A collection of 312 accessions of Burkina Faso rice germplasm was evaluated in Cotonou, Benin and Montpellier, France for resistance using four Rice yellow mottle virus (RYMV) isolates. In Cotonou, the accessions were screened using one isolate from Benin (B27) and two isolates from Niger (Ng122 and Ng144). The experiment was conducted in a screenhouse. Fourteen-day-old plantlets were inoculated and symptoms scored fortnightly from 14 to 56 days after inoculation (dai). Plant height was recorded at 49 dai after all accessions were inoculated with isolates Ng122 and Ng144. Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay (DAS-ELISA) was used to assess RYMV into rice leaves. In Montpellier, aggressive isolate from Burkina Faso BF1 was used to assess the accessions against RYMV. The experiment was conducted in the greenhouse under controlled conditions (28-32°C, 12 h of light per 24 h and 80-90% relative humidity). Inoculation process and symptoms scoring were performed on rice leaves as in Benin and reverse-transcription polymerase chain reaction (RT-PCR) was used to check the presence of the allele rymvl -3 in resistant accessions. The results of both experiments showed that the Oryza sativa accessions of the collection were highly susceptible, except for BM24, which combined partial resistance and tolerance. Twenty-one O. glaberrima accessions out of 48 were resistant to Ng122 and Ng144. When these 21 accessions were subsequently screened with the aggressive RYMV strain BF1, eight of them displayed a delay in the appearance of RYMV symptoms, while two showed resistance. The molecular characterization of the resistant accessions showed that they do not have the allele rymvl-3. These accessions could have new resistance alleles. Therefore, the new sources of resistance identified in this study could be exploited in breeding to tackle the spread of RYMV in Africa.