A survey of aphelenchid nematodes (Nematoda: Aphelenchida) associated with maritime pine, Pinus pinaster, was conducted in Portugal in 1996 and 1999. A Bursaphelenchus species has been identified for the first time in the Iberian Peninsula. B. xylophilus is reported for the first time in Europe. It was found in very high numbers - up to 38 000 per 10 g of pine wood - inside a few declining trees infested with curculionid, cerambycid and scolytid beetles. Morphological observations, including shape of spicules, bursa, vulva, female tail end and stylet as well as morphometrics, were in accordance with the species description. Species-specific DNA fragment patterns were obtained using ITS-RFLP analysis, with five different restriction enzymes. The importance and implications of this finding are discussed. Premiere signalisation de Bursaphelenchus xylophilus au Portugal, at en Europe - Une enquete sur les nematodes Aphelenchides (Nematoda: Aphelenchida) associes au pin maritime (Pinus pinaster) a ete realisee au Portugal de 1996 a 1999. Une espece de Bursaphelenchus a ete identifiee pour la premiere fois dans la Peninsule Iberique. B. xylophilus est signale pour la premiere fois en Europe. Il a ete trouve en tres grand nombre - jusqu'a 38 000 individus pour 10 g de bois de pin - dans des arbres deperissants infestes par des Coleopteres Curculionides, Cerambycides et Scolytides. Les observations concernant la morphologie - en particulier la forme des spicules, la bourse, la vulve, l'extremite de la queue de la femelle et le stylet - de meme que les donnees morphometriques correspondent a la description de l'espece. Des sequences de fragments d'ADN specifique de l'espece ont ete obtenus par analyse ITS-RFLP a l'aide de cinq enzymes de restriction. L'importance et les implications de cette decouverte sont discutees.

The detection of Bursaphelenchus xylophilus in Pinus pinaster trees in Portugal in 1999 led the official services to implement exceptional and urgent control measures to prevent the spread of the pest. Because the pest is vector‐transmitted, special attention was paid to the insects associated with infested trees. A survey comprising 21 species of insects, from six families, with a total of 1367 specimens, was made in order to evaluate their possible role as vectors of the nematode in Portugal. Five insect species were found to carry other Bursaphelenchus spp., but only Monochamus galloprovincialis was associated with B. xylophilus.

Insects associated with maritime pine, Pinus pinaster, in Portugal were collected and screened for the presence of Bursaphelenchus species.

No abstract

Summary -Species of Bursaphelenchus associated with maritime pine, Pinus pinaster, from Portugal -within and outside the quarantine restricted demarcated zone of B. xylophilus -are described and characterised both morphologically (LM and SEM) and with the use of molecular biology (ITS-RFLP). A new staining method for spicules is proposed. Species include B. hellenicus, B. hylobianum, B. leoni, B. pinophilus, B. sexdentati, B. tusciae, B. teratospicularis, B. xylophilus and Bursaphelenchus sp. 1. Bursaphelenchus hylobianum was collected from the insect Hylobius sp. The most frequent species in the demarcated zone, besides B. xylophilus, was Bursaphelenchus sp. 1. Morphological characterisation is compared with the original descriptions and discussed. The differentiation between B. pinophilus and B. sexdentati is not clear in the literature and is discussed. Since differentiation of B. xylophilus (mucronate form) from B. mucronatus, and B. pinophilus from B. sexdentati, as well as their juvenile forms, is almost im possible on the basis of morphological features, a molecular approach based on ITS-RFLPs was used. Ribosomal DNA containing the 5.8S gene, the internal transcribed spacer region 1 and 2, and partial regions of 18S and 28S gene were amplified by PCR. Restriction profiles of the amplified products generated species-specific differences, leading to the unambiguous identification of isolates belonging to B. xylophilus, B. mucronatus, B. sexdentati, B. tusciae and B. hylobianum.

Summary -Species of Bursaphelenchus associated with maritime pine, Pinus pinaster, from Portugal -within and outside the quarantine restricted demarcated zone of B. xylophilus -are described and characterised both morphologically (LM and SEM) and with the use of molecular biology (ITS-RFLP). A new staining method for spicules is proposed. Species include B. hellenicus, B. hylobianum, B. leoni, B. pinophilus, B. sexdentati, B. tusciae, B. teratospicularis, B. xylophilus and Bursaphelenchus sp. 1. Bursaphelenchus hylobianum was collected from the insect Hylobius sp. The most frequent species in the demarcated zone, besides B. xylophilus, was Bursaphelenchus sp. 1. Morphological characterisation is compared with the original descriptions and discussed. The differentiation between B. pinophilus and B. sexdentati is not clear in the literature and is discussed. Since differentiation of B. xylophilus (mucronate form) from B. mucronatus, and B. pinophilus from B. sexdentati, as well as their juvenile forms, is almost im possible on the basis of morphological features, a molecular approach based on ITS-RFLPs was used. Ribosomal DNA containing the 5.8S gene, the internal transcribed spacer region 1 and 2, and partial regions of 18S and 28S gene were amplified by PCR. Restriction profiles of the amplified products generated species-specific differences, leading to the unambiguous identification of isolates belonging to B. xylophilus, B. mucronatus, B. sexdentati, B. tusciae and B. hylobianum.

The tribe Longidorini within the subfamily Longidorinae (Longidorus spp. and Paralongidorus spp.) and the subfamily Xiphineminae (Xiphinema spp.) are two large nematode groups with about 260 and 230 known species, respectively. They are globally two important groups of ectoparasitic nematodes considered to be major pests because of their activity as vectors of important plant nepovirus, with some species included in the list of quarantine pathogenic organisms in many European countries. Knowledge of the biodiversity and occurrence of this nematode group is a prerequisite for the establishment of sound management strategies and control measures. According to data collected from the databases (such as EPPO, FSTA, and Web of Science) and published in specialised literature, a total of 15 Longidorus, 1 Paralongidorus and 40 Xiphinema species have been recorded as present in Portugal. However, the taxonomic status of some species is controversial, and thus needs to be clarified. A comprehensive review for unravelling the biodiversity and occurrence of nematode species of the genus Longidorus, Paralongidorus and Xiphinema in Portugal is herein provided. This review includes an updated checklist of species with information on the localities, host plants and geographical distribution. Additionally, maps on the species distributions of Longidorinae and Xiphineminae nematodes present in Continental Portugal and the Portuguese Macaronesian islands are provided, as well as unpublished data on authors and comments on the current taxonomic status. Finally, new insights and directions for future research on Longidoridae in Portugal are presented.

Fusarium oxysporum f. sp. melonis is an important vascular wilt pathogen of melon. Races 1, 2 and 1–2 of this fungus have been identified in Portugal by pathogenicity tests with appropriate hosts. The aim of this research was to examine the relationships between different races of F. o. melonis of Portuguese and French origin through analysis of random amplified polymorphic DNAs (RAPDs). DNA fingerprint profiles were developed for all the accessions. Each isolate showed 5–10 DNA bands with each of the 16 primers employed. A total of 126 bands was obtained. The size of amplified DNA fragments generated with these primers ranged from 0.5 to 3.2 kb. A phenogram based on the Jaccard coefficient of similarity was computed by the unweighed pair group method using arithmetic averages (UPGMA). It was found that Portuguese race 2 is very similar to French race 1, while French race 2 is the most dissimilar being clearly separated from all other races. The genetic diversity of these isolates is also being studied for vegetative compatibility by using the nit mutant system.