Use of a new multiplex PCR to facilitate the epidemiological study of Cucurbit viruses in Flanders (Belgium)

Beschrijving

In temperate and tropical regions, the cultivation of zucchini (Cucurbita pepo) is under increasing pressure of virus diseases. The most common viruses in Flanders are transmitted by aphids in a non-persistent way and include Cucumber mosaic virus (CMV, Cucumovirus), Zucchini yellow mosaic virus (ZYMV, Potyvirus) and Watermelon mosaic virus (WMV, Potyvirus). They cause yield reduction and unmarketable fruits due to mottling, mosaic, chlorosis and leaf or fruit distortion, symptoms that are highly similar for all four viruses. During an epidemiological survey in the growing seasons from 2007 to 2012, a gradual decrease of ZYMV was observed while CMV became more important and is currently the most prevalent virus on field-grown zucchini in Flanders followed by WMV, ZYMV. Also within a single growing season, shifts in virus prevalence were observed. During this survey we also noticed that in a substantial part of the infections multiple viruses were involved. Several observations that were made during the survey are assessed more in detail through inoculation experiments. This will increase knowledge on introduction pathways, virus survival, epidemiology of mixed infections and cultivar sensitivity.
To ease the processing of high sample numbers (survey and experiments) we developed a sensitive and specific multiplex PCR for the detection of CMV, WMV and ZYMV. The implementation of this multiplex PCR tackles the limitations of ELISA or simplex RT-PCR methods which are currently used for cucurbit virus detection. These include a reduced sensitivity and cross-reactivity for ELISA, and a high cost for the three parallel RT-PCR reactions. This multiplex PCR allows the detection of these viruses in plants with a single or combined infection. No cross reaction was observed and the multiplex RT-PCR can also be used to screen the vectors and seeds. This method will be used in the further epidemiological studies on zucchini viruses in Flanders.

Periode	26-sep-2013
Evenementstitel	International Advances in Plant Virology
Evenementstype	Andere
Locatie	Norwich, Verenigd Koninkrijk
Mate van erkenning	Internationaal