DNA barcoding-at-sea on Polarstern ANTXXXIV-1 TEAM DNA (left to right): Christina Folker, Paola Batta Lona, Leo Blanco Bercial, Lisa Nigro, Rob Jennings,

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Presentation transcript:

DNA barcoding-at-sea on Polarstern ANTXXXIV-1 TEAM DNA (left to right): Christina Folker, Paola Batta Lona, Leo Blanco Bercial, Lisa Nigro, Rob Jennings, Ann Bucklin, CJ Sweetman

Zooplankton Barcodes Sargasso Sea & Eastern Atlantic

DNA Barcoding Steps & Errors 1) Identify specimen 2) Extract DNA 3) PCR mtCOI barcode 4) Sequence DNA 5) Edit / align sequences 6) Validate barcode 7) Submit to database Wrong specimen - specimen incorrectly identified - specimen incorrectly labeled No DNA - bug dead too long before preservation - DNA cooked under microscope - DNA cooked during sample shipment PCR fails - PCR primers don’t match species - PCR protocol not optimal for species - too much or too little DNA Sequence fails - mis-priming in sequencing reaction Incorrect sequence - pseudogene of mtCOI - duplication / translocation of mtCOI - contamination with exogenous DNA - contamination from symbiont, gut

Eastern Atlantic Barcodes 228 individuals, 112 species Neighbor Joining algorithm, Kimura-2-Parameter distances, Bootstrap 1000X Amphipods 9 ind, 9 spp Ostracods 99 ind, 45 spp, 65% Gastropods 46 ind, 17 spp Copepods 26 ind, 15 spp, 95% Gastropods 7 ind, 3 spp, 100% Cephalopods 12 ind, 11 spp, 72% Nemerteans 1 ind, 1 sp Euphausiids 16 ind, 6 spp Chaetognaths 12 ind, 5 spp, 96%

Zooplankton Barcodes Sargasso Sea & Eastern Atlantic 529 individuals, 310 species Neighbor Joining algorithm, Kimura-2-Parameter distances 0.1 Copepods 96 ind, 46 spp, 98% Gastropods 77 ind, 27 spp Gastropods 19 ind, 10 spp, 99% Cephalopods 23 ind, 20 spp, 99 % Polychaetes 5 ind, 3 spp, 99% Nemerteans 3 indiv, 2 spp Amphipods 22 ind, 17 spp, 61% Ostracods 156 ind, 58 spp, 99% Other Crustacea 10 ind, 8 spp, 75% Siphonophores 50 ind, 50 spp Hydromedusae 12 ind, 8 spp Scyphzoa 5 ind, 5 spp Chaetognaths 28 ind, 11 spp, 90% Euphausiids 23 ind, 11 spp, 85%

Barcoding Euphausiids 193 individuals, 40 species Neighbor Joining algorithm, Kimura-2-Parameter distances, bootstrapped 1000X ) E. a m e r i c a n a ( 2 ) E. k r o h n i i ( 1 4 ) 0.05 Euphausia Thysanoessa Nyctiphanes Meganyctiphanes Thysanopoda Stylocheiron Nematobrachion Nematoscelis Bentheuphausia Tessarobrachion % bootstrap

Barcoding Taxonomic Experts Martin Angel Ostracods Francesc Pagès (dec.) Siphonophores Janet Bradford-Grieve Sigrid Schield Copepods Russ Hopcroft Gastropods Uwe Piatkowski Cephalopods Dhugal Lyndsey Amphipods Annelies Pierrot-Bults Chaetognaths Peter Wiebe Fred Buccholtz Euphausiids