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
References
Acuna JR, de Pena M, (1991) Plant regeneration from protoplasts of embryogenic cell suspensions of Coffea arabica L. cv. caturraPlant Cell Rep. 10: 345–348
Acereto-Escoffié POM, BH Chi-Manzanero, S Echeverría- Echeverría, A James, R Grijalva, LC Rodríguez-Zapata, (2005) Agrobacterium-mediated transformation of Musa acuminata cv “Grand Nain” scalps by vacuum infiltrationScientia Horticulturae 105: 359–371
Carneiro MF (1997) Coffee biotechnology and its application in genetic transformationEuphytica 96: 167–172
Hatanaka T, Choi YE, Kusano T, Sano H (1999) Transgenic plants of coffee Coffea canephora from embryogenic callus via Agrobacterium tumefaciens-mediated transformationPlant Cell Rep. 19: 106–110
Gallie DR, Lucas WJ, Walbot V (1989) Visualizing mRNA expression in plant protoplasts: factors influencing efficient mRNA uptake and translationPlant Cell 1: 303–311
Leroy T, Paillard M, Berthouly M, Spiral J, Tessereau S, Legavre T, Altosaar I (1997): Introduction de gènes d’intèrêt agronomique dans l’espèce Coffea canephora Pierre part transformation avec Agrobacterium sp. ASIC, Nairobi, pp 439–446
Leroy T, Henry AM, Royer M, Altosaar I, Frutos R, Duris D, Philippe R, (2000) Genetically modified coffee plants expressing the Bacillus thuringiensis cry1Ac gene for resistance to leaf minerPlant Cell Rep. 19: 382–389
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culturesPhysiol. Plant. 15: 473–497
Ogita S, Uefuji H, Morimoto M, Sano H (2004) Application of RNAi to confirm theobromine as the major intermediate for caffeine biosynthesis in coffee plants with potential for construction of decaffeinated varietiesPlant Mol. Biol. 54: 931–941
Quiroz-Figueroa FR, Fuentes-Cerda CFJ, Rojas-Herrera R, Loyola-Vargas VM (2002) Histological studies on the developmental stages and differentiation of two different somatic embryogenesis systems of Coffea arabicaPlant Cell Rep. 20: 1141–1149
Spiral J, Thierry C, Paillard M, Petiard V, (1993) Regeneration of plantlets of Coffea canephora Pierre (Robusta) transformed by Agrobacterium rhizogenesC. R. Acad. Sci. (Paris) Sér. III 316: 1–6
Yasuda T, Fujii Y, Yamaguchi T (1985) Embryogenic callus induction from Coffea arabica leaf explants by benzyladeninePlant Cell Physiol. 26: 595–597
Zuo J, Niu QW, Frugis G, Chua NH (2002) The WUSCHEL gene promotes vegetative-to-embryonic transition in ArabidopsisPlant J. 30(3):349–359
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