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An intracellular bacterium, strain IAST, was observed to infect several species of the plant-parasitic nematode genus Xiphinema (Xiphinema astaregiense, Xiphinema incertum, Xiphinema madeirense, Xiphinema pachtaicum, Xiphinema parapachydermum and Xiphinema vallense). The bacterium could not be recovered on axenic medium. The 16S rRNA gene sequence of IAST was found to be new, being related to the family Burkholderiaceae, class Betaproteobacteria. Fungal endosymbionts Mycoavidus cysteinexigens B1-EBT (92.9 % sequence identity) and ‘Candidatus Glomeribacter gigasporarum’ BEG34 (89.8 % identity) are the closest taxa and form a separate phylogenetic clade inside Burkholderiaceae. Other genes (atpD, lepA and recA) also separated this species from its closest relatives using a multilocus sequence analysis approach. These genes were obtained using a partial genome of this bacterium. The localization of the bacterium (via light and fluorescence in situ hybridization microscopy) is in the X. pachtaicum females clustered around the developing oocytes, primarily found embedded inside the epithelial wall cells of the ovaries, from where they are dispersed in the intestine. Transmission electron microscopy (TEM) observations supported the presence of bacteria inside the nematode body, where they occupy ovaries and occur inside the intestinal epithelium. Ultrastructural analysis of the bacterium showed cells that appear as mostly irregular, slightly curved rods with rounded ends, 0.8–1.2 µm wide and 2.5–6.0 µm long, possessing a typical Gram-negative cell wall. The peptidoglycan layer is, however, evident only occasionally and not detectable by TEM in most cells. Another irregularly occurring shell surrounding the endosymbiont cells or the cell clusters was also revealed, probably originating from the host cell membrane. Flagella or spore-like cells do not occur and the nucleoid is diffusely distributed throughout the cell. This endosymbiont is transmitted vertically through nema

Three populations of Xiphinema non-americanum group species were detected in agricultural and natural ecosystems, during routine surveys for plant-parasitic nematodes in Spain. Based on morphological and molecular analyses, the species were identified as Xiphinema histriae and Xiphinema lapidosum, being this the first record and molecular characterization of both species in Spain. The morphometrics and morphology of the Spanish populations agree with those of the original description and paratype specimens and the present study provided a first description of the second to fourth juvenile stages of both species. A detailed study on the morphology in the Spanish populations of X. histriae, as well as in paratypes, showed a pseudo-Z-organ with weakly muscularized wall and containing numerous small dense granular bodies, which was different to the original description by Lamberti et al. (1993). This new finding suggests that X. histriae must be considered a member of the morphospecies Group 5 of X. non-americanum. Phylogenetic analysis based on D2 to D3 expansion segments of 28S gene, ITS1 and partial CoxI gene indicated that X. histriae and X. lapidosum are phylogenetically related with other Xiphinema non-americanum group spp. reported from Spain. Considering the pathological and economic importance of this group of nematodes, the combination of morphological characters, measurements, and molecular analysis is crucial for accurate identification of these species.

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated olive (cvs. Chemlali and Chétoui) in Tunisia were investigated. Morphological and morphometrical studies identified three Longidorus and six Xiphinema species, with frequencies of prevalence as following: Longidorus africanus (23.0 %), L. euonymus (4.5 %), L. glycines (13.7 %), Xiphinema conurum (13.7 %), X. italiae (36.4 %), X. meridianum (13.7 %), X. pachtaicum (18.2 %), X. robbinsi (9.1 %), and Xiphinema sp. (4.5 %). The three Longidorus species were reported for the first time in Tunisia, in addition to two species of Xiphinema (viz. X. meridianum and X. robbinsi). Molecular characterisation using D2-D3 expansion regions of 28S rRNA and ITS1-rRNA was carried out and Bayesian inference analysis was used to reconstruct phylogenetic relationships among these species and with other longidorids. Twenty-five new D2-D3 of 28S rRNA gene sequences were obtained in the present study, seven for Longidorus and 18 for Xiphinema spp., as well as 14 new ITS1 rRNA gene sequences (seven for Longidorus and seven for Xiphinema spp.).

Het Strawberry latent ringspot virus (SLRSV, in het Nederlands Latent aardbeikringvlekkenvirus) komt in een groot aantal gewassen voor, waaronder o.a. aardbei, framboos, bessenstruiken, verschillende fruitbomen (kers, pruim, perzik), groenten en kruiden (asperge, selderij, peterselie) en siergewassen (roos, lelie en narcis). Binnen de EU is dit virus een quarantaine organisme in plantmateriaal van aardbei, braam en framboos. In andere delen van de wereld is SLRSV ook een quarantaine organisme in andere gewassen, voor de export van lelie naar o.a. China en de Verenigde Staten geldt een 0-tolerantie. SLRSV wordt overgebracht door een tweetal aaltjes-soorten: Xiphinema diversicaudatum en X. coxi. Er zijn echter vermoedens dat dit virus ook op andere manieren verspreid kan worden dan alleen door de twee genoemde Xiphinema-soorten. Er is namelijk overdracht waargenomen in teelten waarin geen Xiphinema is waargenomen. De vraag is dus op welke manieren SLRSV kan worden verspreid. In de literatuur is al gerapporteerd dat SLRSV in een hoog percentage (~70%) met zaad over kan gaan in bijvoorbeeld selderij, munt en braam. Van zaadoverdracht in het gewas lelie is tot op heden nog niets bekend. Bovendien kan het virus zeer algemeen voorkomende onkruiden als herderstasje en muur infecteren. Het doel van het onderzoek waarover in dit verslag wordt gerapporteerd was het nagaan of de verspreiding van SLRSV in lelie al dan niet via zaadoverdracht kan plaatsvinden. Hiervoor werden experimenten uitgevoerd met een vijftal leliecultivars die via zaad worden vermeerderd. SLRSV kon bij deze cultivars in het zaad, het stuifmeel en in nieuwe zaailingen worden aangetoond. SLRSV kon zo in hoge mate worden aangetoond in zaden afkomstig uit partijen met een zeker percentage SLRSV. Bij één cultivar werd een percentage van 30% infectie gevonden in zaailingen die waren gegroeid uit zaad uit een besmette partij. Het virus kon ook worden aangetoond in het stuifmeel van geïnfecteerde bloemen. De vraag

This work has studied the diversity of plant-parasitic nematodes infecting olive in a wide-region in southern Spain (Andalusia). Our data indicate that there is a high diversity of plant-parasitic nematodes (including root-knot = Meloidogyne spp., root-lesion = Pratylenchus spp., cystforming = Heterodera mediterranea, stubby- root = Trichodorus spp., reniform = Rotylenchulus sp., dagger and needle = Longidorus spp., Xiphinema spp., ring = Criconemoides spp., Ogmo spp., and other migratory ectoparasitic nematodes viz. Amplimerlinius spp., Helicotylenchus spp., Neodolichorhynchus spp., Paratrophurus spp. Paratylenchus spp., Rotylenchus spp., Trophurus sp., Tylenchorhynchus spp.) associated with olive in Southern Spain that can exert different damage to olive roots depending on the olive variety and their abundance. The highest number of species was detected in Xiphinema and Longidorus, with description of eight and seven new species, respectively, apart from several new records for Spain. In addition, seven Meloidogyne species were detected in wi ld and cultivated olive, from which one is being described as a new species. Multivariate analyses allowed determining the agronomic and environmental factors that drive the structure and diversity of plant-parasitic nematodes infecting olive in Southern-Spain, that included: soil physicochemicol factors (texture, pH, sand and doy, and extractable K), climatic variables (mínimum and maximum temperatures) and the olive cultivar. Nematode abundance and diversity indexes (Richness, Shannon, Evenness and plant-parasitic index) were influenced by olive cultivar, and orchard and soil management predices. The plant-parasitic nematode abundance was significantly higher in orchards managed organically compared to those managed conventionally. These findings could be the result of the influence of the herbaceous plants that compase the soil cover, which may affect plant-parasit ic nematodes at the species level.

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of grapevine plants in southern Spain were investigated. Nematode surveys were conducted on 77 vineyards during the spring seasons of 2006, 2007 and 2008 in the main Andalusian grapevine-growing areas, including the provinces of Cádiz, Córdoba, and Huelva. Morphological and morphometrical studies identified two Longidorus and nine Xiphinema species, viz.: Longidorus alvegus, L. magnus, Xiphinema adenohystherum, X. hispidum, X. index, X. italiae, X. lupini, X. nuragicum, X. pachtaicum, X. rivesi, and X. turcicum. Overall, frequencies of infestation were, in decreasing order: X. pachtaicum 90.8%, X. index 30.3%, X. italiae 13.2%, L. magnus 11.8%, X. hispidum 7.9%, X. lupini 3.9%, L. alvegus and X. rivesi 2.6%, and X. adenohystherum, X. nuragicum and X. turcicum 1.3%. Xiphinema hispidum, X. lupini, L. alvegus and L. magnus were compared with nematode type specimens and are reported for the first time in Spain. Furthermore, the male of L. alvegus is described for the first time in the literature. Molecular characterisation of these species using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids. The monophily of the genera Xiphinema and Longidorus was accepted and the genera Paralongidorus and Xiphidorus were rejected by the Shimodaira-Hasegawa test based on tree topologies. © 2010 KNPV.

Poster met onderzoeksinformatie over de detectie van Arabismozaïekvirus (ArMV) en Tabaksratelvirus (TRV) en hun verspreiders Paratrichodorus- en Trichodorus-nematoden in bodemmonsters.