Ann BucklinAnn Bucklin11, Robert M. Jennings, Robert M. Jennings11, Brian D. Ortman, Brian D. Ortman11, Lisa Nigro, Lisa Nigro11, , C.J. SweetmanC.J. Sweetman11, Nancy J. Copley, Nancy J. Copley22, and Peter H. Wiebe, and Peter H. Wiebe22

1Department of Marine Sciences, University of Connecticut, USA2Department of Biology, Woods Hole Oceanographic Institution, USA

Zooplankton images by Russell R. Hopcroft (Univ. of Alaska) Zooplankton images by Russell R. Hopcroft (Univ. of Alaska) and Laurence P. Madin (Woods Hole Oceanographic Inst.)and Laurence P. Madin (Woods Hole Oceanographic Inst.)

DNA barcoding the global marine zooplankton assemblageDNA barcoding the global marine zooplankton assemblage

2nd International Barcode of Life ConferenceTaipei, Taiwan – September 17-21, 2007

We gratefully acknowledge support from:the Alfred P. Sloan Foundation,

NOAA Office of Ocean Exploration, and US National Science Foundation

Barcoding Marine Zooplankton

CMarZ effort to barcode 7,000 species in 15 phyla of holozooplankton is ~25% completed, with >1,500 described species barcoded.

At-sea DNA barcoding of identified specimens, with ship-board team of expert taxonomists, is an effective and efficient approach to barcoding zooplankton.

DNA barcoding will aid species discovery; new species are being discovered in biodiversity hotspots, under-sampled regions (deep sea), among rare and fragile planktonic groups, and within circumglobal taxa.

DNA barcodes will allow rapid, automatable, and remote species identification and biodiversity assessments.

Sapphirina metallina Limacina helicinaHippopodius hippopus Salpa cylindrica

Barcoding Euphausiacea (Crustacea)

Forty of 86 species, including 20 of 31 species of Euphausia, were barcoded by Bucklin, Wiebe et al. (2007).

Barcodes accurately and uniquely identify and discriminate species, and can reveal cryptic species within widespread taxa.

Variation within species 1% - 3%

Atlantic / Pacific cryptic species

Ann Bucklin (UConn), Peter Wiebe (WHOI), et al.

Euphausia

Nematoscelis

Stylocheiron

Thysanoessa

Thysanopoda

0

0.02

0.04

0.06

0.08

0.1

0.12

0.14

0.16

0.18

Me

an

Pa

irw

ise

Dif

fere

nc

e

Within species Between species

Copepods have many sibling species groups, differentiated by subtle morphological or morphometrical characters (secondary sex characters).

MtCOI barcodes provide ancillary characters for species identification; can reveal cryptic species within geographically widespread species.

Overall average difference between species is 23.1% for 91 species of copepods.

Barcoding Copepoda (Crustacea)A. Bucklin (UConn), N.J. Copley (WHOI), L. Nigro (UConn), J. Bradford-Grieve (NIWA)

Congeneric species may not cluster together in a barcode tree

Barcodes resolve some species’ relationships for some genera

Barcoding Siphonophora (Cnidaria, Hydrozoa)

Brian D. Ortman (Ph.D. Disseration at University of Connecticut, USA

DNA barcodes completed for ~80 of 160 species of the Siphonophora; differ by 10% - 40% and clearly resolve all species. Phylogenetic relationships above the genus level are not well-resolved; some evidence of resolution of suborders PHYSONECTA, CYSTONECTA, and CALICOPHORA (names color-coded).

Cla

uso

ph

yes

mo

ser

ae

Ere

nn

a sp

Ste

pha

no

mia

am

ph

itri

tis M

arru

s o

rth

oca

nn

a M

arru

s s

p B

arg

man

nia

sp

Ap

ole

mia

sp

Ph

yso

ph

ora

hy

dro

stat

ica

Fri

llag

alm

a s

p L

ychn

ag

alm

a u

tric

ula

ta N

ect

opy

ram

is n

atan

s P

ray

a re

ticu

lata

Am

ph

icar

yon

po

lifer

a A

mp

hic

aryo

n e

arn

esti

Am

ph

icar

yon

aca

ula

Mar

esea

rsia

pra

ecla

ra F

ors

kalia

tho

loid

es

Ph

ysal

ia p

hys

alis

Rh

izo

ph

ysa

eyse

nha

rdti

Rh

izo

ph

ysa

filifo

rmis

Na

nom

ia b

ijug

a H

alis

tem

ma

sp N

ano

mia

car

a A

gal

ma

eleg

ans

Ag

alm

a ok

eni

Ath

oryb

ia r

osa

cea

Ath

oryb

ia s

p L

ensi

a ca

mpa

nel

la R

osa

cea

cym

bif

orm

is R

osa

cea

sp

Fo

rska

lia c

on

oid

ea H

ipp

op

od

ius

hip

po

pu

s V

og

tia

spi

nos

a L

ilyop

sis

flu

roca

nth

a L

ilyop

sis

ros

ea C

hu

nip

hye

s m

ult

iden

tata

Ke

phy

es o

vata

Dim

op

hye

s ar

ctic

a S

pha

ero

nec

tes

gra

cilis

Len

sia

qu

adri

cu

lata

Len

sia

ho

stile

Pra

ya

sp N

ect

opy

ram

is d

iom

edea

e L

ensi

a aj

ax

Len

sia

mu

ltic

rist

ata

Len

sia

exet

er L

ensi

a g

rim

ald

i L

ensi

a m

eteo

ri L

ensi

a fo

wle

ri F

ors

kalia

asy

met

rica

Len

sia

con

oid

ea L

ensi

a h

osp

ur

Su

lcu

leo

lari

a q

ua

driv

alvi

s L

ensi

a ac

hille

s E

udo

xoi

des

mit

ra E

udo

xoi

des

sp

iral

is C

era

tocy

mb

a s

p1

Dip

hye

s d

isp

ar

Dip

hye

s b

oja

ni

En

neag

on

um

hya

linu

m A

byl

op

sis

esc

holt

zi A

byl

op

sis

tet

rag

on

a B

ass

ia b

asse

nsi

s C

era

tocy

mb

a s

agit

tata

Ce

rato

cym

ba

sp

2

100

100

100

100

100

100

83 5099

539187

75 99

100

100

100

98

70

80

63

74

60

95 69

98

56 95

87

78

7473

92

71

60

75

0

5

10

15

Per

ce

nt

dif

fere

nc

e

Barcodes can both confirm species identifications and reveal errors

------------Species----------- Group Collect Barcode Known------------------------------------------------------------------------------------------ Ctenophora 22 13 90 Cnidaria Hydromedusae 33 12 842

Siphonophora 70 55 160  Scyphozoa 7 4 161 Crustacea Amphipoda 31 12 400  Copepoda 138 38 2,000    Euphausiidae 14 14 86  Ostracoda 58 36 169   Other Crustacea 18 15 23 Mollusca Gastropoda 44 36 144  Other Mollusca 27 12 -- Others   Larvacea 12 5 64  Nemertea 1 1 99  Polychaeta 3 3 110  Thaliacea 14 2 45------------------------------------------------------------------------------------------ Totals 534 258 4869

DNA Sequencing at Sea

UConn Team DNA set up a DNA barcoding laboratory during a CMarZ cruise to the Sargasso Sea in April 2006, extracting and sequencing DNA at sea.