Summary
Tomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transformation and the technique has been useful for the isolation and identification of many genes involved in plant disease resistance, morphology and development. In this method, cotyledon explants from in vitro-grown seedlings are precultured overnight on a tobacco suspension feeder layer. The explants are then inoculated with Agrobacterium and returned to the feeder layer for a 2-d period of cocultivation. After cocultivation, the explants are transferred to an MS-based selective regeneration medium containing zeatin. Regenerated shoots are then rooted on a separate selective medium. This protocol has been used with several tomato cultivars and routinely yields transformation efficiencies of 10–15%.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Horsch, R. B., Fry, J. B., Hoffmann, N. L., et al. (1985) A simple and general method for transferring genes into plants. Science 227, 1229–1231.
McCormick, S., Niedermeyer, J., Fry, J., Barnason, A., Horsch, R., and Fraley, R. (1986) Leaf disc transformation of cultivated tomato (Lycopersicon esculentum) using Agrobacterium tumefaciens. Plant Cell Rep. 5, 81–84.
Toyoda, H., Matsuda, Y., Utsumi, R., and Ouchi, S. (1988) Intranuclear microinjection for transformation of tomato callus cultures. Plant Cell Rep. 7, 293–296.
Xu, Y., Yu, H., and Hall, T.C. (1994) Rice triosephosphate isomerase gene 5′ sequence directs glucuronidase activity in transgenic tobacco but requires an intron for expression in rice. Plant Physiol. 106, 459–467.
Van Eck, J. M., Blowers, A. D., and Earle, E. D. (1995) Stable transformation of tomato cell cultures after bombardment with plasmid and YAC DNA. Plant Cell Rep. 14, 299–304.
Nakata, K., Tanaka, H., Yano, K., and Takagi, M. (1992) An effective transformation system for Lycopersicon peruvianum by electroporation. Jpn. J. Breed. 42, 487–495.
Koornneef, M., Hanhart, C., Jongsma, M., et al. (1986) Breeding of a tomato genotype readily accessible to genetic manipulation. Plant Sci. 45, 201–208.
Tanksley, S. D., Ganal, M. W., Prince, J. P., et al. (1992) High density molecular linkage maps of the tomato and potato genomes. Genetics 132, 1141–1160.
Martin, G. B., Frary, A., Wu, T., et al. (1994) A member of the tomato Pto gene family confers sensitivity to fenthion resulting in rapid cell death. Plant Cell 6, 1543–1552.
Frary, A., Nesbitt, T. C., Frary, A., et al. (2000) fw2.2: a quantitative trait locus key to the evolution of tomato fruit size. Science 289, 85–88.
Jones, D. A., Thomas, C. M., Hammond-Kosack, K. E., Balint-Kurti, P. J., and Jones, J. D. (1994) Isolation of the tomato Cf-9 gene for resistance to Cladosporium fulvum by transposon tagging. Science 266, 789–793.
Milligan, S. B., Bodeau, J., Yaghoobi, J., Kaloshian, I., Zabel, P., and Williamson, V. M. (1998) The root knot nematode resistance gene Mi from tomato is a member of the leucine zipper, nucleotide-binding leucine-rich repeat family of plant genes. Plant Cell 10, 1307–1319.
Liu, J., Van Eck, J., Cong, B., and Tanksley, S. D. (2002) A new class of regulatory genes underlying the cause of pear-shaped tomato fruit. Proc. Natl. Acad Sci. USA 99, 13,302–13,306.
Sheehy, R. E., Kramer, M., and Hiatt, W. R. (1988) Reduction of polygalacturonase activity in tomato fruit by antisense RNA. Proc. Nat. Acad. Sci. USA 85, 8805–8809.
Frary, A. and Hamilton, C. M. (2001) Efficiency and stability of high molecular weight DNA transformation: an analysis in tomato. Transgenic Res. 10, 121–132.
Chyi, Y. S. and Phillips, G. C. (1987) High efficiency Agrobacterium-mediated transformation of Lycopersicon based on conditions favorable for regeneration. Plant Cell Rep. 6, 105–108.
Fillatti, J. J., Kiser, J., Rose, R., and Comai, L. (1987) Efficient transfer of a glyphosate tolerance gene into tomato using a binary Agrobacterium tumefaciens vector. Biotechnology 5, 726–730.
Hamza, S. and Chupeau, Y. (1993) Re-evaluation of conditions for plant regeneration and Agrobacterium-mediated transformation from tomato (Lycopersicon esculentum). J. Exp. Bot. 44, 1837–1845.
Davis, M. E., Miller, A. R., and Lineberger, R. D. (1991) Temporal competence for transformation of Lycopersicon esculentum (L. Mill.) cotyledons by Agrobacterium tumefaciens: relation to wound-healing and soluble plant factors. J. Exp. Bot. 42, 359–364.
van Roekel, J. S., Damm, B., Melchers, L. S., and Hoekema, A. (1993) Factors influencing transformation frequency of tomato (Lycopersicon esculentum). Plan Cell Rep. 12, 644–647.
Frary, A. and Earle, E. D. (1996) An examination of factors affecting the efficiency of Agrobacterium-mediated transformation of tomato. Plant Cell Rep. 16, 235–240.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Frary, A., Van Eck, J. (2005). Organogenesis From Transformed Tomato Explants. In: Peña, L. (eds) Transgenic Plants: Methods and Protocols. Methods in Molecular Biology™, vol 286. Humana Press. https://doi.org/10.1385/1-59259-827-7:141
Download citation
DOI: https://doi.org/10.1385/1-59259-827-7:141
Publisher Name: Humana Press
Print ISBN: 978-1-58829-263-6
Online ISBN: 978-1-59259-827-4
eBook Packages: Springer Protocols