Date of Award
Master of Science
Dr. Donna Becker
Recently the population of Cypripedium arietinum in Grand Sable Dunes, Michigan was estimated at more than 3.5 million plants, greater than 90% of the world’s known population. There is little research on this species’ root associated fungal endophytes. This new population estimate allows the ethical, destructive sampling required to shed light on root associated symbionts and further successful preservation and restoration of the species. Samples were collected of current year’s roots from 75 C. arietinum orchids at 75 locations within 4 jack pine forests in the Grand Sable Dunes, Pictured Rocks National Lakeshore, Michigan. Fungal endophyte cultures were obtained from roots and subcultured for DNA extraction. DNA was PCR amplified with primers that isolate the highly conserved 5.8s rDNA and the highly variable internal transcribed spacer (ITS) regions on either side of it. DNA was sequenced and sequences were identified using NCBI BLAST. Of the 36 DNA samples 27 were positively identified. Phialocephala fortinii and Leptodontidium orchidicola were the most prevalent identifications. These fungi both have saprotrophic abilities and may play an important role in C. arietinum’s lifecycle.
Farkas-Lasich, Kari, "Isolating and Identifying Fungal Endophytes from Roots of Rare Orchid Cypripedium arietinum in Grand Sable Dunes, Michigan" (2018). All NMU Master's Theses. 553.