Co-infection of potyviruses with taxonomically diverse plant viruses results in disease synergism and elevation in the level of accumulation of non-potyviruses involved. In the majority of cases, however, the accumulation level of potyviruses remains essentially unaltered. A few potyviruses, such as Soybean mosaic virus (SMV), naturally infect soybean (Glycine max). Soybean is also a natural host to a number of non-potyviruses including Alfalfa mosaic virus (AMV), which causes mild symptoms often associated with symptom remission. We have now studied the interactions between AMV and SMV on symptom severity and accumulation level of each of the two viruses in soybean. Co-infection of soybean with AMV and SMV was established following mechanical inoculation, irrespective of simultaneous or sequential introduction of the two viruses. In multiple experiments, co-infection of soybean resulted in severe symptoms in doubly infected plants in a strain-independent manner, with enhancement in the level of AMV indicating that the interaction of AMV with SMV is synergistic. Conversely, the level of SMV accumulation was reduced. This suggests that in co-infection with AMV, SMV interacts antagonistically. The observation that co-infection of AMV and SMV results in disease synergism suggests enhancement of potential that AMV may become a serious viral disease of soybean.