The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA

Journal of Bacteriology - Tập 168 Số 3 - Trang 1291-1301 - 1986
Elizabeth E. Hood1,2,3,4,5, Georgia Helmer1,2,3,4,5, Robert T. Fraley1,2,3,4,5, Mary-Dell Chilton
1AGR NOP, AGC OCT, AGR
2Department of Biology, Washington University, St. Louis, Missouri 631301; Ciba-Geigy Biotechnology Facility, Research Triangle Park, North Carolina 277092; and
3Km None Km Cb Cb Km Km Cb, Km Km Cb None Cb, Km None Cb, Km None Cb Km Cb Km Cb Km Cb Km Cb Km Cb, Km Km Cb, Km L,L-SAP, LOP,
4Monsanto Company, Chesterfield, Missouri 631983
5agrocinopine; OCT, octopine.

Tóm tắt

We used a binary-vector strategy to study the hypervirulence of Agrobacterium tumefaciens A281, an L,L-succinamopine strain. Strain A281 is hypervirulent on several solanaceous plants. We constructed plasmids (pCS65 and pCS277) carrying either the transferred DNA (T-DNA) or the remainder of the tumor-inducing (Ti) plasmid (pEHA101) from this strain and tested each of these constructs in trans with complementary regions from heterologous Ti plasmids. Hypervirulence on tobacco could be reconstructed in a bipartite strain with the L,L-succinamopine T-DNA and the vir region on separate plasmids. pEHA101 was able to complement octopine T-DNA to hypervirulence on tobacco and tomato plants. Nopaline T-DNA was complemented better on tomato plants by pEHA101 than it was by its own nopaline vir region, but not to hypervirulence. L,L-Succinamopine T-DNA could not be complemented to hypervirulence on tobacco and tomato plants with either heterologous vir region. From these results we suggest that the hypervirulence of strain A281 is due to non-T-DNA sequences on the Ti plasmid.

Từ khóa


Tài liệu tham khảo

Akiyoshi , D. E. , H. J. Klee , R. M. Amasino , E. W. Nester , and M. R. Gordon . 1984 . T-DNA of Agrobacterium tumefaciens encodes an enzyme of cytokinin biosynthesis . Proc. Natl. Acad. Sci. USA 81 : 5994 - 5998 .

An , G. , B.D. Watson , S. Stachel , M. P. Gordon , and E. W. Nester . 1985 . New cloning vehicles for transformation of higher plants . EMBO J. 4 : 277 - 284 .

Barry , G. F. , S. G. Rogers , R. T. Fraley , and L. Brand . 1984 . Identification of a cloned cytokinin gene . Proc. Natl. Acad. Sci. USA 81 : 4776 - 4780 .

Bevan , M. 1984 . Binary Agrobacterium vectors for plant transformation . Nucleic Acids Res. 12 : 8711 - 8721 .

Bevan , M. W. , R. B. Flavell , and M. -D. Chilton . 1983 . A chimeric antibiotic resistance gene as a selectable marker for plant cell transformation . Nature (London) 304 : 184 - 187 .

Birnboim , H. C. , and J. Doly . 1979 . A rapid alkaline extraction procedure for screening recombinant plasmid DNA . Nucleic Acids Res. 6 : 1513 - 1523 .

Boyer , H. W. , and D. Roulland-Dussoix . 1969 . A complementation analysis of the restriction and modification of DNA in Escherichia coli . J. Mol. Biol. 41 : 459 - 472 .

Chilton M.-D. 1982. Integration and transcription of Ti plasmid fragments p. 299-319. In G. Kahl and J. Schell (ed.) Molecular biology of plant tumors. Academic Press Inc. New York.

Chilton , M. - D. , M. Drummond , D. Merlo , D. Sciaky , A. Montoya , M. Gordon , and E. Nester . 1977 . Stable incorporation of plasmid DNA into higher plant cells: the molecular basis of crown gall tumorigenesis . Cell 11 : 263 - 271 .

Chilton , M. - D. , R. Saiki , N. Yadav , M. Gordon , and F. Quetier . 1980 . T-DNA from Agrobacterium Ti plasmid is in the nuclear DNA fraction of crown gall tumor cells . Proc. Natl. Acad. Sci. USA 77 : 4060 - 4064 .

Chilton , W. S. , E. E. Hood , and M. -D. Chilton . 1985 . L,Lsuccinamopine: an epimeric crown gall opine . Phytochemistry 24 : 2945 - 2948 .

Cohen , S. , A. Chang , and L. Hsu . 1972 . Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA . Proc. Natl. Acad. Sci. USA 69 : 2110 - 2114 .

de Framond , A. J. , E. W. Back , W. S. Chilton , L. Kayes , and M. -D. Chilton . 1986 . Two unlinked T-DNAs can transform the same tobacco plant cell and segregate in the Fl generation . Mol. Gen. Genet. 202 : 125 - 131 .

de Framond , A. J. , K. A. Barton , and M. -D. Chilton . 1983 . Mini Ti: a new vector strategy for plant genetic engineering . Bio/Technology 1 : 262 - 269 .

Ditta , G. , S. Stanfield , D. Corbin , and D. R. Helinski . 1980 . Broad host range DNA cloning system for gram-negative bacteria-construction of a gene bank of Rhizobium meliloti . Proc. Natl. Acad. Sci. USA 77 : 7347 - 7351 .

Douglas , C. J. , R. J. Staneloni , R. A. Rubin , and E. W. Nester . 1985 . Identification and genetic analysis of an Agrobacterium tumefaciens chromosomal virulence region . J. Bacteriol. 161 : 850 - 860 .

Drummond , M. H. , and M. -D. Chilton . 1978 . Tumor-inducing (Ti) plasmids of Agrobacterium share extensive regions of DNA homology . J. Bacteriol. 136 : 1178 - 1183 .

Engler , G. , A. Depicker , R. Maenhaut , R. Villarroel-Mandiola , M. Van Montagu , and J. Schell . 1981 . Physical mapping of DNA base sequence homologies between an octopine and a nopaline Ti-plasmid of Agrobacterium tumefaciens . J. Mol. Biol. 152 : 183 - 208 .

Farrand , S. D. , J. E. Slota , J. -S. Shim , and A. Kerr . 1985 . TnS insertions in the agrocin 84 plasmid: the conjugal nature of pAg 84 and the locations of determinants for transfer and agrocin 84 production . Plasmid 13 : 106 - 117 .

Fraley , R. T. , S. G. Rogers , R. B. Horsch , D. A. Eichholtz , J. S. Flick , C. L. Fink , N. S. Hoffmann , and P. R. Sanders . 1985 . The SEV system: a new disarmed Ti plasmid vector'system for plant transformation . Bio/Technology 3 : 629 - 635 .

Garfinkel , D. J. , R. B. Simpson , L. W. Ream , F. F. White , M. P. Gordon , and E. W. Nester . 1981 . Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis . Cell 27 : 143 - 153 .

Guyon , P. , M. -D. Chilton , A. Petit , and J. Tempe . 1980 . Agropine in "null-type " crown gall tumors: evidence for generality of the opine concept. Proc. Natl. Acad. Sci. USA 77 : 2693 - 2697 .

Hagiya , M. , T. J. Close , R. C. Tait , and C. I. Kado . 1985 . Identification of pTiC 58 plasmid-encoded proteins for virulence in Agrobacterium tumefaciens . Proc. Natl. Acad. Sci. USA 82 : 2669 - 2673 .

Hamilton , R. H. , and M. Z. Fall . 1971 . The loss of tumorinitiating ability in Agrobacterium tumefaciens by incubation at high temperature . Experientia 27 : 229 - 230 .

Hein , M. B. , M. L. Brenner , and W. A. Brun . 1984 . Concentrations of abscisic acid and indole-3-acetic acid in soybean seeds during development . Plant Physiol. 76 : 951 - 954 .

Hille , J. , I. Klasen , and R. Schilperoort . 1982 . Construction and application of R prime plasmids, carrying different segments of an octopine Ti plasmid from Agrobacterium tumefaciens, for complementation of vir genes . Plasmid 7 : 107 - 118 .

Hille , J. , J. van Kan , and R. Schilperoort . 1984 . trans-Acting virulence functions of the octopine Ti plasmid from Agrobacterium tumefaciens . J. Bacteriol. 158 : 754 - 756 .

Hoekema , A. , P. R. Hirsch , P. J. J. Hooykaas , and R. A. Schilperoort . 1983 . A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid . Nature (London) 303 : 179 - 180 .

Hoekema , A. , P. J. Hooykaas , and R. A. Schilperoort . 1984 . 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 .

Hohn , B. , and J. Collins . 1980 . A small cosmid for efficient cloning of large DNA fragments . Gene 11 : 291 - 298 .

Holsters , M. , D. de Waele , A. Depicker , E. Messens , M. Van Montagu , and J. Schell . 1978 . Transfection and transformation of Agrobacterium tumefaciens . Mol. Gen. Genet. 163 : 181 - 187 .

Holsters , M. , B. Silva , F. Van Vliet , C. Genetello , M. DeBlock , P. Dhaese , A. Depicker , D. Inze , G. Engler , R. Villarroel , M. Van Montagu , and J. Schell . 1980 . The functional organization of the nopaline. A. tumefaciens plasmid pTiC58 . Plasmid 3 : 212 - 230 .

. Hood , E. E. , W. S. Chilton , M. -D. Chilton , and R. T. Fraley . 1986 . T-DNA and opine synthetic loci in tumors incited by Agrobacterium tumefaciens A281 on soybean and alfalfa plants . J. Bacteriol. 168 : 1283 - 1290 .

Hood , E. , G. Jen , L. Kayes , J. Kramer , R. T. Fraley , and M. -D. Chilton . 1984 . Restriction endonuclease map of pTiBoS42, a potential Ti plasmid vector for genetic engineering of plants . Bio/Technology 2 : 702 - 708 .

Hooykaas , P. J. J. , M. Hofker , H. den Dulk-Ras , and R. A. Schilperoort . 1984 . A comparison of virulence determinants in an octopine Ti plasmid, a nopaline Ti plasmid, and an Ri plasmid by complementation analysis of Agrobacterium tumefaciens . Plasmid 11 : 195 - 205 .

Horsch , R. B. , J. E. Fry , N. L. Hoffmann , D. Eichholtz , S. G. Rogers , and R. T. Fraley . 1985 . A simple and general method for transferring genes into plants . Science 277 : 1229 - 1231 .

Iyer , V. N. , H. J. Klee , and E. W. Nester . 1982 . Units of genetic expression in the virulence region of a plant tumor-inducing plasmid of Agrobacterium tumefaciens . Mol. Gen. Genet. 188 : 418 - 424 .

Jacoby , G. A. , A. E. Jacob , and R. W. Hedges . 1976 . Recombination between plasmids of incompatibility groups P-1 and P-2 . J. Bacteriol. 127 : 1278 - 1285 .

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

Klee , H. J. , M. P. Gordon , and E. W. Nester . 1982 . Complementation analysis of Agrobacterium tumefaciens Ti plasmid mutations affecting oncogenicity . J. Bacteriol. 150 : 327 - 331 .

Klee , H. J. , F. F. White , V. N. Iyer , M. P. Gordon , and E. W. Nester . 1983 . Mutational analysis of the virulence region of an Agrobacterium tumefaciens Ti plasmid . J. Bacteriol. 153 : 878 - 883 .

Knauf , V. , M. Yanofsky , A. Montoya , and E. Nester . 1984 . Physical and functional map of an Agrobacterium tumefaciens tumor-inducing plasmid that confers a narrow host range . J. Bacteriol. 160 : 564 - 568 .

Komari , T. , W. Halperin , and E. W. Nester . 1986 . Physical and functional map of supervirulent Agrobacterium tumefaciens tumor-inducing plasmid pTiBoS42 . J. Bacteriol. 166 : 88 - 94 .

Lundquist , R. , T. J. Close , and C. I. Kado . 1984 . Genetic complementation of Agrobacterium tumefaciens Ti plasmid mutants in the virulence region . Mol. Gen. Genet. 193 : 1 - 7 .

Maniatis T. E. F. Fritsch and J. Sambrook. 1982. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.

Murai , N. , and J. D. Kemp . 1982 . Octopine synthase mRNA isolated from sunflower crown gall callus is homologous to the Ti plasmid of Agrobacterium tumefaciens . Proc. Natl. Acad. Sci. USA 79 : 86 - 90 .

Murray , N. E. , W. J. Brammer , and K. Murray . 1977 . Lamboid phages that simplify the recovery of in vitro recombinants . Mol. Gen. Genet. 150 : 53 - 61 .

Ooms , G. , A. J. G. Regensburg-Tuink , M. H. Hofker , A. Hoekema , P. J. J. Hooykaas , and R. A. Schilperoort . 1982 . Studies on the structure of cointegrates between octopine and nopaline Ti-plasmids and their tumor-inducing properties . Plant Mol. Biol. 1 : 265 - 276 .

Otten , L. , H. DeGreve , J. Leemans , R. Hain , P. Hooykaas , and J. Schell . 1984 . Restoration of virulence of vir region mutants of Agrobacterium tumefaciens strain B6S3 by coinfection with normal and mutant Agrobacterium strains . Mol. Gen. Genet. 195 : 159 - 163 .

Otten , L. , G. Piotrowiak , P. Hooykaas , M. Dubois , E. Szegedi , and J. Schell . 1985 . Identification of an Agrobacterium tumefaciens pTiB6S3 vir region fragment that enhances the virulence of pTiC58 . Mol. Gen. Genet. 199 : 189 - 193 .

Schroder , G. , S. Waffenschmidt , E. W. Weiler , and J. Schroder . 1984 . The T-region of Ti plasmids codes for an enzyme synthesizing indole-3-acetic acid . Eur. J. Biochem. 138 : 387 - 391 .

Schroder J. , G. Schroder , H. Huisman , R. A. Schilperoort , and J. Schell . 1981 . The mRNA for Iysopine dehydrogenase in plant tumor cells is complementary to a Ti plasmid fragment . FEBS Lett. 129 : 166 - 168 .

Sciaky , D. , A. L. Montoya , and M. -D. Chilton . 1978 . Fingerprints of Agrobacterium Ti plasmids . Plasmid 1 : 238 - 253 .

Thomashow , L. S. , S. Reeves , and M. F. Thomashow . 1984 . Crown gall oncogenesis: evidence that a T-DNA gene from the Agrobacterium Ti plasmid pTi A6 encodes an enzyme that catalyzes synthesis of indoleacetic acid . Proc. Natl. Acad. Sci. USA 81 : 5071 - 5075 .

Van Larebeke , N. , G. Engler , M. Holsters , S. Van den Elsacker , I. Zaenen , R. A. Schilperoort , and J. Schell . 1974 . Large plasmid in Agrobacterium tumefaciens essential for crown gall-inducing ability . Nature (London) 252 : 169 - 170 .

Watson , B. , T. C. Currier , M. P. Gordon , M. -D. Chilton , and E. W. Nester . 1975 . Plasmid required for virulence of Agrobacterium tumefaciens . J. Bacteriol. 123 : 255 - 264 .

Zaenen , I. , N. Van Larabeke , H. Teuchy , M. Van Montagu , and J. Schell . 1974 . Supercoiled circular DNA in crown-gall inducing Agrobacterium strains . J. Mol. Biol. 86 : 109 - 127 .

Thông tin
Thông tin xuất bản

Journal of Bacteriology

Tập 168 Số 3

1291-1301

Thông tin tác giả