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Analysis of genes expressed in second stage juveniles of the potato cyst nematodes Globodera rostochiensis and G. pallida using the expressed sequence tag approach

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Expressed sequence tag (EST) projects offer a rapid route to the discovery of novel genes. Genes expressed in a wide range of parasitic nematodes of medical or veterinary importance have been investigated using EST analysis but these techniques have not yet been applied to plant parasitic nematodes. We describe a small scale EST project using cDNA libraries made from the two species of potato cyst nematode, Globodera rostochiensis and G. pallida, and assess the utility of this approach to identify mRNAs encoding abundantly expressed secreted proteins and other proteins present in the nematode at the onset of parasitism. Approximately 1000 sequences were obtained from G. rostochiensis and 100 from G. pallida. A variety of genes was characterised and approximately 11% of the cDNAs sequenced were apparently PCN specific. Secreted proteins identified included a novel PCN homologue of chorismate mutase, a cDNA recently cloned from the gland cells of Meloidogyne javanica. The results obtained justify a much larger scale application of this technology to these parasites.

Utilisation de L'Expressed Sequence Tag pour l'analyse de gènes s'exprimant chez les juvéniles de deuxième stade des nématodes à kyste de la pomme de terre Globodera rostochiensis et G. pallida - l'Expressed Sequence Tag (EST) ouvre une voie rapide vers la découverte de nouveaux gènes. Des gènes s'exprimant chez un large éventail de nématodes parasites d'importance médicale ou vétérinaire ont été étudiés par analyse EST alors que cette technique n'a pas encore été appliquée aux nématodes phytoparasites. Nous décrivons ici un projet EST à petite échelle utilisant les banques d'ADNc constituées à partir de deux espèces de nématodes à kyste de la pomme de terre (PCN), Globodera rostochiensis et G. pallida et nous évaluons l'utilité de cette approche pour identifier les ARNs codant les protéines abondamment sécrétées - et les autres protéines - présentes chez les nématodes lors de l'attaque parasitaire. Mille séquences environ ont été obtenues chez G. rostochiensis et 100 chez G. pallida. Des gènes variés ont été caractérisés et environ 11% des ADNc séquencés sont apparemment spécifiques des PCN. Parmi les protéines sécrétées identifiées figurent un nouvel homologue PCN de la chorismate mutase ainsi qu'un ADNc récemment cloné à partir de cellules glandulaires de Meloidogyne javanica. Les résultats ainsi obtenus justifient une utilisation à plus grande échelle de l'EST pour l'étude de ces parasites.

Affiliations: 1: Nematology Department, Agricultural University of Wageningen, The Netherlands; 2: Mycology, Bacteriology and Nematology Unit, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK; 3: Genomics Unit, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK

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/content/journals/10.1163/156854100509358
2000-08-15
2016-12-05

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