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. 1991 Jun;96(2):498–506. doi: 10.1104/pp.96.2.498

Transformation of Wall Deficient Cultured Tobacco Protoplasts by Agrobacterium tumefaciens 1

Andrew N Binns 1,2
PMCID: PMC1080798  PMID: 16668214

Abstract

Attachment of virulent Agrobacterium tumefaciens to plant cells is required for transformation. To further study the components of the plant cell wall that may be involved in the attachment process, tobacco (Nicotiana tabacum L.) protoplasts were cultured in the presence of 2,6 dichlorobenzonitrile (DB), an inhibitor of cellulose biosynthesis, and then assayed for their ability to be transformed by Agrobacterium. The DB treated protoplasts were deficient in wall production. Nevertheless, they were transformable at high frequency by wild type Agrobacterium strains but not by mutant strains that lack the ability to bind to normal, walled cells. Small quantities of calcofluor white positive material present on DB treated cells were correlated with their competence to be transformed. Further, the plant:bacterial association that leads to transformation is shown to become stable within 5 hours after bacterial co-cultivation with either control or DB treated cells.

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Selected References

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

  1. An G., Watson B. D., Stachel S., Gordon M. P., Nester E. W. New cloning vehicles for transformation of higher plants. EMBO J. 1985 Feb;4(2):277–284. doi: 10.1002/j.1460-2075.1985.tb03626.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cangelosi G. A., Hung L., Puvanesarajah V., Stacey G., Ozga D. A., Leigh J. A., Nester E. W. Common loci for Agrobacterium tumefaciens and Rhizobium meliloti exopolysaccharide synthesis and their roles in plant interactions. J Bacteriol. 1987 May;169(5):2086–2091. doi: 10.1128/jb.169.5.2086-2091.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Douglas C. J., Staneloni R. J., Rubin R. A., Nester E. W. Identification and genetic analysis of an Agrobacterium tumefaciens chromosomal virulence region. J Bacteriol. 1985 Mar;161(3):850–860. doi: 10.1128/jb.161.3.850-860.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Garfinkel D. J., Simpson R. B., Ream L. W., White F. F., Gordon M. P., Nester E. W. Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis. Cell. 1981 Nov;27(1 Pt 2):143–153. doi: 10.1016/0092-8674(81)90368-8. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Gurlitz R. H., Lamb P. W., Matthysse A. G. Involvement of Carrot Cell Surface Proteins in Attachment of Agrobacterium tumefaciens. Plant Physiol. 1987 Mar;83(3):564–568. doi: 10.1104/pp.83.3.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Matthysse A. G. Characterization of nonattaching mutants of Agrobacterium tumefaciens. J Bacteriol. 1987 Jan;169(1):313–323. doi: 10.1128/jb.169.1.313-323.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Neff N. T., Binns A. N., Brandt C. Inhibitory Effects of a Pectin-Enriched Tomato Cell Wall Fraction on Agrobacterium tumefaciens Binding and Tumor Formation. Plant Physiol. 1987 Mar;83(3):525–528. doi: 10.1104/pp.83.3.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pueppke S. G., Benny U. K. Adsorption of tumorigenic Agrobacterium tumefaciens cells to susceptible potato tuber tissues. Can J Microbiol. 1984 Aug;30(8):1030–1037. doi: 10.1139/m84-160. [DOI] [PubMed] [Google Scholar]
  15. Ruvkun G. B., Ausubel F. M. A general method for site-directed mutagenesis in prokaryotes. Nature. 1981 Jan 1;289(5793):85–88. doi: 10.1038/289085a0. [DOI] [PubMed] [Google Scholar]
  16. Tanimoto E., Douglas C., Halperin W. Factors Affecting Crown Gall Tumorigenesis in Tuber Slices of Jerusalem Artichoke (Helianthus tuberosus, L.). Plant Physiol. 1979 Jun;63(6):989–994. doi: 10.1104/pp.63.6.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Thomashow M. F., Karlinsey J. E., Marks J. R., Hurlbert R. E. Identification of a new virulence locus in Agrobacterium tumefaciens that affects polysaccharide composition and plant cell attachment. J Bacteriol. 1987 Jul;169(7):3209–3216. doi: 10.1128/jb.169.7.3209-3216.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Watson B., Currier T. C., Gordon M. P., Chilton M. D., Nester E. W. Plasmid required for virulence of Agrobacterium tumefaciens. J Bacteriol. 1975 Jul;123(1):255–264. doi: 10.1128/jb.123.1.255-264.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zorreguieta A., Geremia R. A., Cavaignac S., Cangelosi G. A., Nester E. W., Ugalde R. A. Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. Mol Plant Microbe Interact. 1988 Mar;1(3):121–127. doi: 10.1094/mpmi-1-121. [DOI] [PubMed] [Google Scholar]

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