Microsatellite markers were used to determine paternity in 473 open-pollinated progenies from a clonal Eucalyptus nitens seed orchard of 50 trees from 12 different genotypes. The outcrossing rate in this orchard was high, averaging 0.85 (weighted by capsule crop) but variable between trees (ranging from 0.6-1.0). Paternal contribution of each genotype to the open-pollinated seed crop was predicted by the size of the flower crop of each genotype (r = 0.76), but not the number of ramets. While the contamination in our orchard is relatively low (4.5%), this is atypical when compared to other published estimates in eucalypt seed orchards suggesting that with suitable buffering low levels of contamination can be achieved.