Abstract
Estimating the post mortem interval (PMImin) by age determination of blow fly larvae has been well-established for moderate temperatures. Low-temperature developmental data is only available sparsely and usually does not take overwintering strategies into account. The blow fly Calliphora vicina hibernates by diapausing in the third larval stage extending the duration of this developmental stage up to several weeks or even months. As the diagnosis of the diapause status is not possible by morphological characteristics, PMImin estimations might be biased during the cold season if only based on age determination of third instar larvae of C. vicina. Molecular markers were searched for which allows one to identify diapause in larvae. Expression analysis of 19 genes was performed in diapausing and non-diapausing larvae. Three genes encoding for heat shock proteins (hsp23, hsp24 and hsp70) were found to be up-regulated distinctly in diapausing larvae and at 1 day in non-diapausing larvae. If several larvae are subjected to an analysis, a high variance in the expression level of the gene encoding for the anterior fat body protein is a further marker for diapause. The present study proves the potential use of gene expression analysis as a suitable diagnosis tool for diapause in C. vicina.
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Acknowledgments
We thank Friederike Häuser and Petra Böhme for their support in PCR primer design. David Denlinger kindly provided primer sequences of PCNA. Natalie Filmann (Institute of Biostatistics and Mathematical Modelling, JWG-University, Frankfurt a.M.) is acknowledged for checking the statistical analysis and Kirsty Griffin for correcting the language of the manuscript. Finally, we thank two anonymous reviewers for their helpful and critical comments improving the quality of the manuscript.
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Fremdt, H., Amendt, J. & Zehner, R. Diapause-specific gene expression in Calliphora vicina (Diptera: Calliphoridae)—a useful diagnostic tool for forensic entomology. Int J Legal Med 128, 1001–1011 (2014). https://doi.org/10.1007/s00414-013-0920-x
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DOI: https://doi.org/10.1007/s00414-013-0920-x