Title: Isolation of Diapause-Regulated Genes from the Flesh Fly, Sarcophaga crassipalpis by Suppressive Subtractive Hybridization Authors
|Robich, Rebecca -|
|Denlinger, David -|
Submitted to: Journal of Insect Physiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 9, 2009
Publication Date: May 1, 2010
Repository URL: http://parking.nal.usda.gov/shortterm/20083_2010_J_Insect_Physiology_56_603-609.pdf
Citation: Rinehart, J.P., Robich, R.M., Denlinger, D.L. 2010. Isolation of Diapause-Regulated Genes from the Flesh Fly, Sarcophaga crassipalpis by Suppressive Subtractive Hybridization. Journal of Insect Physiology. 56(6):603-609. Interpretive Summary: Winter is a major problem for insects. To be successful in climates with distinct seasons, insects must grow and reproduce only for a few months and then survive the remainder of the year without feeding, while dealing with the stresses that winter brings to them. Most insects survive winter by entering diapause, a condition that is characterized by an arrest in development, decreased metabolism, and increased stress tolerance. In this study, we used molecular techniques to determine what genes are present at higher levels in the flesh fly Sarcophaga crassipalpis. The identification of these genes has given us an increased understanding of how diapause works in this insect species and allows us to compare the molecular basis of diapause in diverse species.
Technical Abstract: Subtractive suppressive hybridization (SSH) was used to characterize the diapause transcriptome of the flesh fly Sarcophaga crassipalpis. Through these efforts, we isolated 97 unique clones which were used as probes in northern hybridization to assess their expression during diapause. Of these, 17 were confirmed to be diapause upregulated and 1 was diapause downregulated, while 12 were shown to be unaffected by diapause in this species. The diapause upregulated genes fall into several broad categories including heat shock proteins, heavy metal responsive genes, neuropeptides, structural genes, regulatory elements, and several genes of unknown function. In combination with other large-scale analyses of gene expression during diapause, this study assists in the characterization of the S. crassipalpis diapause transcriptome, and begins to identify common elements involved in diapause across diverse taxa.