Taxonomic classification and natural history of a population of endangered Cypripedium orchids in eastern Nebraska

The lady’s slipper orchids are a highly diverse group of taxa, Cypripedium parviflorum being one of approximately fifty species currently classified in the genus. However, the taxonomic placement of the Cypripedium orchid in eastern Nebraska was unknown, leaving a gap in the phylogeny of orchids. In order to precisely document the orchid population, this study adequately examined the existing species’ unique floral characteristics, taxonomy, and natural history by conducting a morphological and molecular comparison of Cypripedium parviflorum with that of other species in the Cypripedium genus. A detailed investigation of each individual stem’s morphological characteristics, including, but not limited to, plant, leaf, and stem size, was completed over a sixweek period. Leaf tissue was collected for the extraction of genomic DNA, which was used to sequence and align genes and compare them to closely related species. The analysis of the lady’s slipper orchid was complete after closely inspecting various herbarium specimens and estimating a molecular phylogeny, providing a better understanding of the relationships between the species. Morphologically, the Nebraska orchid population resembles Cypripedium parviflorum var. parviflorum. Both orchids display 4-5 alternating leaves between 9-19 cm x 2.5-9 cm as well as deep burgundy sepals. However, the molecular comparison reveals the species is a close sister to a clade containing C. kentuckiense, C. parviflorum, and C. parviflorum var. pubescens. Collectively, my data suggest the Cypripedium population in eastern Nebraska should be considered a separate species