Abstract
By sequencing the central region of the cucumopine-type T-DNA of Agrobacterium rhizogenes strain 2659, we identified three open reading frames homologous, to different extents, to ORFs 10, 11 and 12 (rolA, B and C) of the agropine-type (1855) T-DNA. Recombinant Agrobacterium strains encompassing the ORFs of 2659 T-DNA-which we refer to as rolα, β and γ-were utilized to infect carrot discs and to obtain transgenic tobacco plants, in order to compare the morphogenetic capabilities to those of the 1855 rol genes. Moreover, a long segment of the 5′ non-coding region of rolα and rolβ was fused to the GUS reporter gene and the pattern of expression and the responsiveness to auxin of the constructs was analysed in transgenic tobacco. Differences in the auxin requirement for root induction between the 2659 rol genes and their respective 1855 counterparts were pinpointed. These differences are not due to gene regulation and presumably reflect functional differences in the proteins encoded. Differences were also observed in the pattern of expression of rolβ in roots of transgenic plants, as compared to rolB. In addition, the pattern of expression of rolα-GUS construct in roots was found to be analogous to that observed for a construct driven by two of the five regulatory domains of the rolB promoter.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahmed A: Use of the transposon-promoted deletions in DNA sequence analysis. Meth Enzymol 155: 177–204 (1987).
Altamura MM, Archiletti T, Arpone I, Costantino P: Histological analysis of the expression of Agrobacterium rhizogenes rolB-GUS gene fusions in transgenic tobacco. New Phytol 118: 69–78 (1991).
Altamura MM, Capitani F, Gazza L, Capone I, Costantino P: The plant oncogene rolB stimulates the root formation on flower and root meristemoids on tobacco thin cell layers. New Phytol 126: 283–293 (1994).
Bevan M: Agrobacterium vectors for plant transformation. Nucl Acids Res. 12: 8711–8721 (1984).
Brevet J, Tempé J: Homology mapping of T-DNA regions of three Agrobacterium rhizogenes Ri plasmids by electron microscope heteroduplex studies. Plasmid 19: 75–83 (1988).
Capone I, Cardarelli M, Mariotti D, Pomponi M, De Paolis A, Costantino P. Different promoter regions control level and tissue specificity of expression of Agrobacterium rhizogenes rolB gene in plants. Plant Mol Biol 16: 427–436 (1991).
Capone I, Cardarelli M, Trovato M, Costantino P: Upstream non-coding region which confers polar expression to Ri plasmid root inducing gene rolB. Mol Gen Genet 216: 239–244 (1989).
Capone I, Spanò L, Cardarelli M, Bellincampi D, Petit A, Costantino P: Induction and growth properties of carrot roots with different complements of Agrobacterium rhizogenes T-DNA. Plant Mol Biol 13: 43–52 (1989).
Capone I, Frugis G, Costantino P, Cardarelli M: Expression in different populations of cells of the root meristem is controlled by different domains of the rolB promoter. Plant Mol Biol 25: 681–691 (1994).
Cardarelli M, Mariotti D, Pomponi M, Spanò L, Capone I, Costantino P: Agrobacterium rhizogenes T-DNA genes capable of inducing hairy root phenotype. Mol Gen Genet 209: 475–480 (1987).
Cardarelli M, Spanò L, De Paolis A, Mauro ML, Vitali G, Costantino P: Identification of the genetic locus responsible for non-polar root induction by Agrobacterium rhizogenes 1855. Plant Mol Biol 5: 385–391 (1985).
Cardarelli M, Spanò L, Mariotti D, Mauro ML, Costantino P: The role of auxin in hairy root induction. Mol Gen Genet 208: 475–480 (1987).
Chilton MD, Tepfer DA, Petit A, David C, Casse-Delbart F, Tempé J: Agrobacterium rhizogenes inserts T-DNA into the genome of host plant root cells. Nature 295: 432–434 (1982).
Costantino P, Mauro ML, Micheli G, Risuleo G, Hooykaas PJJ, Schilperoort RA. Fingerprinting and sequence homology of plasmids from different virulent strains of Agrobacterium rhizogenes. Plasmid 5: 170–182 (1981).
Estruch JJ, Schell J, Spena A: The protein encoded by the rolB plant oncogene hydrolyses indole glycosides. EMBO J 10: 3125–3128 (1991).
Failla MC, Maimone F, De Paolis A, Costantino P, Cardarelli M: The non-conserved region of cucumopinetype Agrobacterium rhizogenes T-DNA is responsible for hairy root induction. Plant Mol Biol 15: 747–753 (1990).
Filetici P, Spanò L, Costantino P: Conserved regions in the T-DNA of different Agrobacterium rhizogenes rootinducing plasmids. Plant Mol Biol 9: 19–26 (1987).
Jefferson RA, Kavanagh TA, Bevan M: GUS fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).
Maurel C, Barbier-Brygoo H, Spena A, Tempé J, Guern J: Single rol genes from the Agrobacterium rhizogenes TL-DNA alter some of the cellular responses to auxin in Nicotiana tabacum. Plant Physiol 97: 212–216 (1991).
Nilsson O, Crozier A, Schmülling T, Sandberg G, Olsson O: Indole-3-acetic acid homeostasis in transgenic tobacco plants expressing the Agrobacterium rhizogenes rolB gene. Plant J 3: 681–689 (1993).
Petit A, David C, Dahl G, Ellis GJ, Guyon P, Casse-Delbart F, Tempé J: Further extension of the opine concept: plasmids in Agrobacterium rhizogenes cooperate for opine degradation. Mol Gen Genet 190: 204–214 (1983).
Ryder MH, Tate ME, Kerr A: Virulence properties of strains of Agrobacterium on the apical and basal surface of carrot root discs. Plant Physiol 77: 215–221 (1985).
Sanger F, Coulson AR, Barrel BG, Smith AJH, Roe BA: Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143: 161–178 (1980).
Schmülling T, Schell J, Spena A: Promoters of the rol A, B, and C genes of Agrobacterium rhizogenes are differentially regulated in transgenic plants. Plant Cell 1: 665–670 (1989).
Schmülling T, Schell J, Spena A: Single genes from Agrobacterium rhizogenes influence plant development. EMBO J 7: 2621–2629 (1988).
Spanò L, Pomponi M, Costantino P, Van Slogteren GMS, Tempé J: Identification of T-DNA in the rootinducing plasmid of the agropine type Agrobacterium rhizogenes 1855. Plant Mol Biol 1: 291–300 (1982).
Spena A, Schmulling T, Koncz C, Schell J: Independent and synergistic activity of rolA, B and C loci in stimulating abnormal growth in plants. EMBO J 6: 3891–3899 (1987).
Vansuyt G, Vilaine F, Tepfer M, Rossignol M: rolA modulates the sensitivity to auxin of the proton translocation catalyzed by the plasma membrane H+ ATPase in transformed tobacco. FEBS Lett 298: 89–92 (1992).
Vilaine F, Charbonnier C, Casse-Delbart F: Further insight concerning the TL region of the Ri plasmid of Agrobacterium rhizogenes strain A4: transfer of a 1.9 kb fragment is sufficient to induce transformed roots on tobacco leaf fragments. Mol Gen Genet 210: 111–115 (1987).
White FF, Ghidossi G, Gordon MP, Nester EW: Tumour induction by Agrobacterium rhizogenes involves the transfer of plasmid DNA to the plant genome. Proc Natl Acad Sci USA 79: 3193–3197 (1982).
Willmitzer L, Sanchez-Serrano J, Buschfeld E, Schell J: DNA from Agrobacterium rhizogenes is transferred to and expressed in axenic hairy root plant tissues. Mol Gen Genet 186: 16–22 (1982).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Serino, G., Clerot, D., Brevet, J. et al. rol genes of Agrobacterium rhizogenes cucumopine strain: sequence, effects and pattern of expression. Plant Mol Biol 26, 415–422 (1994). https://doi.org/10.1007/BF00039550
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00039550