DEVELOPMENT OF AN IC-RT-PCR ASSAY FOR THE DETECTION OF EGGPLANT MOTTLED DWARF VIRUS AND PARTIAL CHARACTERIZATION OF ISOLATES FROM VARIOUS HOSTS IN GREECE

N.I. Katis, E.K. Chatzivassiliou, C.M. Clay, V.I. Maliogka, P. Pappi, K. Efthimiou, C.I. Dovas, A.D. Avgelis

Abstract


In a 10-year survey several Eggplant mottled dwarf virus (EMDV) isolates were collected from diverse hosts from all over Greece. In this study, an IC-RT-PCR assay was developed for the detection of the virus. Degenerate primers were designed on conserved regions of the glycoprotein (G) gene of various plant nucleorhabdoviruses. Based on the initial genetic information obtained, EMDV-specific primers were subsequently designed. The method successfully amplified a 284 bp product from all (76) virus samples originating from tobacco, cucumber, eggplant, Pittosporum tobira and Capparis spinosa (caper). Sequence analysis and Western blot were done for further characterization of representative isolates. Two isolates from tobacco (EMDV-Tob) and cucumber (EMDV-Cu) were also subjected to electron microscopy and host range studies. Sequence analysis of the partial G gene confirmed the classification of EMDV within nucleorhabdoviruses. Genetic comparisons among the EMDV isolates tested indicated low levels of variation with the caper isolate, which is reported for the first time in Greece, being the most deviating isolate. This is the first molecular assay developed for the detection of EMDV and also the first sequence information obtained of its genome.

Keywords


EMDV; IC-RT-PCR detection; cytopathology; electron microscopy; Western blot

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DOI: http://dx.doi.org/10.4454/jpp.v93i2.1190

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