Skip to main content
Log in

Assessment of the efficiency of cotransformation of the T-DNA of disarmed binary vectors derived from Agrobacterium tumefaciens and the T-DNA of A. rhizogenes

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Co-transfer of Agrobacterium rhizogenes T-DNA and T-DNA from the A. tumefaciens binary vector pBin19 (Bevan, 1984) was studied in detail using Nicotiana rustica. High frequencies of co-transfer of T-DNA's were observed, even when no selection pressure was exerted. Increased levels of pBin19 T-DNA were found in hairy root cultures with selection at higher levels of kanamycin sulphate (50–200 μg ml−1). Several other species were also transformed by A. rhizogenes carrying pBin19 and A. rhizogenes harbouring a different binary factor, pAGS125 (Van den Elzen et al., 1985), was used to transform N. rustica hairy roots to confer hygromycin B resistance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bevan M: Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12: 8711–8721 (1984).

    Google Scholar 

  • Cardarelli M, Spano L, DePaolis A, Mauro ML, Vitali G, Costantino P: Identification of the genetic locus responsible for nonpolar root induction by Agrobacterium rhizogenes 1855. Plant Mol Biol 5: 385–391 (1985).

    Google Scholar 

  • Chilton MD, Tepfer DA, Petit A, David C, Casse-Delbart F, Tempe J: Agrobacterium rhizogenes inserts T-DNA into the genomes of the host plant cells. Nature (Lond) 295: 432–434 (1982).

    Google Scholar 

  • D'Amato F: Cytogenetics of plant cell and tissue cultures and their regenerates. CRC Crit Rev Plant Sci 3: 73–112 (1985).

    Google Scholar 

  • DePaolis A, Mauro ML, Pomponi M, Cardarelli M, Spano L, Costantino P: Localization of agropine synthesizing functions in the TR region of the root-inducing plasmid of Agrobacterium rhizogenes 1855. Plasmid 13: 1–7 (1985).

    Google Scholar 

  • Deus-Neuman B, Zenk MH: Instability of indole alkaloid production in Catharanthus roseus cell suspension cultures. Planta Medica 50: 427–431 (1984).

    Google Scholar 

  • Ditta G, Stanfield S, Corbin D, Helinski DR: Broad host range DNA cloning system for gram-negative bacteria: construction of gene bank of Rhizobium melilott. Proc Natl Acad Sci USA 77: 7347–7351 (1980).

    Google Scholar 

  • Flores HE: Use of plant cell organ culture in the production of biological chemicals. In: Lebaron H, Mumma RO, Honeycutt RC, Duesing JH (eds) Application of Biotechnology to Agricultured Chemistry. Proc. 190th Amer. Chem. Soc. Symposium Series 190 (1986).

  • Hamill JD, Parr AJ, Rhodes MJC, Robins RJ, Walton NJ: New routes to secondary products: A review of the potential of ‘hairy root’ tissues transformed with Agrobacterium rhizogenes, for the biotechnological exploitation of plant secondary product formation. Biotechnology 5: 800–804 (1987).

    Google Scholar 

  • Hamill JD, Parr AJ, Robins RJ, Rhodes MJC: Secondary product formation by cultures of Beta vulgaris and Nicotiana rustica transformed with Agrobacterium rhizogenes. Plant Cell Rep 5: 111–114 (1986).

    Google Scholar 

  • Hoekema A, Hooykaas PJ, Schilperoort RA: Transfer of the octopine T-DNA segment to plant cells mediated by different types of Agrobacterium tumor or root inducing plasmids: Generality of virulence systems. J Bacteriol 158: 383–385 (1984).

    Google Scholar 

  • Hooykaas PJJ, Klapwijk PM, Nuti MP, Schilperoort RA, Rörsch A: Transfer of the Agrobacterium tumefaciens Ti plasmid to virulent Agrobacteria and to Rhizobium explanta. J Gen Microbiol 98: 477–484 (1977).

    Google Scholar 

  • Huffman GA, White FF, Gordon M, Nester EW: Hairy-root inducing plasmid: physical map and homology to tumor-inducing plasmids. J. Bacteriol 157: 269–276 (1984).

    Google Scholar 

  • Jouanin L: Restriction of an agropine-type Ri plasmid and its homologies with Ti plasmids. Plasmid 12: 91–102 (1984).

    Google Scholar 

  • Jouanin L, Guerche P, Pamboukdijian N, Tourneur C, Casse-Delbart F, Tourneur J: Structure of T-DNA in plants regenerated from roots transformed by Agrobacterium rhizogenes strain A4. Molec Gen Genet 206: 387–392 (1987).

    Google Scholar 

  • Kamada H, Okamura N, Satake M, Harada H, Shimomura K: Alkaloid production by hairy root cultures in Atropa belladonna. Plant Cell Rep 5: 239–242 (1986).

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning — a Laboratory Manual. Cold Spring Harbour Laboratory, Cold Sping Harbor, NY (1982).

    Google Scholar 

  • Martin C, Carpenter R, Sommer H, Saedler H, Coen ES: Molecular analysis of instability in flower pigmentation of Antirrhinum majus, following isolation of the pallida locus by transposon tagging. EMBO J 4: 1625–1630 (1985).

    Google Scholar 

  • Offringa IA, Melchers LS, Regensburg-Tuink AJG: Costantino P, Schilperoort RA, Hooykaas PJJ: Complementation of Agrobacterium tumefaciens tumor-inducing aux mutants by genes from the TR-region of the Ri plasmid of Agrobacterium rhizogenes. Proc Natl Acad Sci USA 83: 6935–6939 (1986).

    Google Scholar 

  • Ooms G, Karp A, Burrell MM, Twell D, Roberts J: Genetic modification of potato development using Ri T-DNA. Theor Appl Genet 70: 440–446 (1985).

    Google Scholar 

  • Payne J, Hamill JD, Robins RJ, Rhodes MJC: Production of hyoscyamine by hairy root cultures of Datura stramonium. Planta Medica (in press) (1987).

  • Pomponi M, Spano L, Sabbadini MG, Costantino P: Restriction endonuclease mapping of the root inducing plasmid of Agrobacterium rhizogenes 1855. Plasmid 10: 119–129 (1983).

    Google Scholar 

  • Rhodes MJC, Hilton M, Parr AJ, Hamill JD, Robins RJ: Nicotine production by ‘hairy root’ cultures of Nicotiana rustica fermentation and product recovery. Biotech Lett 8: 415–420 (1986).

    Google Scholar 

  • Shahin EA, Sukhapinda K, Simpson RB, Spivey R: Transformation of cultivated tomato by a binary vector in Agrobacterium rhizogenes: transgenic plants with normal phenotypes harbour binary vector T-DNA but no Ri plasmid T-DNA. Theor Appl Genet 72: 770–777 (1986).

    Google Scholar 

  • Simpson RB, Spielmann A, Margossian L, McKnight TD: A disarmed binary vector from Agrobacterium tumefaciens functions in Agrobacterium rhizogenes. Frequent cotransformation of two distinct T-DNAs. Plant Mol Biol 6: 403–415 (1986).

    Google Scholar 

  • Steel RGD, Torrie JH: Principles and Procedures of Statistics. McGraw-Hill Book Company, Inc. New York, Toronto, London (1960).

    Google Scholar 

  • Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517 (1975).

    Google Scholar 

  • Tepfer D: Transformation of several species of higher plants by Agrobacterium rhizogenes: Sexual transmission of the transformed genotype and phenotype. Cell 37: 959–964 (1984).

    Google Scholar 

  • Trulson AJ, Simpson RB, Shahin EA: Transformation of cucumber (Cucumis sativus L.) plants with Agrobacterium rhizogenes. Theor Appl Genet 73: 11–15 (1986).

    Google Scholar 

  • Van denElzen PJM, Townsend J, Lee KY, Bedbrook JR: A chimaeric hygromycin resistance gene as a selectable marker in plant cells. Plant Mol Biol 5: 299–302 (1985).

    Google Scholar 

  • Wahl GM, Stern M, Stark GR: Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethal-paper and rapid hybridization by using dextran sulphate. Proc Natl Acad Sci USA 76: 3683–3687 (1979).

    Google Scholar 

  • 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).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hamill, J.D., Prescott, A. & Martin, C. Assessment of the efficiency of cotransformation of the T-DNA of disarmed binary vectors derived from Agrobacterium tumefaciens and the T-DNA of A. rhizogenes . Plant Mol Biol 9, 573–584 (1987). https://doi.org/10.1007/BF00020534

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00020534

Key words

Navigation