Skip to main content
Log in

Units of genetic expression in the virulence region of a plant tumor-inducing plasmid of Agrobacterium tumefaciens

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The effect of a large number of Tn3 insertions in the vir region of the Ti plasmid pTiA6NC on the virulence of Agrobacterium was determined. The Vir- insertions were mapped in three of the five loci that have been defined previously. Merodiploid Rec- strains carrying one insertion mutation on the Ti plasmid and another insertion mutation (or the homologous wild-type region) on a compatible plasmid were constructed and used in complementation tests for virulence in test plants. This analysis has revealed that there are ten units of gene expression, presumably transcription units in the vir region. Mutation in one of these units is confirmed to be dominant while those in all others are recessive. Co-infection of test plants with pairs of insertion mutants did not restore virulence.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bagdasarian M, Bagdasarian MM, Coleman S, Timmis K (1979) New vector plasmids for gene cloning in Pseudomonas. In: Timmis K, Pühler A (eds) Plasmids of medical, environmental, and commercial importance. Elsevier/North Holland, Amsterdam p 411–422

    Google Scholar 

  • Berg DE, Weiss A, Crossland L (1980) The polarity of Tn5 insertion mutants in Escherichia coli. J Bacteriol 142:438–446

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1513–1525

    Google Scholar 

  • Boyer HW, Rouland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–471

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Currier TC, Nester EW (1976) Isolation of covalently closed circular DNA of high molecular weight from bacteria. Anal Biochem 66:431–441

    Google Scholar 

  • DeCleene M, DeLey J (1976) The host range of crown gall. Bot Rev 42:389–466

    Google Scholar 

  • Engler G, Depicker A, Maenhaut R, Villarroel R, Van Montagu M, Schell J (1981) Physical mapping of DNA base sequence homologies between an octopine and a nopaline Ti-plasmid of Agrobacterium tumefaciens. J Mol Biol 152:183–207

    Google Scholar 

  • Garfinkel DJ, Nester EW (1980) Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J Bacteriol 144:732–743

    Google Scholar 

  • Garfinkel D, Simpson R, Ream L, White F, Gordon M, Nester E (1981) Genetic analysis of crown gall: fine structure mapping of the T-DNA by site-directed mutagenesis. Cell 27:143–153

    Google Scholar 

  • Grunstein M, Hogness DS (1975) Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci USA 72:3961–3965

    Google Scholar 

  • Hille J, Klasen I, Schilperoort R (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

    Google Scholar 

  • Holsters M, deWaele D, Depicker A, Messens E, Van Montagu M, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 163:181–187

    Google Scholar 

  • Klapwijk PM, Scheulderman T, Schilperoort RA (1978) Coordinate regulation of octopine degradation and conjugative transfer of Ti plasmids in Agrobacterium tumefaciens: evidence for a common regulatory gene and separate operons. J Bacteriol 136:775–785

    Google Scholar 

  • Klapwijk P, VanBeelen P, Schilperoort R (1979) Isolation of a recombination deficient. A. tumefaciens mutant. Mol Gen Genet 173:171–175

    Google Scholar 

  • Klee H, Gordon M, Nester E (1982) Complementation analysis of Agrobacterium tumfaciens Ti-plasmid mutations affecting oncogenicity. J Bacteriol 150:327–331

    Google Scholar 

  • Koekman BP, Ooms G, Klapwijk PM, Schilperoort RA (1979) Genetic map of an octopine Ti-plasmid. Plasmid 2:347–357

    Google Scholar 

  • Konarska-Kozlowska M, Iyer VN (1981) Physical and genetic organization of the IncN-group plasmid pCU1. Gene 14:195–204

    Google Scholar 

  • Leemans J, Deblaere R, Willmitzer L, DeGreve H, Hernalsteens J, Van Montagu M, Schell J (1982) Genetic identification of functions of TL-DNA transcripts in octopine grown galls. EMBO J I:147–152

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Olsen RH, Siak J, Gray R (1974) Characteristics of PRD1, a plasmid-dependent broad host range DNA bacteriophage. J Virol 14:689–699

    Google Scholar 

  • Ooms G, Hooykaas PJJ, Moolenaar G, Schilperoort RA (1981) Crown gall plant tumors of abnormal morphology, induced by Agrobacterium tmefaciens carrying mutated octopine Ti plasmids; analysis of T DNA functions. Gene 14:33–50

    Google Scholar 

  • Ooms G, Hooykaas PJJ, Van Veen RJM, Van Beelen P, Regensburg-Tuink TJG, Schilperoort R (1982) Octopine T-plasmid deletion mutants with emphasis on the right side of the T-region. Plasmid 7:15–29

    Google Scholar 

  • Ooms G, Klapwijk PM, Poulis JA, Schilperoort RA (1980) Characterization of Tn904 insertions in octopine Ti plasmid mutants of Agrobacterium tumefaciens. J Bacteriol 144:82–91

    Google Scholar 

  • Prakash RK, Schilperoort RA (1982) Relationship between nif plasmids of fast-growing Rhizobium species and Ti plasmids of Agrobacterium tumefaciens. J Bacteriol 149:1129–1134

    Google Scholar 

  • Risuleo G, Battistoni P, Costantino P (1982) Regions of homology between tumorigenic plasmids from Agrobacterium rhizogenes and Agrobacterium tumefaciens. Plasmid 7:45–51

    Google Scholar 

  • Rubens C, Heffron F, Falkow S (1976) Transposition of a plasmid deoxyribonucleic acid sequence that mediates ampicillin resistance; independence from host rec functions and orientation of insertion. Bacteriol 128:425–434

    Google Scholar 

  • Ruvkun G, Ausubel F (1981) A general method for site-directed mutagenesis in prokaryotes. Nature 289:85–88

    Google Scholar 

  • Sciaky D, Montoya AL, Chilton M-D (1977) Fingerprints of Agrobacterium Ti plasmids. Plasmid 1:238–253

    Google Scholar 

  • Selvaraj G, Iyer VN (1981) Genetic transformation of Rhizobium meliloti by plasmid DNA. Gene 15:279–283

    Google Scholar 

  • Shaw KJ, Berg CM (1979) Escherichia coli autotrophs induced by insertion of the transposable element Tn5. Genetics 92:741–747

    Google Scholar 

  • Thomashow MF, Knauf VC, Nester EW (1981) Relationship between the limited and wide host range octopine-type Ti plasmids of Agrobacterium tumefaciens. J Bacteriol 146:484–493

    Google Scholar 

  • Thomashow MF, Nutter R, Postle K, Chilton M-D, Blattner F, Powell A, Gordon MP, Nester EW (1980) Recombination between higher plant DNA and the Ti plasmid of Agrobacterium tumefaciens. Proc Natl Acad Sci USA 77:6448–6452

    Google Scholar 

  • Van Larabeke N, Engler G, Holsters M, Van den Elsacker S, Laenen T, Schilperoort R, Schell J (1974) Large plasmids in A. tumefaciens essential for crown gall inducing activity. Nature (Lond) 252:169–170

    Google Scholar 

  • Watson B, Currier T, Gordon M, Chilton M-D, Nester E (1975) Plasmid required for virulence of A. tumefaciens. J Bacteriol 123:255–264

    Google Scholar 

  • White F, Nester E (1980) Relationship of plasmids responsible for hairy root and crown gall tumorigenicity. J Bacteriol 144:710–720

    Google Scholar 

  • Willmitzer L, DeBeukeleer M, Lemmers M, Van Montagu M, Schell J (1980) The Ti-plasmid derived T-DNA is present in the nucleus and absent from plastids of plant crown-gall cells. Nature (Lond) 287:359–361

    Google Scholar 

  • Willmitzer L, Simons G, Schell J (1982) The TL-DNA in octopine crown-gall tumours codes for seven well-defined polyadenylated transcripts. The EMBO J 1:139–146

    Google Scholar 

  • Yadav NS, Postle K, Saiki R, Thomashow MF, Chilton M-D (1980) T-DNA of a crown gall teratoma is covalently joined to host plant DNA. Nature (Lond) 287:458–461

    Google Scholar 

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

    Google Scholar 

  • Zambryski P, Holsters M, Kruger K, Schell J, Van Montagu M, Goodman H (1980) Tumor DNA structure in plant cells transformed by A. tumefaciens. Science 209:1385–1391

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by G. O'Donovan

Rights and permissions

Reprints and permissions

About this article

Cite this article

Iyer, V.N., Klee, H.J. & Nester, E.W. Units of genetic expression in the virulence region of a plant tumor-inducing plasmid of Agrobacterium tumefaciens . Mol Gen Genet 188, 418–424 (1982). https://doi.org/10.1007/BF00330043

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330043

Keywords

Navigation