Tag: ITS2

Published paper: Improving mosquito barcoding

I have had the fortune to be involved in a study on the quality of reference material for mosquito barcoding for biodiversity studies. The study, which was led by Maurício Moraes Zenker at the Universidade Federal de São Carlos in Brazil, looked at the availability of public data for mosquitoes in online databases for two widely used DNA barcoding markers in Culicidae: the COI and ITS2 regions (1). Last week, this study was published in Scientific Reports.

The paper shows that around 30% of known species were covered for the COI gene in BOLD  and  GenBank, and 12% of species for ITS2 in GenBank. The Afrotropical, Australian and Oriental biogeographic regions had the lowest coverages, while the Nearctic, Palearctic and Oceanian regions had the highest. Countries with a higher diversity of mosquitoes tended to have lower coverage, which was surprisingly also the case for countries with higher numbers of medically important species. At the same time, countries with a higher number of endemic species tended to have a higher species coverage in the databases.

With this study, we would like to advocate for better curatorship of voucher specimens representing sequences in the databases. Also, an integrative taxonomic approach that combines various genetic markers with morphological analyses is important to allow a better use of DNA barcoding and metabarcoding in a diverse array of applications, including vector species detection and biodiversity monitoring.

Importantly, this work underscores how reliant DNA barcoding is on proper taxonomic foundations, including morphological characterisations. Molecular identification of species cannot happen in a vacuum! I would like to extend a big thanks for Maurício who invited me to take part in this study and who have done an excellent job putting it all together!


  • Moraes Zenker M, Pineda Portella T, Costa Pessoa FA, Bengtsson-Palme J, Galetti PM: Low coverage of species constrains the use of DNA barcoding to assess mosquito biodiversity. Scientific Reports, 14, 7432 (2024). doi: 10.1038/s41598-024-58071-1 [Paper link]

Published paper: Is ITS1 a better barcode than ITS2?

Another paper I have made a contribution to have just recently been published in Molecular Ecology Resources. The paper (1), which is lead-authored by Xin-Cun Wang and Chang Liu at the Institute of Medicinal Plant Development in Beijing, investigates the usability of the ITS1 and ITS2 as separate barcodes across the Eukaryotes. The study is a large scale meta-analysis comparing available high-quality sequence data in as many taxonomic groups at possible from three different aspects: PCR amplification, DNA sequencing efficiency and species discrimination ability. Specifically, we have looked for the presence of DNA barcoding gaps, species discrimination efficiency, sequence length distribution, GC content distribution and primer universality, using bioinformatic approaches. We found that the ITS1 had significantly higher efficiencies than the ITS2 in 17 of 47 families and 20 of 49 investigated genera, which was markedly better than the performance of ITS2. We conclude that, in general, ITS1 represents a better DNA barcode than ITS2 for a majority of eukaryotic taxonomic groups. This of course doesn’t mean that using the ITS2 or the ITS region in its entirety should be dismissed, but our results can serve as a ground for making informed decisions about which region to choose for your amplicon sequencing project. The results complement what have previously been observed for e.g. fungi, where the difference between ITS1 and ITS2 were much less pronounced (2).


  1. Wang X-C, Liu C, Huang L, Bengtsson-Palme J, Chen H, Zhang J-H, Cai D, Li J-Q: ITS1: A DNA barcode better than ITS2 in eukaryotes? Molecular Ecology Resources. Early view. doi: 10.1111/1755-0998.12325 [Paper link]
  2. Blaalid R, Kumar S, Nilsson RH, Abarenkov K, Kirk PM, Kauserud H: ITS1 versus ITS2 as DNA metabarcodes for fungi. Molecular Ecology Resources. Volume 13, Issue2, Page 218-224. doi: 10.1111/1755-0998.12065 [Paper link]