Summary
A 1.9 kb clone of the T-DNA region of the Agrobacterium tumefaciens Ti plasmid Bo542 which exhibited homology to the isopentenyl transferase (ipt) locus of pTiA6 was identified by low stringency DNA hybridization. Introduction of this segment of pTiBo542 DNA into cells of Nicotiana tabacum or N. glauca caused tumor formation in vivo, and allowed hormone independent growth in vitro. Furthermore, this DNA segment complemented ipt mutant strains of A. tumefaciens, restoring their ability to cause tumors on Kalanchöe leaves and tomato stems. The complete DNA sequence of this segment has been determined, revealing an open reading frame homologous to other known Agrobacterium ipt genes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amasino RM (1986) Acceleration of nucleic acid hybridization rate by polyethylene glycol. Anal Biochem 152:304–307
Amasino RM, Miller CO (1982) Hormonal control of tobacco tumor morphology. Plant Physiol 69:389–392
An G (1987) Binary Ti vectors for plant transformation and promoter analysis. Methods Enzymol 153:292–305
Barker RF, Idler KB, Thompson DV, Kemp JD (1983) Nucleotide sequence of the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid pTi15955. Plant Mol Biol 2:335–350
Chen EY, Seeburg PH (1985) Supercoil sequencing: A fast and simple method for sequencing plasmid DNA DNA 4:165–170
Chilton M-D, Currier TC, Farrand SK, Bendich AJ, Gordon MP, Nester EW (1974) Agrobacterium DNA and PS8 bacteriophage DNA not detected in crown gall tumors. Proc Natl Acad Sci USA 71:3673–3676
Chilton M-D, Drummond MH, Merlo DJ, Sciaky D (1978) Highly conserved DNA of Ti plasmids overlaps T-DNA maintained in plant tumours. Nature 275:147–149
de Pater BS, Klinkhamer MP, Amesz PA, de Kam RJ, Memelink J, Hoge JHC, Schilperoort RA (1987) Plant expression signals of the Agrobacterium T-cyt gene. Nucleic Acids Res 15:8267–8281
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Garfinkel DJ, Nester EW (1980) Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J Bacteriol 144:732–743
Garfinkel DJ, Simpson RB, Ream LW, White FF, Gordon MP, Nester EW (1981) Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis. Cell 27:143–153
Gielen J, de Beuckeleer M, Seurinck J, Deboeck F, de Greve H, Lemmers M, van Montagu M, Schell J (1984) The complete nucleotide seuquence of the TL-DNA of the Agrobacterium tumefaciens plasmid pTiAch5. EMBO J 3:835–846
Hanahan D (1985) Techniques for transformation of E. coli. In: Glover DM (ed) DNA cloning, a practical approach, vol I. IRL Press, Oxford, pp 109–135
Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303:179–180
Hood EE, Jen G, Kayes L, Kramer J, Fraley RT, Chilton M-D (1984) Restriction endonuclease map of pTiBo542, a potential Ti plasmid vector for genetic engineering of plants. Bio Technology 2:702–709
Hood EE, Chilton WS, Chilton M-D, Fraley RT (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 EE, Fraley RT, Chilton M-D (1987) Virulence of Agrobacterium tumefaciens strain A281 on legumes. Plant Physiol 83:529–534
John MC, Amasino RM (1988) Expression of an Agrobacterium Ti plasmid gene involved in cytokinin biosynthesis is regulated by virulence loci and induced by plant phenolic compounds. J Bacteriol 170:790–795
Klee HJ, Yanofsky MF, Nester EW (1985) Vectors for transformation of higher plants. Bio Technology 3:637–642
Komari T, Halperin W, Nester EW (1986) Physical and functional map of supervirulent Agrobacterium tumefaciens tumor-inducing plasmid pTiBo542. J Bacteriol 166:88–94
Leong SA, Ditta GS, Helinski DR (1982) Heme biosynthesis in Rhizobium. J Biol Chem 257:8724–8730
Lichtenstein C, Klee H, Montoya A, Garfinkel D, Fuller S, Flores C, Nester EW, Gordon MP (1984) Nucleotide sequence and transcript mapping of the tmr gene of the pTiA6NC octopine Ti plasmid: A bacterial gene involved in plant tumorigenesis. J Mol Appl Genet 2:354–362
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Miller CO (1968) Naturally-occurring cytokinins. In: Wightman F, Setterfield G (eds) Biochemistry and physiology of plant growth substances. The Runge Press Ltd, Ottawa, pp 33–45
Montoya AL, Chilton M-D, Gordon MP, Sciaky D, Nester EW (1977) Octopine and nopaline metabolism in Agrobacterium tumefaciens and crown gall tumor cells: role of plasmid genes. J Bacteriol 129: 101–107
Morris RO (1986) Genes specifying auxin and cytokinin biosynthesis in phytopathogens. Annu Rev Plant Physiol 37:509–538
Nester EW, Gordon MP, Amasino RM, Yanofsky MF (1984) Crown gall: a molecular and physiological analysis. Annu Rev Plant Physiol 35:387–413
Reid RA, John MC, Amasino RM (1988) Deoxyribonuclease I sensitivity of the T-DNA ipt gene is associated with gene expression. Biochemistry 27:5748–5754
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Vieira J, Messing J (1987) Production of single-stranded plasmid DNA. Methods Enzymol 153:3–11
Weiler EW, Schröder J (1987) Hormone genes and crown gall disease. Trends Biochem Sci 12:271–275
Yanofsky M, Lowe B, Montoya A, Rubin R, Krul W, Gordon M, Nester E (1985) Molecular and genetic analysis of factors controlling host range in Agrobacterium tumefaciens. Mol Gen Genet 201:237–246
Author information
Authors and Affiliations
Additional information
Communicated by E. Meyerowitz
Rights and permissions
About this article
Cite this article
Strabala, T.J., Bednarek, S.Y., Bertoni, G. et al. Isolation and characterization of an ipt gene from the Ti plasmid Bo542. Mol Gen Genet 216, 388–394 (1989). https://doi.org/10.1007/BF00334380
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00334380