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Annals of Botany 89: 491-497, 2002
© 2002 Annals of Botany Company

Green Fluorescent Protein as a Visual Marker in Somatic Hybridization

O. OLIVARES-FUSTER1, L. PEÑA1, N. DURAN-VILA1 and L. NAVARRO*,1

1Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias, Apartado Oficial 46113, Moncada (Valencia), Spain

* For correspondence. Fax 34 96 139 0240, e-mail lnavarro{at}ivia.es

Received: 4 June 2001; Returned for revision: 10 October 2001; Accepted: 14 December 2001.

Using a transgenic citrus plant expressing Green Fluorescent Protein (GFP) as a parent in somatic fusion experiments, we investigated the suitability of GFP as an in vivo marker to follow the processes of protoplast fusion, regeneration and selection of hybrid plants. A high level of GFP expression was detected in transgenic citrus protoplasts, hybrid callus, embryos and plants. It is demonstrated that GFP can be used for the continuous monitoring of the fusion process, localization of hybrid colonies and callus, and selection of somatic hybrid embryos and plants.

Key words: Protoplast fusion, genetic transformation, green fluorescent protein, GFP, woody plant, vital marker, Citrus reticulata, Citrus sinensis, Poncirus trifoliata, Agrobacterium tumefaciens.


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