NSF BS&I 0333722

NSF BS&I Award 0344722 (at University of Kansas):

“Biodiversity of cryptic fungal symbionts, Harpellales, living within the guts of aquatic insects in North America”

$400,000 5/1/04 to 4/30/08.

The objectives were to study the species richness of Harpellales gut fungi in North America (Canada, USA, and Mexico), to explore new types of hosts and habitats, and to use the newly discovered species to strengthen phylogenetic hypotheses of the symbionts. Major surveys and field collections were carried out in Canada, including Algonquin Provincial Park and vicinity (Ontario, May 2004) and in the Kootenay Mountain Range (British Columbia, July 2006); in the USA, collections were in the Great Smoky Mountains National Park (July-August 2004) and Cascade range of Oregon (August-September 2005). The first surveys, ever, for these fungi in Mexico were conducted in the state of Veracruz (near Xalapa, November 2005 and Los Tuxtlas, July-August 2006).

Our surveys were not intended to collect every larval aquatic insect or gut fungus in North America, indeed not even for any given region we visited. Dissecting gut fungi requires about 2 days of microscopy per 1 day of collecting and many hosts are not present year round. Our approach was to sample the diversity of likely suitable habitats in regions of North America known to be rich in host taxa but not well sampled from a fungal or ecological perspective. Expanding our surveys geographically and at the most suitable time of year, were two strategies we employed to this end.

We discovered an unusual new genus (Barbatospora) in black flies, rediscovered long-missing taxa (e.g. Pteromaktron, in mayflies) and will have described more than 40 new species of Harpellales in a variety of insect hosts, some new and unusual. Every survey has revealed new taxa of gut fungi and as this project concludes numerous species descriptions are being prepared for several papers simultaneously. Several axenic cultures were obtained, and dozens of fungal specimens were preserved in DNA buffer and others have been sequenced already and will contribute to the ongoing molecular systematics program at Boise State University.

Contributors and participants in our research were:

1. Senior collaborators Dr. Doug Strongman (Canada) and Dr. Matías Cafaro (Puerto Rico), and visiting scholar Laia Valle (Spain), who also trained at the KU laboratory for 7 weeks.

2. Graduate student Augusto Siri, Universidad de La Plata (Argentina) studied in our Kansas laboratory for 3 months and attended the Mycological Society of America (MSA) meeting in Asheville, NC. He has now completed his Ph.D. dissertation research on gut fungi and is submitting chapters for publication.

3. Undergraduate Laboratory Assistants: Alex Garrett, Kelly Morton, Sara Pittman, and Sheila Tsau were trained in techniques from culturing gut fungi to preparing DNA sequencing reactions to analyzing data. One is enrolled in graduate school, one is considering this option, one will teach high school science and the last is still with the project.

4. Undergraduate trainee Jason Koontz, a student at the Haskell Indian Nations University, has been working in the University of Kansas laboratory on an NIH RISE program for over two years. Koontz (2006) was his 1st paper on the effects of temperature, pH, and nutritional preference of a culture of Smittium culisetae that he isolated from mosquito larvae. He is continuing his studies, expects to publish a 2nd paper, and aspires to conduct environmental/conservation graduate studies.

5. A number of Mexican biologists facilitated the research in Mexico. Graduate student Ricardo García-Sandoval, currently a postdoctoral researcher at Clark University, Massachusetts, was trained in collecting and identifying gut fungi and has expressed interest in conducting his own surveys in Mexico.

Field work in Canada has been carried out by White and Strongman, in Mexico by White, Valle and Cafaro and in the USA by both PI’s, and with Siri in the Great Smoky Mountains National Park. Additionally, Strongman also has been collecting in the Maritime Provinces, eastern Canada through the grant period.

Networking and Cooperation:

Both PIs also participated in the NSF funded DEEP HYPHA (#0228657) and Assembling the Fungal Tree of Life, AFToL (#0090301) projects. Some samples of gut fungi donated to the cause of these other projects have been used in broader publications.

Outreach:

White, Bob Lichtwardt, and Hiroki Sato (Japan) organized a workshop (symposium) entitled “Phylogenetic studies of Trichomycetes,” and presented papers at the joint meeting of the MSA and the Mycol. Soc. Japan in Hilo, Hawaii, 30 July to 5 August 2005; 2) Cafaro, Valle and White gave a joint presentation, “Gut Biota of Arthropods (Trichomycetes)“ and round table discussion for researchers and students at Los Tuxtlas Mexico in August 2006; 3) in Mexico and at other collecting sites, we conversed with curious local children and adults about the general practice of collecting aquatic insects, the biology of the hosts and our interest to look inside for “organisms related to mushrooms” etc.); 4) on surveys, we were joined by local scientists (including entomologist’s, aquatic biologists, a fisheries biologist, park rangers, a geologist, bird enthusiast, and government researcher) in each country, who acquainted us with local habitats and fauna; 5) the PIs and Strongman contributed a section on gut fungi that wasincluded in George Barron’s popular “MycoAlbum CD for Introductory Mycology” (ISBN 0-920370-01-2).

Meetings and Dissemination:

One or both PIs attended and presented papers at the final DEEP HYPHA meeting, MSA meetings, and the 8th International Mycological Congress (Cairns, Australia); 2) An illustrated interactive key (Lucid software) was developed for traditional Trichomycetes (www.nhm.ku.edu/~fungi). This product and the online monograph have been updated continuously, as new data are obtained; 3) A searchable database of Trichomycetes (Specify software) is now online with 2400+ entries available; 4) A DVD and web-based video clips are available to anyone interested in the “process, methods and reasons why” we are conducting this research. These videos are now online to serve as possible tutorials for undergraduate and grade school students or anyone interested in learning more about the gut fungi.