Alternative preservatives of insect DNA for citizen science and other low-cost applications
Sedonia Steininger A B , Caroline Storer B , Jiri Hulcr A B and Andrea Lucky A CA Entomology and Nematology Department, University of Florida-IFAS, Steinmetz Hall, 1881 Natural Area Drive, Gainesville, FL 32611-0620, USA.
B School of Forest Resources and Conservation, University of Florida-IFAS, 317 Newins-Ziegler Hall, Gainesville, FL 32611-0410, USA.
C Corresponding author. Email: alucky@ufl.edu
Invertebrate Systematics 29(5) 468-472 https://doi.org/10.1071/IS15003
Submitted: 29 January 2015 Accepted: 4 September 2015 Published: 30 October 2015
Abstract
Prevention of DNA degradation is essential to conducting molecular analyses of field-captured specimens. This is especially important for projects that incorporate participation of non-specialists in research, such as agency monitoring of pests, or citizen science, where standard methods of preservation may be inaccessible. We examined efficacy of three common alternative products as a substitute for 95% ethanol or pure propylene glycol in preserving DNA: alcohol-based hand sanitiser and propylene and ethylene glycol-based automobile antifreeze. We subjected Xylosandrus compactus ambrosia beetles (Coleoptera : Curculionidae : Scolytinae) to each preservative for two or seven days under direct outdoor exposure and assessed relative quantity of intact DNA by performing real-time polymerase chain reaction amplification of a single-copy nuclear marker. Amplification was observed in all treatments and electrophoresis of the amplified product showed clear bands of the appropriate weight. Successful amplification of the target gene was verified by sequencing the amplified control. No statistically significant differences were found between the cycle threshold values of any treatment. Our results suggest that alcohol-based hand sanitiser and automobile antifreeze can successfully preserve DNA for short-term storage and serve as effective substitutes for laboratory-grade preservatives in citizen science projects, large-scale trapping projects or by professionals.
Additional keywords: biodiversity, Coleoptera, genetics.
References
Altschul, S. F., Gish, W., Miller, W., Myers, E. W., and Lipman, D. J. (1990). Basic local alignment search tool. Journal of Molecular Biology 215, 403–410.| 1:CAS:528:DyaK3MXitVGmsA%3D%3D&md5=94d19a2ce5b001071ce80235322d981eCAS | 2231712PubMed |
Ball, S. L., and Armstrong, K. F. (2006). DNA barcodes for insect pest identification: a test case with tussock moths (Lepidoptera: Lymantriidae). Canadian Journal of Forest Research 36, 337–350.
| DNA barcodes for insect pest identification: a test case with tussock moths (Lepidoptera: Lymantriidae).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XkvF2lt78%3D&md5=7229adcd00caf56da501b55b02c9e3deCAS |
Bass, C., Nikou, D., Vontas, J., Donnelly, M. J., Williamson, M. S., and Field, L. M. (2010). The vector population monitoring tool (VPMT): high-throughput DNA-based diagnostics for the monitoring of mosquito vector populations. Malaria Research and Treatment 2010, 190434.
| 22347668PubMed |
Clark, W. H., and Blom, P. E. (1992). An efficient and inexpensive pitfall trap system. Entomological News 103, 55–59.
Dole, S. A., Jordal, B. H., and Cognato, A. I. (2010). Polyphyly of Xylosandrus Reitter inferred from nuclear and mitochondrial genes (Coleoptera: Curculionidae: Scolytinae). Molecular Phylogenetics and Evolution 54, 773–782.
| Polyphyly of Xylosandrus Reitter inferred from nuclear and mitochondrial genes (Coleoptera: Curculionidae: Scolytinae).Crossref | GoogleScholarGoogle Scholar | 19925873PubMed |
Ferro, M. L., and Park, J. (2013). Effect of propylene glycol concentration on mid-term DNA preservation of Coleoptera. Coleopterists Bulletin 67, 581–586.
| Effect of propylene glycol concentration on mid-term DNA preservation of Coleoptera.Crossref | GoogleScholarGoogle Scholar |
Fisher, R. A. (1925). ‘Statistical Methods for Research Workers.’ (Oliver and Boyd: London.)
Fox, J. (2005). The R Commander: a basic statistics graphical user interface to R. Journal of Statistical Software 14, 1–42.
Frampton, M., Droege, S., Conrad, T., Prager, S., and Richards, M. H. (2008). Evaluation specimen preservatives for DNA analysis of bees. Journal of Hymenoptera Research 17, 195–200.
Hulcr, J., and Dunn, R. R. (2011). The sudden emergence of pathogenicity in insect-fungus symbioses threatens naive forest ecosystems. Proceedings of the Royal Society B 278, 2866–2873.
| 21752822PubMed |
King, J. R., and Porter, S. D. (2004). Recommendations on the use of alcohols for preservation of ant specimens (Hymenoptera, Formicidae). Insectes Sociaux 51, 197–202.
| Recommendations on the use of alcohols for preservation of ant specimens (Hymenoptera, Formicidae).Crossref | GoogleScholarGoogle Scholar |
Lucky, A., Savage, A. M., Nichols, L. M., Castracani, C., Shell, L., Grasso, D., Mori, A., and Dunn, R. R. (2014). Ecologists, educators, and writers collaborate with the public to assess backyard diversity in The School of Ants Project. Ecosphere 5, 78.
| Ecologists, educators, and writers collaborate with the public to assess backyard diversity in The School of Ants Project.Crossref | GoogleScholarGoogle Scholar |
Moreau, C. S., Wray, B. D., Czekanski-Moir, J. E., and Rubin, B. E. R. (2013). DNA preservation: a test of commonly used preservatives for insects. Invertebrate Systematics 27, 81–86.
| DNA preservation: a test of commonly used preservatives for insects.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXktVOgs78%3D&md5=5fad9c33791a20d02ca9e81e30937169CAS |
Quicke, D. L. J., Belshaw, R., and Lopez-Vaamonde, C. (1999). Preservation of hymenopteran specimens for subsequent molecular and morphological study. Zoologica Scripta 28, 261–267.
| Preservation of hymenopteran specimens for subsequent molecular and morphological study.Crossref | GoogleScholarGoogle Scholar |
R Core Team (2013). ‘R: A language and environment for statistical computing.’ (R Foundation for Statistical Computing: Vienna, Austria.) Available at http://www.R-project.org/ [Accessed May 2014].
Rabaglia, R. J., Duerr, D., Acciavatti, R., and Ragenovich, I. (2008). ‘Early Detection and Rapid Response for Non-native Bark and Ambrosia Beetles.’ US Department of Agriculture Forest Service, Forest Health Protection.
Schmidt, M. H., Clough, Y., Schulz, W., Westphalen, A., and Tscharntke, T. (2006). Capture efficiency and preservation attributes of different fluids in pitfall traps. The Journal of Arachnology 34, 159–162.
| Capture efficiency and preservation attributes of different fluids in pitfall traps.Crossref | GoogleScholarGoogle Scholar |
Stevens, M. M., Warren, G. N., Mo, J., and Schlipalius, D. I. (2011). Maintaining DNA quality in stored-grain beetles caught in Lindgren funnel traps. Journal of Stored Products Research 47, 69–75.
| Maintaining DNA quality in stored-grain beetles caught in Lindgren funnel traps.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXjtFSks7o%3D&md5=892a620f0d1759b94a9bb94820cdd403CAS |
Stoeckle, B. C., Dworschak, K., Gossner, M. M., and Kuehn, R. (2010). Influence of arthropod sampling solutions on insect genotyping reliability. Entomologia Experimentalis et Applicata 135, 217–223.
| Influence of arthropod sampling solutions on insect genotyping reliability.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXmslCit7k%3D&md5=c6215780dab960d29943bf663f40c13dCAS |
Thomas, D. B. (2008). Nontoxic antifreeze for insect traps. Entomological News 119, 361–365.
| Nontoxic antifreeze for insect traps.Crossref | GoogleScholarGoogle Scholar |
US Forest Service (2014). Invasive Species Program, Early Detection and Rapid Response (EDRR). Available at http://www.fs.fed.us/invasivespecies/earlydetection.shtml [Accessed September 2014].
Vink, C. J., Thomas, S. M., Paquin, P., Hayashi, C. Y., and Hedin, M. (2005). The effects of preservatives and temperature on arachnid DNA. Invertebrate Systematics 19, 99–104.
| The effects of preservatives and temperature on arachnid DNA.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXls1SlsL8%3D&md5=6ed6ee5b2fa1cbfcc7447b8c229a28d3CAS |
