Shape and Size Variability of the Gynostemium in Epipactis helleborine (L.) Crantz (Orchidaceae)

Epipactis helleborine (L.) Crantz is considered a challenging and phenotypically difficult species to identify due to its wide range of morphological variability. This variability is mainly observed in the perianth parts but also extends to the gynostemium structure, which has so far been considered one of the most useful diagnostic characteristics. As a result, a simple graphic illustrating the structural pattern of gynostemium morphology has appeared in 10 different forms in available European taxonomic keys, which significantly complicates the identification of this species. A total of 122 flowers of E. helleborine were collected from four natural populations in the Lower Silesia region (Poland) between 2017 and 2019 and analysed for gynostemium morphological variation. Geometric morphometric analyses, including Procrustes ANOVA, PCA, and CVA, were used to examine gynostemium shape, with statistical tests assessing variation in size and stigma inclination angle among populations, individual plants (ramets), and years of research. Statistical analysis revealed significant positive correlations between gynostemium width and height, with significant variation in size and angle of stigma inclination, primarily driven by population, while ramet and year of research had a lesser impact. Geometric morphometric analyses indicated significant population-level variation in gynostemium shape, with principal component analysis identifying the ventral view as the most informative for discriminating these differences. The first two principal components explained the major shape variation, and canonical variate analysis confirmed that this view is most important for species identification.