Summary
A method (termed co-cultivation) for transforming plant cells in vitro with A. tumefaciens strains, which was originally developed by Marton et al. (1978) Nature 277: 129–131, has been modified by the incorporation of a novel feeder plate culture system and been extended to use with petunia protoplasts. Using efficient cell plating and selection conditions for phytohormone-independent growth, large numbers of independent transformed calli can be obtained efficiently (∼10-1) and in less than 3 weeks following protoplast isolation. Southern hybridization analysis has confirmed that the majority of the resulting in vitro transformants contain a single copy of full length T-DNA.
The high efficiency of this procedure allows simple screening to identify plant cells transformed by Ti plasmids attenuated by deletion of internal T-DNA regions. Results are presented that demonstrate the co-cultivation method can be used in conjunction with short term assays for monitoring plant gene expression.
Similar content being viewed by others
References
Bevan M, Chilton M-D: Ann Rev Genet 16: 357–384, 1982.
Kahl G, Schell J: Molecular Biology of Plant Tumors. Academic Press, New York, NY, 1982.
Gelvin S, Gordon M, Nester E, Aronson A: Plasmid 6: 17–29, 1981.
Bevan M, Chilton M-D: J Mol Appl Genet 1539–546, 1982.
Willmitzer L, Dhaese P, Schreier P, Schmalenbach W, Van Montague M, Schell J: Cell 32: 1045–1056, 1983.
Willmitzer L, Simons G, Schell J: The EMBO Journal 1: 139–146, 1982.
Bevan M, Flavell R, Chilton M-D: Nature 304: 184–187, 1983.
Herrera-Estrella L, Depicker A, Van Montagu M, Schell J: Nature 303: 184–187, 1983.
Fraley R, Rogers S, Horsch R, Sanders P, Flick J, Adams S, Bittner M, Brand L, Fink C, Fry J, Galluppi G, Goldberg S, Hoffman N, Woo S: Proc Natl Acad Sci USA 80: 4803–4807, 1983.
Herrera-Estrella L, De Block M, Messens E, Hernalsteens J-P, Van Montague M, Schell J: The EMBO Journal 2: 987–995, 1983.
Marton L, Wullems G, Molendijk L, Schilperoort R: Nature 277: 129–131, 1979.
Wullems G, Molendijk L, Ooms G, Schilperoort R: Proc Natl Acad Sci USA 78: 4344–4348, 1981.
Matzke A, Chilton M-D: J Molec Appl Genet 1: 39–49, 1981.
Ausubel F, Bahnsen K, Hanson M, Michell A, Smith H: PMB Newsletter 1: 26–32, 1980.
Horsch R, Jones G: In vitro 16: 103–108, 1980.
Otten L, Schilperoort R: Biochem Biophys Acta 527: 497–500, 1978.
Nagao R, Shah D, Eckenrode V, Meagher R: DNA 1: 1–9, 1981.
Southern E: J Mol Biol 98: 503–517, 1975.
Jia J-F, Shillito R, Potrykus I: Z Pflanzemphysid BD 112S: 1–6, 1983.
Fraley R, Horsch R: in Genetic Engineering of Plants (eds T. Kosuge, C. Meredith and H. Hollaender) Plenum Publishing Corp NY, NY pp 177–194, 1983.
Wullems G, Molendijk L, Ooms G, Schilperoort R: Cell 24: 719–727, 1981.
Joos H, Inze D, Caplan A, Sormann M, Van Montague M, Schell J: Cell 32: 1057–1067, 1983.
Hanold D: Plant Science Lett 30: 177–183, 1983.
Gorman C, Padmanabhan R, Howard B: Science 221: 551–553, 1983.
Horowitz M, Cepko C, Sharp P: J Mol Appl Genet 2: 147–159, 1983.
DeGreve H, Decraemer H, Seurinck J, Van Montagu M, Schell J: Plasmid 6: 235–248, 1981.
Sciaky D, Montoya A, Chilton M-D: Plasmid 1: 238–253, 1978.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fraley, R.T., Horsch, R.B., Matzke, A. et al. In vitro transformation of petunia cells by an improved method of co-cultivation with A. tumefaciens strains. Plant Mol Biol 3, 371–378 (1984). https://doi.org/10.1007/BF00033384
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00033384