Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Sep;83(18):6935–6939. doi: 10.1073/pnas.83.18.6935

Complementation of Agrobacterium tumefaciens tumor-inducing aux mutants by genes from the TR-region of the Ri plasmid of Agrobacterium rhizogenes

I A Offringa *, L S Melchers *, A J G Regensburg-Tuink *, P Costantino †, R A Schilperoort *, P J J Hooykaas *,‡
PMCID: PMC386625  PMID: 16593762

Abstract

In this paper we provide information indicating that the agropine-type root-inducing (Ri) plasmid pRi1855 of Agrobacterium rhizogenes contains functional genes for auxin production (aux) in the right transferred DNA (T-DNA) region (TR-region). These genes were cloned and introduced into the T-region of the tumor-inducing (Ti) plasmids of mutants of Agrobacterium tumefaciens carrying an aux mutation. Depending on the Ri aux gene present, the oncogenicity of the Ti aux-1 and/or aux-2 mutations was restored, showing that the Ri aux genes are able to complement the Ti aux genes. Agrobacterium strains with an agropine-type Ri plasmid not only cause hairy root on certain plant species, but they also induce tumors on other plant species. In this paper it is shown that a mutation in either of the aux genes in the Ri plasmid leads to a total loss of tumorigenicity and a strongly diminished rhizogenicity of the host bacterium, revealing that the aux genes are important for tumor and root induction. Agrobacterium strains containing the TR-region but not the TL (left)-region of the Ri plasmid are still tumorigenic on certain plant species but are no longer capable of hairy-root induction.

Keywords: hairy root disease, phytohormones

Full text

PDF
6935

Images in this article

6937Image
on p.6937
6937Image
on p.6937
6937Image
on p.6937
6938Image
on p.6938
6938Image
on p.6938
6938Image
on p.6938

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akiyoshi D. E., Klee H., Amasino R. M., Nester E. W., Gordon M. P. T-DNA of Agrobacterium tumefaciens encodes an enzyme of cytokinin biosynthesis. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5994–5998. doi: 10.1073/pnas.81.19.5994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barry G. F., Rogers S. G., Fraley R. T., Brand L. Identification of a cloned cytokinin biosynthetic gene. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4776–4780. doi: 10.1073/pnas.81.15.4776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braun A. C. A Physiological Basis for Autonomous Growth of the Crown-Gall Tumor Cell. Proc Natl Acad Sci U S A. 1958 Apr;44(4):344–349. doi: 10.1073/pnas.44.4.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buchholz W. G., Thomashow M. F. Comparison of T-DNA oncogene complements of Agrobacterium tumefaciens tumor-inducing plasmids with limited and wide host ranges. J Bacteriol. 1984 Oct;160(1):319–326. doi: 10.1128/jb.160.1.319-326.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buchholz W. G., Thomashow M. F. Host range encoded by the Agrobacterium tumefaciens tumor-inducing plasmid pTiAg63 can be expanded by modification of its T-DNA oncogene complement. J Bacteriol. 1984 Oct;160(1):327–332. doi: 10.1128/jb.160.1.327-332.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Buchmann I., Marner F. J., Schröder G., Waffenschmidt S., Schröder J. Tumour genes in plants: T-DNA encoded cytokinin biosynthesis. EMBO J. 1985 Apr;4(4):853–859. doi: 10.1002/j.1460-2075.1985.tb03710.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chilton M. D., Drummond M. H., Merio D. J., Sciaky D., Montoya A. L., Gordon M. P., Nester E. W. Stable incorporation of plasmid DNA into higher plant cells: the molecular basis of crown gall tumorigenesis. Cell. 1977 Jun;11(2):263–271. doi: 10.1016/0092-8674(77)90043-5. [DOI] [PubMed] [Google Scholar]
  10. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Costantino P., Hooykaas P. J., den Dulk-Ras H., Schilperoort R. A. Tumor formation and rhizogenicity of Agrobacterium rhizogenes carrying Ti plasmids. Gene. 1980 Oct;11(1-2):79–87. doi: 10.1016/0378-1119(80)90088-8. [DOI] [PubMed] [Google Scholar]
  12. Costantino P., Spanò L., Pomponi M., Benvenuto E., Ancora G. The T-DNA of Agrobacterium rhizogenes is transmitted through meiosis to the progeny of hairy root plants. J Mol Appl Genet. 1984;2(5):465–470. [PubMed] [Google Scholar]
  13. De Paolis A., Mauro M. L., Pomponi M., Cardarelli M., Spanò L., Costantino P. Localization of agropine-synthesizing functions in the TR region of the root-inducing plasmid of Agrobacterium rhizogenes 1855. Plasmid. 1985 Jan;13(1):1–7. doi: 10.1016/0147-619x(85)90050-2. [DOI] [PubMed] [Google Scholar]
  14. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Garfinkel D. J., Simpson R. B., Ream L. W., White F. F., Gordon M. P., Nester E. W. Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis. Cell. 1981 Nov;27(1 Pt 2):143–153. doi: 10.1016/0092-8674(81)90368-8. [DOI] [PubMed] [Google Scholar]
  16. Hedges R. W. R factors from Proteus mirabilis and P. vulgaris. J Gen Microbiol. 1975 Apr;87(2):301–311. doi: 10.1099/00221287-87-2-301. [DOI] [PubMed] [Google Scholar]
  17. Hille J., van Kan J., Klasen I., Schilperoort R. Site-directed mutagenesis in Escherichia coli of a stable R772::Ti cointegrate plasmid from Agrobacterium tumefaciens. J Bacteriol. 1983 May;154(2):693–701. doi: 10.1128/jb.154.2.693-701.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hoekema A., Hooykaas P. J., Schilperoort R. A. Transfer of the octopine T-DNA segment to plant cells mediated by different types of Agrobacterium tumor- or root-inducing plasmids: generality of virulence systems. J Bacteriol. 1984 Apr;158(1):383–385. doi: 10.1128/jb.158.1.383-385.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hoekema A., Roelvink P. W., Hooykaas P. J., Schilperoort R. A. Delivery of T-DNA from the Agrobacterium tumefaciens chromosome into plant cells. EMBO J. 1984 Nov;3(11):2485–2490. doi: 10.1002/j.1460-2075.1984.tb02160.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hoekema A., de Pater B. S., Fellinger A. J., Hooykaas P. J., Schilperoort R. A. The limited host range of an Agrobacterium tumefaciens strain extended by a cytokinin gene from a wide host range T-region. EMBO J. 1984 Dec 20;3(13):3043–3047. doi: 10.1002/j.1460-2075.1984.tb02255.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hooykaas P. J., Hofker M., den Dulk-Ras H., Schilperoort R. A. 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. 1984 May;11(3):195–205. doi: 10.1016/0147-619x(84)90026-x. [DOI] [PubMed] [Google Scholar]
  22. Hooykaas P. J., den Dulk-Ras H., Ooms G., Schilperoort R. A. Interactions between octopine and nopaline plasmids in Agrobacterium tumefaciens. J Bacteriol. 1980 Sep;143(3):1295–1306. doi: 10.1128/jb.143.3.1295-1306.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Huffman G. A., White F. F., Gordon M. P., Nester E. W. Hairy-root-inducing plasmid: physical map and homology to tumor-inducing plasmids. J Bacteriol. 1984 Jan;157(1):269–276. doi: 10.1128/jb.157.1.269-276.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jouanin L. Restriction map of an agropine-type Ri plasmid and its homologies with Ti plasmids. Plasmid. 1984 Sep;12(2):91–102. doi: 10.1016/0147-619x(84)90055-6. [DOI] [PubMed] [Google Scholar]
  25. Kostriken R., Morita C., Heffron F. Transposon Tn3 encodes a site-specific recombination system: identification of essential sequences, genes, and actual site of recombination. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4041–4045. doi: 10.1073/pnas.78.7.4041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ooms G., Hooykaas P. J., Moolenaar G., Schilperoort R. A. Grown gall plant tumors of abnormal morphology, induced by Agrobacterium tumefaciens carrying mutated octopine Ti plasmids; analysis of T-DNA functions. Gene. 1981 Jun-Jul;14(1-2):33–50. doi: 10.1016/0378-1119(81)90146-3. [DOI] [PubMed] [Google Scholar]
  27. Ooms G., Hooykaas P. J., Van Veen R. J., Van Beelen P., Regensburg-Tuïnk T. J., Schilperoort R. A. Octopine Ti-plasmid deletion mutants of agrobacterium tumefaciens with emphasis on the right side of the T-region. Plasmid. 1982 Jan;7(1):15–29. doi: 10.1016/0147-619x(82)90023-3. [DOI] [PubMed] [Google Scholar]
  28. Pomponi M., Spanò L., Sabbadini M. G., Costantino P. Restriction endonuclease mapping of the root-inducing plasmid of Agrobacterium rhizogenes 1855. Plasmid. 1983 Sep;10(2):119–129. doi: 10.1016/0147-619x(83)90064-1. [DOI] [PubMed] [Google Scholar]
  29. Risuleo G., Battistoni P., Costantino P. Regions of homology between tumorigenic plasmids from Agrobacterium rhizogenes and Agrobacterium tumefaciens. Plasmid. 1982 Jan;7(1):45–51. doi: 10.1016/0147-619x(82)90025-7. [DOI] [PubMed] [Google Scholar]
  30. SKOOG F., MILLER C. O. Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol. 1957;11:118–130. [PubMed] [Google Scholar]
  31. Salomon F., Deblaere R., Leemans J., Hernalsteens J. P., Van Montagu M., Schell J. Genetic identification of functions of TR-DNA transcripts in octopine crown galls. EMBO J. 1984 Jan;3(1):141–146. doi: 10.1002/j.1460-2075.1984.tb01774.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schröder G., Waffenschmidt S., Weiler E. W., Schröder J. The T-region of Ti plasmids codes for an enzyme synthesizing indole-3-acetic acid. Eur J Biochem. 1984 Jan 16;138(2):387–391. doi: 10.1111/j.1432-1033.1984.tb07927.x. [DOI] [PubMed] [Google Scholar]
  33. Tepfer D. Transformation of several species of higher plants by Agrobacterium rhizogenes: sexual transmission of the transformed genotype and phenotype. Cell. 1984 Jul;37(3):959–967. doi: 10.1016/0092-8674(84)90430-6. [DOI] [PubMed] [Google Scholar]
  34. Thomashow L. S., Reeves S., Thomashow M. F. Crown gall oncogenesis: evidence that a T-DNA gene from the Agrobacterium Ti plasmid pTiA6 encodes an enzyme that catalyzes synthesis of indoleacetic acid. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5071–5075. doi: 10.1073/pnas.81.16.5071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thomashow M. F., Nutter R., Montoya A. L., Gordon M. P., Nester E. W. Integration and organization of Ti plasmid sequences in crown gall tumors. Cell. 1980 Mar;19(3):729–739. doi: 10.1016/s0092-8674(80)80049-3. [DOI] [PubMed] [Google Scholar]
  36. White F. F., Nester E. W. Relationship of plasmids responsible for hairy root and crown gall tumorigenicity. J Bacteriol. 1980 Nov;144(2):710–720. doi: 10.1128/jb.144.2.710-720.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Willmitzer L., Simons G., Schell J. The TL-DNA in octopine crown-gall tumours codes for seven well-defined polyadenylated transcripts. EMBO J. 1982;1(1):139–146. doi: 10.1002/j.1460-2075.1982.tb01137.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yanofsky M., Montoya A., Knauf V., Lowe B., Gordon M., Nester E. Limited-host-range plasmid of Agrobacterium tumefaciens: molecular and genetic analyses of transferred DNA. J Bacteriol. 1985 Jul;163(1):341–348. doi: 10.1128/jb.163.1.341-348.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES