Genetic transformation of Coffea arabica cv. Red caturra by electroporation of cell suspensions embryogenic
Journal Title: Biotecnología Vegetal - Year 2011, Vol 11, Issue 1
Abstract
Plant regeneration by somatic embryogenesis from cell suspensions is considered the most efficient method for mass production of plants. The objective of this work was to achieve the genetic transformation of Coffea arabica cv. Caturra rojo by electroporation of embryogenic cell suspensions. The effect of different electroporation conditions (capacitance and field strength) on the cell suspensions without the application of plasmid DNA, by analysis of cell vitality at 24, 48 and 72 hours after electroporation was evaluated. The effect of electroporation buffer on cell vitality and transient expression of the gene for beta-glucuronidase was also determined. Besides, the transient expression of GUS gene in embryogenic cell suspensions electroporated with plasmid PDB-GUS-INT was studied. In the electroporated cell suspensions, under different electroporation conditions, electric shocks significantly affect cell vitality compared to the control group. In the Electroporated cell suspensions with buffer of Dekeyser et al. (1990) and applying a discharge time of 500 msec was obtained the highest percentage of vitality, with a value of 32.14, compared to other treatments. While in the electroporated cell aggregates with a capacitance of 100 and 1200 uF there was a higher transient GUS gene activity (59.37 and 61.01%) compared to electroporated treatment with a capacitance of 490 uF, where values obtained were 50.0%.
Authors and Affiliations
R. Barbón, E. Jiménez, A. Capote, V. Gil, B. Ocaña
Keywords
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