The aim of this study was to evaluate the viability and longevity of the pollen grains of the olive cultivar ‘Picual’, the most extensive and important olive cultivar in the Iberian Peninsula. Over a 3-year period (2007–2009), viability of the pollen grains of 15 ‘Picual’ cultivar trees was measured using the Fluorochromatic Reaction (FCR). These olive trees were distributed along different altitudinal areas of the province of Jaen (southeast Iberian Peninsula). The percentages of viability did not show any geographical variability. In general, the percentage of viability did not vary from one year to another. With an average value of 49%, the viability of the pollen in the cultivar ‘Picual’ could be classified as medium-range. ‘Picual’ pollen grains have the highest viability after 24 hours of anther dehiscence. Viability declines rapidly with time. The longevity of the pollen grains was about seven days. The first three days after anthesis are the most important for fertilisation. A rapid decline in pollen viability may considerably reduce effective fertilisation and could negatively influence fruit production. For this reason, these findings should be considered for the improvement of olive fruit yields.