title: Identification of Reference Genes for RT-qPCR Data Normalization in Cannabis sativa Stem Tissues.
abstract: Gene expression profiling via quantitative real-time PCR is a robust technique widely used in the life sciences to compare gene expression patterns in, e.g., different tissues, growth conditions, or after specific treatments. In the field of plant science, real-time PCR is the gold standard to study the dynamics of gene expression and is used to validate the results generated with high throughput techniques, e.g., RNA-Seq. An accurate relative quantification of gene expression relies on the identification of appropriate reference genes, that need to be determined for each experimental set-up used and plant tissue studied. Here, we identify suitable reference genes for expression profiling in stems of textile hemp (Cannabis sativa L.), whose tissues (isolated bast fibres and core) are characterized by remarkable differences in cell wall composition. We additionally validate the reference genes by analysing the expression of putative candidates involved in the non-oxidative phase of the pentose phosphate pathway and in the first step of the shikimate pathway. The goal is to describe the possible regulation pattern of some genes involved in the provision of the precursors needed for lignin biosynthesis in the different hemp stem tissues. The results here shown are useful to design future studies focused on gene expression analyses in hemp.
authors.affiliation: Environmental Research and Innovation (ERIN), Luxembourg Institute of Science and Technology (LIST), L-4362 Esch/Alzette, Luxembourg. lauralie.mangeot@yahoo.fr.
authors.forename: Lauralie
authors.initials: L
authors.lastname: Mangeot-Peter
authors.affiliation: Environmental Research and Innovation (ERIN), Luxembourg Institute of Science and Technology (LIST), L-4362 Esch/Alzette, Luxembourg. sylvain.legay@list.lu.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Legay
authors.affiliation: Environmental Research and Innovation (ERIN), Luxembourg Institute of Science and Technology (LIST), L-4362 Esch/Alzette, Luxembourg. jean-francois.hausman@list.lu.
authors.forename: Jean-Francois
authors.initials: JF
authors.lastname: Hausman
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Via Cinthia, I-80126 Napoli, Italy. sergio.esposito@unina.it.
authors.forename: Sergio
authors.initials: S
authors.lastname: Esposito
authors.affiliation: Environmental Research and Innovation (ERIN), Luxembourg Institute of Science and Technology (LIST), L-4362 Esch/Alzette, Luxembourg. gea.guerriero@list.lu.
authors.forename: Gea
authors.initials: G
authors.lastname: Guerriero
mesh_terms: D002188:Cannabis / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D010940:Plant Proteins; D018749:RNA, Plant
keywords: Cannabis sativa; cell wall; gene expression; lignification; reference genes