EAPSI: Innovative Molecular Methods for Low Cost DNA Barcoding of Tropical Arthropods

Meierotto Sarah L, Lexington KY

Investigators

Abstract

Preservation of biodiversity is essential to an ecosystem?s ability to withstand disturbances like climate change and to provide services to human society. Traditional methods of measuring insect and other arthropod diversity are time consuming and expensive which impedes creation of effective conservation policies. In collaboration with Dr. Rudolf Meier at the National University of Singapore, this project will improve and share the knowledge of new techniques developed in the Meier lab for generating DNA barcodes for insect identification. DNA barcodes are a specific short sequence of DNA which can be used to identify an organism, much like how the barcode on the back of a cereal box is scanned at a grocery store. Paired with the researcher?s larger thesis project, this work will improve biodiversity evaluation and monitoring methods while uncovering relationships in arthropod forest communities. This project will use new direct PCR protocols and next-generation sequencing methods to DNA barcode thousands of insect specimens. A biodiversity assessment and comparison between the forests of Kentucky (United States) and French Guiana using metabarcoding is underway. Groups of interest, including parasitoids and caterpillars, as well as representatives of all arthropod orders present will be removed from the metabarcoded samples for use as voucher specimens. To evaluate the metabarcoding process and discover parasitoid-host relationships, DNA barcodes linked to these specimens will be generated using a low cost method developed by Rudolf Meier's lab. The proportion of voucher specimen sequences found within the metabarcoding sequence datasets will allow for the adjustment of diversity estimates to account for biases in the sequencing process. Direct PCR and next-generation sequencing (NGS) methods can produce barcodes at less than $1 per specimen but require training and experience to perform successfully and consistently. Dr. Meier?s protocols were designed for use with Diptera and will be optimized for use with hymenopteran parasitoids. The data generated during this project will be used to compare tropical and temperate arthropod biodiversity, to compare morphological and molecular identification techniques, and discover new parasitoid-host relationships. This award under the East Asia and Pacific Summer Institutes program supports summer research by a U.S. graduate student and is jointly funded by NSF and the National Research Foundation of Singapore.