1. 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

2. The clock is ticking …
Global disasters:
continuous loss of radioactivity

3. The clock is ticking …

4. Fascinating biodiversity
Urgently to be assessed!

5. Time‘s running out for Biodiversity

6. Continuous loss of habitats

7. Continuous loss of habitats

8. Continuous loss of habitats

9. The clock is ticking …
1. Correct and reliable analysis
of biodiversity
Knowledge / Identification
2. Effective tools allowing rapid action

10. Reliable analysis: easy identification?

11. Who is who?
undeterminable sibling species:
Eupithecia pulchellata – Eupithecia pyreneata

12. Who is who?
undeterminable insect larvae

13. Main gap:
Megadiverse groups of tropical Insects
fresh samples of geometrids from Peru

14. The clock is ticking …
1. Correct and reliable analysis
of biodiversity
Knowledge / Identification
2. Effective tools allowing rapid action

15. Traditional assessment:
species descriptions

16. Traditional assessment:
species descriptions
Geometridae

17. Traditional assessment:
species descriptions
Geometridae

18. Traditional assessment
versus DNA barcoding
Geometridae
2011: 14,500 species!

19. Traditional assessment
versus DNA barcoding
Geometridae
2018: 25,000 species!

20. The clock is ticking!
Geometridae

21. ____ 63% of the world’s geometrid species
55%
are DNA barcoded
14,500 geometrid species
barcoded (63%)

22. Bavarian State Collection of Zoology, Munich

23. Sampling Strategies
Reference Collections Europe North America
9%

24. Sampling Strategies
Reference Collection ANIC
9%

25. Sampling Strategies
Reference Collection Staude (ZA)
4%

26. Sampling Strategies
Reference Collection Herbulot
22 %

27. Reliably identified collection material
versus fresh samplings
Europe & North America (2000)
ANIC Australia (2000)
coll. Staude, ZA (1000)
Rest
(13,000)
coll. Herbulot, world (5000)

28. Sampling Strategies
Unsorted fresh samplings
ZSM: 500K geometrids worldwide

29. Sampling Strategies

30. Selection
of study
sites:
Ghana,
Kakum NP
Eudaemonia argus
Canopy Access
by Walkway

31. Sampling under tropical conditions
Ammonia syringe
Pinned in the field
(Larger specimens in
Killing jar small envelopes)
Back home: Labelling & leg sampling
Barcode success rate: >95%
Relaxing (max. 2 days) & spreading later on!

32. Sampling Strategies
95well morphospecies selection

33. Sampling Strategies
95well morphospecies selection

34. A priori identification
of samples from tropical countries
to (sub)family
(e.g. ‘Larentiinae')
5% to species
20% (e.g. ‘Eois aurata')
45%
to genus
30%
(e.g. ‘Eois AH01Ge')
to species group, tentatively
(e.g. ' Eois aurataAH01Ge')

35. Optimizing the procedure
Taking photographs

36. Optimizing the procedure
Preparing the insects for the PCR plates 96

37. Optimizing the procedure
Databasing and quality control

38. Optimizing the procedure
Large-scale production

39. Calibration
Taxonomic expertise
Forum Herbulot

40. Calibration
Type specimens
coll. Herbulot, 100 years, 658 bp MNHN, >150 years, 658 bp
>200 geometrid holotypes
>500 paratypes

41. Continental assessments
European campaign

42. Continental assessments European campaign
pending
9%
barcoded
European geometrids
91%
 total of 970 species (Hausmann 2007)
 barcoded by April 2011: 880 species (91 %)
 accessible on BOLD with image and georeferencing

43. Continental assessments
African geometrids

44. Continental assessments African geometrids
3168 species described

45. Continental assessments African geometrids
649 Ma-
dagascar
1543 Rest
914 South
Africa
3168 species described

46. Continental assessments African geometrids
 > 8,500 specimens, 5,750 barcodes,
 > 2,500 species (“BIN”s)
 calibration: Hausmann, Staude & Lenz

47. Continental assessments African geometrids
 goal: 2.8-3K species (almost complete)
 data release publication late 2012 (PLoS one)

48. 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

49. Distances between species
14
Minimum pairwise distances between species
> 2%
1-2%
12
0-1%
10
8
6
4
2
0
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- 11- 12- 13-
% % % % % % % % % % 10,5 11,5 12,5 13,5
% % % %
99 % diagnostic barcodes!

50. Distances between species
‘Barcode sharing’: Thera obeliscata, T. v. mugo and T. cembrae
SYNONYMY

51. Synonymy
Kuchleria garciapitai Exposito, 2005 syn.n. to
Kuchleria insignata Hausmann, 1995
TAXONOMY
Kuchleria insignata Kuchleria garciapitai
Holotype, E Spain from type locality, C Spain

52. Variation within species
70
Variation (maximum pairwise distance) within species
>2%
1-2%
60
0-1%
50
40
30
20
10
0
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
5 % with cryptic diversity in COI (15+5 species, >2%)
9/20 with double clusters, 11/20 with out-sticking singletons

53. TAXONOMY Cryptic species detection

54. TAXONOMY Cryptic species detection

55. A new pest for the Fauna of North
Africa: The winter moth O. brumata
PEST CONTROL
photo: E. Friedrich
map: BOLD database

56. Agents for the control of invasive plants
Eueupithecia cisplatensis for the control
QUARANTAINE
of Parkinsonia aculeata in Australia
photos: CSIRO, Australia by the courtesy of Tim Heard

57. ECOLOGY Agents for the control of invasive plants

58. ECOLOGY Analysis of gut content
COI Sequence
unknown larva (Dead Sea)
Identification moth:
Semiothisa aestimaria

59. ECOLOGY Analysis of gut content
matK/rbcL Sequence COI Sequence
unknown larva (Dead Sea)
Identification plant: Identification moth:
Tamarisk Semiothisa aestimaria

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