Classical Morphometrics in V. arvensis and V. kitaibeliana (Viola sect. Melanium) Reveals Intraspecific Variation with Implications for Species Delimitation: Inferences from a Case Study in Central Italy

The high morphological variability of Viola arvensis may hinder the proper identification of the closely related species with an implication for biodiversity surveys. Variation in floral and vegetative morphology was explored in V. arvensis, compared to V. kitaibeliana, based upon 14 wild Italian populations, to provide new insights into their diagnostic features. Species were characterized using 32 morphological descriptors assessed on 272 flowers and as many leaves and scored as quantitative and categorical variables. Statistical methods, including Linear Discriminant Analysis (LDA), were applied to test species delimitation. Data highlighted variations in sepal size, petal size, leaves shape, stylar dark spot, and pollen magazine higher within V. arvensis than between V. arvensis and V. kitaibeliana. LDA partitioned the V. arvensis samples into two distinct clusters; no clear distinction was found between the cluster combining individuals from grasslands and V. kitaibeliana. The separation of V. arvensis and V. kitaibeliana from V. tricolor, included as a reference, was noticeable. Correlations were found in all species between the flower/leaf position on the stem and some floral and vegetative features. The shape and margin of the lower sepal, the stylar flap, and the lamina margin and apex were diagnostic in field identification. The results support the recognition of an undescribed V. arvensis eco-phenotype linked to seminatural dry grasslands, easily distinguishable from the field-grown type of V. arvensis but hardly distinguishable from the dwarf pansy. Data further corroborate the assumption of general deep-rooted confusion in ascribing poorly developed individuals of V. arvensis to the rare and locally threatened V. kitaibeliana, leading to potential implications for its conservation.