How to assess the ‘Megabiodiversity’ of Invertebrates: 55% of the world’s geometrid species are DNA barcoded

Axel Hausmann(1), Rodolphe Rougerie(2), Paul D. N. Hebert(3)

(1) Zoologische Staatssammlung München, Germany (2) Université de Rouen, France (3) Biodiversity Institute of Ontario, University of Guelph, Canada

The clock is ticking …

Global disasters: continuous loss of radioactivity

The clock is ticking …

Fascinating biodiversity Urgently to be assessed!

Time‘s running out for Biodiversity

Continuous loss of habitats

Continuous loss of habitats

Continuous loss of habitats

1. Correct and reliable analysis of biodiversity Knowledge / Identification 2. Effective tools allowing rapid action

The clock is ticking …

Reliable analysis: easy identification?

undeterminable sibling species: Eupithecia pulchellata – Eupithecia pyreneata

Who is who?

Who is who?

undeterminable insect larvae

Main gap: Megadiverse groups of tropical Insects

fresh samples of geometrids from Peru

1. Correct and reliable analysis of biodiversity Knowledge / Identification 2. Effective tools allowing rapid action

The clock is ticking …

Traditional assessment: species descriptions

Traditional assessment: species descriptions

Geometridae

Traditional assessment: species descriptions

Geometridae

Traditional assessment versus DNA barcoding

2011: 14,500 species!

Geometridae

Traditional assessment versus DNA barcoding

2018: 25,000 species!

Geometridae

The clock is ticking!

Geometridae

55% 63% of the world’s geometrid species are DNA barcoded

____

14,500 geometrid species

barcoded (63%)

Bavarian State Collection of Zoology, Munich

Sampling Strategies

Reference Collections Europe North America

9 %

Sampling Strategies

Reference Collection ANIC

9 %

Sampling Strategies

Reference Collection Staude (ZA)

4 %

Sampling Strategies

Reference Collection Herbulot

22 %

ANIC Australia (2000)

Europe & North America (2000)

Reliably identified collection material

coll. Staude, ZA (1000)

coll. Herbulot, world (5000)

Rest (13,000)

versus fresh samplings

Sampling Strategies

Unsorted fresh samplings

ZSM: 500K geometrids worldwide

Sampling Strategies

Selection of study sites: Ghana, Kakum NP

Canopy Access by Walkway

Eudaemonia argus

Sampling under tropical conditions

Killing jar

Ammonia syringe Pinned in the field (Larger specimens in small envelopes)

Back home: Labelling & leg sampling Barcode success rate: >95% Relaxing (max. 2 days) & spreading later on!

Sampling Strategies

95well morphospecies selection

Sampling Strategies

95well morphospecies selection

A priori identification of samples from tropical countries

to species (e.g. ‘Eois aurata')

to species group, tentatively (e.g. ' Eois aurataAH01Ge')

to genus (e.g. ‘Eois AH01Ge')

to (sub)family (e.g. ‘Larentiinae')

20%

30%

45%

5%

Optimizing the procedure

Taking photographs

Optimizing the procedure

Preparing the insects for the PCR plates 96

Optimizing the procedure

Databasing and quality control

Optimizing the procedure

Large-scale production

Calibration

Taxonomic expertise

Forum Herbulot

Calibration

Type specimens

>200 geometrid holotypes >500 paratypes

MNHN, >150 years, 658 bp coll. Herbulot, 100 years, 658 bp

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European campaign

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European campaign

European geometrids total of 970 species (Hausmann 2007) barcoded by April 2011: 880 species (91 %) accessible on BOLD with image and georeferencing

barcoded 91%

pending 9%

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African geometrids

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African geometrids

3168 species described

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African geometrids

3168 species described

649 Ma- dagascar

1543 Rest

914 South Africa

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African geometrids

> 8,500 specimens, 5,750 barcodes, > 2,500 species (“BIN”s) calibration: Hausmann, Staude & Lenz

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African geometrids

goal: 2.8-3K species (almost complete) data release publication late 2012 (PLoS one)

Library of Life – made in Germany

Data release geometrids of Bavaria after less than 2 years 406 of 408 species barcoded (99%) 1815 specimens, 1393 barcodes

99 % diagnostic barcodes!

Distances between species

0

2

4

6

8

10

12

14

0-0,5

%

1-1,5

%

2-2,5

%

3-3,5

%

4-4,5

%

5-5,5

%

6-6,5

%

7-7,5

%

8-8,5

%

9-9,5

%

10-

10,5

%

11-

11,5

%

12-

12,5

%

13-

13,5

%

> 2 %

1 - 2 %

0 - 1 %

Minimum pairwise distances between species

‘Barcode sharing’: Thera obeliscata, T. v. mugo and T. cembrae SYNONYMY

Distances between species

Synonymy TA

XON

OM

Y

Kuchleria garciapitai Exposito, 2005 syn.n. to Kuchleria insignata Hausmann, 1995

Kuchleria insignata Holotype, E Spain

Kuchleria garciapitai from type locality, C Spain

5 % with cryptic diversity in COI (15+5 species, >2%) 9/20 with double clusters, 11/20 with out-sticking singletons

Variation within species

0

10

20

30

40

50

60

70

0-0,5 1-1,5 2-2,5 3-3,5 4-4,5 5-5,5 6-6,5 7-7,5 8-8,5 9-9,5 10-10,5 11-11,5 12-12,5 13-13,5

> 2 %

1 - 2 %

0 - 1 %

Variation (maximum pairwise distance) within species

Cryptic species detection TA

XON

OM

Y

Cryptic species detection TA

XON

OM

Y

P

EST

CO

NTR

OL

A new pest for the Fauna of North Africa: The winter moth O. brumata

map: BOLD database

photo: E. Friedrich

Q

UA

RA

NTA

INE Eueupithecia cisplatensis for the control

of Parkinsonia aculeata in Australia

Agents for the control of invasive plants

photos: CSIRO, Australia by the courtesy of Tim Heard

ECO

LOG

Y Agents for the control of invasive plants

unknown larva (Dead Sea)

Identification moth: Semiothisa aestimaria

COI Sequence

Analysis of gut content EC

OLO

GY

unknown larva (Dead Sea)

Identification moth: Semiothisa aestimaria

Identification plant: Tamarisk

matK/rbcL Sequence

COI Sequence

Analysis of gut content EC

OLO

GY

Acknowledgements Paul Hebert for a wonderful and successful partnership! Rodolphe Rougerie, University of Rouen for iBOL 1.9 co-lead Sujeevan Ratnasingham (BOLD), and the teams of BIO and CCDB BFB-team: G. Haszprunar, S. Schmidt, M. Balke, L. Hendrich, A. Segerer Bayerisches Staatsministerium für Wissenschaft und Kunst Hundreds of enthusiastic and active partners all over the world

Funding agencies

How to assess the ‘Megabiodiversity’ of Invertebrates: 63% of the world’s geometrid species are DNA barcoded