Exploring new nuclear markers for phylogeographic study in beetles

The recent growth of the fields of phylogenetics, phylogeography, population genetics, and conservation genetics creates a great demand for new nuclear markers that provide insights into the evolution of the beetle genome. Mitochondrial markers are traditionally used for these types of studies but they reveal only a small part of the evolutionary history. In our research we tested two protein-coding genes Phosphoenolpyruvate carboxykinase (PEPCK) and rudimentary (CAD) along with four novel introns synaptojanin, guftagu, chitin synthase 1 (krotzkopf verkehrt in Apis), and 66533157(need name). We selected three different individuals from each of the five beetle species Geodercodes latipennis, Nyctoporis carinata, Zarhipis integripennis, Phyllophaga fraterna, and Calathus ruficollis and amplified each individual’s genetic genome with the six targeted protein- coding and non-coding markers. CAD and PEPCK yielded the best amplifying results across all the diversity of beetle species. We reached optimum results with CAD by using a nested PCR technique. As for PEPCK and the novel introns we had to use a variety of different PCR methods ranging from touch-up amplifications, to gel plugs and nested PCR’s. The nuclear DNA sequence data can be used to create more species specific primer sets along with the ability to compare beetles within species and among species.