Fruit set in the muscadine grape (Vitis rotundifolia Michx.) depended on insect cross-pollination, although flowers were well adapted for selling. Pollinizer cultivars produced about half of their optimal fruit set when selfed, but cross-pollination was needed to reach an optimal fruit set of 33.7%. Eighty-one percent of the overall fruit set in pistillate vines was attributed to insect cross-pollination; wind played only a small role. Diminished fruit sets and fewer seeds per berry occurred in cultivars receiving no effective cross-pollination. Components of fruit quality were not profoundly affected by the pollination treatments, although seed set and berry weight in pistillate cultivars were lower in the absence of cross-pollination. Parthenocarpy was rare, except in ‘FrySeedless’. Muscadine production throughout the southeastern United States depends on cross-pollination by indigenous insects, particularly bees. To ensure consistently high yields, bees must have safe access to flowers and their nesting sites must be preserved.