Abstract
Agrobacterium tumefaciens-mediated transformation has been widely used in molecular characterization of genes inArabidopsis thaliana. A number of procedures have been developed for transformation ofArabidopsis explants usingAgrobacterium. This paper describes an improved protocol for transformation ofArabidopsis thaliana root explants. Most significantly, using this protocol one can achieve efficient root regeneration of transformation in Landsbergerecta, an ecotype which is widely used in genetic and molecular analyses and which has been difficult to transform in the past. Additional modifications allow easy production of roots for transformation and regeneration of large numbers of transformation t shoots.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- 2ip Ade:
-
N6-(2-isopentenyl)-adenine
- B5:
-
Gamborg’s B5 salts
- CIM:
-
callus-inducing medium
- DMSO:
-
dimethyl sulfoxide
- GM:
-
seed germination and seedling growth medium
- GUS:
-
β-glucuronidase
- IAA:
-
Indole-3-acetic acid
- MES:
-
2-(N-morpholino)-ethanesulfonic acid
- MS:
-
Murashige and Skoog salts
- NAA:
-
α-naphthalene acetic acid
- RIM:
-
root-inducing medium
- SIM:
-
shoot-inducing medium
- TC:
-
tissue culture
- uidA :
-
gene encoding β-glucuronidase
References
An, G., B.D. Watson, and C.C. Chiang. 1986. Transformation of tobacco, tomato, andArabidopsis thaliana using a binary Ti vector system. Plant Physiol. 81:301–305.
Jefferson, R.A.. 1987. Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol. Biol. Reptr. 5:387–405.
Koornneef, M. 1987. Linkage map ofArabidopsis thaliana (2n=10). InGenetic Maps 1987: A Compliation of Linkage and Restriction Maps of Genetically Studied Organisms, ed. S.J. O'Brien. Cold Spring Harbor, pp. 742–745.
Lazo, G.R., P.A. Stein, and R.A. Ludwig. 1991. A DNA transformation-competentArabidopsis genomic library inAgrobacterium.Bio/Technology 9:963–968.
Lloyd, A.M., A.R. Barnason S.G. Rogers, M.C. Byrne, R.T. Fraley, and R.B. Horsch. 1986. Transformation ofArabidopsis thaliana withAgrobacterium tumefaciens. Science 234:464–466.
Ma, H., Yanofsky, M.F., and E.M. Meyerowitz. 1990. Molecular cloning and characterization ofGPA1, a G protein α subunit gene fromArabidopsis thaliana. Proc. Natl. Acad. Sci. USA. 87:3821–3825.
Meyerowitz, E.M.. 1989.Arabidopsis, a useful weed. Cell 56:263–269.
Rogers, S.G., H.J. Klee, R.B. Horsch, and R.T. Fraley. 1987. Improved vectors for plant transformation: expression cassette vectors and new selectable markers. Methods Enzymol. 153:253–277.
Sheikholeslam, S.N., and D.P. Weeks. 1987. Acetosyringone promotes high efficiency transformation ofArabidopsis thaliana explants byAgrobacterium tumefaciens. Plant Mol. Biol. 8:291–298.
Valvekens, D., M. Van Montagu, and M. Van Lijsebettens. 1988.Agrobacterium tumefaciens-mediated transformation ofArabidopsis thaliana root explants by using kanamycin selection. Proc. Natl. Acad. Sci. USA. 85: 5536–5540.
Van Sluys, M., J. Tempé, and N. Fedoroff, 1987. Studies on the introduction and mobility of the maize activator element inArābidopsis thaliana andDaucus carota. EMBO. 6:3881–3889.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Huang, H., Mā, H. An improved procedure for transformingArabidopsis thaliana (Landsbergerecta) root explant. Plant Mol Biol Rep 10, 372–383 (1992). https://doi.org/10.1007/BF02668914
Issue Date:
DOI: https://doi.org/10.1007/BF02668914