Abstract
We show that among ecotypes of Arabidopsis, there is considerable variation in their susceptibility to crown gall disease. Differences in susceptibility are heritable and, in one ecotype, segregate as a single major contributing locus. In several ecotypes, recalcitrance to tumorigenesis results from decreased binding of Agrobacterium to inoculated root explants. The recalcitrance of another ecotype occurs at a late step in T-DNA transfer. Transient expression of a T-DNA-encoded beta-glucuronidase gusA gene is efficient, but the ecotype is deficient in crown gall tumorigenesis, transformation to kanamycin resistance, and stable GUS expression. This ecotype is also more sensitive to gamma radiation than is a susceptible ecotype. DNA gel blot analysis showed that after infection by Agrobacterium, less T-DNA was integrated into the genome of the recalcitrant ecotype than was integrated into the genome of a highly susceptible ecotype.
Full Text
The Full Text of this article is available as a PDF (4.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aufsatz W., Grimm C. A new, pathogen-inducible gene of Arabidopsis is expressed in an ecotype-specific manner. Plant Mol Biol. 1994 May;25(2):229–239. doi: 10.1007/BF00023240. [DOI] [PubMed] [Google Scholar]
- Bent A. F., Innes R. W., Ecker J. R., Staskawicz B. J. Disease development in ethylene-insensitive Arabidopsis thaliana infected with virulent and avirulent Pseudomonas and Xanthomonas pathogens. Mol Plant Microbe Interact. 1992 Sep-Oct;5(5):372–378. doi: 10.1094/mpmi-5-372. [DOI] [PubMed] [Google Scholar]
- Cangelosi G. A., Hung L., Puvanesarajah V., Stacey G., Ozga D. A., Leigh J. A., Nester E. W. Common loci for Agrobacterium tumefaciens and Rhizobium meliloti exopolysaccharide synthesis and their roles in plant interactions. J Bacteriol. 1987 May;169(5):2086–2091. doi: 10.1128/jb.169.5.2086-2091.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Citovsky V., Warnick D., Zambryski P. Nuclear import of Agrobacterium VirD2 and VirE2 proteins in maize and tobacco. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3210–3214. doi: 10.1073/pnas.91.8.3210. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Citovsky V., Zupan J., Warnick D., Zambryski P. Nuclear localization of Agrobacterium VirE2 protein in plant cells. Science. 1992 Jun 26;256(5065):1802–1805. doi: 10.1126/science.1615325. [DOI] [PubMed] [Google Scholar]
- Dangl J. L., Ritter C., Gibbon M. J., Mur L. A., Wood J. R., Goss S., Mansfield J., Taylor J. D., Vivian A. Functional homologs of the Arabidopsis RPM1 disease resistance gene in bean and pea. Plant Cell. 1992 Nov;4(11):1359–1369. doi: 10.1105/tpc.4.11.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davies C., Howard D., Tam G., Wong N. Isolation of Arabidopsis thaliana mutants hypersensitive to gamma radiation. Mol Gen Genet. 1994 Jun 15;243(6):660–665. doi: 10.1007/BF00279575. [DOI] [PubMed] [Google Scholar]
- Deng W., Pu X. A., Goodman R. N., Gordon M. P., Nester E. W. T-DNA genes responsible for inducing a necrotic response on grape vines. Mol Plant Microbe Interact. 1995 Jul-Aug;8(4):538–548. doi: 10.1094/mpmi-8-0538. [DOI] [PubMed] [Google Scholar]
- Dong X., Mindrinos M., Davis K. R., Ausubel F. M. Induction of Arabidopsis defense genes by virulent and avirulent Pseudomonas syringae strains and by a cloned avirulence gene. Plant Cell. 1991 Jan;3(1):61–72. doi: 10.1105/tpc.3.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Douglas C. J., Halperin W., Nester E. W. Agrobacterium tumefaciens mutants affected in attachment to plant cells. J Bacteriol. 1982 Dec;152(3):1265–1275. doi: 10.1128/jb.152.3.1265-1275.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Filichkin S. A., Gelvin S. B. Formation of a putative relaxation intermediate during T-DNA processing directed by the Agrobacterium tumefaciens VirD1,D2 endonuclease. Mol Microbiol. 1993 May;8(5):915–926. doi: 10.1111/j.1365-2958.1993.tb01637.x. [DOI] [PubMed] [Google Scholar]
- Frisch D. A., Harris-Haller L. W., Yokubaitis N. T., Thomas T. L., Hardin S. H., Hall T. C. Complete sequence of the binary vector Bin 19. Plant Mol Biol. 1995 Jan;27(2):405–409. doi: 10.1007/BF00020193. [DOI] [PubMed] [Google Scholar]
- Gelvin S. B. Crown gall disease and hairy root disease : a sledgehammer and a tackhammer. Plant Physiol. 1990 Feb;92(2):281–285. doi: 10.1104/pp.92.2.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gheysen G., Villarroel R., Van Montagu M. Illegitimate recombination in plants: a model for T-DNA integration. Genes Dev. 1991 Feb;5(2):287–297. doi: 10.1101/gad.5.2.287. [DOI] [PubMed] [Google Scholar]
- Gurlitz R. H., Lamb P. W., Matthysse A. G. Involvement of Carrot Cell Surface Proteins in Attachment of Agrobacterium tumefaciens. Plant Physiol. 1987 Mar;83(3):564–568. doi: 10.1104/pp.83.3.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harlow G. R., Jenkins M. E., Pittalwala T. S., Mount D. W. Isolation of uvh1, an Arabidopsis mutant hypersensitive to ultraviolet light and ionizing radiation. Plant Cell. 1994 Feb;6(2):227–235. doi: 10.1105/tpc.6.2.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herrera-Estrella A., Van Montagu M., Wang K. A bacterial peptide acting as a plant nuclear targeting signal: the amino-terminal portion of Agrobacterium VirD2 protein directs a beta-galactosidase fusion protein into tobacco nuclei. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9534–9537. doi: 10.1073/pnas.87.24.9534. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hood E. E., Fraley R. T., Chilton M. D. Virulence of Agrobacterium tumefaciens Strain A281 on Legumes. Plant Physiol. 1987 Mar;83(3):529–534. doi: 10.1104/pp.83.3.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howard E. A., Zupan J. R., Citovsky V., Zambryski P. C. The VirD2 protein of A. tumefaciens contains a C-terminal bipartite nuclear localization signal: implications for nuclear uptake of DNA in plant cells. Cell. 1992 Jan 10;68(1):109–118. doi: 10.1016/0092-8674(92)90210-4. [DOI] [PubMed] [Google Scholar]
- Janssen B. J., Gardner R. C. Localized transient expression of GUS in leaf discs following cocultivation with Agrobacterium. Plant Mol Biol. 1990 Jan;14(1):61–72. doi: 10.1007/BF00015655. [DOI] [PubMed] [Google Scholar]
- Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koch E., Slusarenko A. Arabidopsis is susceptible to infection by a downy mildew fungus. Plant Cell. 1990 May;2(5):437–445. doi: 10.1105/tpc.2.5.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koukolíková-Nicola Z., Raineri D., Stephens K., Ramos C., Tinland B., Nester E. W., Hohn B. Genetic analysis of the virD operon of Agrobacterium tumefaciens: a search for functions involved in transport of T-DNA into the plant cell nucleus and in T-DNA integration. J Bacteriol. 1993 Feb;175(3):723–731. doi: 10.1128/jb.175.3.723-731.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee S., Stenger D. C., Bisaro D. M., Davis K. R. Identification of loci in Arabidopsis that confer resistance to geminivirus infection. Plant J. 1994 Oct;6(4):525–535. doi: 10.1046/j.1365-313x.1994.6040525.x. [DOI] [PubMed] [Google Scholar]
- Leisner S. M., Turgeon R., Howell S. H. Effects of host plant development and genetic determinants on the long-distance movement of cauliflower mosaic virus in Arabidopsis. Plant Cell. 1993 Feb;5(2):191–202. doi: 10.1105/tpc.5.2.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lloyd A. M., Barnason A. R., Rogers S. G., Byrne M. C., Fraley R. T., Horsch R. B. Transformation of Arabidopsis thaliana with Agrobacterium tumefaciens. Science. 1986 Oct 24;234(4775):464–466. doi: 10.1126/science.234.4775.464. [DOI] [PubMed] [Google Scholar]
- Matsumoto S., Ito Y., Hosoi T., Takahashi Y., Machida Y. Integration of Agrobacterium T-DNA into a tobacco chromosome: possible involvement of DNA homology between T-DNA and plant DNA. Mol Gen Genet. 1990 Dec;224(3):309–316. doi: 10.1007/BF00262423. [DOI] [PubMed] [Google Scholar]
- Matthysse A. G. Characterization of nonattaching mutants of Agrobacterium tumefaciens. J Bacteriol. 1987 Jan;169(1):313–323. doi: 10.1128/jb.169.1.313-323.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matthysse A. G., White S., Lightfoot R. Genes required for cellulose synthesis in Agrobacterium tumefaciens. J Bacteriol. 1995 Feb;177(4):1069–1075. doi: 10.1128/jb.177.4.1069-1075.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mayerhofer R., Koncz-Kalman Z., Nawrath C., Bakkeren G., Crameri A., Angelis K., Redei G. P., Schell J., Hohn B., Koncz C. T-DNA integration: a mode of illegitimate recombination in plants. EMBO J. 1991 Mar;10(3):697–704. doi: 10.1002/j.1460-2075.1991.tb07999.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Narasimhulu S. B., Deng X. B., Sarria R., Gelvin S. B. Early transcription of Agrobacterium T-DNA genes in tobacco and maize. Plant Cell. 1996 May;8(5):873–886. doi: 10.1105/tpc.8.5.873. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neff N. T., Binns A. N. Agrobacterium tumefaciens Interaction with Suspension-Cultured Tomato Cells. Plant Physiol. 1985 Jan;77(1):35–42. doi: 10.1104/pp.77.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohba T., Yoshioka Y., Machida C., Machida Y. DNA rearrangement associated with the integration of T-DNA in tobacco: an example for multiple duplications of DNA around the integration target. Plant J. 1995 Jan;7(1):157–164. doi: 10.1046/j.1365-313x.1995.07010157.x. [DOI] [PubMed] [Google Scholar]
- Owens L. D., Cress D. E. Genotypic variability of soybean response to agrobacterium strains harboring the ti or ri plasmids. Plant Physiol. 1985 Jan;77(1):87–94. doi: 10.1104/pp.77.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robbs S. L., Hawes M. C., Lin H. J., Pueppke S. G., Smith L. Y. Inheritance of Resistance to Crown Gall in Pisum sativum. Plant Physiol. 1991 Jan;95(1):52–57. doi: 10.1104/pp.95.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rossi L., Hohn B., Tinland B. The VirD2 protein of Agrobacterium tumefaciens carries nuclear localization signals important for transfer of T-DNA to plant. Mol Gen Genet. 1993 Jun;239(3):345–353. doi: 10.1007/BF00276932. [DOI] [PubMed] [Google Scholar]
- Sangwan R. S., Bourgeois Y., Sangwan-Norreel B. S. Genetic transformation of Arabidopsis thaliana zygotic embryos and identification of critical parameters influencing transformation efficiency. Mol Gen Genet. 1991 Dec;230(3):475–485. doi: 10.1007/BF00280305. [DOI] [PubMed] [Google Scholar]
- Sciaky D., Montoya A. L., Chilton M. D. Fingerprints of Agrobacterium Ti plasmids. Plasmid. 1978 Feb;1(2):238–253. doi: 10.1016/0147-619x(78)90042-2. [DOI] [PubMed] [Google Scholar]
- Smarrelli J., Watters M. T., Diba L. H. Response of various cucurbits to infection by plasmid-harboring strains of agrobacterium. Plant Physiol. 1986 Oct;82(2):622–624. doi: 10.1104/pp.82.2.622. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sonti R. V., Chiurazzi M., Wong D., Davies C. S., Harlow G. R., Mount D. W., Signer E. R. Arabidopsis mutants deficient in T-DNA integration. Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11786–11790. doi: 10.1073/pnas.92.25.11786. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Squire L. R., Ojemann J. G., Miezin F. M., Petersen S. E., Videen T. O., Raichle M. E. Activation of the hippocampus in normal humans: a functional anatomical study of memory. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1837–1841. doi: 10.1073/pnas.89.5.1837. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stachel S. E., Nester E. W. The genetic and transcriptional organization of the vir region of the A6 Ti plasmid of Agrobacterium tumefaciens. EMBO J. 1986 Jul;5(7):1445–1454. doi: 10.1002/j.1460-2075.1986.tb04381.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomashow M. F., Karlinsey J. E., Marks J. R., Hurlbert R. E. Identification of a new virulence locus in Agrobacterium tumefaciens that affects polysaccharide composition and plant cell attachment. J Bacteriol. 1987 Jul;169(7):3209–3216. doi: 10.1128/jb.169.7.3209-3216.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson D. V., Melchers L. S., Idler K. B., Schilperoort R. A., Hooykaas P. J. Analysis of the complete nucleotide sequence of the Agrobacterium tumefaciens virB operon. Nucleic Acids Res. 1988 May 25;16(10):4621–4636. doi: 10.1093/nar/16.10.4621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tinland B., Hohn B., Puchta H. Agrobacterium tumefaciens transfers single-stranded transferred DNA (T-DNA) into the plant cell nucleus. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8000–8004. doi: 10.1073/pnas.91.17.8000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Valvekens D., Van Montagu M., Van Lijsebettens M. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536–5540. doi: 10.1073/pnas.85.15.5536. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whalen M. C., Innes R. W., Bent A. F., Staskawicz B. J. Identification of Pseudomonas syringae pathogens of Arabidopsis and a bacterial locus determining avirulence on both Arabidopsis and soybean. Plant Cell. 1991 Jan;3(1):49–59. doi: 10.1105/tpc.3.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winans S. C. Two-way chemical signaling in Agrobacterium-plant interactions. Microbiol Rev. 1992 Mar;56(1):12–31. doi: 10.1128/mr.56.1.12-31.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yu G. L., Katagiri F., Ausubel F. M. Arabidopsis mutations at the RPS2 locus result in loss of resistance to Pseudomonas syringae strains expressing the avirulence gene avrRpt2. Mol Plant Microbe Interact. 1993 Jul-Aug;6(4):434–443. doi: 10.1094/mpmi-6-434. [DOI] [PubMed] [Google Scholar]
- Zupan J. R., Zambryski P. Transfer of T-DNA from Agrobacterium to the plant cell. Plant Physiol. 1995 Apr;107(4):1041–1047. doi: 10.1104/pp.107.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]