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.
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Abbreviations
- EDTA:
-
ethylenediaminetetraacetic acid
- MS:
-
Murashige and Skoog
- PCR:
-
polymerase chain reaction
- SDS:
-
sodium dodecyl sulfate
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Acknowledgements
We wish to thank Francisco Quiroz and Mirian Monforte for their technical help and Luis Joel Figeroa from Accesolab for the contribution of reagents for this study. Dr Castano is partially funded by CONACYT Grant 39731-z.
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Canche-Moo, R., Ku-Gonzalez, A., Burgeff, C. et al. Genetic transformation of Coffea canephora by vacuum infiltration. Plant Cell Tiss Organ Cult 84, 373–377 (2006). https://doi.org/10.1007/s11240-005-9036-4
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DOI: https://doi.org/10.1007/s11240-005-9036-4