Coding changes in the 3a cell-to-cell movement gene can extend the host range of brome mosaic virus systemic infection.

The M1 and M2 strains of brome mosaic virus (BMV) both systemically infect the monocot host barley, but only the M2 strain systemically infects the dicot cowpea line TVu-612. We have shown previously that this difference in host range maps primarily to RNA3. To further characterize the role of RNA3 in host specificity, a series of RNA3 hybrids were tested, in inoculations with M1 RNA1 and RNA2, for ability to systemically infect TVu-612 cowpea. Although all hybrids were amplified well in cowpea protoplasts and all supported systemic infection in barley plants, only those with the 3a cell-to-cell movement gene of BMV-M2 supported systemic infection of cowpea. The sequences of the M1 and M2 3a proteins differ at four positions. Introducing these four coding differences individually or in various combinations into M1 RNA3 revealed that all four influenced BMV adaptation to cowpea and that these four differences were sufficient to account for the difference in ability between M1 and M2 RNA3s to support systemic infection of this legume. These coding changes were also associated with faster spread of infection in inoculated cowpea leaves, suggesting that they influence the ability to systemically infect TVu-612 cowpea through effects on the rate of cell-to-cell spread.