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Genetic transformation of Coffea canephora by vacuum infiltration |
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| Authors: | RLR Canche-Moo A Ku-Gonzalez C Burgeff VM Loyola-Vargas LC Rodríguez-Zapata E Castaño |
| Institution: | (1) Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, Calle 43, No. 130, Colonia Chuburná de Hidalgo, C.P. 97200, Mérida, Yucatán, México;(2) Unidad de Biotecnología, Centro de Investigación Científica de Yucatán, Calle 43, No. 130, Colonia Chuburná de Hidalgo, C.P. 97200, Mérida, Yucatán, México |
| Abstract: | Agrobacterium-mediated plant transformation protocol was evaluated as a fast method to obtain genetically modified Coffea canephora plantlets. Leaf explants were used as source material for Agrobacterium tumefaciens-mediated transformation involving a vacuum infiltration protocol, followed by a step of somatic embryogenesis induction and a final selection of the transformed plants. A. tumefaciens strain C58CI containing the binary vector pER10W-35SRed was used. PCR amplification of DsRFP gene and visual detection of the red fluorescent protein demonstrated 33% transformed embryos. The protocol presented here produces reliable transgenic coffee embryos in two months. |
| Keywords: | A grobacterium transformation coffee somatic embryogenesis vacuum infiltration |
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