Tangalooma Diptera Workshops
August 23-25, 2004
Tangalooma, Moreton Island, South East Queensland

Notes dated 25 Oct 2004
A 2.5 day series of workshops was held just after the International Congress of Entomology in Brisbane (August 2004) to track issues across three collaborative research projects in Diptera. Aims of the workshops were as follows:

1. Discuss goals and strategies for the final year of funding of the NSF PEET grant “Towards a World Monograph of Therevidae”. This workshop also addressed planning for the upcoming PEET V meeting hosted by grant participants in Illinois in September 2004.  Full NSF Therevid PEET participant list available on website.

2. First meeting of participants in the NSF Diptera ATOL (FLYTREE) grant to discuss progress to date, taxon sampling for the first tier, and issues pertaining to development of the morphological dataset for the first and second tier.  Full ATOL FLYTREE participant list available on website.

3. Discussions to explore the integration of the Biosystematic Database of World Diptera into other on-going projects, such as PEET and FLYTREE.

These notes from the workshops were prepared by Gail Kampmeier, Chris Lambkin, Brian Wiegmann and David Yeates, with input from the other attendees. Most of these notes refer to discussions about the FLYTREE project (item 2 above).

Present at Meeting

Principal investigators

Brian Wiegmann, Rudolf Meier, David Yeates, Greg Courtney,
F. Christian Thompson, Michael Irwin, Gail Kampmeier.

Major Collaborators

Christine Lambkin, Thomas Pape, Jeffrey Skevington, Evert Schlinger, Shaun Winterton.

Grad Students

Michelle Trautwein – Bombyliidae/ lower Brachycera
Matt Bertone – Lower Diptera
Frederik Petersen - Hippoboscoidea
Sujatha Kutty -Calyptrata

Introduction and News

News from Brian Wiegmann, Chief PI of FLYTREE

• Converted PhD assistantships into postdoctoral funding at 2:1 at Iowa State University ( FLYTREE Postdoc -- Dr. Vladimir Blagoderov - start date 1 July 2004). NCSU - FLYTREE postdocs were interviewed and 2 were hired (Dr. Norman Barr - start date 1 Oct 2004, and Jun Wook Kim - start date 6 Dec 2004)
• New year of funding granted
• Complete mitochondrial genome sequences for first tier – 3 finished; 25 will be done within the next year.
• Nuclear gene sequencing – genes for ones we have already will be easy to. Meaningful and beneficial cooperation w/Rob DeSalle and Jerry Regier established. Working on timely, early successes.
• Michael Eisen at UC Berkeley is leading effort to sequence genomic Bac Libraries for 18 non-drosophilid dipterans --6 tephritids; 6 diopsids; 6 additional Cyclorrhapha – is coordinating efforts with FLYTREE; will make molecular data available – open access of information.
• Brian Cassel is sequencing 3 genes while workshop is underway.
• Sonja Scheffer (USDA Beltsville) working on 3rd tier Opomyzoidea w/grad students.
• Brian Wiegmann has taken on administrative role in Genome Sequencing Laboratory (GRL) at NC State.

News from Greg Courtney – Coordinator of Lower Diptera

• Revisit published data matrices – need to confirm character states looking at exemplars – questionable character information
• Expand what we know about larval morphology, especially mouthparts – character list with up to 80 characters
• Expand phylogenetic information on pupae – 7 characters on preliminary list (Martin Hauser, Illinois, may be interested)
• Possibility to construct good key to Diptera pupae (as well as other life stages) – supplemental NSF funding?
• Expand character matrix to include more recent classifications and analyses (e.g., for Sciaroidea, Tipuoidea)

Challenges:

• need all larval stages, especially first instars and mature larvae and pupae and pupal exuviae
• first series goes into 95% ethanol for molecular work; second goes for electron microscopy SEM – put in hot water to get rid of fats etc. for 10 seconds, then into 95% ethanol
• changing species because of availability of life stages for tier 1
• tracking down immature stages of sciaroid specimens – not common, difficult to associate with adults
• character systems – larval in particular; looking at characters of anal division (respiratory siphons); mouthpart morphology; chaetotaxy (antennal and mouthpart setae) Art Borkent developing characters of feeding apparatus; Art wants to work on pupal characters as well
• what to do if adults of one species are available but not other stages
• 1st instars a real problem
  • Cyclorrhapha – all inapplicable for Lower Diptera characters?
  • improbable for many taxa, many unknown

News from David Yeates – Coordinator of Lower Brachycera

• Second tier should be well served by the Therevid PEET grant expeditions
• Will need a lot more information on 1st tier taxa – getting larvae and pupae, major challenge for parasitoids in first tier.
• May need to score from literature
• Larvae and pupae will be even more of a problem for the 2nd tier
• The mitochondrial group – genome by genome (taxon by taxon) – need material now
• Tabanid already partially done – need to get larvae of European tabanid
• Many 2nd tier and 3rd tier have already been done for lower Brachycera
• Getting immatures for parasitoids will be very difficult unless we in put a lot of resources
• One suggestion for difficult larvae is to go to Dipterists Day in England and set challenges for amateurs

News from Rudolph Meier – Coordinator of Cyclorrhapha

• No character matrix – for Schizophora – preliminary list will be prepared by RM and Tom Pape by end of workshop
• Scoring by end of year? RM will be busy teaching until May
• Molecular work – Sujatha's project
• Some challenges getting larvae, especially first instars
• half a dozen families for 2nd tier for which we lack material
• 250 species of Cyclorrhapha in 2nd tier
• Phytomyza replaced Opomyza, conopid added to first tier.

First Tier Taxon List

• Need to provide final list of first tier taxa to BMW by end of September 2004.

Bioinformatics

There was a broad ranging discussion on bioinformatics needs for FLYTREE.

• The moment you start working on a specimen/species it needs to be registered – ATOL#
• Family genus, species, gene, - list genes being sequenced, life stages that we have material for; sex/es; lab location; status of work.
• Give recognition of data owner - Many scientists very willing to share
• Archive & share information for each taxon in our program – Genbank serves as an historical record
• Source of funding recognized for work
• Material available for sequencing; stage (1st instar and mature larva); how it is stored
• Automatic notifier to whomever is coordinating group if sequencing started
• Species identificatons essential – vouchers – specimens need unique identifier – lot identifier not enough – primary voucher in the tail
• Vouchering & voucher location; and databasing of sequences – text file
• Chromatograms – proprietary image file – not included
• View as spreadsheet as well as searchable
• Mark tier of project
• Three stages-planning, working, completed
• Gail Kampmeier to set up in FileMaker Pro 7 in October 2004.

Species Pages

The FLYTREE grant will develop species pages for all 42 species in the first tier. Gail Kampmeier demonstrated a fully populated species page mock-up from the Therevidae. The species page should include information on:

• Source of biological material
• Part of the initiative to develop an Encyclopedia of the Earth's species
• Image(s)
• Brief diagnosis
• “Story”
• Explanation of biogeographic regions
• Thumbnails
• Metadata
• Links to world – nomenclature and synonymies – Diptera.org
• Intent is to serve in FileMaker Pro 7 to web
• Mock up to stimulate discussion (obviously need to add Family for FLYTREE)

Characters and Character States – a Morphobank for Diptera

The FLYTREE grant gives us the opportunity to develop a morphological character matrix for the entire order. The first step in this process is to develop a character list.

Prior to the workshop Christine Lambkin and David Yeates developed a character list based on the Lower Brachycera, and began integrating this with a character list for the Lower Diptera provided by Greg Courtney. This will be integrated by CL with a character list for the Cyclorrhapha (provided by RM and TP) to provide a first draft of an overall character list for the Diptera. It is difficult to assess how long this will take-2 months??

• Jeff Skevington has 142 characters for larvae, behavior, adults (Cumming & Sinclair, unpublished) – permission granted already for use; incorporate into Schizophora list.
• Which character sets should be included or removed?
• Musculature, spermatozoa, embryology and nervous system characters sets too difficult for us to examine in all taxa, taxa currently scored for many of these datasets are very limited
• We should approach those who are working on these difficult but potentially informative character sets and encourage their involvement in the 1st tier character scoring for FLYTREE and possibly supply them with specimens of taxa from the first tier
• Amalgamated character list will be emailed to all, in MacClade format**
• Any additions/changes/corrections/deletions to characters/states must be quickly emailed to CL, who will make changes to the data matrix and inform all.
• The current character list/data matrix may be added to the website.
• Scoring should start as soon as the data matrix is available
  • Character list includes characters for 1st and 2nd tier taxa
  • 2nd tier characters/states will be maintained in the matrix for the 1st tier scoring and, and may be inapplicable in the 1st tier analysis
  • RM considered that inapplicables should be scored as missing data, rather than an extra state
• Morphological character scoring
  • Exemplar approach – first choice
  • Entries from literature – if exemplars unavailable
  • Extrapolation from understanding – if specific taxon not included in literature, but other related taxa are described

Adult-male genitalia

There is a problem with homology interpretations in the Schizophora. RM suggested that the Schizophora may require the development of 2 sets of characters for male genitalia, depending on homology statements. The two sets could be tested for congruence with the remaining data (a la Wiegmann et al. 1993)

SEM atlas of Diptera Character State Images

Rudolf Meier introduced the idea of a SEM atlas photos of characters:

• 1st tier atlas (SEM/photo) of all 1st tier characters
• Views of all taxa for 1st tier
• Expand on Manual of Nearctic Diptera
• Look for existing images, find undergraduates to complete this work?
• Important way to discover new characters
• Greg Courtney would try to incorporate for larvae of Lower Diptera

RM, TP Added list of suggested views required for atlas

SEM of first and last instar [WILL be expanded for lower Diptera]

1-2. head (anterior region)
3-4. ventral view
5-6. lateral view
7-8. dorsal view
9-10. dissection of cephaloskeleton
11-12. SEM of anterior spiracle
13-14. SEM of posterior spiracle
15-16. SEM of segmental view (habitus)

SEM of first and last instar, terminal segment [may be expanded for lower Diptera]

17-18. ventral view
19-20. lateral view
21-22. dorsal view

Adult-body

23. Lateral habitus SEM
24. Lateral habitus, colour photograph

Adult-head

25. Antenna, lateral view, SEM
26. Antenna with flagellum removed, SEM
27. Scape articulation with head capsule, SEM

Adult, head morphology

29. Head, lateral view SEM
30. Head, ventral view SEM
31. Head, dorsal view SEM
32. Head, frontal view SEM
33. Head, posterior view SEM

Head, proboscis

34. Proboscis dissected off, boiled in KOH, photographed in alcohol/glycerine

Adult-thorax (wings and legs, incl. coxae, removed)

36. Lateral SEM
37. Frontal SEM
38. Dorsal SEM
39. Ventral SEM
40. Notopleural area, SEM
41. Anterior spiracular area, SEM
42. Metathoracic spiracular area, SEM
43. Lateral view, colour photograph
44. Dorsal view, colour photograph

Wing

45. Entire wing, white background, light from below; photograph
46. Costal - r1 section; photograph

Legs (male legs; incl. coxae). [may be less important for lower Diptera]

47-49. Legs, anterior view, SEM
50-52. Legs, posterior view, SEM
53-55. Legs, lateral view, SEM
56. Mid leg (if suitable), tarsal apex, dorsal view, SEM
57. Mid leg (if suitable), tarsal apex, ventral view, SEM

Adult-abdomen, non-genitalic (KOH treated; stretched)

58. Male abdomen, dorsal view; photograph
59. Male abdomen, ventral view; photograph
60. Female abdomen, dorsal view; photograph
61. Female abdomen, ventral view; photograph

+ genitalia

Publications from FLYTREE

• 1st tier paper will be multiauthored – major players … acknowledgements section have clear statement of each participants' role – challenge of first authorship is whoever pulls this together as a first draft
• smaller papers – call FLYTREE products, but bring in authorship as appropriate – not be too controlling of decisions of individual authors.

Meeting next year:

Meeting planned July 2005 in Copenhagen to discuss morphological character scorings for first tier

SUMMARY OF MAJOR ACTIONS AND OUTCOMES FROM MEETING

• Provide final list of first tier taxa to Brian Wiegmann by end of September 2004.
• Gail Kampmeier to set up FLYTREE specimen tracking database in FileMaker Pro 7 in October 2004.
• Chris Lambkin to circulate draft of character list for first and second tier in McClade format by December 2004.
• Meeting planned July 2005 in Copenhagen to discuss morphological character scorings for first tier.