Abstract: Rice Yellow Mottle Virus (RYMV) genus Sobemovirus is a highly variable pathogen that is very infectious to rice plant. This variability hinders rice breeding for durable resistance to the virus and effective deployment of improved cultivars in Southwest Nigeria. Disease surveys in 5 Southwest states (Lagos, Oyo, Ogun, Ekiti and Ondo) revealed RYMV disease incidence of between 15-70% in farmers’ fields and serological indexing confirmed 92% of collected leaf samples positive to RYMV with 24% from rice and 76% from weeds. The weed with 76% RYMV positive suggests being the main reservoir of RYMV in Southwest Nigeria. Biological test on collected fields leaf samples identified 3 groups (GroupA, GroupD and GroupE) of Resistance Breaking (RB) RYMV isolates and 2 groups (GroupB and GroupC) of normal isolates. Pathotyping 20 RYMV isolates against 10 differential varieties identified 17 isolates as Highly Pathogenic Isolates (HPI) and 3 as Mildly Pathogenic Isolates (MPI) while 4 rice varieties were Highly Resistant (HR), 2 were Moderately Resistant (MR) and 4 were susceptible. HPI isolates present in five states and MPI isolates in two states. Serological study using the same 20 RYMV isolates revealed two major Nigeria serogroup (NSg1 and NSg2) and four subgroups (NSg1a, NSg1b, NSg2a and NSg2b). NSg1a and NSg1b comprised both normal and RB isolates while NSg2a and NSg2b were typical of RB isolates only. This information would assist rice breeding programs to develop durable resistant cultivars to RYMV disease in Southwest Nigeria.