Molecular and General Genetics MGG

, Volume 243, Issue 1, pp 32–38 | Cite as

Efficiency of the tetracycline-dependent gene expression system: complete suppression and efficient induction of the rolB phenotype in transgenic plants

  • Frank T. Röder
  • Thomas Schmülling
  • Christiane Gatz
Original Paper


We have investigated the use of the tetracycline-dependent gene expression system to regenerate and propagate tobacco plants transformed with a gene whose product — when highly expressed — interferes with regeneration and/or further reproduction. Plants transformed with the Agrobacterium rhizogenes rolB gene under the control of the tetracycline-dependent expression system were phenotypically indistinguishable from wild type owing to efficient repression of the promoter. Induction of the rolB gene with tetracycline led to high-level expression of the rolB mRNA, which resulted in extremely stunted plants with necrotic and wrinkled leaves that did not develop a floral meristem. Upon cessation of tetracycline treatment healthy shoots developed even from severely affected meristems. Data on the dose response of the rolB phenotype as a function of tetracycline concentration demonstrate that the tetracycline-dependent gene expression system can be used to modulate the manifestation of a particular phenotype.

Key words

Regulated expression rolB gene Tetracycline Tet repressor 


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  1. Benfey PN, Ren L, Chua N-H (1989) The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue specific expression patterns. EMBO J 8:2195–2202Google Scholar
  2. Deblaere R, Bytebier B, De Greve H, Debroeck F, Schell J, van Montagu M, Leemans J (1985) Efficient octopine Ti plasmid derived vectors for Agrobacterium-mediated gene transfer to plants. Nucleic Acids Res 13:4777–4788Google Scholar
  3. Estruch JJ, Schell J, Spena A (1991) The protein encoded by the rolB plant oncogene hydrolyses indole glucosides. EMBO J 10:3125–3128Google Scholar
  4. Jefferson RA, Kavanagh TA, Bevan M (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 13:3901–3907Google Scholar
  5. Gatz C, Kaiser A, Wendenburg R (1991) Regulation of a modified CaMV 35S promoter by the Tn10-encoded Tet repressor in transgenic tobacco. Mol Gen Genet 227:229–237Google Scholar
  6. Gatz C, Frohberg C, Wendenburg R (1992) Stringent repression and homogeneous derepression by tetracycline of a modified CaMV 35S promoter in intact transgenic tobacco plants. Plant J 2:397–404Google Scholar
  7. Höfgen R, Willmitzer L (1990) Biochemical and genetic analysis of different patatin isoforms expressed in various organs of potato (Solanum tuberosum). Plant Sci 66:221–230Google Scholar
  8. Maurel C, Barbier-Brygoo H, Spena A, Tempe J, Guern J (1991) Single rol genes from the Agrobacterium rhizogenes TL-DNA alter some of the cellular responses to auxin in Nicotiana tabacum. Plant Physiol 97:212–216Google Scholar
  9. Mett VL, Lochhead LP, Reynolds PHS (1993) Copper controllable gene expression system for whole plants. Proc Natl Acad Sci USA 90:4567–4571Google Scholar
  10. Nilsson O, Crozier A, Schmiilling T, Sandberg G, Olsson O (1993) Indole-3-acetic acid homeostasis in transgenic tobacco plants expressing the Agrobacterium rhizogenes rolB gene. Plant J 3:681–689Google Scholar
  11. Schena M, Lloyd AM; Davis RW (1991) A steroid-inducible gene expression system for plant cells. Proc Natl Acad Sci USA 88:10421–10425Google Scholar
  12. Schmülling T, Schell J, Spena A (1988) Single genes from Agrobacterium rhizogenes influence plant development. EMBO J 7:2621–2629Google Scholar
  13. Schmülling T, Fladung M, Grossmann K, Schell J (1993) Hormonal content and sensitivity of transgenic tobacco and potato plants expressing single rol genes of Agrobacterium rhizogenes T-DNA. Plant J 3:371–382Google Scholar
  14. Spena A, Schmülling T, Koncz C, Schell J (1987) Independent and synergistic activities of the rolA, B and C loci in stimulating abnormal growth in plants. EMBO J 6:3891–3899Google Scholar
  15. Spena A, Estruch JJ, Prinsen E, Nacken W, Van Onckelen H, Sommer H (1992) Anther-specific expression of the rolB gene of Agrobacterium rhizogenes increases IAA content in anthers and altes anther development and whole flower growth. Theor Appl Genet 84:520–527Google Scholar
  16. Takahashi M, Altschmied L, Hillen W (1986) Kinetic and equilibrium characterization of the tet repressor-tetracycline complex by fluorescence measurements. Evidence for divalent metal ion requirement and energy transfer. J Mol Biol 187:341–348Google Scholar
  17. Tepfer DA (1984) Transformation of several species of higher plants by Agrobacterium rhizogenes. Mol Gen Genet 206:17–23Google Scholar
  18. White RR, Taylor BH, Huffman GA, Gordon MP, Nester EW (1985) Molecular and genetic analysis of the transferred DNA regions of the root-inducing plasmid of Agrobacterium rhizogenes. J Bacteriol 164:33–44Google Scholar
  19. Wilde RJ, Shufflebottom D, Cooke S, Jasinska I, Merryweather A, Beri R, Brammar WJ; Bevan M, Schuch W (1992) Control of gene expression in tobacco cells using a bacterial operator-repressor system. EMBO J 11:1251–1259Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Frank T. Röder
    • 1
  • Thomas Schmülling
    • 1
  • Christiane Gatz
    • 1
  1. 1.Institut für Genbiologische Forschung Berlin GmbHBerlinGermany

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