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A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites

Felix Moran; Antonio Olmos; Leonidas Lotos; Lukas Predajňa; Nikolaos Katis; Miroslav Glasa; Varvara Maliogka; Ana B. Ruiz-Garcia


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  <identifier identifierType="URL">https://zenodo.org/record/1248646</identifier>
  <creators>
    <creator>
      <creatorName>Felix Moran</creatorName>
      <affiliation>Centro de Proteccion Vegetal y Biotecnologia, Instituto Valenciano de Investigaciones Agrarias (IVIA), Moncada, Valencia, Spain</affiliation>
    </creator>
    <creator>
      <creatorName>Antonio Olmos</creatorName>
      <affiliation>Centro de Proteccion Vegetal y Biotecnologia, Instituto Valenciano de Investigaciones Agrarias (IVIA), Moncada, Valencia, Spain</affiliation>
    </creator>
    <creator>
      <creatorName>Leonidas Lotos</creatorName>
      <affiliation>Aristotle University of Thessaloniki, Faculty of Agriculture, Forestry and Natural Environment, School of Agriculture, Plant Pathology Laboratory, Thessaloniki, Greece</affiliation>
    </creator>
    <creator>
      <creatorName>Lukas Predajňa</creatorName>
      <affiliation>Institute of Virology, Biomedical Research Centre, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, Slovakia</affiliation>
    </creator>
    <creator>
      <creatorName>Nikolaos Katis</creatorName>
      <affiliation>Aristotle University of Thessaloniki, Faculty of Agriculture, Forestry and Natural Environment, School of Agriculture, Plant Pathology Laboratory, Thessaloniki, Greece</affiliation>
    </creator>
    <creator>
      <creatorName>Miroslav Glasa</creatorName>
      <affiliation>Institute of Virology, Biomedical Research Centre, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, Slovakia</affiliation>
    </creator>
    <creator>
      <creatorName>Varvara Maliogka</creatorName>
      <affiliation>Aristotle University of Thessaloniki, Faculty of Agriculture, Forestry and Natural Environment, School of Agriculture, Plant Pathology Laboratory, Thessaloniki, Greece</affiliation>
    </creator>
    <creator>
      <creatorName>Ana B. Ruiz-Garcia</creatorName>
      <affiliation>Centro de Proteccion Vegetal y Biotecnologia, Instituto Valenciano de Investigaciones Agrarias (IVIA), Moncada, Valencia, Spain</affiliation>
    </creator>
  </creators>
  <titles>
    <title>A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites</title>
  </titles>
  <publisher>Zenodo</publisher>
  <publicationYear>2018</publicationYear>
  <subjects>
    <subject>GPGV, Grapevine, Real-time RT-PCR, Quantification</subject>
  </subjects>
  <dates>
    <date dateType="Issued">2018-05-15</date>
  </dates>
  <language>en</language>
  <resourceType resourceTypeGeneral="JournalArticle"/>
  <alternateIdentifiers>
    <alternateIdentifier alternateIdentifierType="url">https://zenodo.org/record/1248646</alternateIdentifier>
  </alternateIdentifiers>
  <relatedIdentifiers>
    <relatedIdentifier relatedIdentifierType="DOI" relationType="IsIdenticalTo">10.1371/journal.pone.0197237</relatedIdentifier>
  </relatedIdentifiers>
  <rightsList>
    <rights rightsURI="https://creativecommons.org/licenses/by/4.0/legalcode">Creative Commons Attribution 4.0 International</rights>
    <rights rightsURI="info:eu-repo/semantics/openAccess">Open Access</rights>
  </rightsList>
  <descriptions>
    <description descriptionType="Abstract">&lt;p&gt;&lt;em&gt;Grapevine Pinot gris virus&lt;/em&gt; (GPGV) is a widely distributed grapevine pathogen that has been associated to the grapevine leaf mottling and deformation disease. With the aim of better understanding the disease epidemiology and providing efficient control strategies a specific and quantitative duplex TaqMan real-time RT-PCR assay has been developed. This method has allowed reliable quantitation of the GPGV titer ranging from 30 up to 3 x 10&lt;sup&gt;8&lt;/sup&gt; transcript copies, with a detection limit of 70 viral copies in plant material. The assay targets a grapevine internal control that reduces the occurrence of false negative results, thus increasing the diagnostic sensitivity of the technique. Viral isolates both associated and non-associated to symptoms from Greece, Slovakia and Spain have been successfully detected. The method has also been applied to the absolute quantitation of GPGV in its putative transmission vector &lt;em&gt;Colomerus vitis&lt;/em&gt;. Moreover, the viral titer present in single mites has been determined. In addition, in the current study a new polymorphism in the GPGV genome responsible for a shorter movement protein has been found. A phylogenetic study based on this genomic region has shown a high variability among Spanish isolates and points to a different evolutionary origin of this new polymorphism. The methodology here developed opens new possibilities for basic and epidemiological studies as well as for the establishment of efficient control strategies.&lt;/p&gt;</description>
    <description descriptionType="Other">This publication reflects only the authors' view. The Agency is not responsible for any use that may be made of the information it contains.</description>
  </descriptions>
  <fundingReferences>
    <fundingReference>
      <funderName>European Commission</funderName>
      <funderIdentifier funderIdentifierType="Crossref Funder ID">10.13039/501100000780</funderIdentifier>
      <awardNumber awardURI="info:eu-repo/grantAgreement/EC/H2020/734736/">734736</awardNumber>
      <awardTitle>Virus free Fruit Nurseries</awardTitle>
    </fundingReference>
  </fundingReferences>
</resource>
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