A root-knot nematode parasitizing rice (Oryza sativa L.) in Santa Catarina state (Brazil) was identified as Meloidogyne oryzae Maas, Sanders and Dede, 1978 using different approaches. The specimens studied from this Brazilian population were compared with the type description of M. oryzae from Suriname, with additional morphological, biochemical and molecular characterization. The female has a longer stylet (15.0 μm) when compared with M. graminicola (11.2 μm) with irregularly shaped knobs, vulva offset and slightly protruding in posterior region. The lip region was distinct from first body annuli, and labial disc and the medial lips form an anchor-shaped structure. Perineal patterns were similar to M. graminicola. The male has a lip region offset and the presence of few short and irregular lines; medial lips divided, not fused with labial disc and stylet (18.2 μm) longer than in M. graminicola (16.8 μm). Second-stage juvenile (J2) tail (75.8 μm) was longer than in M. graminicola (70.9 μm) with a very long narrow hyaline portion (22 μm in M. oryzae and 17.9 μm in M. graminicola). Biochemically, it presented a distinct esterase profile (Est O1=R1), differentiating it from M. graminicola (Est VS1). The number of chromosomes was 3n = 50–56, and in DNA sequences of ITS1–5.8S–ITS2 rRNA the two populations of M. oryzae clustered together with other mitotic parthenogenetic species, differentiating them from M. graminicola with n = 18 chromosomes and clustered with meiotic species. Phylogenetic analysis using neutral markers (AFLP and RAPD) showed that both M. oryzae populations form a coherent, closely related cluster separately from M. graminicola isolates. This study represented the first detection of M. oryzae in Brazil and the second in the world after the species description in 1971.