Reference gene selection for normalization of RT-qPCR gene expression data from Actinidia deliciosa leaves infected with Pseudomonas syringae pv. actinidiae
Identifieur interne : 001047 ( Main/Merge ); précédent : 001046; suivant : 001048Reference gene selection for normalization of RT-qPCR gene expression data from Actinidia deliciosa leaves infected with Pseudomonas syringae pv. actinidiae
Auteurs : Milena Petriccione [Italie] ; Francesco Mastrobuoni [Italie] ; Luigi Zampella [Italie] ; Marco Scortichini [Italie]Source :
- Scientific Reports [ 2045-2322 ] ; 2015.
Abstract
Normalization of data, by choosing the appropriate reference genes (RGs), is fundamental for obtaining reliable results in reverse transcription-quantitative PCR (RT-qPCR). In this study, we assessed
Url:
DOI: 10.1038/srep16961
PubMed: 26581656
PubMed Central: 4652207
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PMC:4652207Le document en format XML
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Reference gene selection for normalization of RT-qPCR gene expression data from <italic>Actinidia deliciosa</italic>
leaves infected with <italic>Pseudomonas syringae</italic>
pv. <italic>actinidiae</italic>
</title>
<author><name sortKey="Petriccione, Milena" sort="Petriccione, Milena" uniqKey="Petriccione M" first="Milena" last="Petriccione">Milena Petriccione</name>
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, Via Torrino 3, I-81100 Caserta,<country>Italy</country>
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<country xml:lang="fr">Italie</country>
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<author><name sortKey="Mastrobuoni, Francesco" sort="Mastrobuoni, Francesco" uniqKey="Mastrobuoni F" first="Francesco" last="Mastrobuoni">Francesco Mastrobuoni</name>
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<author><name sortKey="Zampella, Luigi" sort="Zampella, Luigi" uniqKey="Zampella L" first="Luigi" last="Zampella">Luigi Zampella</name>
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<author><name sortKey="Scortichini, Marco" sort="Scortichini, Marco" uniqKey="Scortichini M" first="Marco" last="Scortichini">Marco Scortichini</name>
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<series><title level="j">Scientific Reports</title>
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<front><div type="abstract" xml:lang="en"><p>Normalization of data, by choosing the appropriate reference genes (RGs), is fundamental for obtaining reliable results in reverse transcription-quantitative PCR (RT-qPCR). In this study, we assessed <italic>Actinidia deliciosa</italic>
leaves inoculated with two doses of <italic>Pseudomonas syringae</italic>
pv. <italic>actinidiae</italic>
during a period of 13 days for the expression profile of nine candidate RGs. Their expression stability was calculated using four algorithms: geNorm, NormFinder, BestKeeper and the deltaCt method. Glyceraldehyde-3-phosphate dehydrogenase (<italic>GAPDH</italic>
) and protein phosphatase 2A (<italic>PP2A</italic>
) were the most stable genes, while β-tubulin and 7s-globulin were the less stable. Expression analysis of three target genes, chosen for RGs validation, encoding the reactive oxygen species scavenging enzymes ascorbate peroxidase (APX), superoxide dismutase (SOD) and catalase (CAT) indicated that a combination of stable RGs, such as <italic>GAPDH</italic>
and <italic>PP2A</italic>
, can lead to an accurate quantification of the expression levels of such target genes. The <italic>APX</italic>
level varied during the experiment time course and according to the inoculum doses, whereas both <italic>SOD</italic>
and <italic>CAT</italic>
resulted down-regulated during the first four days, and up-regulated afterwards, irrespective of inoculum dose. These results can be useful for better elucidating the molecular interaction in the <italic>A. deliciosa</italic>
/<italic>P. s</italic>
. pv. <italic>actinidiae</italic>
pathosystem and for RGs selection in bacteria-plant pathosystems.</p>
</div>
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