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 Δ Ct 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.
| Original language | English |
|---|---|
| Article number | 18201 |
| Journal | Scientific Reports |
| Volume | 5 |
| DOIs | |
| State | Published - Dec 10 2015 |
Bibliographical note
Funding Information:Authors are grateful to Drs. Hui Li and Zhen Li for their assistance with the data analysis. This is publication No. 15-08-044 of the Kentucky Agricultural Experiment Station and is published with the approval of the Director. This work is supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture, the Biotechnology Risk Assessment Grant under Award Agreement No.: 3048108827, a Special Fund for Agroscience Research in the Public Interest from China under Award Agreement No.: 201303028, and Hunan Provincial Natural Science Foundation of China No.: 14JJ6058. The granting agencies have no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
ASJC Scopus subject areas
- General