Pollen from six southern highbush blueberry cultivars derived from Vaccinium corymbosum L. and one or more other species (V. darrowi Camp, V. ashei Reade, and V. angustifolium Aiton) was incubated on nutrient agar to determine tetrad viability, pollen tube growth rates, and incidence of multiple pollen tube germinations. `Avonblue' pollen had a significantly lower tetrad germination percentage than `Georgiagem', `Flordablue', `Sharpblue', `Gulfcoast', or `O'Neal', all of which had >90% viable tetrads. The in vitro growth rate of `O'Neal' pollen tubes was significantly higher than the growth rates of `Sharpblue' and `Georgiagem pollen tubes. Of those tetrads that were viable, more than two pollen tubes germinated from 83% and 91% of the `Gulfcoast' and `Sharpblue' tetrads, respectively, while only 11% of the `Flordablue' tetrads produced more than two pollen tubes. The total number of pollen tubes germinated per 100 tetrads ranged from 157 (`Flordablue') to 324 (`Sharpblue'), resulting in actual pollen grain viabilities ranging from 39% to 81%. Genetic differences in pollen vigor, as indicated by pollen viability, pollen tube growth rates, and multiple pollen tube germinations, may influence blueberry growers' success in optimizing the beneficial effects of cross-pollination on fruit development.