Evaluation of the uniformity and stability of microsatellite markers in grapevine

Resumen

In this study, more than 4,000 plants have been sampled to evaluate the uniformity and stability of nine microsatellites (VVS2, VVMD5, VVMD27, VVMD28, ssrVrZAG29, ssrVrZAG62, ssrVrZAG67, ssrVrZAG83, and ssrVrZAG112) in 19 different cultivars of Vitis vinifera L. ('Airén', 'Cabernet sauvignon', 'Cardinal', 'Chardonnay', 'Crimson seedless', 'Flame seedless', 'Garnacha', 'Italia', 'Merlot', 'Monastrell', 'Muscat d'Alexandrie', 'Napoleón', 'Ohanes', 'Palomino Fino', 'Red Globe', 'Sauvignon blanc', 'Sugraone', 'Tempranillo', and 'Thompson seedless'). The markers are publicly available, genetically independent, and allowed their multiplexing. The selection of varieties included variability for a number of traits, like use, color, time of ripening, time of culture, origin, etc. For the uniformity study, three plantations per variety and 50 plants per plantation were analyzed, while for the stability study, seven additional plantations per variety and five plants per plantation were analyzed. The plant material comes from different countries in different continents (12 varieties) or from different places of Spain (seven varieties). DNA extraction, multiplex PCR, and capillary electrophoresis in an automatic sequencer were used to produce more than 29,000 genotypes. The differences found between the values determined for each allele of each variety were studied to evaluate the uniformity and stability of the microsatellites. The results obtained demonstrate that the selected microsatellites are uniform and stable markers, although some cautions could be necessary.