title: Identification of reference genes suitable for qRT-PCR in grapevine and application for the study of the expression of genes involved in pterostilbene synthesis.
abstract: The recent publication of the grapevine genome sequence facilitates the use of qRT-PCR to study gene expression changes. For this approach, reference genes are commonly used to normalize data and their stability of expression should be systematically validated. Among grapevine defenses is the production of the antimicrobial stilbenic phytoalexins, notably the highly fungitoxic pterostilbene, which plays a crucial role in grapevine interaction with Plasmopara viticola and Botrytis cinerea. As a resveratrol O-methyltransferase (ROMT) gene involved in pterostilbene synthesis was recently identified, we investigated the accumulation of the corresponding transcripts to those of two other stilbene biosynthesis related genes phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) in response to pathogen infection. Using three computer-based statistical methods and C(t) values or LRE method generated values as input data, we have first identified two reference genes (VATP16 and 60SRP) suitable for normalization of qPCR expression data obtained in grapevine leaves and berries infected by P. viticola and B. cinerea, respectively. Next, we have highlighted that the expression of ROMT is induced in P. viticola-infected leaves and also in B. cinerea-infected berries, confirming the involvement of pterostilbene in grapevine defenses.
authors.affiliation: Unité Mixte de Recherche INRA 1088/CNRS 5184, Université de Bourgogne Plante-Microbe-Environnement, 17 rue Sully, BP 86510, 21065 Dijon cedex, France.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Gamm
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Héloir
authors.forename: Jani
authors.initials: J
authors.lastname: Kelloniemi
authors.forename: Benoît
authors.initials: B
authors.lastname: Poinssot
authors.forename: David
authors.initials: D
authors.lastname: Wendehenne
authors.forename: Marielle
authors.initials: M
authors.lastname: Adrian
mesh_terms: D017931:DNA Primers / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012984:Software; D013223:Statistics as Topic; D013267:Stilbenes / Q000378:metabolism*; D027843:Vitis / Q000201:enzymology / Q000235:genetics*
chemical_list: D017931:DNA Primers; D013267:Stilbenes; C107773:pterostilbene