Summary
We have precisely localized virulent (vir) genes of the hairy root-inducing plasmid pRiA4b on the basis of sequence similarity with the tumor-inducing plasmid pTiA6NC, and shown that the overall organizations of vir genes in both plasmids are fairly analogous, although sizes and spacer lengths in some genes differ from each other. Among the vir genes thus mapped, the virC and virD loci were characterized in detail. Transposon insertions in virD led to loss of tumorigenicity on Kalanchoe stems and carrot discs, and one within virC exhibited an attenuated pathogenicity. The avirulent phenotype of the virD2 strain among these mutants was due to the lack of ability to recombine T-DNA border repeats in Agrobacterium cells. The nucleotide sequence of most parts of the virCD loci were similar in both plasmids. The virCD genes of these two plasmids, therefore, seem comparable both functionally and structurally. Phylogeny of pRi and pTi has also been discussed from the sequence data.
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References
Close TJ, Tait RC, Rempel HC, Hirooka T, Kim L, Kado CI (1987) Molecular characterization of the virC genes of the Ti plasmid. J Bacteriol 169:2336–2344
Das A, Stachel S, Ebert P, Allenza P, Montoya A, Nester E (1986) Promoters of Agrobacterium tumefaciens Ti-plasmid virulence genes. Nucleic Acids Res 14:1355–1364
Hagiya M, Close TJ, Tait RC, Kado CI (1985) Identification of pTiC58 Plasmid-encoded proteins for virulence in Agrobacterium tumefaciens. Proc Natl Acad Sci USA 82:2669–2673
Harley CB, Reynolds RP (1987) Analysis of E. coli promoter sequences. Nucleic Acids Res 15:2343–2361
Holsters M, Silva B, Van Vliet F, Genetello C, DeBlock M, Dhaese P, Depicker A, Inzé D, Engler G, Villarroel R, Van Montagu M, Schell J (1980) The functional organization of the nopaline A. tumefaciens plasmid pTiC58. Plasmid 3:212–230
Hooykaas PJJ, Hofker M, Den Dulk-Ras H, Schilperoort RA (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 mutants. Plasmid 11:195–205
Huffman GA, White FF, Gordon MP, Nester EW (1984) Hairyroot-inducing plasmid: physical map and homology to tumor-inducing plasmids. J Bacteriol 157:269–276
Jayaswal RK, Veluthambi K, Gelvin SB, Slightom JL (1987) Double-stranded cleavage of T-DNA and generation of single-stranded T-DNA molecules in Escherichia coli by a virD-encoded border-specific endonuclease from Agrobacterium tumefaciens. J Bacteriol 169:5035–5045
Klee HJ, White FF, Iyer VN, Gordon MP, Nester EW (1983) Mutational analysis of the virulence region of an Agrobacterium tumefaciens Ti plasmid. J Bacteriol 153:878–883
Machida Y, Sakurai M, Kiyokawa S, Ubasawa A, Suzuki Y, Ikeda J (1984) Nucleotide sequence of the insertion sequence found in the T-DNA region of mutant Ti plasmid pTiA66 and distribution of its homologues in octopine Ti plasmid. Proc Natl Acad Sci USA 81:7495–7499
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Nester EW, Gordon MP, Amasino RM, Yanofsky MF (1984) Crown gall: a molecular and physiological analysis. Annu Rev Plant Physiol 35:387–413
Nishiguchi R, Oka A (1986) Structure of the hairy-root-inducing plasmid and identification of its replicator region. Bull Inst Chem Res (Kyoto Univ) 64:79–87
Nishiguchi R, Takanami M, Oka A (1987) Characterization and sequence determination of the replicator region in the hairy-root-inducing plasmid pRiA4b. Mol Gen Genet 206:1–8
Porter SG, Yanofsky MF, Nester EW (1987) Molecular characterization of the virD operon from Agrobacterium tumefaciens. Nucleic Acids Res 15:7503–7517
Sanger F (1981) Determination of nucleotide sequences in DNA. Science 214:1205–1210
Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346
Stachel SE, Nester EW (1986) The genetic and transcriptional organization of the vir region of the A6 Ti plasmid of Agrobacterium tumefaciens. EMBO J 5:1445–1454
Stachel SE, Messens E, Van Montagu M, Zambryski P (1985) Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318:624–629
Stachel SE, Timmerman B, Zambryski P (1987) Activation of Agrobacterium tumefaciens vir gene expression generates multiple single-stranded T-strand molecules from the pTiA6 T-region: requirement for 5′ virD gene products. EMBO J 6:857–863
Tanaka N, Harayama M, Mano Y, Ohkawa H, Matsui C (1985) Infection of turnip and radish storage roots with Agrobacterium rhizogenes. Plant Cell Rep 4:74–77
Van Putten AJ, Jochems GJ, De Lang R, Nijkamp HJJ (1987) Structure and nucleotide sequence of the region encoding the mobilizing proteins of plasmid CloDF13. Gene 51:171–178
Veluthambi K, Jayaswal RK, Gelvin SB (1987) Virulence genes A, G, and D mediate the double-stranded border cleavage of T-DNA from the Agrobacterium Ti plasmid. Proc Natl Acad Sci USA 84:1881–1885
Von Heijne G (1986) A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 14:4683–4690
Winans SC, Ebert PR, Stachel SE, Gordon MP, Nester EW (1986) A gene essential for Agrobacterium virulence is homologous to a family of positive regulatory loci. Proc Natl Acad Sci USA 83:8278–8282
Yamamoto A, Iwahashi M, Yanofsky MF, Nester EW, Takebe I, Machida Y (1987) The promoter proximal region in the virD locus of Agrobacterium tumefaciens is necessary for the plant-inducible circularization of T-DNA. Mol Gen Genet 206:174–177
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
Yanofsky MF, Nester EW (1986) Molecular characterization of a host-range-determining locus from Agrobacterium tumefaciens. J Bacteriol 168:244–250
Yanofsky MF, Porter SG, Young C, Albright LM, Gordon MP, Nester EW (1986) The virD operon of Agrobacterium tumefaciens encodes a site-specific endonuclease. Cell 47:471–477
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Hirayama, T., Muranaka, T., Ohkawa, H. et al. Organization and characterization of the virCD genes from Agrobacterium rhizogenes . Mol Gen Genet 213, 229–237 (1988). https://doi.org/10.1007/BF00339586
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DOI: https://doi.org/10.1007/BF00339586