New EPIC nuclear DNA sequence markers to improve the resolution of phylogeographic studies of coenagrionids and other odonates

Abstract

While phylogeographic data provide valuable information to inform conservation plans, there are comparatively few Odonata phylogeographic studies. This lack of research is partially due to a lack of independent DNA markers with appropriate levels of polymorphism that PCR-amplify in a range of species. We followed an exon-primed, intron-crossing (EPIC) PCR strategy to develop five new, polymorphic nuclear DNA sequence loci (six distinct DNA fragments) for the southern damselfly Coenagrion mercuriale. These markers were: cell division cycle 5 protein (CDC5), arginine methyltransferase (PRMT), acetylglucosaminyl-transferase (AgT), myosin light chain (MLC) and phosphoglucose isomerase (PGI). Between three and five of these new markers could be PCR-amplified in five other species from the genus Coenagrion; one locus (PRMT) can be used in 26 other species of odonates that we examined, including three species of Anisoptera belonging to the genus Onychogomphus. These new nuclear genetic markers will be useful for phylogeographic studies in a range of odonate species, but also for phylogenetic studies, providing a particularly useful complement to the existing mitochondrial and nuclear loci.

Keywords:

• Coenagrionidae
• Coenagrion mercuriale
• dragonfly
• genetic diversity
• introns
• multilocus approach
• non-model organism
• nuclear DNA loci
• Odonata
• phylogeny

Acknowledgements

The authors are grateful to their colleagues who sent samples for the genetic analysis: Jean-Pierre Boudot, Sonke Hardersen, Elisa Riservato and Janne Swaegers. SF would particularly like to acknowledge Bruno Carvalho, Arie van der Meijden and Diana Pedroso for productive brainstorming discussions during the early stages of this work. Sample collection benefited from fieldwork funded by the project “Coenagrion mercuriale: distribución, diversidad genética y estado de conservación” of Instituto de Estudios Ceutíes (Ceuta – Spain). SF is supported by Fundação para a Ciência e Tecnologia (SFRH/BD/65038/2009). YTC was supported by a project “Ciencia y Conciencia” funded by Universidad de Oriente, Cuba (project code 9617) and a WDA Conservation and Research Grant. ACR and MOLC were supported by grants from the Spanish Ministry with competence in Science, which included FEDER funds (CGL2010-11959-E and CGL2011-22629).

Disclosure statement

The authors declare that no financial interest or benefit exists arising from the direct applications of their research.