Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Feb 20;93(4):1613–1618. doi: 10.1073/pnas.93.4.1613

Agrobacterium tumefaciens-mediated transformation of yeast.

K L Piers 1, J D Heath 1, X Liang 1, K M Stephens 1, E W Nester 1
PMCID: PMC39990  PMID: 8643679

Abstract

Agrobacterium tumefaciens transfers a piece of its Ti plasmid DNA (transferred DNA or T-DNA) into plant cells during crown gall tumorigenesis. A. tumefaciens can transfer its T-DNA to a wide variety of hosts, including both dicotyledonous and monocotyledonous plants. We show that the host range of A. tumefaciens can be extended to include Saccharomyces cerevisiae. Additionally, we demonstrate that while T-DNA transfer into S. cerevisiae is very similar to T-DNA transfer into plants, the requirements are not entirely conserved. The Ti plasmid-encoded vir genes of A. tumefaciens that are required for T-DNA transfer into plants are also required for T-DNA transfer into S. cerevisiae, as is vir gene induction. However, mutations in the chromosomal virulence genes of A. tumefaciens involved in attachment to plant cells have no effect on the efficiency of T-DNA transfer into S. cerevisiae. We also demonstrate that transformation efficiency is improved 500-fold by the addition of yeast telomeric sequences within the T-DNA sequence.

Full text

PDF
1613

Images in this article

1614Fig. 1
on p.1614
1617Fig. 2
on p.1617

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blackburn E. H. Telomeres: no end in sight. Cell. 1994 Jun 3;77(5):621–623. doi: 10.1016/0092-8674(94)90046-9. [DOI] [PubMed] [Google Scholar]
  3. Bundock P., den Dulk-Ras A., Beijersbergen A., Hooykaas P. J. Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae. EMBO J. 1995 Jul 3;14(13):3206–3214. doi: 10.1002/j.1460-2075.1995.tb07323.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cangelosi G. A., Best E. A., Martinetti G., Nester E. W. Genetic analysis of Agrobacterium. Methods Enzymol. 1991;204:384–397. doi: 10.1016/0076-6879(91)04020-o. [DOI] [PubMed] [Google Scholar]
  5. Cangelosi G. A., Martinetti G., Leigh J. A., Lee C. C., Thienes C., Theines C., Nester E. W. Role for [corrected] Agrobacterium tumefaciens ChvA protein in export of beta-1,2-glucan. J Bacteriol. 1989 Mar;171(3):1609–1615. doi: 10.1128/jb.171.3.1609-1615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Christie P. J., Ward J. E., Winans S. C., Nester E. W. The Agrobacterium tumefaciens virE2 gene product is a single-stranded-DNA-binding protein that associates with T-DNA. J Bacteriol. 1988 Jun;170(6):2659–2667. doi: 10.1128/jb.170.6.2659-2667.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Citovsky V., Zupan J., Warnick D., Zambryski P. Nuclear localization of Agrobacterium VirE2 protein in plant cells. Science. 1992 Jun 26;256(5065):1802–1805. doi: 10.1126/science.1615325. [DOI] [PubMed] [Google Scholar]
  8. Dente L., Cortese R. pEMBL: a new family of single-stranded plasmids for sequencing DNA. Methods Enzymol. 1987;155:111–119. doi: 10.1016/0076-6879(87)55011-x. [DOI] [PubMed] [Google Scholar]
  9. Douglas C. J., Halperin W., Nester E. W. Agrobacterium tumefaciens mutants affected in attachment to plant cells. J Bacteriol. 1982 Dec;152(3):1265–1275. doi: 10.1128/jb.152.3.1265-1275.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gheysen G., Villarroel R., Van Montagu M. Illegitimate recombination in plants: a model for T-DNA integration. Genes Dev. 1991 Feb;5(2):287–297. doi: 10.1101/gad.5.2.287. [DOI] [PubMed] [Google Scholar]
  11. Glogowski W., Galsky A. G. Agrobacterium tumefaciens Site Attachment as a Necessary Prerequisite for Crown Gall Tumor Formation on Potato Discs. Plant Physiol. 1978 Jun;61(6):1031–1033. doi: 10.1104/pp.61.6.1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heinemann J. A. Genetics of gene transfer between species. Trends Genet. 1991 Jun;7(6):181–185. doi: 10.1016/0168-9525(91)90433-q. [DOI] [PubMed] [Google Scholar]
  13. Heinemann J. A., Sprague G. F., Jr Bacterial conjugative plasmids mobilize DNA transfer between bacteria and yeast. Nature. 1989 Jul 20;340(6230):205–209. doi: 10.1038/340205a0. [DOI] [PubMed] [Google Scholar]
  14. Inomata K., Nishikawa M., Yoshida K. The yeast Saccharomyces kluyveri as a recipient eukaryote in transkingdom conjugation: behavior of transmitted plasmids in transconjugants. J Bacteriol. 1994 Aug;176(15):4770–4773. doi: 10.1128/jb.176.15.4770-4773.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Janda M., Ahlquist P. RNA-dependent replication, transcription, and persistence of brome mosaic virus RNA replicons in S. cerevisiae. Cell. 1993 Mar 26;72(6):961–970. doi: 10.1016/0092-8674(93)90584-d. [DOI] [PubMed] [Google Scholar]
  16. Jin S. G., Komari T., Gordon M. P., Nester E. W. Genes responsible for the supervirulence phenotype of Agrobacterium tumefaciens A281. J Bacteriol. 1987 Oct;169(10):4417–4425. doi: 10.1128/jb.169.10.4417-4425.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Koukolíková-Nicola Z., Raineri D., Stephens K., Ramos C., Tinland B., Nester E. W., Hohn B. Genetic analysis of the virD operon of Agrobacterium tumefaciens: a search for functions involved in transport of T-DNA into the plant cell nucleus and in T-DNA integration. J Bacteriol. 1993 Feb;175(3):723–731. doi: 10.1128/jb.175.3.723-731.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lessl M., Lanka E. Common mechanisms in bacterial conjugation and Ti-mediated T-DNA transfer to plant cells. Cell. 1994 May 6;77(3):321–324. doi: 10.1016/0092-8674(94)90146-5. [DOI] [PubMed] [Google Scholar]
  19. Lippincott B. B., Lippincott J. A. Bacterial attachment to a specific wound site as an essential stage in tumor initiation by Agrobacterium tumefaciens. J Bacteriol. 1969 Feb;97(2):620–628. doi: 10.1128/jb.97.2.620-628.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mazodier P., Davies J. Gene transfer between distantly related bacteria. Annu Rev Genet. 1991;25:147–171. doi: 10.1146/annurev.ge.25.120191.001051. [DOI] [PubMed] [Google Scholar]
  21. Nishikawa M., Suzuki K., Yoshida K. Structural and functional stability of IncP plasmids during stepwise transmission by trans-kingdom mating: promiscuous conjugation of Escherichia coli and Saccharomyces cerevisiae. Jpn J Genet. 1990 Oct;65(5):323–334. doi: 10.1266/jjg.65.323. [DOI] [PubMed] [Google Scholar]
  22. Porter S. G., Yanofsky M. F., Nester E. W. Molecular characterization of the virD operon from Agrobacterium tumefaciens. Nucleic Acids Res. 1987 Sep 25;15(18):7503–7517. doi: 10.1093/nar/15.18.7503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Robertson J. L., Holliday T., Matthysse A. G. Mapping of Agrobacterium tumefaciens chromosomal genes affecting cellulose synthesis and bacterial attachment to host cells. J Bacteriol. 1988 Mar;170(3):1408–1411. doi: 10.1128/jb.170.3.1408-1411.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rossi L., Hohn B., Tinland B. The VirD2 protein of Agrobacterium tumefaciens carries nuclear localization signals important for transfer of T-DNA to plant. Mol Gen Genet. 1993 Jun;239(3):345–353. doi: 10.1007/BF00276932. [DOI] [PubMed] [Google Scholar]
  25. Sciaky D., Montoya A. L., Chilton M. D. Fingerprints of Agrobacterium Ti plasmids. Plasmid. 1978 Feb;1(2):238–253. doi: 10.1016/0147-619x(78)90042-2. [DOI] [PubMed] [Google Scholar]
  26. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stachel S. E., Nester E. W. The genetic and transcriptional organization of the vir region of the A6 Ti plasmid of Agrobacterium tumefaciens. EMBO J. 1986 Jul;5(7):1445–1454. doi: 10.1002/j.1460-2075.1986.tb04381.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stinchcomb D. T., Mann C., Davis R. W. Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol. 1982 Jun 25;158(2):157–190. doi: 10.1016/0022-2836(82)90427-2. [DOI] [PubMed] [Google Scholar]
  29. Tinland B., Koukolíková-Nicola Z., Hall M. N., Hohn B. The T-DNA-linked VirD2 protein contains two distinct functional nuclear localization signals. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7442–7446. doi: 10.1073/pnas.89.16.7442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tsang T., Copeland V., Bowden G. T. A set of cassette cloning vectors for rapid and versatile adaptation of restriction fragments. Biotechniques. 1991 Mar;10(3):330–330. [PubMed] [Google Scholar]
  31. Uttaro A. D., Cangelosi G. A., Geremia R. A., Nester E. W., Ugalde R. A. Biochemical characterization of avirulent exoC mutants of Agrobacterium tumefaciens. J Bacteriol. 1990 Mar;172(3):1640–1646. doi: 10.1128/jb.172.3.1640-1646.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ward J. E., Akiyoshi D. E., Regier D., Datta A., Gordon M. P., Nester E. W. Characterization of the virB operon from an Agrobacterium tumefaciens Ti plasmid. J Biol Chem. 1988 Apr 25;263(12):5804–5814. [PubMed] [Google Scholar]
  33. Wellinger R. J., Wolf A. J., Zakian V. A. Saccharomyces telomeres acquire single-strand TG1-3 tails late in S phase. Cell. 1993 Jan 15;72(1):51–60. doi: 10.1016/0092-8674(93)90049-v. [DOI] [PubMed] [Google Scholar]
  34. Winans S. C. Two-way chemical signaling in Agrobacterium-plant interactions. Microbiol Rev. 1992 Mar;56(1):12–31. doi: 10.1128/mr.56.1.12-31.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yanofsky M. F., Porter S. G., Young C., Albright L. M., Gordon M. P., Nester E. W. The virD operon of Agrobacterium tumefaciens encodes a site-specific endonuclease. Cell. 1986 Nov 7;47(3):471–477. doi: 10.1016/0092-8674(86)90604-5. [DOI] [PubMed] [Google Scholar]
  36. Zupan J. R., Zambryski P. Transfer of T-DNA from Agrobacterium to the plant cell. Plant Physiol. 1995 Apr;107(4):1041–1047. doi: 10.1104/pp.107.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. de Iannino N. I., Ugalde R. A. Biochemical characterization of avirulent Agrobacterium tumefaciens chvA mutants: synthesis and excretion of beta-(1-2)glucan. J Bacteriol. 1989 May;171(5):2842–2849. doi: 10.1128/jb.171.5.2842-2849.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES