Multivariate morphometrics and an assessment of genetic diversity obtained using amplified fragment length polymorphism (AFLP) were used to determine the variability of the polymorphic group Cyanus triumfetti in Central Europe. The ploidy level of the populations studied was also determined; all individuals from the C. triumfetti group were diploid (2n ~ 2x ~ 22) and all those of the related C. montanus group were tetraploid (2n ~ 4x ~ 44). A multivariate morphometric study of 71 populations revealed that three species from the C. triumfetti group occur in Central Europe, namely ‘Cyanus axillaris’, C. strictus and C. dominii. Three subspecies are recognized within the latter species, namely C. dominii subsp. dominii, C. dominii subsp. slovenicus and C. dominii subsp. sokolensis. Morphological characters of leaves are the best features for delimiting these taxa; a shift in characters caused by cultivation did not affect the value of key characters and differences among the taxa remained. AFLP analysis of 38 populations from the C. triumfetti group and two from the C. montanus group revealed a contrasting pattern of genetic variation that was related to the geographic distribution of the populations rather than the morphological variation in the C. triumfetti group. The AFLP data revealed the following three genetically differentiated and allopatric groups: (i) C. triumfetti s.s. and C. montanus from the Western Alps, (ii) ‘C. axillaris’ from Austria and the Czech Republic (except the Carpathians) and (iii) ‘C. axillaris’, C. strictus and C. dominii from the Western Carpathians and Pannonia. The striking genetic gap between the Austrian-Czech and the W Carpathian-Pannonia groups and the high genetic diversity and weak genetic differentiation within the latter group are discussed in the light of potential glacial refuges, postglacial migration routes and/or the probability of hybridization events occurring during the evolutionary history of this group. An identification key for the taxa of the C. triumfetti group in Central Europe is presented.
Gymnadenia densiflora was recently either misinterpreted or not accepted as a distinct taxon by several authors. To resolve its taxonomic position and differentiation from the related G. conopsea, a detailed study of the morphology, chromosome numbers and distribution of these two taxa in the Czech Republic, Slovakia and neighbouring areas was carried out. Chromosome counts showed an invariable diploid chromosome number (2n = 40) for G. densiflora, while G. conopsea is diploid, tetraploid and rarely also pentaploid (2n = 40, 80, 100). Results of morphometric analyses (principal component analysis, cluster analysis, classificatory and canonical discriminant analysis) confirmed a good morphological separation between G. densiflora and G. conopsea. Characters such as the width of the second lowermost leaf, height of the plant, number of flowers in the inflorescence, number of leaves, and the ratio of height of the plant and distance from the stem base to the base of the uppermost sheathed leaf contributed most to this separation. Our study supports the recognition of G. densiflora as a distinct species.