To see Jessica’s PP Presentation  on this subject, go to Introduction to Mycoflora: The Power of Citizen Science.

DNA sequencing through the Mycoflora project adds another piece to the identification puzzle

by Jessica Benson Evans
Remember that beautiful Amanita that I collected at the NEMF gathering in Geneseo, NY? Based on its morphology – striate cap margins and a sack-like volva at the base – I called it “Amanita sect. Vaginatae” and dropped it off at the display tables. While I was away on an afternoon collecting walk, Rod Tulloss tentatively identified it as
Amanita luzernensis, a provisional name he had given to a species collected in Luzerne County, Pennsylvania. 

The physical characteristics seemed to be a great match, but we could obtain another piece of the puzzle in the species determintation by utilizing DNA sequencing. In this case, my Amanita was saved and dried at the NEMF foray so that it could go for sequencing through the North American Mycoflora project. The results determined that it is not actually A. luzernensis; instead, it is another as-yet-unnamed species within Amanita sect. Vaginatae! Click here to see the observation on I-naturalist, posted by Mycoflora project coordinator Stephen Russell. For Rod Tulloss’ page on this unnamed Amanita species click here.

This coming winter, we will be offering a workshop on how to participate in the Mycoflora project, including how to write great descriptions. Each of us can contribute to our club’s citizen science project: photographing, collecting, describing, and drying specimens for herbarium storage and/or DNA sequencing!

PVMA Citizen Science Workshops

Jessica, Sue, and Paul will be introducing our club’s Citizen Science project; you’ll learn about our project goals, discover how to collect and document specimens for the project, and participate in a walk on the trails at Dianna’s to put your new knowledge into practice. We’ll walk you through all the necessary steps for participation in this incredible project.

Topics covered during the workshop will include:
- Documenting fungal specimens in the field through note-taking and photography - Collecting specimens responsibly
- Creating thorough descriptions of collected specimens
- Using Mushroom Observer to record information on specimens
- Using the Mycoportal and GenBank to decide which specimens to submit
- Preserving specimens for herbarium storage

Citizen Science Project Update By Sue Lancelle (January 2019)

Earlier in the fall, we applied to the North American Mycoflora Project for a grant to pay for DNA sequencing of 30 fungal specimens, and in late November, we were notified that we received the grant. Jess and I had already done most of the work of documenting, drying and storing the specimens, now we had to decide for certain which ones we wanted to have sequenced and which ones would be submitted to the New York Botanical Garden as voucher specimens only. How to decide? There are two very useful sources of information that we used. The first is the Mycoportal. This is the first step we used to determine if a specimen should be vouchered. Here you can search for herbarium specimens by genus and species and designate which geographical location you are interested in, whether it be country, state, county or town. We tried to determine if, how many and how recently our various specimens had been collected in our area. It was often interesting to discover that even fairly common fungi had never been vouchered or hadn’t been vouchered in a very long time. Once we determined what to voucher, we decided which ones to sequence by accessing GenBank. Here you can search the “Nucleotide” section to see how many and what type of DNA sequences have already been performed for the species of interest. Those specimens for which we could not determine a species at all were

given top priority. Then we ranked our collections based on how much information seemed to be already available. If there were few or no sequences done for a species, we put that in our priority pile. In the end, it wasn’t hard at all to come up with 30 specimens to have sequenced! The DNA sequencing data will eventually be available to anyone interested in accessing it through GenBank. Those specimens will also be vouchered at The New York Botanical Garden, along with an additional 28 that will be vouchered only and not sequenced. The vouchers, even if not sequenced, provide important information for researchers who are interested in distribution and timing of fruiting of fungi. The specimens are also then available for scientists who may wish to borrow them for further study, including doing sequencing if it hasn’t already been done.

Now that we have been through the whole process once, we are very excited to continue collecting, documenting, vouchering and sequencing our local fungi! We hope that more of you might be interested in contributing next year.

We will give you an update on the results once the sequences have been returned to us. Here are the specimens we submitted: 

For DNA Sequencing and Vouchering:

Amanita submaculata Peck
Armillaria gemina Bérubé & Dessur.
Boletopsis grisea (Peck) Bondartsev & Singer
Boletus vermiculosus group
Cantharellus cibarius group
Clitocybe robusta Peck
Conocybe macrospora (G.F. Atk.) Hauskn. Cuphophyllus lacmus group
Cystodermella granulosa (Batsch) Harmaja Flammulaster erinaceellus (Peck) Watling
Gloiodon strigosus (Sw.) P. Karst.
Gomphidius smithii Singer
Inocybe tahquamenonensis D.E. Stuntz
Lactarius atroviridis Peck
Lactarius sordidus Peck
Lepiota (Pers.) Gray
Mycena epipterygia var. lignicola A.H. Sm. Neoalbatrellus caeruleoporus (Peck) Audet
Neolentinus lepideus (Fr.) Redhead & Ginns
Pholiota lenta (Pers.) Singer
Pholiota limonella (Peck) Sacc.
Protostropharia alcis (Kytöv.) Redhead, Thorn & Malloch Psathyrella conissans (Peck) A.H. Sm.
Rimbachia Pat.
Tricholoma caligatum (Viv.) Ricken
Tricholoma davisiae Peck
Tricholoma odorum Peck
Tricholoma pullum Ovrebo
Tricholoma subresplendens (Murrill) Murrill Xanthoconium Singer

Vouchers Only:

Amanita banningiana Tulloss nom. prov.

Amanita cokeri (E.-J. Gilbert & Kühner) E.-J. Gilbert
Amanita frostiana (Peck) Sacc. (two specimens from different locales)

Amanita onusta (Howe) Sacc.
Armillaria gallica Marxm. & Romagn.
Boletus ferrugineus Schaeff.
Boletus longicurvipes Snell & A.H. Sm. (two specimens from different locales)

Butyriboletus brunneus (Peck) D. Arora & J.L. Frank Butyriboletus roseopurpureus (Both, Bessette & Roody) K. Zhao, Z.L. Yang & Halling, comb. nov.

Clitopilus prunulus (Scop.) P. Kumm.
Cortinarius torvus (Fr.) Fr.
Cyptotrama asprata (Berk.) Redhead & Ginns
Hypholoma lateritium (Schaeffer) P. Kummer
Lactarius gerardii (Peck) Kuntze
Leccinellum albellum (Peck) Bresinsky & Manfr. Binder Leccinum holopus (Rostk.) Watling
Leccinum rubropunctum (Peck) Singer
Oudemansiella furfuracea (Peck) Zhu L. Yang, G.M. Muell., G. Kost & Rexer

Psathyrella delineata (Peck) A.H. Sm. 

Pseudomerulius curtisii (Berk.) Redhead & Ginns 

Retiboletus griseus (Frost) Manfr. Binder & Bresinsky Suillus subaureus (Peck) Snell
Tricholoma davisiae Peck
Tricholomopsis decora (Fr.) Singer
Tylopilus plumbeoviolaceus (Snell & E.A. Dick) Singer 

Tylopilus violatinctus T.J. Baroni & Both