Stable Reference Gene Selection for RT-qPCR Analysis in Nonviruliferous and Viruliferous Frankliniella occidentalis

• Published in: PloS one 2015-08, Vol.10 (8), p.e0135207-e0135207
• Main Authors: Yang, Chunxiao, Li, Hui, Pan, Huipeng, Ma, Yabin, Zhang, Deyong, Liu, Yong, Zhang, Zhanhong, Zheng, Changying, Chu, Dong
• Format: Article
• Language: English
• Subjects: Agronomy; Algorithms; Animals; Biological activity; Chaperonin 60 - genetics; Computational Biology - methods; DNA polymerases; Elongation; Flowers & plants; Flowers - parasitology; Frankliniella occidentalis; Gene expression; Gene Expression Profiling - methods; Genes; Genes, Insect - genetics; Genomics; Heat; Heat shock proteins; Hemiptera; Host-Pathogen Interactions; Hsp60 protein; HSP70 Heat-Shock Proteins - genetics; Hsp70 protein; Hsp90 protein; Insect Proteins - genetics; Introduced species; Invasive species; Methods; Molecular chains; Molecular modelling; Peptide Elongation Factor 1 - genetics; Polymerase chain reaction; Reference Standards; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction - methods; Reverse Transcriptase Polymerase Chain Reaction - standards; Ribosomal Proteins - genetics; RNA-directed DNA polymerase; Software; Studies; Thripidae; Thysanoptera; Thysanoptera - genetics; Thysanoptera - virology; Tomatoes; Tospovirus - physiology; Viruses; Wilt
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0135207

Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is a reliable technique for measuring and evaluating gene expression during variable biological processes. To facilitate gene expression studies, normalization of genes of interest relative to stable reference genes is crucial. The western flower thrips Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), the main vector of tomato spotted wilt virus (TSWV), is a destructive invasive species. In this study, the expression profiles of 11 candidate reference genes from nonviruliferous and viruliferous F. occidentalis were investigated. Five distinct algorithms, geNorm, NormFinder, BestKeeper, the ΔCt method, and RefFinder, were used to determine the performance of these genes. geNorm, NormFinder, BestKeeper, and RefFinder identified heat shock protein 70 (HSP70), heat shock protein 60 (HSP60), elongation factor 1 α, and ribosomal protein l32 (RPL32) as the most stable reference genes, and the ΔCt method identified HSP60, HSP70, RPL32, and heat shock protein 90 as the most stable reference genes. Additionally, two reference genes were sufficient for reliable normalization in nonviruliferous and viruliferous F. occidentalis. This work provides a foundation for investigating the molecular mechanisms of TSWV and F. occidentalis interactions.