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Nondestructive sampling of insect DNA from defensive secretion.
H M Donald1, C W Wood, K M Benowitz
1Mountain Lake Biological Station, Department of Biology, University of Virginia, Charlottesville, VA 22904, USA.
Molecular Ecology Resources
|May 18, 2012
Summary
This study introduces a nondestructive DNA collection method using insect defensive secretions. This technique successfully obtained DNA for genetic analysis without harming the insects, proving effective for population studies.
Area of Science:
- Ecology and Evolutionary Biology
- Molecular Genetics
- Entomology
Background:
- Nondestructive DNA sampling is crucial for genetic analyses like diversity and fitness estimation without harming populations.
- Existing methods (frass, exuviae, clippings) are not always feasible, especially for insects sensitive to disturbance or tissue removal.
Purpose of the Study:
- To evaluate the impact and efficacy of a novel nondestructive technique for DNA extraction from insect defensive secretions.
- To assess the feasibility of using haemolymph from defensive secretions for microsatellite DNA amplification.
Main Methods:
- Haemolymph was collected from the defensive secretion of Bolitotherus cornutus (forked fungus beetle) using a capillary tube.
- A laboratory experiment assessed the sampling method's impact on insect mortality, reproductive success, and gland expression.
- Field sampling involved collecting haemolymph from 187 individuals for genotyping at nine microsatellite loci.
Main Results:
- The sampling methodology showed no negative impact on insect mortality, reproductive success, or defensive gland expression.
- DNA was successfully extracted and genotyped from 95.7% of field-collected haemolymph samples across nine microsatellite loci.
- Haemolymph-rich defensive secretions serve as a viable source of DNA for genetic analyses.
Conclusions:
- Extracting haemolymph from defensive secretions is a safe and effective nondestructive DNA sampling technique for insects.
- This method supports genetic studies, including population genetics and conservation, by enabling DNA collection without adverse effects on individuals.
