A single codon change in a conserved motif of a bromovirus movement protein gene confers compatibility with a new...

Brome mosaic virus (BMV) and cowpea chlorotic mottle virus (CCMV) are closely related bromoviruses with tripartite RNA genomes, but distinct host ranges: BMV systemically infects the monocot barley, while CCMV systemically infects the dicot cowpea. We have previously shown that in approximately 10% of inoculated cowpea plants, a CCMV hybrid [CCMV(B3a)] with the 3a cell-to-cell movement protein gene replaced by that of cowpea-nonadapted BMV directs systemic infections, which are caused by secondary mutation(s) of the hybrid virus. Here, to further analyze the role of RNA3 in adaptation to a new host, RNA3 cDNA clones were constructed from total RNA recovered from the uninoculated upper leaves of systemically infected cowpea plants inoculated with CCMV(B3a). Sequence and mutational analysis of two such RNA3 clones revealed that a single codon change (A776-->C) in a conserved motif of the 3a movement protein gene conferred compatibility for systemic infection of a new host, cowpea, suggesting that this site in the 3a gene is directly or indirectly involved in crucial host interactions associated with host-range specificity. The adaptive hybrid viruses carrying this mutation induced exacerbated symptoms, while wt CCMV appeared nearly symptomless, showing that the bromovirus 3a movement protein gene can significantly contribute to regulating symptom development. However, introducing this cowpea-adaptive mutation into the BMV genome had little effect on the ability of BMV to systemically infect barley.