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A T-DNA transfer stimulator sequence in the vicinity of the right border of pRi8196

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Abstract

An 8 bp sequence repeated 6 times is present to the right of the mannopine type pRi8196 T-DNA righ-border sequence. Experiments were designed to test whether these repeats have a role in T-DNA transfer. Several constructs in which different lengths of pRi8196 right-border region were linked to the cucumopine synthesis gene on anAgrobacterium-Escherichia coli shuttle vector were made. The recombinant plasmids were tested for their efficiency to act as a source of T-DNA in a binary system in which a wild-type Ri plasmid provided virulence and root-inducing functions. The T-DNA transfer efficiency of the constructs was assessed by computing the relative frequency of roots containing cucumopine. Depending on the Ri plasmid used as source of virulence functions, a high level of T-DNA transfer was observed only if 6 (pRi8196) or 5 (pRiA4) repeats were present. These results were confirmed by looking for single-stranded T-DNA molecules (T-strands) in bacteria induced for virulence. The repetition of the 8 bp unit was named ‘T-DNA transfer stimulator sequence’ (TSS).

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References

  1. Ahmed A: Use of transposon-promoted deletions in DNA sequence analysis. Meth Enzymol 155: 177–204 (1987).

    PubMed  Google Scholar 

  2. Albright LM, Yanofsky MF, Leroux B, Ma D, Nester EW: Processing of the T-DNA ofAgrobacterium tumefaciens generates border nicks and linear, single-stranded T-DNA. J Bact 169: 1046–1055 (1987).

    PubMed  Google Scholar 

  3. Alt-Moerbe J, Rak B, Schröder J: A 3.6-kbp segment from thevir region of Ti plasmids contains genes responsible for border sequence-directed production of T region circles inE. coli. EMBO J 5: 1129–1135 (1986).

    Google Scholar 

  4. Brevet J, Borowski D, Tempé J: Identification of the region encoding opine synthesis and of a region involved in host specificity on the T-DNA of cucumber-type Ri plasmid. Mol Plant-Microbe Interact 1: 75–79 (1988).

    Google Scholar 

  5. Chilton M-D, Tepfer DA, Petit A, David C, Casse-Delbart F, Tempé J:Agrobacterium rhizogenes inserts T-DNA into the genomes of the host-plant root cells. Nature 295: 432–434 (1982).

    Google Scholar 

  6. Christie PJ, Ward JE, Winans SC, Nester EW: TheAgrobacterium tumefaciens virE2 gene product is a single stranded DNA binding protein that associates with T-DNA. J Bact 170: 2659–2667 (1988).

    PubMed  Google Scholar 

  7. David C, Chilton M-D, Tempé J: Conservation of T-DNA in plants regenerated from hairy root cultures. Bio/technology 2: 73–76 (1984).

    Article  Google Scholar 

  8. Depicker A, Herman L, Jacobs L, Schell J, Van Montagu M: Frequencies of simultaneous transformation with different T-DNAs and their relevance to theAgrobacterium/plant cell interaction. Mol Gen Genet 201: 477–484 (1985).

    Article  Google Scholar 

  9. Dessaux Y, Petit A, Tempé J: Opines inAgrobacterium biology. In: Verma DPS (ed) Molecular Signals in Plant-Microbe Communications, pp. 109–136, CRC Press, Boca Raton, FL (1992).

    Google Scholar 

  10. De Vos G, Zambryski P: Expression ofAgrobacterium nopaline-specific VirD1, VirD2, and VirC1 proteins and their requirement for T-stand production inE. coli. Mol Plant-Microbe Interact 2: 43–52 (1989).

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  12. Ditta G, Schmidhauser T, Yakobson E, Lu P, Liang XW, Finlay DR, Guiney D, Helinski DR: Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid 13: 149–153 (1985).

    PubMed  Google Scholar 

  13. Gielt C, Koukolikova Z, Hohn B: Mobilization of T-DNA fromAgrobacterium to plant cells involve a protein that binds single-stranded DNA. Proc Natl Acad Sci USA 84: 9006–9010 (1987).

    PubMed  Google Scholar 

  14. Hansen G, Larribe M, Vaubert D, Tempé J, Biermann BJ, Montoya AL, Chilton M-D, Brevet J:Agrobacterium rhizogenes pRi8196 T-DNA: Mapping and DNA sequence of functions involved in mannopine synthesis and hairy root differentiation. Proc Natl Acad Sci USA 88: 7763–7767 (1991).

    PubMed  Google Scholar 

  15. Jen GC, Chilton M-D: The right border region of pTiT37 T-DNA is intrinsically more active than the left border region in promoting T-DNA transformation. Proc Natl Acad Sci USA 83: 3895–3899 (1986).

    PubMed  Google Scholar 

  16. Joos H, Inze D, Caplan A, Sormann M, Van Montagu M, Schell J: Genetic analysis of T-DNA transcripts in nopaline crown galls. Cell 32: 1057–1067 (1983).

    Article  PubMed  Google Scholar 

  17. Jouanin L, Bouchez D, Drong RF, Tepfer D, Slightom JL: Analysis of TR-DNA/plant junctions in the genome of aConvolvulus arvensis clone transformed withAgrobacterium rhizogenes strain A4. Plant Mol Biol 12: 75–85 (1989).

    Article  Google Scholar 

  18. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  19. Melchers LS, Regensburg-Tuink JG, Bourret RB, Sedee NJA, Schilperoort RA, Hooykaas PJJ: Membrane topology and functional analysis of the sensory protein VirA ofAgrobacterium tumefaciens. EMBO J 8: 1919–1925 (1989).

    PubMed  Google Scholar 

  20. Peralta GE, Helmiss R, Ream W: ‘Overdrive’, a T-DNA transmission enhancer on theAgrobacterium tumefaciens tumour-inducing plasmid. EMBO J 5: 1137–1142 (1986).

    Google Scholar 

  21. Petit A, Berkaloff A, Tempé J: Multiple transformation of plant cells byAgrobacterium may be responsible for the complex organization of T-DNA in crown gall and hairy root. Mol Gen Genet 202: 388–393 (1986).

    Article  Google Scholar 

  22. Sanger F, Coulson AR, Barell BG, Smith AJH, Roe B: Cloning in single strand bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143: 161–178 (1980).

    PubMed  Google Scholar 

  23. Shurviton CE, Ream W: Stimulation ofAgrobacterium tumefaciens T-DNA transfer by overdrive depends on a flanking sequence but not on helical position with respect to the border repeat. J Bact 173: 5558–5563 (1991).

    PubMed  Google Scholar 

  24. Slightom JL, Durand-Tardif M, Jouanin L, Tepfer D: Nucleotide sequence analysis of TL-DNAAgrobacterium rhizogenes type plasmid. Identification of open reading frames. J Biol Chem 261: 108–121 (1986).

    PubMed  Google Scholar 

  25. Stachel SE, Zambryski PC: VirA and virG control the plant induced activation of the T-DNA transfer process ofAgrobacterium tumefaciens. Cell 46: 325–333 (1986).

    Article  PubMed  Google Scholar 

  26. Stachel SE, Nester EW, Zambryski PC: A plant cell factor inducesAgrobacterium tumefaciens vir gene expression. Proc Natl Acad Sci USA 83: 379–383 (1986).

    Google Scholar 

  27. Stachel SE, Timmerman B, Zambryski PC: Generation of single stranded T-DNA molecules during the initial stages of T-DNA transfer fromAgrobacterium tumefaciens to plant cells. Nature 322: 706–712 (1986).

    Google Scholar 

  28. Toro N, Datta A, Yanofsky M, Nester E: Role of the overdrive sequence in T-DNA border cleavage inAgrobacterium. Proc Natl Acad Sci USA 85: 8558–8562 (1988).

    PubMed  Google Scholar 

  29. Toro N, Datta A, Carmi OA, Young C, Prusti RK, Nester EW: TheAgrobacterium tumefaciens virC1 gene product binds to overdrive, a T-DNA transfer enhancer. J Bact 171: 6845–6849 (1989).

    PubMed  Google Scholar 

  30. Van Haaren MJJ, Sedee NJA, Schilperoort RA, Hooykaas PJJ: Overdrive is a T-region transfer enhancer which stimulates T-strand production inAgrobacterium tumefaciens. Nucl Acids Res 15: 8983–8997 (1987).

    PubMed  Google Scholar 

  31. Van Haaren MJJ, Pronk JT, Schilperoort RA, Hooykaas PJJ: Functional analysis of theAgrobacterium tumefaciens octopine Ti-plasmid left and right T-region border fragments. Plant Mol Biol 8: 95–104 (1987).

    Google Scholar 

  32. Van Haaren MJJ, Sedee NJA, de Boer HA, Schilperoort RA, Hooykaas PJJ: Mutational analysis of the conserved domains of a T-region border repeat ofAgrobacterium tumefaciens. Plant Mol Biol 13: 523–531 (1989).

    PubMed  Google Scholar 

  33. Van Larebeke N, Engler G, Holsters M, Van den Elsacker S, Zaenen I, Schilperoort RA, Schell J: Large plasmid inAgrobacterium tumefaciens essential for crown gall-inducing ability. Nature 252: 169–170 (1974).

    PubMed  Google Scholar 

  34. Wang K, Herrera-Estrella L, Van Montagu M, Zambryski P: Right 25 bp terminus sequence of the nopaline T-DNA is essential for and determines direction of DNA transfer fromAgrobacterium to the plant genome. Cell 38: 455–462 (1984).

    Article  PubMed  Google Scholar 

  35. Yanish-Perron C, Vieira J, Messing J: Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33: 103–119 (1985).

    Article  PubMed  Google Scholar 

  36. Young C, Nester EW: Association of the VirD2 protein with the 5′ end of T-strands inAgrobacterium tumefaciens. J Bact 170: 3367–3374 (1988).

    PubMed  Google Scholar 

  37. Zambryski P, Tempé J, Schell J: Transfer and function of T-DNA genes fromAgrobacterium Ti and Ri plasmids in plants. Cell 56: 193–201 (1989).

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. CNRS, Institut des Sciences Végétales, 91198-, Gif sur Yvette, France

    Jean Brevet

  2. Institut National Agronomique Paris-Grignon, 75231, Paris 05, France

    Geneviève Hansen

Authors
  1. Geneviève Hansen
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  2. Jacques Tempé
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  3. Jean Brevet
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Hansen, G., Tempé, J. & Brevet, J. A T-DNA transfer stimulator sequence in the vicinity of the right border of pRi8196. Plant Mol Biol 20, 113–122 (1992). https://doi.org/10.1007/BF00029154

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  • DOI: https://doi.org/10.1007/BF00029154

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