A cDNA clone containing the complete coding region of RNA2 of the newly described TPA56 isolate of pea early-browning virus (English serotype, PEBV-E) has been obtained. A plasmid (pFLA56) containing this clone together with the 5′ and 3′ non-coding regions of PEBV isolate SP5 (the type isolate of PEBV-E) and flanked by the CaMV 35S promoter and NOS terminator is infectious when co-inoculated onto plants with pCaN1, an infectious clone of PEBV SP5 RNA1. Virus containing RNA2 derived from the cDNA clone of the TPA56 isolate is transmitted by nematodes, whereas virus containing RNA2 from a clone of the SP5 isolate is not transmitted. Sequencing revealed 11 differences out of 3374 nucleotides between the transmissible TPA56 and non-transmissible SP5 clones. However, only three of the base changes affected the amino acid sequences of virus gene products. A single, conservative amino acid change was present in the virus coat protein. Two non-conservative changes occurred in the protein of molecular mass 29.6 kDa expressed from an open reading frame located down-stream of the coat protein gene, suggesting that it has a function in nematode transmission and that changes in this protein prevent vector transmission of the SP5 isolate of PEBV.


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