In species where the female mates more than once, sperm priority patterns have important implications for the mating behavior of both sexes. The shape of female sperm storage organs may play a role in determining priority patterns, although the extent of this effect has been controversial. Spiders provide an interesting test of this hypothesis because 2 phylogenetically distinct groups differ in the structure of sperm storage organs. In entelegyne spiders, the female sperm-storage organs have separate ducts for sperm entrance and exit. The male inseminates the female through the external fertilization duct. Sperm then travel through a ''conduit'' and are released from an internal fertilization duct to fertilize eggs. This arrangement has been hypothesized to lead to 1st-male priority. Six species in 3 entelegyne families have been shown to have 1st-male sperm priority with some mixing; 1 entelegyne species, however shows last-male priority. Haplogyne females, in contrast, have a sperm-storage area with only a single opening, which may be more likely to lead to last-male sperm priority because the last sperm to enter the storage area would be likely to be the 1st to leave. Few data are available for this group. We used a balanced sterile-male design to test the sperm priority pattern in Females mated once with each of a pair of males of a haplogyne spider Holocnemus pluchei Scopoli. Egg sacs from 72.5% of females were sired predominantly by 2nd males, 7.5% were predominantly sired by 1st males, and 20% showed mixed paternity (n = 40); these frequencies differed significantly from random expectations. The average percentage of eggs fathered by the 2nd male, or P-2, was 74%. These data clearly differ from the predominantly 1st-male pattern generally found in entelegynes, thereby supporting the hypothesized relationship between female reproductive morphology and sperm use patterns. Mating behavior of spiders that exhibit end-male priority should differ from that exhibiting 1st-male priority, and evidence from H. pluchei and other haplogynes supports this inference.