Temperature and Development Impacts on Housekeeping Gene Expression in Cowpea Aphid, Aphis craccivora (Hemiptera: Aphidiae)

Chunxiao Yang; Huipeng Pan; Yong Liu; Xuguo Zhou

doi:10.1371/journal.pone.0130593

Temperature and Development Impacts on Housekeeping Gene Expression in Cowpea Aphid, Aphis craccivora (Hemiptera: Aphidiae)

PLoS One. 2015 Jun 19;10(6):e0130593. doi: 10.1371/journal.pone.0130593. eCollection 2015.

Authors

Chunxiao Yang¹, Huipeng Pan², Yong Liu³, Xuguo Zhou²

Affiliations

¹ Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Changsha, Hunan, China; Department of Entomology, University of Kentucky, Lexington, KY, United States of America.
² Department of Entomology, University of Kentucky, Lexington, KY, United States of America.
³ Hunan Academy of Agricultural Sciences, Institute of Plant Protection, Changsha, Hunan, China.

Abstract

Quantitative real-time PCR (qRT-PCR) is a powerful technique to quantify gene expression. To standardize gene expression studies and obtain more accurate qRT-PCR analysis, normalization relative to consistently expressed housekeeping genes (HKGs) is required. In this study, ten candidate HKGs including elongation factor 1 α (EF1A), ribosomal protein L11 (RPL11), ribosomal protein L14 (RPL14), ribosomal protein S8 (RPS8), ribosomal protein S23 (RPS23), NADH-ubiquinone oxidoreductase (NADH), vacuolar-type H+-ATPase (ATPase), heat shock protein 70 (HSP70), 18S ribosomal RNA (18S), and 12S ribosomal RNA (12S) from the cowpea aphid, Aphis craccivora Koch were selected. Four algorithms, geNorm, Normfinder, BestKeeper, and the ΔCt method were employed to evaluate the expression profiles of these HKGs as endogenous controls across different developmental stages and temperature regimes. Based on RefFinder, which integrates all four analytical algorithms to compare and rank the candidate HKGs, RPS8, RPL14, and RPL11 were the three most stable HKGs across different developmental stages and temperature conditions. This study is the first step to establish a standardized qRT-PCR analysis in A. craccivora following the MIQE guideline. Results from this study lay a foundation for the genomics and functional genomics research in this sap-sucking insect pest with substantial economic impact.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Aphids / genetics*
Computational Biology / methods
Gene Expression Profiling
Gene Expression Regulation*
Genes, Essential*
Genes, Insect*
Temperature*
Transcriptome

Grants and funding

This research was supported by a start-up fund from the University of Kentucky to XGZ, a grant from USDA BRAG grant (Award Agreement No.: 3048108827) to XGZ, and a Special Fund for Agroscience Research in the Public Interest (Award Agreement No.: 201303028) to YL. The granting agencies have no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.