Use of squash-PCR to study tomato yellow leaf curl bigeminivirus transmission by Bemisia tabaci
A squash-PCR method has been developed in Israel to detect tomato yellow leaf curl bigeminivirus (TYLCV - EPPO A2 quarantine pest). With this method, the virus can be detected on samples of infected tissues (leaves, roots, stems) as small as 1 mm² squashed onto nylon membrane. TYLCV can also be detected in individual viruliferous Bemisia tabaci (EPPO A2 quarantine pest). This squash-PCR method was used to study whitefly transmission of TYLCV. Tomato plants were inoculated by placing a single viruliferous insect in the centre of a young leaflet. TYLCV could be detected at the site of inoculation on certain plants as early as 5 min after the beginning of the access feeding, and in all plants after 30 min. Using this technique, it was also possible to detect the virus in the head of B. tabaci as early as 5 min after the beginning of the access feeding on infected tomato plants, after 10 min in the thorax and after 25 min in the abdomen.
Atzmon, G.; van Oss, H.; Czosnek, H. (1998) PCR-amplification of tomato yellow leaf curl virus (TYLCV) DNA from squashes of plants and whitefly vectors: Application to the study of TYLCV acquisition and transmission.
European Journal of Plant Pathology, 104(2), 189-194.