Rice root knot nematode, Meloidogyne graminicola is a concern to global rice production, yet it is underexplored in many regions where it is cultivated. To gain a better understanding of M. graminicola prevalence and incidence in Tamil Nadu (India), M. graminicola isolates were obtained from soil and root samples and identified using perineal patterns and rDNA ITS-based sequencing. Galling index, root-knot nematode juveniles per root system and juveniles per 200 cc of soil were used to assess the severity of nematode infestation on rice roots and infested fields in various regions. Our findings show that rice is severely infested by a genetically varied and aggressive M. graminicola, demanding the successful implementation control strategies in rice. Both the conventional posterior cuticular pattern method as well as the molecular characterization method were used to validate the identity of the root-knot nematode (M. graminicola). Molecular study revealed that M. graminicola produced a single band of 790 bp for Meloidogyne graminicola isolate TNAU004 cytochrome c oxidase subunit I (COX1) gene and 764 bp Meloidogyne graminicola isolate TNAU003 large subunit ribosomal RNA gene. Each genus is given a detailed description, including morphometrics, illustrations, and key traits.