A new diagnostic tool for the identification of four beet yellows viruses by multiplex RT-qPCR
Résumé
Virus yellows disease (VY) of sugar beet is caused in Europe by a complex of four aphid-transmissible virus species present as mono- or co-infections: beet yellows virus (BYV), beet chlorosis virus (BChV), beet mild yellowing virus (BMYV) and beet mosaic virus (BtMV). VY can impact yields by up to 50%. The losses were limited with neonicotinoids (NNI), that are now banned in UE.
To find alternatives to NNI, the National Plan of Research and Innovation has financed the Yellows Resistbeet project led by GEVES and involved in the evaluation of varietal resistance. As the ELISA method does not distinguish BChV from BMYV detection, BioGEVES with INRAE Colmar developed primer and probe pairs to detect and identify the four viruses simultaneously in a same sample.
To ensure viral diagnosis, GEVES validated a multiplex RT-qPCR method by evaluating performance criteria. Our first results showed that all primers and probes are specific to the four targeted viruses with 100% inclusivity and exclusivity and that RT-qPCR multiplex is 100 to 10,000 times more sensitive than ELISA.
Our new diagnostic tool is used for varietal resistance testing to control inocula and to provide in one step the viral composition as well as semi-quantitative data on the viral load in sugar beets tested. This detection method at a lower cost has been validated and offered as a service by GEVES. Later, it can be used in epidemiological surveillance to monitor the evolution of VY in the field depending on the year.
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