Abstract
Several conventional PCR tests have been developed for the identification of the European quarantine root-knot nematodes Meloidogyne chitwoodi and M. fallax but data are lacking for the evaluation of their performance in terms of sensitivity, repeatability, reproducibility and specificity against a large range of populations. This study evaluated the performance criteria of three conventional PCR tests recommended by the consensus diagnostic protocol for Meloidogyne chitwoodi and Meloidogyne fallax published by the European and Mediterranean Plant Protection Organization (EPPO): a species-specific PCR (IGS target), a SCAR PCR, and a rDNA ITS PCR-RFLP. Evaluation was carried out with DNA extracts from juveniles, males and females according to EPPO recommendations for test validation. A minimum of 34 populations of target and non target nematode species were tested to check the specificity of these three PCR assays. The three PCR tests were ranked according to their specificity (with regard to cross reaction with other nematodes species or genus) and their sensitivity (detection of a single juvenile or mixed with other species). The species-specific PCR proved to be more sensitive but less specific than the SCAR PCR. The PCR-RFLP enables the identification of several Meloidogyne species but profile analysis can be difficult when several species are present in the mixture. Specific PCR products and RFLP profiles were also observed for M. arenaria and M. enterolobii, and described for M. minor and M. artiellia.
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Acknowledgments
This assessment study was both funded by the French Federation of Seed Potato Growers (FN3PT) and the French Ministry in charge of agriculture. We would like to thank the French National Institute for Agricultural Research (INRA) unit 1303 based in Sophia Antipolis (France), the Oregon State University, the University of Idaho in Florida or Florida Department of Agriculture & Consumer Services and the Belgian Institute for Agricultural and Fisheries Research for providing us with nematode populations or DNA solutions used in this study. The authors also wish to thank Dr R. Ioos and Dr E. Grenier for their helpful review.
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Gamel, S., Huchet, E., Le Roux-Nio, AC. et al. Assessment of PCR-based tools for the specific identification of some temperate Meloidogyne species including M. chitwoodi, M. fallax and M. minor . Eur J Plant Pathol 138, 807–817 (2014). https://doi.org/10.1007/s10658-013-0355-8
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DOI: https://doi.org/10.1007/s10658-013-0355-8