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 species 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 damselflies What we wish…
Species A
Species B
Species C Species D
Species F The GOOD The GOOD
Ceriagrion tenellum Lestes viridis Crocothemis erythraea Phyrrhosoma nymphula Lestes macrostigma Diplacodes lefebvrii Ischnura pumilio Lestes sponsa Libellula depressa Erythromma lindenii Lestes virens Libellula quadrimaculata Erythromma viridulum Lestes barbarous Libelula fusca Coenagrion caerulescens Sympecma fusca Orthetrum caerulescens Coenagrion scitulum Orthetrum cancellatum Coenagrion mercuriale Orthetrum trinacria (…) >80% of the species of Odonata 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 Platycnemididae Gomphidae Calopteryx virgo Platycnemis 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