Abstract
Studies on the expression of genes in different contexts are essential to our understanding of the functioning of organisms and their adaptations to the environment. Gene expression studies require steps of normalization, which are done using the stable expression pattern of reference genes. For many different eusocial bees reference genes have been discovered, but not for the primitively eusocial euglossine bees.We used available genomic resources of euglossine species and the gene information of Apis melliferato develop a set of reference genes for the primitive eusocial bee Euglossaviridissima. We tested nine genes in distinct developmental stages three different algorithms to infer the stability of gene expression. The Tata binding protein(Tbp) and 14-3-3epsilon were the most stable genes across all different stages. The strongest deviation in gene expression pattern occurred in pupae, which require a different set of genes for normalizing gene expression.
Highlights
Candidates for suitable reference genes have been identified in several species of the Apidae, e.g. honeybees (Lourenço et al, 2008), bumble bees (Horňáková et al, 2009; Niu et al, 2014), and stingless bees (Dallacqua et al, 2007)
Reference genes have not been described for orchid bees, transcriptome and genome data are available for a few of these species (Woodard et al, 2011; Kapheim et al, 2015; Brand et al, 2017)
We identified a set of candidate reference genes for the orchid bee Euglossa viridissima Friese based on the transcriptome reference for Euglossa cordata (Linnaeus) (Woodard et al, 2011) and annotated genes of Apis mellifera Linnaeus (Elsik et al, 2015)
Summary
Candidates for suitable reference genes have been identified in several species of the Apidae, e.g. honeybees (Lourenço et al, 2008), bumble bees (Horňáková et al, 2009; Niu et al, 2014), and stingless bees (Dallacqua et al, 2007). We tested for stability of gene expression in individuals across different developmental stages (including adults). We choose nine candidate reference genes, namely Ribosomal protein 28S (28S), Actin, Arginine Kinase (ArgK), Elongation Factor 1a (EF-1a), 14-3-3 epsilon, Glutathione S-transferase-1 (Gst1), Inositol 1,4,5-triphosphate receptors (Itpr), Ribosomal protein S18 (Rps18) and TATA-binding protein (TBP).
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