A field population of Heterodera glycines was inbred by a combination of controlled male-female matings and inoculation of soybean with second-stage juveniles (J2) from single cysts. The initial and four F inbred populations were subjected to random amplified polymorphic DNA analysis and were also tested for their ability to reproduce on race differentials. The RAPD patterns of the inbred populations had a lower number of total bands and a lower percentage of polymorphic bands among individual cysts than the initial population. The estimated number of polymorphic loci detected by RAPD analysis was about 25% for the initial population and 4% to 7% for the inbred lines. Reproduction of H. glycines decreased for 6 of 24 inbred-soybean combinations. In particular, reproduction of three inbred populations on PI 90763 was greatly reduced. Inbreeding did not decrease variance of cyst number on soybean genotypes. The inbreeding coefficient calculated from RAPD data was greater than that derived from the known inbreeding pedigree.