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Development and validation of species-specific primers that provide a molecular diagnostic for virus-vector longidorid nematodes and related species in German viticulture

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Abstract

The nematode species Longidorus attenuatus, L. elongatus, L. macrosoma and Paralongidorus maximusare economically important pests to the viticulture industry due to their ability to vector two nepoviruses (Raspberry Ringspot Virus and Tomato Black Ring Virus) to grapevines. In Germany, these species occur in vineyard soil with other non-vector but morphologically similar longidorid species, L. helveticus, L.␣profundorum and L. sturhani. Species-specific primers were designed from ribosomal DNA for all seven species to facilitate taxonomic identification for non-specialists. Primers were assessed for their reliability by screening, where possible, a number of populations of each species. Furthermore, their selectivity and sensitivity were determined when challenged with closely related longidorid species and general nematode communities typical of vineyard soil. A multiplex approach using a common forward primer combined with species-specific reverse primers enabled three target nematode species to be detected in the same PCR reaction. All primers were highly specific, detecting all nematode developmental forms from disparate populations and were sufficiently sensitive to detect a single target nematode within a whole nematode community typical of a vineyard soil comprising of a range of non-target species. Given their specificity, sensitivity and reliability, these diagnostic primers should be of great benefit to both phytosanitary/quarantine services related to the viticulture industry and also as a decision management tool for growers.

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Authors and Affiliations

  1. Dienstleistungszentrum Ländlicher Raum Rheinpfalz, Neustadt/Weinstrasse, Germany

    Judith Hübschen, Lilo Kling & Ulrike Ipach

  2. Lehrstuhl für Phytopathologie, Technische Universität München, Freising-Weihenstephan, Germany

    Volker Zinkernagel

  3. Central Laboratory of General Ecology, 2 Gagarin Street, Bulgaria

    Derek Brown

  4. Invergowrie, Scottish Crop Research Institute, Dundee, DD2 5DA, UK

    Roy Neilson

Authors
  1. Judith Hübschen
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  2. Lilo Kling
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  3. Ulrike Ipach
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  4. Volker Zinkernagel
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  5. Derek Brown
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  6. Roy Neilson
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Hübschen, J., Kling, L., Ipach, U. et al. Development and validation of species-specific primers that provide a molecular diagnostic for virus-vector longidorid nematodes and related species in German viticulture. European Journal of Plant Pathology 110, 883–891 (2004). https://doi.org/10.1007/s10658-004-4841-x

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  • DOI: https://doi.org/10.1007/s10658-004-4841-x

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