title: Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae).
abstract: Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is a reliable technique for quantifying gene expression across various biological processes, of which requires a set of suited reference genes to normalize the expression data. Coleomegilla maculata (Coleoptera: Coccinellidae), is one of the most extensively used biological control agents in the field to manage arthropod pest species. In this study, expression profiles of 16 housekeeping genes selected from C. maculata were cloned and investigated. The performance of these candidates as endogenous controls under specific experimental conditions was evaluated by dedicated algorithms, including geNorm, Normfinder, BestKeeper, and ΔCt method. In addition, RefFinder, a comprehensive platform integrating all the above-mentioned algorithms, ranked the overall stability of these candidate genes. As a result, various sets of suitable reference genes were recommended specifically for experiments involving different tissues, developmental stages, sex, and C. maculate larvae treated with dietary double stranded RNA. This study represents the critical first step to establish a standardized RT-qPCR protocol for the functional genomics research in a ladybeetle C. maculate. Furthermore, it lays the foundation for conducting ecological risk assessment of RNAi-based gene silencing biotechnologies on non-target organisms; in this case, a key predatory biological control agent.
authors.affiliation: Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Hunan, China.|Department of Entomology, University of Kentucky, Lexington, KY, USA.
authors.forename: Chunxiao
authors.initials: C
authors.lastname: Yang
authors.affiliation: Department of Entomology, University of Kentucky, Lexington, KY, USA.
authors.forename: Huipeng
authors.initials: H
authors.lastname: Pan
authors.affiliation: Department of Entomology, University of Kentucky, Lexington, KY, USA.
authors.forename: Jeffrey Edward
authors.initials: JE
authors.lastname: Noland
authors.affiliation: Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Hunan, China.
authors.forename: Deyong
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Hunan Vegetable Institute, Hunan Academy of Agricultural Sciences, Hunan, China.
authors.forename: Zhanhong
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Hunan, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Entomology, University of Kentucky, Lexington, KY, USA.
authors.forename: Xuguo
authors.initials: X
authors.lastname: Zhou
mesh_terms: D000818:Animals; D061046:Biological Control Agents; D001517:Coleoptera / Q000235:genetics*; D019295:Computational Biology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D017344:Genes, Insect; D020871:RNA Stability; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D061046:Biological Control Agents