The presence of a small open reading frame embedded in the P3 cistron of potyvirus turnip mosaic virus, termed "pipo," was recently discovered. We have now studied the putative pipo of soybean mosaic virus (SMV). Introduction of single, or multiple, stop codon mutations at different locations within pipo, without substitution in polyprotein amino acids, did not abolish replication, but restricted the virus to small cluster of cells within the inoculated leaves. Furthermore, extensive mutagenesis of the conserved GA(6) motif at the 5' end of pipo also generated two out of five mutants that remained restricted to small foci of infected cells within the inoculated leaves. Long-distance movement function of the movement-defective PIPO-mutants was not restored following co-inoculation with competent SMV strains. Taken together, the data suggest that the putative pipo of SMV is essential for the virus movement; however, knock out of its expression does not abolish replication.
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