My student succeeded in isolating and purifying the Iris Yellow Spot Virus. Actually, no one isolated this virus from onion before and also there is no good image for this virus.We need a virologist to confirm what we have obtained these results. Any advice please?
You can try to amplify a fragment of the viral genome by PCR and sequence it. Then, BLAST your sequence to confirm that the PCR product you amplified corresponds to that virus.
A diagnosis cannot be made from looking at the EM images of the purified virions as tospoviruses can be variable in size, they share a similar size range, and are irregular in shape assuming that the membrane isn't lost during purification (http://apsjournals.apsnet.org/doi/abs/10.1094/PDIS.2000.84.11.1185). The best way to confirm infection is to either use ELISA or RT-PCR followed by sequencing (as suggested by Benjamin). As an aside, it has been well established that Iris yellow spot virus infects onion for many years now (http://apsjournals.apsnet.org/doi/abs/10.1094/PDIS-92-8-1247A; http://apsjournals.apsnet.org/doi/abs/10.1094/PDIS-93-10-0976; http://link.springer.com/article/10.1007%2Fs10658-012-0026-1) with IYSV being considered one of the most important pathogens of onion worldwide.
Indeed, specific antiserum for DAS ELISA is available since 1994; RT-PCR targeting the viral CP could also work well for subsequent sequencing. Conserved stretches of sequences between all tospoviruses might help amplification and make the sequence data diagnostic.
BTW, N. benthamiana is a good host for IYSV with distinct necrotic spots systemically.
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