Suitable reference gene selection for different strains and developmental stages of the carmine spider mite, Tetranychus cinnabarinus , using quantitative real-time PCR
1Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing
400716, P. R. China
2Institute for Control of Agrochemicals, Shandong Province, Jinan 250100, P. R. China
Abstract
Reference genes are used as internal controls in gene expression studies, but their expression levels vary according to tissue types and experimental treatments. Quantitative real-time PCR (qPCR) is the most sensitive technique for transcript quantification provided that gene transcription patterns are normalized to an evaluated reference gene. In this study, the suitability of eight commonly used genes (β-actin, 5.8SrRNA, α-TUB, GAPDH, RPL13a, RPS18, TBP, SDHA) were cloned and investigated to find the most stable candidates for normalizing real-time PCR data generated from the four different strains (abamectin-resistant, fenpropathrin-resistant, omethoate-resistant, and susceptible strains) and different developmental stages (eggs, protonymphs, nymphs, and adults) of carmine spider mite, Tetranychus cinnabarinus (Boisduval) (Acarina: Tetranychidae). The stability of gene expression was assessed using two different analysis programs, geNorm and NormFinder. Using these analyses, RPS18 and 5.8SrRNA had the most stable expression regardless of the four different strains, whereas RPS18 and α-TUB were expressed most stably in different developmental stages.
Keywords: normalization
Abbreviations: AbR, abamectin resistant; FeR, fenpropathrin resistant; OmR, omethoate resistant; qPCR, Quantitative real-time
PCR
Correspondence: asw1984@163.com, bhelinok@tom.com, cluwencai201@yahoo.com.cn, dpiaopiaogg@yahoo.cn
Received: 24 May 2009 | Accepted: 24 September 2009 | Published: 16 December 2010
Copyright: This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.
ISSN: 1536-2442 | Volume 10, Number 208
Sun W, Jin Y, He L, Lu W-C, Li M. 2010. Suitable reference gene selection for different strains and developmental stages of the carmine spider mite, Tetranychus cinnabarinus, using quantitative real-time PCR. Journal of Insect Science 10:208 available online: insectscience.org/10.208
 Figure 1 |
 Figure 2 |