We evaluated pollen contamination, supplemental mass pollination efficacies, and natural selfing in a Douglas-fir (Pseudotsuga menziesii(Mirb.) Franco) clonal-row seed orchard using a genetic marker on the paternally inherited chloroplast (cp) genome. A primer pair for the polymerase chain reaction amplification of a variable region on the cpDNA in Douglas-fir was developed. The amplified DNA product was highly variable in size, yielding 13 different haplotype bands from 20 orchard genotypes growing in the clonal-row seed orchard. Observed band sizes ranged from 859 to 1110 base pairs (bp). To estimate variation levels in the orchard background pollen pool, 96 assayed genotypes from surrounding stands gave rise to 36 different haplotypes, ranging from 367 to 1119 bp in size, resulting in a gene diversity estimate of 0.91. Most orchard clones' haplotypes were also present in the background. After adjusting for the presence of orchard-type haplotypes in the background, contamination was found to be 40%. Natural selfing in six individual clones ranged from 0 to 19% with an average of 6%. Supplemental mass pollination efficacy was estimated to be 55%, ranging from 39 to 73%, depending on the maternal clone and flowering phenology. This DNA marker proved to be very useful in assessing seed orchard mating dynamics and orchard management efficacies for Douglas-fir.