Development of EST-SSR markers in Tricyrtis sp., which reproduce through vegetative propagation via bulbils

The genus Tricyrtis Wall. (Liliaceae) comprises approximately twenty species of short perennial herbs that occur in temperate to subtropical mesic forest biomes. These plants are distributed from the Himalayas to East Asia, mainly distributed in Japan (13 species), and include many local endemics and endangered species. The genus Tricyrtis is divided into four sections, sect. Flavae, Brachycyrtis, Hirtae, and Tricyrtis. The largest section Tricyrtis is mainly distributed in Japan and China and includes approximately 8 species. Recently, an entity of undescribed Tricyrtis plants belonging to sect. Tricyrtis was noticed during botanical research in northern Kyushu Island, Japan. It used to be identified as T. macropoda because of its close morphological resemblances to the species, but it can be distinguished from T. macropoda by producing bulbils on its axils. This new taxon and Japanese T. macropoda are shown to be a sister group based on molecular data and morphological characters. Any Tricyrtis species that reproduce both sexually and through vegetative propagation via bulbils have never reported. Genetic markers can provide insights into the relative contribution of sexual vs. clonal reproduction within population. To date, however, there have been few genetic markers available for sect. Tricyrtis. To investigate the genetic structure and the spatial extent of genets within populations, additional microsatellite SSR (simple sequence repeat) markers that can be stably PCR (polymerase chain reaction) amplified were needed.