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

Authors

Chunxiao Yang, Institute of Plant Protection, China
Hui Li, Qingdao Agricultural University, China
Huipeng Pan, University of KentuckyFollow
Yabin Ma, Qingdao Agricultural University, China
Deyong Zhang, Institute of Plant Protection, China
Yong Liu, Institute of Plant Protection, China
Zhanhong Zhang, Hunan Vegetable Institute, China
Changying Zheng, Qingdao Agricultural University, China
Dong Chu, Qingdao Agricultural University, China

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.

Document Type

Article

Publication Date

8-5-2015

Notes/Citation Information

Published in PLOS One, v. 10, no. 8, article e0135207, p. 1-11.

© 2015 Yang et al.

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Digital Object Identifier (DOI)

http://dx.doi.org/10.1371/journal.pone.0135207

Funding Information

This work was supported by the Special Fund for Agroscience Research in the Public Interest (201303028), the Shandong Modern Agricultural Technology & Industry System (SDAIT-02-021-11), the Science and Technology Development Planning Program of Qingdao (13-1-3-108-nsh), and the Taishan Mountain Scholar Constructive Engineering Foundation of Shandong, China. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Repository Citation

Yang, Chunxiao; Li, Hui; Pan, Huipeng; Ma, Yabin; Zhang, Deyong; Liu, Yong; Zhang, Zhanhong; Zheng, Changying; and Chu, Dong, "Stable Reference Gene Selection for RT-qPCR Analysis in Nonviruliferous and Viruliferous Frankliniella occidentalis" (2015). Entomology Faculty Publications. 91.
https://uknowledge.uky.edu/entomology_facpub/91