Development of a SNP-based tool for the identification and discrimination of <i>Melolontha melolontha</i> and <i>Melolontha hippocastani</i>

Abstract The European ( Melolontha melolontha L.) and Forest M. hippocastani F.) cockchafer are widespread pests throughout Central Europe. Both species exhibit a 3–5-year life cycle occur in temporally shifted populations, which have been monitored documented for more than 100 years. Visual identification of adults larvae belonging to these morphologically similar requires expertise and, particularly the case larvae, is challenging equivocal. goal study was development an efficient fast molecular genetic tool discrimination . We established collection both from Switzerland, Austria Northern Italy 2016, 2017 2018. An approximately 1550 bp long fragment cytochrome c oxidase subunit 1 (CO1) mitochondrial gene amplified sequenced 13 beetles. Alignment new sequences with reference (NCBI GenBank BOLDSYSTEMS databases) subsequent phylogenetic analysis revealed consistent clustering two species. After species-specific single nucleotide polymorphisms (SNPs) CO1 alignment, we developed effective SNP based on ABI PRISM® SNaPshot™ Multiplex Kit rapid accurate larvae.