In Europe the most devastating phytoplasma associated with grapevine yellows (GY) diseases is a quarantine pest, flavescence dorée (FDp), from the 16SrV taxonomic group. The on-site detection of FDp with an affordable device would contribute to faster and more efficient decisions on the control measures for FDp. Therefore, a real-time isothermal LAMP assay for detection of FDp was validated according to the EPPO standards and MIQE guidelines. The LAMP assay was shown to be specific and extremely sensitive, because it detected FDp in all leaf samples that were determined to be FDp infected using quantitative real-time PCR. The whole procedure of sample preparation and testing was designed and optimized for on-site detection and can be completed in one hour. The homogenization procedure of the grapevine samples (leaf vein, flower or berry) was optimized to allow direct testing of crude homogenates with the LAMP assay, without the need for DNA extraction, and was shown to be extremely sensitive.

The larvae of the Old World genera Leucinodes Guenée, 1854 and Sceliodes Guenée, 1854 are internal feeders in the fruits of Solanaceae, causing economic damage to cultivated plants like Solanum melongena and Solanum aethiopicum. In sub-Saharan Africa five nominal species of Leucinodes and one of Sceliodes occur. One of these species, the eggplant fruit and shoot borer Leucinodes orbonalis Guenée, 1854, is regarded as regularly intercepted from Africa and Asia in Europe, North and South America and is therefore a quarantine pest on these continents. We investigate the taxonomy of African Leucinodes and Sceliodes based on morphological characters in wing pattern, genitalia and larvae, as well as mitochondrial DNA, providing these data for identification of all life stages. The results suggest that both genera are congeneric, with Sceliodes syn. n. established as junior subjective synonym of Leucinodes. Leucinodes orbonalis is described from Asia and none of the samples investigated from Africa belong to this species. Instead, sub-Saharan Africa harbours a complex of eight endemic Leucinodes species. Among the former nominal species of Leucinodes (and Sceliodes) from Africa, only Leucinodes laisalis (Walker, 1859), comb. n. (Sceliodes) is confirmed, with Leucinodes translucidalis Gaede, 1917, syn. n. as a junior subjective synonym. The other African Leucinodes species were unknown to science and are described as new: Leucinodes africensis sp. n., Leucinodes ethiopica sp. n., Leucinodes kenyensis sp. n., Leucinodes malawiensis sp. n., Leucinodes pseudorbonalis sp. n., Leucinodes rimavallis sp. n. and Leucinodes ugandensis sp. n. An identification key based on male genitalia is provided for the African Leucinodes species. Most imports of Leucinodes specimens from Africa into Europe refer to Leucinodes africensis, which has been frequently imported with fruits during the last 50 years. In contrast, Leucinodes laisalis has been much less frequently recorded, and Leucinodes pseudorbonalis as well as Leucinodes rimavallis only very recently in fruit imports from Uganda. Accordingly, interceptions of Leucinodes from Africa into other continents will need to be re-investigated for their species identity and will likely require, at least in parts, revisions of the quarantine regulations. The following African taxa are excluded from Leucinodes: Hyperanalyta Strand, 1918, syn. rev. as revised synonym of Analyta Lederer, 1863; Analyta apicalis (Hampson, 1896), comb. n. (Leucinodes); Lygropia aureomarginalis (Gaede, 1916), comb. n. (Leucinodes); Syllepte hemichionalis Mabille, 1900, comb. rev., Syllepte hemichionalis idalis Viette, 1958, comb. rev. and Syllepte vagans (Tutt, 1890), comb. n. (Aphytoceros). Deanolis iriocapna (Meyrick, 1938), comb. n. from Indonesia is originally described and misplaced in Sceliodes, and Leucinodes cordalis (Doubleday, 1843), comb. n. (Margaritia) from New Zealand, Leucinodes raondry (Viette, 1981), comb. n. (Daraba) from Madagascar as well as Leucinodes grisealis (Kenrick, 1912), comb. n. (Scelio...

The yellow-legged or Asian hornet (Vespa velutina colour form nigrithorax) was introduced into France from China over a decade ago. Vespa velutina has since spread rapidly across Europe, facilitated by suitable climatic conditions and the ability of a single nest to disperse many mated queens over a large area. Yellow-legged hornets are a major concern because of the potential impact they have on populations of many beneficial pollinators, most notably the western honey bee (Apis mellifera), which shows no effective defensive behaviours against this exotic predator. Here, we present the first report of this species in Great Britain. Actively foraging hornets were detected at two locations, the first around a single nest in Gloucestershire, and the second a single hornet trapped 54 km away in Somerset. The foraging activity observed in Gloucestershire was largely restricted to within 700 m of a single nest, suggesting highly localised movements. Genetic analyses of individuals from the Gloucestershire nest and the single hornet from Somerset suggest that these incursions represent an expansion of the European population, rather than a second incursion from Asia. The founding queen of the Gloucestershire nest mated with a single male, suggesting that sexual reproduction may have occurred in an area of low nest density. Whilst the nest contained diploid adult males, haploid ‘true’ males were only present at the egg stage, indicating that the nest was detected and removed before the production of queens. Members of the public reported additional dead hornets associated with camping equipment recently returned from France and imported timber products, highlighting possible pathways of incursion. The utility of microsatellites to inform surveillance during an incursion and the challenge of achieving eradication of this damaging pest are discussed.

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Biological Journal of the Linnean Society 69 (2000) 1-20. doi:10.1006/bijl.1999.0345Received by publisher: 1998-06-02Harvest Date: 2016-01-04 12:20:29DOI: 10.1006/bijl.1999.0345Page Range: 1-2

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...

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This is the first LAMP assay for Xcm which provides a significant improvement compared to existing diagnostics.