Determination of suitable insect part for non‐lethal DNA sampling: case study of DNA quality and regeneration capability of dragonflies

• Published in: Insect conservation and diversity Vol. 13; no. 4; pp. 319 - 327
• Main Authors: Ožana, Stanislav, Pyszko, Petr, Dolný, Aleš
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.07.2020; Wiley Subscription Services, Inc
• Subjects: Conservation genetics; Deoxyribonucleic acid; DNA; DNA fingerprints; DNA isolation method; DNA sampling; Endangered species; Genetic testing; insects; Larvae; Leg; non‐invasive; non‐lethal; Protected species; regeneration; Sampling; Wings
• ISSN: 1752-458X; 1752-4598
• DOI: 10.1111/icad.12400

Genetics has been widely used in insect ecology and conservation. To minimise the effect of DNA sampling on organisms as much as possible, it would be ideal to use non‐invasive or non‐lethal DNA sources. Therefore, it is also very important to determine the responses of organisms to DNA sampling. In this study, the quality and quantity of genomic DNA samples from three types of insect tissues (exuvia, mid‐legs and wings) were evaluated. As model organisms, we used two dragonfly species of different sizes (Leucorrhinia dubia, Anax imperator). We also tested the regenerative ability of dragonfly larvae as a repair mechanism after mid‐leg cut‐off, with respect to factors such as size and quantity of diet. We found that DNA of sufficient quality for analyses was obtained from all tested tissues. Nonetheless, isolates from exuviae were conclusively less useful for sequencing than those from the mid‐legs and wings. The highest quantity of DNA was obtained from the mid‐legs. The survival of larvae is not affected by removing the legs, which can usually regenerate. All the tested tissues could be a source of adequate DNA; however, we concluded that primarily the legs should be used because they provided the best DNA samples in terms of quantity and quality of DNA. Furthermore, their exploitation would not affect individuals seriously if young larvae with sufficient time (at least 6 months) for regeneration are sampled. The exuviae should be used for absolutely non‐invasive studies involving endangered or protected species. Our research has focused on determining the optimal source of DNA that would allow the acquisition of high‐quality DNA samples with minimal effects on individuals. Three types of insect tissues (exuvia, mid‐legs and wings) were evaluated. DNA of sufficient quality and quantity for analyses was obtained from all tested tissues. Source material such as the dragonfly larval leg is ideal, because it provides a considerable amount of DNA, and there is an excellent possibility for regeneration of the amputated legs.