[Establisment of a genetic transformation method of coffee (Coffea arabica cv. Catimor) and incorporation of bar gene for ammonium glufosinate resistance].
In order to establish a successful method of genetic transformation in Coffea arabica cv. Catimor, different conditions of generation and electroporation were evaluated on different plant tissues. Cell suspension system was improved using one hormone only (BA), obtaining high yields of primary and secondary somatic embryo production. For selection of viable and potentially transformed cells, MTT (1%) method and ammonium glufosinate concentration (1 mg/L in leaf, callus and embryos; and 5 mg/L in cells) were established. Different conditions were evaluated to electroporate different explants (embryogenic callus, vitroplants leaves, globular and torpedo embryos). The highest gus gene expression percentage by explant were found on enzymatic treated tissues at 375 V/cm in callus, and at 625 V/cm in leaves and embryos. Torpedo embryos cultured on liquid medium were the only type of tissue that could regenerate into plants, where secondary somatic embryos were obtained. Those embryos were positive to the gus gene histochemical test and to the gus and bar genes amplification on a PCR reaction.
Fernández Da Silva
Rafael
R
Post-grado en Botánica, Facultad de Ciencias, Universidad Central de Venezuela, Apartado Postal 47114, Caracas 1041, Venezuela. rafaelfer@tecel.net.ve
D000613:Aminobutyrates / Q000494:pharmacology*; D003069:Coffee / Q000187:drug effects / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics*; D006540:Herbicides / Q000494:pharmacology*; D030821:Plants, Genetically Modified / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic / Q000235:genetics*
D000613:Aminobutyrates; D003069:Coffee; D006540:Herbicides; C003121:phosphinothricin