Biologia plantarum 49:493-497, 2005 | DOI: 10.1007/s10535-005-0038-1

Genetic transformation of Coffea canephora by particle bombardment

A. F. Ribas1, A. K. Kobayashi2, L. F. P. Pereira3, L. G. E. Vieira1,*
1 IAPAR - Laboratorio de Biotecnologia, Londrina, PR, Brazil
2 Embrapa - Mandioca e Fruticultura (CNPMF), Cruz das Almas, BA, Brazil
3 Embrapa - Cafe, Laboratorio de Biotecnologia, IAPAR, Londrina, PR, Brazil

Stable transformation of Coffea canephora P. was obtained by particle bombardment of embryogenic tissue. Leaf explants were cultured on medium supplemented with 5 µM isopentenyl-adenosine to induce direct embryogenesis. Explants with somatic embryos were transferred to half strength MS medium with 9 µM 2,4 dichlorophenoxyacetic acid. After 2 weeks, the explants with somatic embryos and embryogenic tissue were bombarded with tungsten particles (M-25) carrying the plasmid pCambia3301 (containing the bar and uidA genes) using a high pressure helium microprojectile device. The bombarded explants were submitted to selection on medium containing 5 µM ammonium glufosinate herbicide as selective agent. After 6 months, putative transgenic embryos were transferred to a growth regulator-free medium for germination. The regenerated plantlets were β-glucuronidase (GUS) positive whereas no GUS activity was observed in non-transgenic controls. Incorporation of the bar gene into the genome was confirmed by PCR and Southern blot analysis of the regenerated transformed plants. Greenhouse grown transgenic coffee plants were found to withstand the recommended level of the herbicide Finale™ for weed control.

Keywords: biolistic; transgenic coffee; ammonium glufosinate
Subjects: ammonium glufosinate; Coffea canephora; cytokinins; embryo development, somatic; β-glucuronidase; in vitro culture, somatic embryogenesis; phosphinothricin acetyltransferase; polymerase chain reaction (PCR); Southern blot analysis; transformation, particle bombardment

Received: April 13, 2004; Accepted: April 22, 2005; Published: December 1, 2005Show citation

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Ribas, A.F., Kobayashi, A.K., Pereira, L.F.P., & Vieira, L.G.E. (2005). Genetic transformation of Coffea canephora by particle bombardment. Biologia plantarum, 49(4), 493-497. doi: 10.1007/s10535-005-0038-1.
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    References

    1. Aragao, F.J.L., Vianna, G.R., Albino, M.M.C., Rech, E.L.: Transgenic dry bean tolerant to the herbicide glufosinate ammonium.-Crop Sci. 43: 1298-1302, 2002. Go to original source...
    2. Bayer, E., Gugel, K.H., Haegele, K., Hagenmaier, H., Jessipow, S., Koenig, W.A., Zaehner, H.: Phosphinothricin and phosphinothricyl-alanyl-alanin.-Helv. chim. Acta. 55: 224-239, 1972. Go to original source...
    3. Bower, R., Elliot, A.R., Pottier, B.A.M., Birch, R.G.: High-efficiency, microprojectile-mediated co-transformation of sugarcane, using visible or selectable markers.-Mol. Breed. 2: 239-249, 1996. Go to original source...
    4. Cabrera-Ponce, J.L., Vegas-Garcia, A., Herrera-Estrella, L.: Herbicide resistant transgenic papaya plants produced by an efficient particle bombardment transformation method.-Plant Cell Rep. 15: 1-7, 1995. Go to original source...
    5. Cao, J., Duan, X., McElroy, D., Wu, R.: Regeneration of herbicide resistant transgenic rice plants following microprojectile-mediated transformation of suspension culture cells.-Plant Cell Rep. 11: 586-591, 1992. Go to original source...
    6. Carneiro, M.F.: Advances in coffee biotechnology.-AgBiotechNet 1: 1-7, 1999.
    7. Falco, M.C., Tulmann Neto, A., Ulian, E.C.: Transformation and expression of a gene for herbicide resistance in Brazilian sugarcane.-Plant Cell Rep. 19: 1188-1194, 2000. Go to original source...
    8. Gray, D.J., Finer, J.J.: Development and operation of five particle guns for introduction of DNA into plant cells.-Plant Cell Tissue Organ Cult. 33: 219, 1993. Go to original source...
    9. Ha, S.B., Lee, S.B., Lee, D.E., Guh, J.O., Back, K.: Transgenic rice plants expressing Bacillus subtilis protoporphyrinogen oxidase gene show herbicide oxyfluorfen resistence.-Biol. Plant. 47: 277-280, 2003/4.
    10. Hatanaka, J., Arakawa, O., Yasuda, T., Ushida, N., Yamaguchi, I.: Effect of plant growth regulators on somatic embryogenesis in leaf cultures of Coffea canephora.-Plant Cell Rep. 10: 179-182, 1991. Go to original source...
    11. Hatanaka, T., Choi, Y.E., Kusano, T., Sano, H.: Transgenic plants of Coffea canephora from embryogenic callus via Agrobacterium tumefaciens-mediated transformation.-Plant Cell Rep. 19: 106-110, 1999. Go to original source...
    12. Klein, T.M., Wolf, E.D., Wu, R., Sanford, J.C.: High-velocity microprojectiles for delivering nucleic acids into living cells.-Nature 327: 70-73, 1987. Go to original source...
    13. Kosugi, S., Ohashi, Y., Nakajima, K., Arai, Y.: An improved assay for β-glucuronidase in transformed cells: methanol almost completely supresses a putative endogenous β-glucuronidase activity.-Plant Sci. 70: 133-140, 1990. Go to original source...
    14. Leroy, T., Henry, A. M., Royer, M., Altosar, I., Frutos, R., Duris, D., Philippe, R.: Genetically modified coffee plants expressing the Bacillus thuringiensis cry1Ac gene for resistance to leaf miner.-Plant Cell Rep. 19: 382-389, 2000. Go to original source...
    15. Murashige, T., Skoog, F.: A revised medium for rapid growth and bio-assays with tobacco tissue culture.-Plant Physiol. 15: 473-497, 1962. Go to original source...
    16. Rasco-Gaunt, S., Riley, A., Lazzeri, P., Barcelo, P.: A facile method for screening for phosphinothricin (PPT)-resistant transgenic wheats.-Mol. Breed. 5: 255-262, 1999. Go to original source...
    17. Pfister, K., Steinback, K.E., Gardner, E., Arntzen, C.J.: Photoaffinity-labeling of an herbicide receptor protein in chloroplast membranes.-Proc. nat. Acad. Sci. USA. 78: 981-985, 1981. Go to original source...
    18. Rosillo, A. G., Acuna, J.R., Gaitan, A.L., Pena, M.: Optimised DNA delivery into Coffea arabica suspension culture cells by particle bombardment.-Plant Cell Tissue Organ Cult. 74: 45-49, 2003. Go to original source...
    19. Sambrook, J., Fritsch, E.F., Maniatis, T. (ed.): Molecular Cloning: a Laboratory Manual.-Cold Spring Harbour Laboratory Press, New York 1989.
    20. Spiral, J., Leroy, T., Paillard, M., Petiard, V.: Transgenic coffee (Coffea sp.).-In: Bajaj, Y.P.S. (ed.): Transgenic Trees. (Biotechnology in Agriculture and Forestry. Vol. 44). Pp. 55-76. Spring-Verlag, Berlim-Heidelberg 1999. Go to original source...
    21. Spiral, J., Thierry, C., Paillard, M., Petiard, V.: Obtention de plantules de Coffea canephora Pierre (Robusta) transformees par Agrobacterium rhizogenes.-Compt. rend. Acad. Sci. Paris 316: 1-6. 1993.
    22. Sripaoraya, S., Marchant, R., Power, J.B., Davey, M.R.: Herbicide-tolerant transgenic pineapple (Ananas comosus) produced by microprojectile bombardment.-Ann. Bot. 88: 597-603, 2001. Go to original source...
    23. Sugiyama, M., Matsuoka, C., Takagi, T.: Transformation of Coffea with Agrobacterium rhizogenes.-In: ASIC, 16eme Colloque Scientifique sur le Cafe. Pp. 853-859. Paris 1995.
    24. Van Boxtel, J., Berthouly, M., Carasco, C., Dufuor, M., Eskes, A.: Transient expresssion of β-glucoronidase following biolistic delivery of foreign DNA into coffee tissue.-Plant Cell Rep. 14: 748-752, 1995a. Go to original source...
    25. Van Boxtel, J., Eskes, A., Berthouly, M.: Glufosinate as an efficient inhibitor of callus proliferation in coffee tissue.-In Vitro cell. dev. Biol. Plant 33: 6-12, 1995b.
    26. Visarada, K.B.R.S., Sarma, N.P.: Transformation of indica rice throught particle bombardment: factors influencing transient expression and selection.-Biol. Plant. 48: 25-31, 2004. Go to original source...

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