Plant Cell Reports

, Volume 16, Issue 11, pp 745–753

Factors affecting Agrobacterium-mediated transformation in Citrus and production of sour orange (Citrus aurantium L.) plants expressing the coat protein gene of citrus tristeza virus

  • M. A. Gutiérrez-E.
  • D. Luth
  • G. A. Moore

DOI: 10.1007/s002990050313

Cite this article as:
Gutiérrez-E., M., Luth, D. & Moore, G. Plant Cell Reports (1997) 16: 745. doi:10.1007/s002990050313

Abstract

Factors influencing transformation frequencies using the Agrobacterium-mediated protocol developed for Citrus seedling internodal stem segments in this laboratory were evaluated, with particular emphasis on decreasing the numbers of ``escape'' shoots produced. Although the use of a wild-type ``shooty'' Agrobacterium strain allowed relatively high frequencies of β-glucuronidase positive (GUS+) shoots to be produced, none of the shoots were free of wild-type T-DNA and would not root. Both use of a liquid medium/kanamycin overlay and horizontal placement of stem segments increased the efficiency of kanamycin selection. Wounding via particle bombardment prior to Agrobacterium inoculation did not increase transformation frequencies. The concentration of benzyladenine (BA) in the regeneration/selection medium inversely influenced the numbers of shoots that regenerated and the subsequent ability of the shoots to root. Regeneration in the presence of kanamycin also influenced the ability of shoots to root. Many of the shoots that regenerated on selection medium were chimeric for GUS expression, and plants established from such shoots ranged from non-staining to solidly staining for GUS. However, solidly transformed plants with integrated T-DNA were obtained, and these plants have maintained the expression of transgenes over several years. The transgenic plants include ones of sour orange (C. aurantium L.) and Key lime (C. aurantifolia (Christm.) Swing.), two species not previously transformed, and have integrated and express the coat protein gene of citrus tristeza virus. This is the first report of a potentially agriculturally important transgene being expressed in Citrus.

Key words Genetic transformation Key lime Carrizo citrange 

Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • M. A. Gutiérrez-E.
    • 1
  • D. Luth
    • 1
  • G. A. Moore
    • 1
  1. 1.Horticultural Sciences Department, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32611, USAUS

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