A new primer pair for barcoding of bees (Hymenoptera: Anthophila) without amplifying the orthologous coxA gene of Wolbachia bacteria

BMC Res Notes. 2021 Nov 25;14(1):427. doi: 10.1186/s13104-021-05845-9.

Abstract

Objectives: DNA barcoding became an effective method for the identification and monitoring of bees. However, standard primer pairs used for barcoding often result in (co-) amplification of bacterial endosymbionts of the genus Wolbachia, which are widespread among bee species. Here we designed a new primer pair and compared it with the performance of the standard Folmer-primers for a small sample set of bees representing the main taxonomic groups of bees.

Results: The newly designed primer pair (BeeCox1F1/BeeCox1R2) outperformed the standard barcoding primer (LCO1490/HCO2198). By generating barcodes for a small test set of bees we found that the new primer pair produced high-quality sequences in all cases for unambiguous species identification using BOLD. Conversely, the standard barcoding primers often co-amplified the homologous Wolbachia gene and resulted in mixed chromatogram signals. These sequences showed high similarity with the bacterial endosymbiont instead of the host.

Keywords: Apoidea; Biomonitoring; DNA barcoding; Misamplification.

MeSH terms

  • Animals
  • Bacteria
  • Bees / genetics
  • DNA Barcoding, Taxonomic
  • Hip
  • Hymenoptera*
  • Wolbachia* / genetics