Abstract: Virus isolates from Crotalaria plants in germplasm regeneration plots in Griffin, GA were found to produce an unexpected wilting and necrosis on Nicotiana benthamiana. DAC-ELISA tests produced positive results for alfalfa mosaic virus (AlMV) and a mixture of viruses including cucumber mosaic, blackeye cowpea mosaic, peanut stunt and white clover mosaic viruses. After serial transfer of an isolate from C. juncea in N. benthamiana in the greenhouse, only AlMV could be detected by ELISA while the wilting and necrosis symptoms remained. In addition, RT-PCR tests produced a product of the expected size with the crotalaria isolate RNA as it did with a known isolate of AlMV. Sequencing of the 300 bp PCR-band indicated that there was 99% identity between the Crotalaria isolate and strain 425 of AlMV. These results show that this isolate from Crotalaria is AlMV and possibly a new strain of AlMV.