F: The legumin boxes and the 3′ part of a soybean β-conglycinin promoter are involved in seed gene expression in transgenic tobacco plants. Plant Mol Biol 19: 937–949 (1992).

    Google Scholar 

  14. 14.

    Dickinson CD, Evans RP, Nielsen NC: RY repeats are conserved in the 5′ flanking region of legume seed protein genes. Nucl Acids Res 16: 371 (1988).

    Google Scholar 

  15. 15.

    Fiedler U: Funktionsanalyse eines samenspezifischen Promotors der Ackerbohne Vicia faba. Diploma Thesis, Halle University (1992).

  16. 16.

    Filistein R: Deletionsanalyse eines samenspezifischen Promotors der Ackerbohne Vicia faba L.. Diploma Thesis, Halle University (1991).

  17. 17.

    Galau GA, Jakobsen KS, Hughes DW: The controls of late dicot embryogenesis and early germination. Physiol Plant 81: 280–288 (1991).

    Google Scholar 

  18. 18.

    Goldberg R: Plants: Novel developmental processes. Science 240: 1460–1467 (1988).

    Google Scholar 

  19. 19.

    Goldberg R, Baker SJ, Perez-Grau L: Regulation of gene expression during plant embryogenesis. Cell 56: 149–160 (1989).

    Google Scholar 

  20. 20.

    Hahn S: The Yin and Yang of mammalian transcription. Curr Biol 2: 152–154 (1992).

    Google Scholar 

  21. 21.

    Hattori T, Vasil V, Rosenkrans L, Hannah LC, McCarty DR, Vasil IK: The viviparous-1 gene and abscisic acid activates the C1 regulatory gene for anthocyanin biosynthesis during seed maturation in maize. Genes Devel 6: 609–618 (1992).

    Google Scholar 

  22. 22.

    Herman LMF, VanMontagu M, Depicker AG,: Isolation of tobacco DNA segments with plant promoter activity. Mol Cell Biol 6: 4486–4492 (1986).

    Google Scholar 

  23. 23.

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

    Google Scholar 

  24. 24.

    Kuhlemeier C, Fluhr R, Green PJ, Chua NH: Sequences in the pea rbcS-3A gene have homology to constitutive mammalian enhancers but function as negative regulatory elements. Genes Devel 1: 247–255 (1987).

    Google Scholar 

  25. 25.

    Lelievre JM, Oliveira LO, Nielsen NC: 5′-CATG-CATG-3′ elements modulate the expression of glycinin genes. Plant Physiol 98: 387–391 (1992).

    Google Scholar 

  26. 26.

    Mariani BD, Lingappa JR, Kafatos FC: Temporal regulation in development: Negative and positive cis regulators dictate the precise timing of expression of a Drosophila chorion gene. Proc Natl Acad Sci USA 85: 3029–3033 (1988).

    Google Scholar 

  27. 27.

    Nagy I. Untersuchungen zur Expression und Regulation samenspezifischer Gene von Vicia faba. Thesis, Halle University (1990).

  28. 28.

    Peleman J, Boerjan W, Engler G, Seurinck J, Bottermann J, Alliotte T, VanMontagu M, Inze D: Strong cellular preference in the expression of a housekeeping gene of Arabidopsis thaliana encoding S-adenosylmethionine synthetase. Plant Cell 1: 81–93 (1989).

    Google Scholar 

  29. 29.

    Reiss B, Sprengel R, Will H, Schaller H: A new sensitive method for quantitative and qualitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30: 211–218 (1984).

    Google Scholar 

  30. 30.

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

    Google Scholar 

  31. 31.

    Schmidt RJ, Kedudat M, Aukerman MJ, Hoschek G: Opaque-2 is a transcriptional activator that recognizes a specific target site in 22-kD zein genes. Plant Cell 4: 689–700 (1992).

    Google Scholar 

  32. 32.

    Shirsat AH: Control of gene expression in the developing seed. In: Grierson D (ed) Developmental Regulation of Plant Gene Expression, pp. 153–181. Chapman and Hall, New York (1991).

    Google Scholar 

  33. 33.

    Weising K, Kahl G: Towards and understanding of plant gene regulation: the action of nuclear factors. Z Naturforsch 46c: 1–11 (1991).

    Google Scholar 

  34. 34.

    Wobus U, Bäumlein H, Inze D, Nagy I: Vicia faba storage protein genes and their promoter activity in transgenic tobacco plants. In: Galling G (ed) Applied Plant Molecular Biology, pp. 98–103. Technical University, Braunschweig (1989).

    Google Scholar 

  35. 35.

    Zenke M, Grundström T, Matthes H, Wintzerith M, Schatz C, Wildeman A, Chambon P: Multiple sequence motifs are involved in SV40 enhancer function. EMBO J 5: 387–397 (1986).

    Google Scholar 

Download references

Author information



Rights and permissions

Reprints and Permissions

About this article

Cite this article

Fiedler, U., Filistein, R., Wobus, U. et al. A complex ensemble of cis-regulatory elements controls the expression of a Vicia faba non-storage seed protein gene. Plant Mol Biol 22, 669–679 (1993).

Download citation

Key words

  • cis-regulatory elements
  • Vicia faba
  • seed protein gene
  • promoter deletions
  • seed-specific expression
  • transgenic tobacco