Results of NewHybrids - engiadina group

Results of the hybrid test conducted in New Hybrids based on 466 amplified fragment-length polymorphism markers for the M. engiadina species group. Individuals were defined as parental species (z1 or z0) or putative hybrids (unknowns; highlighted red). Columns 4 to 9 represent six admixture classes and values are the posterior probabilities of belonging to one of these classes for each individual. P1 = parental species 1; P2 = parental species 2; F1 = first-generation hybrid; F2 = second-generation hybrid; BX1 = backcross to parental species 1; BX2 = backcross to parental species 2. (a) Machilis rubrofusca and sexual M. ticinensis as parental species and just M. engiadina as unknowns. (b) Machilis rubrofusca and sexual M. ticinensis as parental species and all other members of the M. engiadina species group as unknowns

Reanalysis of TM characters including type specimens

Reanalysis of traditional morphometric characters for discriminating among final species hypotheses and classifying type specimens (when available). Classification results based on discriminant functions calculated from an optimal combination of characters (see Table S6 for details), separately for four species groups. We defined these groups according to ovipositor length (i.e., number of articles on gonapophysis VIII). This corresponded to the first steps in the Machilis identification key of Palissa (1964). Type specimens were not included in the training set. Classification success is given as the actual number (absolute) and the percentage (%) of individuals classified in original and leave-one-out cross-validated (LOOCV) discriminant analyses. Correct and wrong classifications are highlighted in green and red, respectively. dis = M. distincta, eng = M. engiadina, rub = M. rubrofusca, tic = M. ticinensis, tir = M. tirolensis, vag = M. vagans, spE = M. sp. E, spF = M. sp. F

Supplementary_material1_TM_dataset

Traditional morphometrics datase

Geographical distributions of M. helleri species group.

Geographical distributions of M. helleri species group. AT = Austria, CZ = Czech Republic, DE = Germany, IT = Italy

Reanalysis of traditional morphometric characters.

Principal component analysis (PCA) scatterplots based on optimal character combinations for each of the four species groups. Whenever available, type specimens were included and are represented by stars

TM character definitions

Definition of characters used in traditional morphometrics analysi

Supplementary_material2_CO1_alignment

Cytochrome Oxidase 1 haplotype alignmen

NJ trees of individual AFLP primer combinations

NJ trees for each of the four primer combinations (each represented by the four dyes FAM, HEX, NED, and PET) used to produce amplified fragment-length polymorphism profiles. Sample names include the following information: ’plate number’_’well number’_’specimen ID’_’species code’

Optimized AFLP scoring parameters

Optimized parameters used for the final scoring of the amplified fragment-length polymorphism profiles. The four dyes (FAM, HEX, NED and PET) represent four different primer combinations

Estimating the number of K from STRUCTURE results.

Graphical representation of Evanno’s ∆K (circles) and the mean marginal likelihood across K (diamonds) from each 10 replicated runs from K=5 to K=27