Abstract
Supramembrane structures that connect conjugating agrobacterial cells were visualized for the first time by transmission electron microscopy. The primary contact of cells during conjugation was shown to occur through the formation of long pili containing no VirB1 protein. Pretreatment of agrobacterial cells with acetosyringone resulted in a six-to tenfold increase in the transfer frequency of plasmid pTd33 at 19–25°C and had almost no effect at 30°C. The transfer of plasmid pTd33 fromA. tumefaciens strain GV3101 to plasmid-freeA. tumefaciens strain UBAPF-2 was 16 times decreased after the centrifugation of cells. The transfer efficiency of plasmid pTd33 fromA. tumefaciens strain LBA2525 (virB2::lacZ) to plasmid-freeA. tumefaciens strain UBAPF-2 was one order of magnitude lower than the transfer from the wild-typeA. tumefaciens strain GV3101. Treatment of donor cells with 0.01% SDS before mating decreased the transfer efficiency by a factor of 26. The role of pili in the establishment of contact between conjugating cells of agrobacteria is discussed.
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Jones, A.L., Lai, E. M., Shirasu, K., and Kado, C.I., VirB2 Is a Processed Pilin-like Protein Encoded by theAgrobacterium tumefaciens Ti Plasmid,J. Bacteriol., 1996, vol. 178, pp. 5706–5711.
Fullner, K.J., Lara, J.C., and Nester, E.W., Temperature Affects the T-DNA Transfer Machineryof Agrobacterium tumefaciens, Science, 1996, vol. 273, pp. 1107–1109.
Baron, C., Llosa, M., Zhou, S., and Zambryski, P., VirB 1, a Component of the T-Complex Transfer Machinery ofAgrobacterium tumefaciens, Is Processed to a C-Terminal Secreted Product, VirBl,J. Bacterioi., 1997, vol. 179, pp. 1203–1210.
Stachel, S.E. and Zambryski, P.C.,virA andvirG Control the Plant-Induced Activation of the T-DNA Transfer Process ofAgrobacterium tumefaciens, Cell, 1986, vol. 47, pp. 155–157.
Engstrom, P., Zambryski, P., van Montague, M., and Stachel, S., Characterization ofAgrobacterium tumefaciens Virulence Proteins Induced by the Plant Factor Acetosyringone,J. Mol. Biol., 1987, vol. 197, pp. 635–645.
Lessl, M., Balzer, D., Pansegrau, W., and Lanka, E., Sequence Similarities between the RP4, Tra2, and the Ti VirB Region Strongly Support the Conjugation Model for T-DNA Transfer,J. Biol. Chem., 1992, vol. 267, pp. 20471–20480.
Lessl, M. and Lanka, E., Common Mechanisms in Bacterial Conjugation and Ti-Mediated T-DNA Transfer to Plant Cells,Cell, 1994, vol. 77, pp. 321–324.
Steck, T.R. and Kado, C.I., Virulence Genes Promote Conjugative Transfer of the Ti Plasmid betweenAgrobacterium Strains,J. Bacterioi., 1990, vol. 172, pp. 2191–2193.
Gelvin, S.B. and Habeck, L.L., Vir Genes Influence the Conjugal Transfer of the Ti Plasmid ofAgrobacterium tumefaciens, J. Bacterioi., 1990, vol. 172, pp. 1600–1608.
Bodman, S.B., McCutchan, J.E., and Farrand, S., Characterization of Conjugal Transfer Functions ofAgrobacterium tumefaciens Ti Plasmid pTiC58,J. Bacteriol., 1989, vol. 171, pp. 5281–5289.
Cook, D.M., Li, P. L., Ruchaud, F., Padden, S., and Farrand, S.K., Ti Plasmid Conjugation Is Independent ofvir: Reconstruction of thetra Functions from pTiC58 as a Binary System,J Bacteriol., 1997, vol. 179, pp. 1291–1297.
Chumakov, M.I., Gorban’, V.V., Kovler, L.A., Solovova, G.K., Krivopalov, Yu.V., Vasil’ev, A.Yu., Frolova, V.D., Muronets, E.M., and Kameneva, S.V., A New Associative Diazotroph,Agrobacterium radiobacter, from Wheat Rhizosphere,Mikrobiologiya, 1992, vol. 61, pp. 92–102.
McMichael, J.C. and Ou, J.T., Structure of Common Pili fromEscherichia coli, J. Bacterioi., 1979, vol. 138, pp. 969–975.
Tempe, J., Petit, A., Holsters, M., Montagu, M., and Shell, J., Thermosensitivity Step Associated with Transfer of the Ti Plasmid during Conjugation: Possible Relation to Transformation in Crown Gall,Proc. Natl. Acad. Sci. USA, 1977, vol. 711, pp. 2848–2849.
Fullner, K.J. and Nester, E.W., Temperature Affects the T-DNA Transfer Machinery ofAgrobacterium tumefaciens, J. Bacterioi., 1996, vol. 178, pp. 1498–1504.
Shirasu, K. and Kado, C.I., Membrane Location of the Ti Plasmid VirB Proteins Involved in the Biosynthesis of a Pilin-like Conjugative Structure ofAgrobacterium tumefaciens, FEMS Microbiol. Lett, 1993, vol. 111, pp. 287–294.
Christie, P.J., Ward, J.E., Jr., Gordon, M.O., and Nester, E. W., A Gene Required for Transfer of T-DNA to Plant Encodes an ATPase with Antiphosphorylating Activity,Proc. Natl. Acad. Sci. USA, 1989, vol. 86, pp. 9677–9681.
Sabel’nikov, A.G. and Borovyagin, V.L., A Model for Cation-Induced Transmembrane DNA Transfer in Bacteria,Biol. Membr., 1988, T7 5. S. 743–751.
Willets, N. and Skurray, R.,Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology, Neihard, F.C.et al., Eds., Washington, DC, 1987, vol. 2, pp. 1110–1133.
Durrenberger, M.B., Villiger, W., and Bachi, T., Conjugation Junction: Morphology of Specific Contact in ConjugatingEscherichia coli Bacteria,J. Struct. Biol., 1991, vol. 107, pp. 146–156.
Sabel’nikov, A.G., Berezkina, N.E., Chupin, V.V., Gongadze, G.M., and Borovyagin, V.L., Structural Changes inEscherichia coli Membranes during Conjugation,Biol. Membr., 1994, vol. 4, pp. 393–401.
Pansgrau, W. and Lanka, E., Enzymology of DNA Transfer by Conjugation Mechanisms,Prog. Nucleic Acids Res. Mol. Biol., 1996, vol. 54, pp. 197–251.
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Chumakov1, M.I., Kurbanova, I.V. Conjugative transfer of plasmid pTd33 in agrobacteria. Microbiology 69, 68–74 (2000). https://doi.org/10.1007/BF02757260
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DOI: https://doi.org/10.1007/BF02757260