title: Analysis of reference gene stability after Israeli acute paralysis virus infection in bumblebees Bombus terrestris.
abstract: To date, there are no validated internal reference genes for the normalization of RT-qPCR data from virus infection experiments with pollinating insects. In this study we evaluated the stability of five candidate internal reference genes: elongation factor-1-alpha (ELF1α), peptidylprolyl isomerase A (PPIA), 60S ribosomal protein L23 (RPL23), TATA-binding protein (TBP) and polyubiquitin (UBI), in relation to Israeli acute paralysis virus (IAPV) infection of Bombus terrestris. We investigated the stability of these genes: in whole bodies and individual body parts, as well as in whole bodies collected at different time intervals after infection with IAPV. Our data identified PPIA as the single, most-optimal internal reference gene and the combination of PPAI-RPL23-UBI as a fully-sufficient multiple internal reference genes set for IAPV infection experiments in B. terrestris.
authors.affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium.
authors.forename: Jinzhi
authors.initials: J
authors.lastname: Niu
authors.affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium.
authors.forename: Kaat
authors.initials: K
authors.lastname: Cappelle
authors.affiliation: Department of Ecology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Joachim R
authors.initials: JR
authors.lastname: de Miranda
authors.affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium.
authors.forename: Guy
authors.initials: G
authors.lastname: Smagghe
authors.affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium. Electronic address: ivan.meeus@ugent.be.
authors.forename: Ivan
authors.initials: I
authors.lastname: Meeus
mesh_terms: D000818:Animals; D001516:Bees / Q000235:genetics* / Q000821:virology*; D056629:Dicistroviridae; D017344:Genes, Insect / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction
keywords: Bee virus; Dicistroviruses; Gene expression; RT-qPCR; qBase(PLUS)