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  • 1
    Digitale Medien
    Digitale Medien
    The virA promoter is a host-range determinant in Agrobacterium tumefaciens (1993)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 7 (1993), S. 0 
    Details
    ISSN: 1365-2958
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie , Medizin
    Notizen: The limited host range (LHR) Agrobacterium tumefaciens strain Ag162 is an isolate with a narrow host range. Introduction of the wide host range (WHR) virA gene is essential for extending the host range to Kalanchoë daigremontiana. In this report we show that the region upstream of the ATG start codon is responsible for the LHR phenomenon and that this is probably due to the non-inducibility of the LHR virA promoter. By comparing the characteristics of the LHR and WHR VirA receptor proteins, it was found that the LHR VirA protein is able to activate the WHR VirG protein in the presence of acetosyringone and that this acetosyringone-dependent vir-induction is enhanced by the presence of d-glucose, as in the case of WHR VirA proteins. These results indicate that the domains, acting as receptors for sugars and phenolic signals, must be conserved between the LHR and WHR VirA receptor proteins.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1111/j.1365-2958.1993.tb01162.x
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  • 2
    Digitale Medien
    Digitale Medien
    Symbiotic phenotypes and translocated effector proteins of the Mesorhizobium loti strain R7A VirB/D4 type IV secretion system (2004)
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 54 (2004), S. 0 
    Details
    ISSN: 1365-2958
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie , Medizin
    Notizen: The symbiosis island of Mesorhizobium loti strain R7A contains genes with strong similarity to the structural vir genes (virB1-11; virD4) of Agrobacterium tumefaciens that encode the type IV secretion system (T4SS) required for T-DNA transfer to plants. In contrast, M. loti strain MAFF303099 lacks these genes but contains genes not present in strain R7A that encode a type III secretion system (T3SS). Here we show by hybridization analysis that most M. loti strains contain the VirB/D4 T4SS and not the T3SS. Strikingly, strain R7A vir gene mutants formed large nodules containing bacteroids on Leucaena leucocephala in contrast to the wild-type strain that formed only uninfected tumour-like structures. A rhcJ T3SS mutant of strain MAFF303099 also nodulated L. leucocephala, unlike the wild type. On Lotus corniculatus, the vir mutants were delayed in nodulation and were less competitive compared with the wild type. Two strain R7A genes, msi059 and msi061, were identified through their mutant phenotypes as possibly encoding translocated effector proteins. Both Msi059 and Msi061 were translocated through the A. tumefaciens VirB/D4 system into Saccharomyces cerevisiae and Arabidopsis thaliana, as shown using the Cre recombinase Reporter Assay for Translocation (CRAfT). Taken together, these results suggest that the VirB/D4 T4SS of M. loti R7A plays an analogous symbiotic role to that of T3SS found in other rhizobia. The heterologous translocation of rhizobial proteins by the Agrobacterium VirB/D4 T4SS is the first demonstration that rhizobial effector proteins are translocated into plant cells and confirms functional conservation between the M. loti and A. tumefaciens T4SS.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1111/j.1365-2958.2004.04292.x
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  • 3
    Digitale Medien
    Digitale Medien
    VirD4-independent transformation by CloDF13 evidences an unknown factor required for the genetic colonization of plants via Agrobacterium (2003)
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 47 (2003), S. 0 
    Details
    ISSN: 1365-2958
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie , Medizin
    Notizen: Agrobacterium uses a mechanism similar to conjugation for trans -kingdom transfer of its oncogenic T-DNA. A defined VirB/VirD4 Type IV secretion system is responsible for such a genetic transfer. In addition, certain virulence proteins as VirE2 can be mobilized into host cells by the same apparatus. VirE2 is essential to achieve plant but not yeast transformation. We found that the limited host range plasmid CloDF13 can be recruited by the virulence apparatus of Agrobacterium for transfer to eukaryotic hosts. As expected the VirB transport complex was required for such trans -kingdom DNA transfer. However, unexpectedly, the coupling factor VirD4 turned out to be necessary for transfer to plants but not for transport into yeast. The CloDF13 encoded coupling factor (Mob) was essential for transfer to both plants and yeast though. This is interpreted by the different specificities of Mob and VirD4. Hence, Mob being required for the transport of the CloDF13 transferred DNA (to both plants and yeast) and VirD4 being required for transport of virulence proteins such as VirE2. Nevertheless, the presence of the VirE2 protein in the host plant was not sufficient to restore the deficiency for VirD4 in the transforming bacteria. We propose that Mob functions encoded by the plasmid CloDF13 are sufficient for DNA mobilization to eukaryotic cells but that the VirD4-mediated pathway is essential to achieve DNA nuclear establishment specifically in plants. This suggests that other Agrobacterium virulence proteins besides VirE2 are translocated and essential for plant transformation.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1046/j.1365-2958.2003.03328.x
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  • 4
    Digitale Medien
    Digitale Medien
    Root lectin as a determinant of host–plant specificity in the Rhizobium –legume symbiosis (1989)
    [s.l.] : Nature Publishing Group
    Nature 338 (1989), S. 579-581 
    Details
    ISSN: 1476-4687
    Quelle: Nature Archives 1869 - 2009
    Thema: Biologie , Chemie und Pharmazie , Medizin , Allgemeine Naturwissenschaft , Physik
    Notizen: [Auszug] Rhizobium leguminosarum bv. viciae nodulates the cross-inoculation group of pea, vetch and lentil, whereas clover is nodulated by R. leguminosarum bv. trifolii. The involvement of root lectin in determining this host-plant specificity was testedby introducing the pea lectin gene (psl) into the ...
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1038/338579a0
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  • 5
    Digitale Medien
    Digitale Medien
    Localization of the VirA domain involved in acetosyringone-mediatedvir gene induction inAgrobacterium tumefaciens (1994)
    Springer
    Plant molecular biology 25 (1994), S. 899-907 
    Details
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium ; gene regulation ; sensor protein ; signal transduction ; VirA protein ; acetosyringone
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The VirA protein ofAgrobacterium tumefaciens is thought to be a receptor for plant phenolic compounds such as acetosyringone. Although it is not known whether the interaction between VirA and the phenolics is direct or requires other phenolic-binding proteins, it is shown in this study that the first 280 amino acids of the VirA protein are not essential for the acetosyringone mediatedvir gene induction response. Considering the fact that the cytoplasmic region between the amino acids 283 and 304 is highly conserved between the different VirA proteins, and that deletion of this region abolishes VirA activity, we suggest that the acetosyringone receptor domain is located in this cytoplasmic domain of the VirA protein.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00028884
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  • 6
    Digitale Medien
    Digitale Medien
    Transformation of plant cells via Agrobacterium (1989)
    Springer
    Plant molecular biology 13 (1989), S. 327-336 
    Details
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium ; plant cell transformation ; plant tumor induction ; plant vector systems
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00025321
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  • 7
    Digitale Medien
    Digitale Medien
    Octopine and nopaline strains of Agrobacterium tumefaciens differ in virulence; molecular characterization of the virF locus (1990)
    Springer
    Plant molecular biology 14 (1990), S. 249-259 
    Details
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium tumefaciens ; virulence gene ; host range ; plant tumour induction
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Octopine and nopaline strains of Agrobacterium tumefaciens were found to differ in virulence on Nicotiana glauca. This difference is due to the absence of a functional virF locus, which is necessary for efficient tumorigenesis on N. glauca, from the nopaline Ti plasmids. Genetic studies and DNA sequence analysis of the virF locus revealed that virF embraces one open reading frame coding for a hydrophilic protein with a molecular mass of 22437 Da. Transcription of virF is directed from left to right, towards the T region, and is strongly induced by the phenolic compound acetosyringone. We established that virA and virG, two genes known to be essential for induction of the vir regulon, are necessary for acetosyringoneinduced virF expression, implying that virF is a member of this vir regulon. Agrobacterium virF mutants can be complemented for tumor induction by co-infection with avirulent Agrobacterium ‘helper’ strains. We found that such ‘helper’ strains must express not only the virF gene but also the vir operons virA, virB, virD and virG.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00018565
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  • 8
    Digitale Medien
    Digitale Medien
    Electroporation of megaplasmids into Agrobacterium (1991)
    Springer
    Plant molecular biology 16 (1991), S. 917-918 
    Details
    ISSN: 1573-5028
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00015085
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  • 9
    Digitale Medien
    Digitale Medien
    Non-recombinant background in gene targeting: illegitimate recombination between a hpt gene and a defective 5′ deleted nptII gene can restore a Kmr phenotype in tobacco (1994)
    Springer
    Plant molecular biology 25 (1994), S. 721-733 
    Details
    ISSN: 1573-5028
    Schlagwort(e): direct gene transfer ; gene targeting ; homologous recombination ; illegitimate recombination ; Nicotiana tabacum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Previously we have demonstrated gene targeting in plants after Agrobacterium-mediated transformation. In these initial experiments a transgenic tobacco line 104 containing a T-DNA insertion with a defective neomycin phosphotransferase (nptII) gene was transformed with a repair construct containing an otherwise defective nptII gene. Homologous recombination between the chromosomally located target and the incoming complementary defective nptII construct generated an intact nptII gene and led to a kanamycin-resistant (Kmr) phenotype. The gene targeting frequency was 1×10−5. In order to compare direct gene transfer and Agrobacterium-mediated transformation with respect to gene targeting we transformed the same transgenic tobacco line 104 via electroporation. A total of 1.35×108 protoplasts were transformed with the repair construct. Out of nearly 221 000 transformed cells 477 Kmr calli were selected. Screening the Kmr calli via PCR for recombination events revealed that in none of these calli gene targeting had occurred. To establish the origin of the high number of Kmr calli in which gene targeting had not occurred we analysed plants regenerated from 24 Kmr calli via PCR and sequence analysis. This revealed that in 21 out of 24 plants analysed the 5′-deleted nptII gene was fused to the hygromycin phosphotransferase (hpt) gene that was also present on the repair construct. Sequence analysis of 7 hpt/nptII gene fusions showed that they all contained a continuous open reading frame. The absence of significant homology at the fusion site indicated that fusion occurred via a process of illegitimate recombination. Therefore, illegitimate recombination between an introduced defective gene and another gene present on the repair construct or the chromosome has to be taken into account as a standard byproduct in gene targeting experiments.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00029609
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  • 10
    Digitale Medien
    Digitale Medien
    Isolation and characterization of an auxin-inducible glutathione S-transferase gene of Arabidopsis thaliana (1996)
    Springer
    Plant molecular biology 30 (1996), S. 839-844 
    Details
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis thaliana ; auxin ; gene expression ; glutathione S-transferase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Genes homologous to the auxin-inducible Nt103 glutathione S-transferase (GST) gene of tobacco, were isolated from a genomic library of Arabidopsis thaliana. We isolated a λ clone containing an auxin-inducible gene, At103-1a, and part of a constitutively expressed gene, At103-1b. The coding regions of the Arabidopsis genes were highly homologous to each other and to the coding region of the tobacco gene but distinct from the GST genes that have been isolated from arabidopsis thusfar. Overexpression of a cDNA clone in Escherichia coli revealed that the AT103-1A protein had GST activity.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00019016
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