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Microsatellite and AFLP Analysis of Autochthonous Grapevine Cultivars from Galicia (Spain)

¹ Departamento de Fisiología Vegetal, Universidad de Santiago de Compostela, Campus Sur, 15872 Santiago de Compostela, Spain; ² Estación de Viticultura y Enología de Galicia, Xunta de Galicia, Leiro, 32427 Ponte San Clodio, Spain and (current address) Centro Superior de Hostelería de Galicia, Crta Santiago-Noia, km 1, 15896 A Barcia, Santiago de Compostela, Spain; ³ Tragsa, Vivero de Ourense, Crta Maceda-Valdrey, km 2, 32700 Maceda, Spain; and ⁴ Departamento de Biología Vegetal y Ciencia del Suelo, Universidad de Vigo, Campus Universitario, 36310 Vigo, Spain.

Acknowledgments: Marián López and Nuria Cid contributed equally to this work and hence should be regarded as joint senior authors. This research was funded by Xunta de Galicia (project PGIDIT03RAG07E). This paper is a contribution of the Interuniversity Research Group in Biotechnology and Reproductive Biology of Woody Plants.

The authors thank Marina Novo for her help with the modified AFLP markers, Javier Ibáñez (IMIA, Madrid) and José Miguel Martínez Zapater (CNB-CSIC, Madrid) for their help with grapevine microsatellites, and Verónica López and Olga Lortes for their excellent technical assistance. The authors also thank Paulino Martínez, Jaime Castro, and María L. Villar (Departamento de Genética, Universidad de Santiago de Compostela) for support with fragment analysis at their sequencing facilities.

^* Corresponding author (email: mrey{at}uvigo.es; fax 34 986 812 556)

Selected clones belonging to the most important autochthonous grapevine cultivars from Galicia (northwestern Spain) were analyzed using microsatellites and two AFLP methods. The goal was molecular identification of the analyzed germplasm using microsatellite and standard AFLPs analyses. The usefulness of a modified AFLP methodology for grapevine fingerprinting was also evaluated. Microsatellite analysis supported our cultivar assignment in accordance with other databases available, although the cultivar named Torrontés did not match with previously reported data, so further research will be necessary to clarify its genetic relationships and origin. Standard AFLP analysis on a subset of samples including at least one representative clone of each of the seven autochthonous cultivars allowed a clear differentiation among them, while no differentiation among clones of the same cultivar was obtained. To test a modified AFLP procedure, six selective primers were used to analyze the same subset of samples as for standard AFLPs. Although few scorable bands were obtained, a high percentage were polymorphic, allowing the unequivocally differentiation of all studied cultivars. In addition, 11 cultivar-specific bands were identified, which distinguished all cultivars except for Treixadura. This modified AFLP technique allowed us to obtain intravarietal fragments in the cultivars Albariño and Mencía. To our knowledge, this is the first report of application of this methodology to molecular characterization of grapevine cultivars.

Key words: Vitis vinifera, microsatellite, AFLP, cultivar identification, grapevine, clone identification