Validation and selection of reference genes for quantitative Real-time PCR of GLRaV-3 in Vitis vinifera L. (Malvasia cv.) for different seasons and tissues samples (Abstract/Poster in atti di convegno)

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  • Validation and selection of reference genes for quantitative Real-time PCR of GLRaV-3 in Vitis vinifera L. (Malvasia cv.) for different seasons and tissues samples (Abstract/Poster in atti di convegno) (literal)
Anno
  • 2013-01-01T00:00:00+01:00 (literal)
Alternative label
  • Montero R., Elaououad H.M., Pacifico D., Marzachì C., Flexas J., Bota J. (2013)
    Validation and selection of reference genes for quantitative Real-time PCR of GLRaV-3 in Vitis vinifera L. (Malvasia cv.) for different seasons and tissues samples
    in International Advances in Plant Virology, Norwich, UK, 25th-27nd September 2013
    (literal)
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  • Montero R., Elaououad H.M., Pacifico D., Marzachì C., Flexas J., Bota J. (literal)
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  • Atti del Convegno International Advances in Plant Virology (literal)
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  • 1 (literal)
Note
  • Abstract (literal)
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  • MR, BJ: Institut de Recerca i Formació Agrària i Pesquera (IRFAP), Conselleria d'Agricultura, Medi Ambient i Territori. Govern de les Illes Balears. Palma de Mallorca, Spain EH, FJ: 2Grup de Recerca en Biologia de les Plantes en Condicions Mediterrànies, Departament de Biologia, Universitat de les Illes Balears, Palma de Mallorca, Balears, Spain PD, MC: Istituto di Virologia Veghetale, CNR, Torino, Italy (literal)
Titolo
  • Validation and selection of reference genes for quantitative Real-time PCR of GLRaV-3 in Vitis vinifera L. (Malvasia cv.) for different seasons and tissues samples (literal)
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  • Association of Applied Biologists (literal)
Abstract
  • Grapevine Leafroll virus is one of the most important virus diseases of grapevines. Grapevine Leafroll associated-3 (GLRaV-3), as the other members of the Closteroviridae family are systemic in the vine, although they are generally localized in the vascular plant tissue (phloem). This virus are generally more concentrated in leaves. Real-time reverse transcription PCR (RT-qPCR) has become a routine technique for gene expression analysis, allowing accurate high-throughput RNA quantification over a wide dynamic range at a relatively low cost. Normalization is essential to control for experimental error between samples that can be introduced at a number of stages throughout the procedure. It is essential to validate potential reference genes to establish whether they are appropriate for a specific experimental purpose. UBIQUITIN, NAD5 and GAPDH were selected as candidate housekeeping genes for normalization of gene expression in reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) in different tissue samples and throughout the growing season. The present work was conducted using 'Malvasia de Banyalbufar', a grapevine variety from Mallorca (Balearic Islands, Spain). Leaf samples were collected in July and August and wood samples were collected in January. Using the geNorm program the most stable housekeeping throughout the growing season and different tissues was UBIQUITIN, therefore the best one for GLRaV-3 quantification. UBIQUITIN gene showed the lowest M value. GAPDH and NAD5 genes gave the highest M value and hence considered as having lowest stability under different experimental conditions. (literal)
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