ODONATA BARCODING: The GOOD, the BAD and the UGLY S. Ferreira, A. Cordero‐Rivera, J. Paupério, J. Veríssimo, F.M.S.Martins, V.A. Mata , A. Muñoz‐Merida, P.C. Alves, S. Jarman , P. Beja Monitoring freshwater diversity Diets and food webs

Detecting target InBIO Barcoding Initiative European Odonates

138 species only 1 new species has been described in the last 20 years Barcoding Odonates Methods

Aim

to produce a Barcoding database of odonates to assist eDNA studies

Material and Methods

Northwestern Iberia: 413 specimens of 68 species

Sanger sequencing (LCO1490 ‐ HCO2198) (Folmer et al. 1994)

Illumina sequencing (LCO1490 ‐ Ill_C_R; Ill_B_F ‐ HCO2198) (Shokralla et al. 2015) Results

Dragonflies and What we wish…

Species A

Species B

Species C Species D

Species F The GOOD The GOOD

Ceriagrion tenellum viridis Crocothemis erythraea Phyrrhosoma nymphula Diplacodes lefebvrii Ischnura pumilio Libellula depressa Erythromma lindenii Libellula quadrimaculata Erythromma viridulum Lestes barbarous Libelula fusca caerulescens fusca Orthetrum caerulescens Coenagrion scitulum Orthetrum cancellatum Orthetrum trinacria (…) >80% of the species of sequenced Results

Dragonflies and damselflies The BAD The BAD

Number of observations (…) Genetic distance (%) The BAD

Coenagrion ornatum is included in the Annex II of the European Habitats Directive Overlapping range and similar habitat, often co‐occur (syntopy) Very distinct morphology

Coenagrion puella C. ornatum C. pulchellum The BAD

mtDNA mtDNA is not adequate for Barcoding/Metabarcoding these species

nDNA

16S+COI Results

Dragonflies and damselflies The UGLY The UGLY Number of observations Genetic distance (%) The UGLY

Calopterygidae Gomphidae Calopteryx virgo acutipennis Gomphus graslinii Calopteryx haemorrhoidalis Platycnemis latipes Gomphus pulchellus Calopteryx xanthostoma Gomphus simillimus

>10% of the species of Odonata sequenced

Use of distinct primers might yield a single sequence Numts nuclear copies of mtDNA exist If we are able to recover them they are also in the eDNA

make the best of it? why not deposit Numts data in databases

Opportunity to improve taxa assignation rate: ‐ Avoid overestimation of taxa (biodiversity inventories, diet studies, etc…) Increase detection rate (targeted species: conservation concern or invasive) Numts

Let´s not throw the baby out with the bath water Conclusions

We obtained COI sequences for 413 specimens of 68 species

55 species had unique Barcodes

5 species shared COI haplotypes with at least another species

8 species exhibited Numts

more results soon… Thank you for listening!

Biodiversity Chair