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Insects 2018, 9(1), 18; https://doi.org/10.3390/insects9010018

Gene Expression Profile Analysis is Directly Affected by the Selected Reference Gene: The Case of Leaf-Cutting Atta Sexdens

1
Central Laboratory of Molecular Biology, Federal University of Lavras (UFLA), Lavras, MG 37200-000, Brazil
2
Entomology Department, Federal University of Lavras (UFLA), Lavras, MG 37200-000, Brazil
3
Chemistry Department, Federal University of Lavras (UFLA), Lavras, MG 37200-000, Brazil
*
Author to whom correspondence should be addressed.
Received: 8 January 2018 / Revised: 31 January 2018 / Accepted: 2 February 2018 / Published: 8 February 2018
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Abstract

Although several ant species are important targets for the development of molecular control strategies, only a few studies focus on identifying and validating reference genes for quantitative reverse transcription polymerase chain reaction (RT-qPCR) data normalization. We provide here an extensive study to identify and validate suitable reference genes for gene expression analysis in the ant Atta sexdens, a threatening agricultural pest in South America. The optimal number of reference genes varies according to each sample and the result generated by RefFinder differed about which is the most suitable reference gene. Results suggest that the RPS16, NADH and SDHB genes were the best reference genes in the sample pool according to stability values. The SNF7 gene expression pattern was stable in all evaluated sample set. In contrast, when using less stable reference genes for normalization a large variability in SNF7 gene expression was recorded. There is no universal reference gene suitable for all conditions under analysis, since these genes can also participate in different cellular functions, thus requiring a systematic validation of possible reference genes for each specific condition. The choice of reference genes on SNF7 gene normalization confirmed that unstable reference genes might drastically change the expression profile analysis of target candidate genes. View Full-Text
Keywords: ant; endogenous controls; normalization; quantitative RT-PCR; RefFinder; SNF7; validation ant; endogenous controls; normalization; quantitative RT-PCR; RefFinder; SNF7; validation
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Livramento, K.G.; Freitas, N.C.; Máximo, W.P.F.; Zanetti, R.; Paiva, L.V. Gene Expression Profile Analysis is Directly Affected by the Selected Reference Gene: The Case of Leaf-Cutting Atta Sexdens. Insects 2018, 9, 18.

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