Four biologically active cDNA clones were derived from the Alternanthera mosaic virus (AltMV; genus Potexvirus) isolate, AltMV-SP, which differ in symptoms in infected Nicotiana benthamiana plants. Two clones induced necrosis and plant death; a mixture of all four clones induced milder symptoms than AltMV-SP. Replication of all clones was enhanced by a minimum of fourfold at 15 6C. A mixture of clones 4-7 (severe) and 3-1 (mild) was indistinguishable from AltMV-SP, but the ratio of 4-7 to 3-1 differed at 25 and 15 6C. RNA copy numbers of mixed infections were always below those of 4-7 alone. Determinants of symptom severity were identified in both Pol and TGB1; the mildest (4-1) and most severe (3-7) clones differed at three residues in the 'core' Pol domain [R(1110)P, K(1121)R, R(1255)K] and one [S(1535)P] in the C-terminal Pol domain of RNA-dependent RNA polymerase, and one in TGB1 [P(88)L]. Pol [P 1110 ,R 1121 ,K 1255 ]+TGB1 L(88) ] always induced systemic necrosis at 15 6C. Gene exchanges of Pol and TGB1 each affected replication and symptom expression, with TGB1 P(88) significantly reducing silencing suppression. The difference in silencing suppression between TGB1 P(88) and TGB1 L(88) was confirmed by an agroinfiltration assay. Further, co-expression of TGB1 P(88) and TGB1 L(88) resulted in interference in the suppression of silencing by TGB1 L(88) . Yeast two-hybrid analysis confirmed that TGB1 P(88) and TGB1 L(88) interact. These results identify a TGB1 residue that significantly affects replication and silencing suppression, but maintains full movement functions.
A potexvirus was isolated from creeping phlox (Phlox stolonifera) plants from a commercial nursery in Pennsylvania. The virus was serologically related to clover yellow mosaic virus, plantain virus X, potato virus X, and potato aucuba mosaic virus, and was most closely related to papaya mosaic virus (PapMV). The sequence of a PCR fragment obtained with potexvirus group-specific primers was distinct from that of PapMV; the coat protein (CP) gene and 3' untranslated region (UTR) were closely related to Alternanthera mosaic virus (AltMV), previously reported only from Australia. The host range was similar to that of the Australian isolate (AltMV-Au), and the phlox isolate reacted strongly with antiserum to AltMV-Au. The full sequence of the phlox isolate was more closely related to PapMV throughout the genome than to any potexvirus other than AltMV-Au, for which only the CP and 3'UTR sequences are available. The phlox isolate was therefore named AltMV-PA (for Pennsylvania), and the full 6607 nt sequence is presented(1). Additional AltMV isolates from creeping phlox (AltMV-BR and AltMV-SP) and trailing portulaca (Portulaca grandiflora; AltMV-Po) were also isolated, suggesting that AltMV may be widespread, and may have been mis-diagnosed in the past as PapMV. AltMV has the potential to spread to other ornamental crops.
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