The fruit of several Opuntia species (prickly pear) are a good source of calcium, potassium, and ascorbic acid and are consumed fresh or processed as juices or preserves. Plants of Opuntia may be grown in arid and semiarid environments on marginal soils. Various cultivars, particularly in the species Opuntiaficus-indica, are grown commercially in the United States, Israel, Italy, Mexico, and South Africa. There is a need for new sources of genetic diversity and subsequent germplasm evaluation, and until recently, no publicly maintained germplasm collection of Opuntia existed in the United States. The purpose of this study was to evaluate fruit quality of 25 Opuntia accessions, originating from six countries, and maintained at the USDA collection at the National Arid Land Plant Genetic Resource Unit, Parlier, Calif. The largest fruits were harvested from plants of accessions PARL 201, 202, and 228 (227.6, 247.3, and 231.3 g/fruit, respectively). The hardest peel was on fruits of PARL 225 and 234 (both 3.7 kg), and fruit pulp of the same two accessions had the highest firmness (2.3 and 2.4 kg, respectively). Soluble solids in mature fruit varied from 6.1% (PARL 231) to 15.0% (PARL 254). The fruit color ranged from light yellow through orange, pink to dark purple. These characteristics and other traits such as fruit acidity, presence of spines, and seed mass/fruit indicated that the material represents a diverse germplasm collection, usable for future cultivar development.