During nematode surveys in southern Spain and Italy 14 populations of Xiphinema species tentatively identified as Xiphinema americanum-group were detected. Morphological and morphometrical studies identified three new species and six known Xiphinema americanumgroup species, viz.: Xiphinema parabrevicolle n. sp., Xiphinema parapachydermum n. sp., Xiphinema paratenuicutis n. sp., Xiphinema duriense, Xiphinema incertum, Xiphinema opisthohysterum, Xiphinema pachtaicum, Xiphinema rivesi, and Xiphinema santos. The Xiphinema americanum-group is the most difficult Xiphinema species group for diagnosis since the morphology is very conservative and morphometric characters often overlap. This group includes vectors of several important plant pathogenic viruses that cause significant damage to a wide range of agricultural crops. Molecular characterisation of these species using D2-D3 expansion regions of 28S rRNA, 18S rRNA, ITS1-rRNA and the protein-coding mitochondrial gene, cytochrome oxidase c subunit 1 was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other Xiphinema americanum-group species.

For many organisms, there is agreement on the specific genomic region used for developing barcode markers. With nematodes, however, it has been found that the COI region designated for most animals lacks the taxonomic coverage (ability to amplify a diverse group of taxa) required of a metabarcoding marker. For that reason, studies on metabarcoding of nematodes thus far have utilized primarily regions within the highly conserved 18S ribosomal DNA. Two popular markers within this region are the ones flanked by the primer pairs NF1-18Sr2b and SSUF04-SSUR22. The NF1-18Sr2b primer pair, especially, has been critiqued as not being specific enough for nematodes leading to suggestions for other candidate markers while the SSUF04-SSUR22 region has hardly been tested on soil nematodes. The current study aimed to evaluate these two markers against other alternative ones within the 28S rDNA and the COI region for their suitability for nematode metabarcoding. The results showed that the NF1-18Sr2b marker could offer wide coverage and good resolution for characterizing soil nematodes. Sufficient availability of reference sequences for this region was found to be a significant factor that resulted in this marker outperforming the other markers, particularly the 18S-based SSUFO4-SSUR22 marker. None of the other tested regions compared with this marker in terms of the proportion of the taxa recovered. The COI-based marker had the lowest number of taxa recovered, and this was due to the poor performance of its primers and the insufficient number of reference sequences in public databases. In summary, this study highlights how dependent the success of metabarcoding is on the availability of a good reference sequence collection for the marker of choice as well as its taxonomic coverage.

Since its conception, DNA barcoding has seen a rapid uptake within the research community. Nevertheless, as with many new scientific tools, progression towards the point of routine deployment within diagnostic laboratories has been slow. In this paper, we discuss the application of DNA barcoding in the Defra plant health diagnostic laboratories, where DNA barcoding is used primarily for the identification of invertebrate pests. We present a series of case studies that demonstrate the successful application of DNA barcoding but also reveal some potential limitations to expanded use. The regulated plant pest, Bursephalenchus xylophilus, and one of its vectors, Monochamus alternatus, were found in dining chairs. Some traded wood products are potentially high risk, allowing the movement of longhorn beetles; Trichoferus campestris, Leptura quadrifasciata, and Trichoferus holosericeus were found in a wooden cutlery tray, a railway sleeper, and a dining chair, respectively. An outbreak of Meloidogyne fallax was identified in Allium ampeloprasum and in three weed species. Reference sequences for UK native psyllids were generated to enable the development of rapid diagnostics to be used for monitoring following the release of Aphalara itadori as a biological control agent for Fallopia japonica.Key words: plant health, regulated quarantine pest, DNA barcoding, diagnostics, invertebrate.Résumé : Depuis son développement, le codage à barres de l'ADN a connu une adoption rapide au sein de la communauté scientifique. Néanmoins, comme pour plusieurs nouveaux outils scientifiques, les avancées en vue d'un usage routinier au sein de laboratoires de diagnostic ont été lentes. Dans ce travail, les auteurs discutent de l'emploi du codage à barres au sein des laboratoires de diagnostique phytosanitaire de DEFRA, où le codage à barres est principalement employé pour l'identification des ravageurs invertébrés. Les auteurs présentent une série d'études de cas le codage à barres a été mis en oeuvre avec succès tout en révélant des limitations potentielles à son usage accru. Le parasite réglementé, Bursephalenchus xylophilus, et l'un de ses vecteurs, Monochamus alternatus, ont été retrouvés dans des chaises de salle à manger. Certains produits du bois sont potentiellement à haut risque en facilitant le mouvement des longicornes; Trichoferus campestris, Leptura quadrifasciata et Trichoferus holosericeus ont été trouvés, respectivement, dans un range-couvert en bois, une traverse ferroviaire, et une chaise de salle à manger. Une épidémie du Meloidogyne fallax a été identifiée chez l'Allium ampeloprasum et chez trois adventices. Des séquences de référence pour des psylles indigènes du Royaume-Uni ont été générées pour permettre le développe-ment d'outils diagnostiques rapides en vue de surveiller l'Aphalara itadori suite à son introduction en tant qu'agent de lutte biologique contre le Fallopia japonica. [Traduit par la Rédaction] Mots-clés : santé des végétaux, organisme nuisible réglementé, codage à barres de l'ADN, diagnostique...

No abstract

Root‐lesion nematodes of the genus Pratylenchus are migratory endoparasites with worldwide economic impact on several important crops including potato, where certain species like P. penetrans, P. neglectus, and P. scribneri reduce the yield and quality of potato tubers. Morphological identification of Pratylenchus spp. is challenging, and recent advancements in molecular techniques provide robust and rapid diagnostics to differentiate species without the need of specialist skills. However, the fact that molecular diagnostics are not available for all Pratylenchus species means that there are limitations in worldwide application. In general, root‐lesion nematodes are difficult to manage once introduced into agricultural land and damage can be related to pathogenicity and population densities. In addition, root‐lesion nematodes interact with fungi such as Verticillium dahliae, resulting in disease complexes that enhance the damage inflicted on the potato crop. Management interventions are often focused on limiting nematode reproduction before planting crops and include the application of nematicides, and cultural practices such as crop rotation, cover crops, biofumigation, and biological control. Understanding the limitations of the available crop protection strategies is important and there are many gaps for further study. This review discusses the status of the diagnosis, distribution, pathogenicity, and management of the main species of root‐lesion nematodes, reported to infect potatoes worldwide, and highlights areas for potential future research.