de Haan E.G.,Dutch General Inspection Service for agricultural seeds and seed potatoes NAK |
Dekker C.C.E.M.,Dutch General Inspection Service for agricultural seeds and seed potatoes NAK |
Tameling W.I.L.,Dutch General Inspection Service for agricultural seeds and seed potatoes NAK |
den Nijs L.J.M.F.,National Reference Center |
And 2 more authors.
EPPO Bulletin | Year: 2014
Potato is one of the many important hosts for the root-knot nematodes Meloidogyne chitwoodi and M. fallax that can infest roots as well as tubers. In the latter they may cause surface galls and necrotic spots below the skin. In the EU these pathogens are categorized as quarantine organisms and are therefore regulated. Phytosanitary measures (PMs) are implemented and one aspect involves diagnostic procedures to detect these pathogens. To date, visual screening of external and internal symptoms is combined with the specific identification of these pests, either through microscopy, biochemical or molecular tests. A disadvantage of all these tests is the requirement of the prior extraction of nematodes from the tubers, which is not suitable for high-throughput screening. This paper describes the MeloTuber Test developed to simultaneously detect M. chitwoodi and M. fallax by triplex real-time TaqMan® PCR directly in secondary potato tuber peelings. DNA extraction is carried out in a 96-well plate of pooled secondary peelings from one hundred tubers. The analytical sensitivity is such that a single female can be readily detected in such a sample size. The validation data, described here, prove the suitability of this molecular test for the detection of M. chitwoodi and M. fallax in large scale screening tests. © 2014 OEPP/EPPO.