Identification of Auchenorrhyncha Nymphs Using DNA Barcoding and Phylogenetic Analysis of the Most Common Genera Collected in Olive Fields

Due to the potential role of Auchenorrhyncha in the transmission of the bacterium Xylella fastidiosa in a wide variety of cultivations, during recent years in Europe, many studies have focused on species composition, abundance and seasonal appearance of Auchenorrhyncha. However, females and nymphs are difficult to identify, as species-level identification relies primarily on male genitalia morphology. Sampling was conducted over four years in olive fields in Lesvos Island, in the Northeast Aegean, Greece, using sweep nets and Malaise traps. Both adults and nymphs were collected, with males identified to species level, while females and nymphs were separated on different morphotypes. Representatives from each morphotype and identified adults were sequenced using the mitochondrial cytochrome oxidase subunit I (COI) gene. Using a classical morphological approach, 58 species were identified to species level, and using DNA barcoding, nymph morphotypes and females were successfully identified within the families Cicadellidae, Aphrophoridae, Delphacidae and Issidae. A phylogenetic tree was generated, clustering nymphs together with the corresponding adults. Our results demonstrate the utility of combining morphological and molecular methods for accurate species identification and highlight the importance of enriching online databases with additional species records.