title: Stable Reference Gene Selection for RT-qPCR Analysis in Nonviruliferous and Viruliferous Frankliniella occidentalis.
abstract: 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.
authors.affiliation: Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Changsha, Hunan, China.
authors.forename: Chunxiao
authors.initials: C
authors.lastname: Yang
authors.affiliation: College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Entomology, University of Kentucky, Lexington, Kentucky, United States of America.
authors.forename: Huipeng
authors.initials: H
authors.lastname: Pan
authors.affiliation: College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, China.
authors.forename: Yabin
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Changsha, Hunan, China.
authors.forename: Deyong
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Changsha, Hunan, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Hunan Academy of Agricultural Sciences, Hunan Vegetable Institute, Changsha, Hunan, China.
authors.forename: Zhanhong
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, China.
authors.forename: Changying
authors.initials: C
authors.lastname: Zheng
authors.affiliation: College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Chu
mesh_terms: D000465:Algorithms; D000818:Animals; D018834:Chaperonin 60 / Q000235:genetics; D019295:Computational Biology / Q000379:methods; D035264:Flowers / Q000469:parasitology; D020869:Gene Expression Profiling / Q000379:methods*; D017344:Genes, Insect / Q000235:genetics*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D054884:Host-Pathogen Interactions; D019476:Insect Proteins / Q000235:genetics; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012269:Ribosomal Proteins / Q000235:genetics; D012984:Software; D061149:Thysanoptera / Q000235:genetics* / Q000821:virology; D017865:Tospovirus / Q000502:physiology
chemical_list: D018834:Chaperonin 60; D018840:HSP70 Heat-Shock Proteins; D019476:Insect Proteins; D020648:Peptide Elongation Factor 1; D012269:Ribosomal Proteins; C041806:ribosomal protein L32