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Validation of reference genes for gene expression studies in Aphis glycines (Hemiptera: Aphididae)

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Quantitative real-time PCR (qRT-PCR) is a common tool for quantifying mRNA transcripts. To normalize results, a reference gene is mandatory. Aphis glycines is a significant soybean pest, yet gene expression and functional genomics studies are hindered by a lack of stable reference genes. We evaluated 7 potential reference genes (SDFA, succinate dehydrogenase flavoprotein subunit a, EF1a, elongation factor-1 alpha, HEL, Helicase, GAPDH, glyceraldehyde-3 phosphate dehydrogenase, RPS9, ribosomal protein S9, TBP, TATA-box binding protein, and UBQ, ubiquitin conjugating protein) to determine the most efficient reference genes that have stable expression among tissues, developmental stages and aphids fed on susceptible and host-plant resistant plants. Stability was determined using GeNorm and NormFinder. Results revealed high stability of TBP compared to the other reference genes profiled across all samples. Stable expression was shown with RPS9 among aphids on susceptible and resistant plants and EF1a tissues and developmental stages. Therefore, we recommend the gene TBP as a suitable reference gene for normalization of A. glycines gene expression studies. Additionally, RPS9 may be used for host-plant resistance experiments and EF1a could be considered for testing differential expression across tissues or developmental stages. These results will enable a more accurate normalization of qRT-PCR data in A. glycines.
Raman Bansal , Preveen Mamidala , M. A. Rouf Mian , Omprakash Mittapalli , Andy P. Michel
Aphis glycines , Hemiptera , binding proteins , developmental stages , gene expression , gene expression regulation , genes , glyceraldehyde-3-phosphate dehydrogenase , insect pests , messenger RNA , quantitative polymerase chain reaction , ribosomal proteins , soybeans , succinate dehydrogenase , transcription factors
Journal of economic entomology 2012 v.105 no.4
Journal Articles, USDA Authors, Peer-Reviewed
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