Hybridization between two meiotic parthenogenetic species of root-knot nematodes, Meloidogyne chitwoodi and M. fallax, was investigated in two different crossing experiments on tomato plants grown in sand. The first experiment was a controlled cross between the two species. The second experiment was a bulk mating in a 1:1 mixture of two isolates. The haploid chromosome number of the parental isolates was n = 18. Successful interspecific hybridization was obtained, and the resulting hybrids produced egg masses. In eggs, cell division was observed, but most of them were without clear differentiation and consequently were sterile. Hatched F2 juveniles were small in number, not viable, and showed morphological distortions. In the progeny of the isolate mixture of the bulk mating experiment, parental-type females of the two isolates were present in equal numbers, and 10% of all females were nonviable hybrids. Similar ratios of parental-type and hybrid females were detected in roots of test plants grown in soil from a field sample that contained a mixture of M. chitwoodi and M. fallax populations. In the controlled cross experiment, isozyme electrophoresis of malate dehydrogenase was applied to distinguish the two species and their hybrids. In the bulk mating experiment, malate dehydrogenase, esterase, and glucose 6-phosphate dehydrogenase were used as markers, two by two simultaneously on the same individual females, providing conclusive evidence for the occurrence of hybrids. This is the first report on interspecific hybridization in Meloidogyne. The possible role of interspecific hybridization in species differentiation and interspecific exchange of genetic material within Meloidogyne is discussed.