The root-knot nematode Meloidogyne incognita is widely distributed and a major pathogen of cotton (Gossypium spp.) worldwide. The objectives of this study were to assess the genetic variability and aggressiveness of Brazilian populations of M. incognita in cotton. Five populations of M. incognita and one isolate of M. enterolobii (outgroup) were used in the molecular analysis. Our results showed that only 2.7 % of the RAPD and AFLP fragments were polymorphic. Despite the existence of two races (races 3 and 4) and two esterase phenotypes (I1 and I2), a low genetic variability among populations was observed, which might be due to the mitotic parthenogenetic mode of reproduction of this pathogen. The aggressiveness/virulence among populations towards different cotton genotypes was also studied. None of the populations was virulent to the resistant cotton genotypes M-315 RNR, TX-25, CIR1343, Wild Mexican Jack Jones and CIR1348 (reproduction factor <1). Two populations of M. incognita from the states of Mato Grosso do Sul and Parana (Umuarama) (races 4 and 3, respectively) were highly aggressive to the susceptible control FM966 and virulent to the accessions LA-887 and Clevewilt-6 that showed moderate resistance to other populations tested.