Summary
A method is described for detecting plant metabolites capable of inducing the virulence genes of Agrobacterium tumefaciens. The method uses A. tumefaciens containing a plasmid with an inducible virulence gene fused to a galactosidase gene (virE::lacZ). Thin layer chromatography plates are overlayed with agar containing the indicator bacterium and a chromogenic galactoside (X-gal). Virulence gene inducing plant metabolites induce galactosidase which releases an aglycone readily oxidized by air to a blue pigmented zone at the Rf of the inducer. The method has been used to demonstrate the presence of virulence gene inducers in corn, wheat and rye. The uninduced background level of galactosidase also permits detection of bacterial growth inhibitors after a longer incubation period.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- X-gal:
-
5-Bromo-4-chloro-3-indoxyl ß-D-galactopyranoside
- TLC:
-
Thin Layer Chromatography
- AS:
-
Acetosyringone
- DIMBOA:
-
2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one
- MBOA:
-
6-Methoxy-2(3H)-benzoxazolone
- T-DNA:
-
Transfer DNA
- Ti Plasmid:
-
Tumor-inducing Plasmid
References
Bolton GW, Nester E W and Gordon M P (1986) Science 232, 983–985.
Chilton M-D, Currier T, Farrand S, Bendich A, Gordon M and Nester E W (1974) Proc. Natl. Acad. Sci. (USA) 71, 6708–6712.
Chilton M-D, Saiki R K, Yadav N, Gordon M P and Quetier F (1980) Proc. Natl. Acad. Sci. (USA) 77, 4060–4064.
Corcuera LJ, Woodward MD, Helgeson JP, Kelman CD and Upper CD (1978) Plant Physiol. 61, 791–795.
Grimsley N, Hohn B, Ramos C, Kado C and Rogowsky P (1989) Mol. Gen. Genet. 217, 309–316.
Hooykaas PJJ (1989) Plant Mol. Biol. 13, 327–336.
Huang Y, Morel P, Powell B and Kado CI (1990) J. Bacteriol. 172, 1142–1144.
Jin S, Roitsch T, Ankenbauer RG, Gordon MP and Nester EW (1990) J. Bacteriol. 172, 525–530.
Melchers LE, Regensburg-Tuink AJG, Schilperoort RA and Hooykaas PJJ (1989) Mol. Microbiol. 3, 969–977.
Messens E, Dekeyser R and Stachel SE (1990) Proc. Natl. Acad. Sci. (USA) 87, 4368–4372.
Morris JW, and Morris RO (1990) Proc. Natl. Acad. Sci. (USA) 87, 3614–3618.
Miller JH (1972) Experiments in Molecular Genetics (Cold Spring Harbor Lab., Cold Spring Harbor, NY).
Sahi SV, Chilton M-D and Chilton W S (1990) Proc. Natl. Acad. Sci. (USA) 87, 3879–3883.
Spencer PA, Tanaka A and Towers GHN (1990) Phytochem. 29, 3785–3788.
Spencer PA and Towers GHN (1988) Phytochem. 27, 2781–2785.
Stachel SE, Messens E, Van Montagu M and Zambryski P (1985) Nature 318, 624–629.
Stachel SE and Nester EW (1986) EMBO J. 5, 1445–1454.
Stachel SE, Nester EW and Zambryski PC (1986) Proc. Natl. Acad. Sci. (USA) 83, 379–383.
Stachel SE and Zambryski PC (1986) Cell 46, 325–333.
Usami S, Okamoto S, Takebe I and Machida Y (1988) Proc. Natl. Acad. Sci. (USA) 85, 3748–3752.
Winans SC, Kerstetter RA and Nester EW (1988) J. Bacteriol. 170, 4047–4054.
Zambryski P (1988) Ann. Rev. Genet. 22, 1–30.
Zerback R, Dressler K and Hess D (1989) Plant Sci. 62, 83–91.
Author information
Authors and Affiliations
Additional information
Communicated by W. Parrott
Rights and permissions
About this article
Cite this article
Sahi, S.V., Gagliardo, R.W., Chilton, MD. et al. A thin layer chromatographic technique for detecting inducers of Agrobacterium virulence genes in corn, wheat and rye. Plant Cell Reports 13, 489–492 (1994). https://doi.org/10.1007/BF00232942
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00232942