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A novel species, Gymnadenia winkeliana, has been identified in the Bucegi Natural Park ROSCI0013, located in the Southern Carpathians of Central Romania. Two moderately sized populations of Gymnadenia winkeliana, totalling 120–140 individuals, were discovered inhabiting the alpine grasslands of the park, situated 2.000 m above sea level. To describe this newly found population as comprehensively as possible, 44 vegetative and floral organs/organ parts were directly studied and measured from living plants. Special attention was focused on the characteristics that proved to have taxonomic significance, particularly those involving distinctive details in the morphology of the leaves, perianth, labellum and gynostemium. A total of 223 characteristics were analysed encompassing the morphology of every organ of the plant, cytology and breeding system. Furthermore, comprehensive taxonomic treatment and description, accompanied by colour photographs illustrating the holotype, are provided. Voucher specimens were deposited at the Herbarium of the University of Agriculture and Veterinary Medicine, Bucharest (USAMVB Herbarium barcode: 40102, NEA); Gymnadenia winkeliana, a (micro)endemic species, is characterized as a putative allogamous, facultatively apomict that significantly differs from other Gymnadenia R.Br. species found in Romania. Notably, it distinguishes itself through its smaller habitus (reaching heights of up to 8–10 cm), its two-coloured, rounded/hemispherical inflorescence displaying a gradient of pink hues in an acropetal fashion (ranging from whitish-pink at the base to vivid-pink at the topmost flowers), and its limited distribution in high-altitude areas, encompassing approximately 8–10 km2 in the central area of the Bucegi Natural Park. This species has been under observation since 2005, with observed population numbers showing a significant increase over time, from ca. 50–55 (counted at the time of its discovery) to 120–140 individuals (counted in June 2023). Additionally, comprehensive information regarding the habitat, ecology, phenology and IUCN conservation assessments of Gymnadenia winkeliana are provided, including maps illustrating its distribution. [ABSTRACT FROM AUTHOR]

This study presents an updated analysis of cytogenetic data for several species within the 40,42-chromosome genera of the subtribe Orchidinae. The research includes insights into the distribution of heterochromatin obtained using C-banding and fluorochrome techniques. Our investigation confirmed variation in the distribution of heterochromatin and repetitive DNA sequences among species pertaining to Neotinea s.l. and Orchis s.str. These variations also potentially contribute to the diversification of these species. Cytogenetic analyses of the Neotinea group demonstrated that both H33258 and DAPI staining result in blocks of fluorescent regions on numerous chromosomes. Particular attention was paid to the cytological composition of the polyploid Neotinea commutata, focusing on its potential origin. Based on the karyological results acquired, a hypothesis concerning the origin of N. commutata is proposed. The most noteworthy revelations regard the O. mascula complex. In these species, the telomeric areas of all chromosome sets display extensive heterochromatin. Fluorochrome staining revealed telomeric blocks on many chromosomes that were not seen with Giemsa staining. This highlighted a distinct feature of O. mascula, where particularly large C-bands surrounding the centromeric regions of multiple chromosomes were found. However, in O. mascula, O. provincialis, O. pauciflora, and O. patens, C+ chromatin may not show a significant response to fluorochrome Hoechst or DAPI+ staining. The unique cytomorphological arrangement observed in the O. mascula species, unlike other members of the O. mascula complex, suggest epigenetic phenomena. Additional data are presented for the genera Dactylorhiza and Gymnadenia. A deeper understanding of the diversity of chromosomal structures among these orchids promises to shed light on the mechanisms underlying speciation, adaptation, and the remarkable diversity characteristic of the Orchidaceae family. [ABSTRACT FROM AUTHOR]

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Copyright of Flora Montibérica is the property of Flora Montiberica and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

A novel species, Gymnadenia winkeliana, has been identified in the Bucegi Natural Park ROSCI0013, located in the Southern Carpathians of Central Romania. Two moderately sized populations of Gymnadenia winkeliana, totalling 120–140 individuals, were discovered inhabiting the alpine grasslands of the park, situated 2.000 m above sea level. To describe this newly found population as comprehensively as possible, 44 vegetative and floral organs/organ parts were directly studied and measured from living plants. Special attention was focused on the characteristics that proved to have taxonomic significance, particularly those involving distinctive details in the morphology of the leaves, perianth, labellum and gynostemium. A total of 223 characteristics were analysed encompassing the morphology of every organ of the plant, cytology and breeding system. Furthermore, comprehensive taxonomic treatment and description, accompanied by colour photographs illustrating the holotype, are provided. Voucher specimens were deposited at the Herbarium of the University of Agriculture and Veterinary Medicine, Bucharest (USAMVB Herbarium barcode: 40102, NEA); Gymnadenia winkeliana, a (micro)endemic species, is characterized as a putative allogamous, facultatively apomict that significantly differs from other Gymnadenia R.Br. species found in Romania. Notably, it distinguishes itself through its smaller habitus (reaching heights of up to 8–10 cm), its two-coloured, rounded/hemispherical inflorescence displaying a gradient of pink hues in an acropetal fashion (ranging from whitish-pink at the base to vivid-pink at the topmost flowers), and its limited distribution in high-altitude areas, encompassing approximately 8–10 km2 in the central area of the Bucegi Natural Park. This species has been under observation since 2005, with observed population numbers showing a significant increase over time, from ca. 50–55 (counted at the time of its discovery) to 120–140 individuals (counted in June 2023). Additionally, comprehensive information regarding the habitat, ecology, phenology and IUCN conservation assessments of Gymnadenia winkeliana are provided, including maps illustrating its distribution.

On the basis of numerous own observations, occasionally with information from third parties and with selected information from relevant literature, all orchid species of the island of Krk mentioned or recorded so far are described in detail. The basis for this is more than 2,000 entries of documented occurrences on the island of Krk, which were taken from our „data treasure“ with more than 12,000 entries on the orchid flora of the northeastern Adriatic (period of the years 1999 to 2022). The distribution maps, also based mainly on our own data, were supplemented by reports of findings in the Flora of Istria and, in some cases, with information from the Flora Croatica Database, thus providing an almost complete representation of the currently known distribution picture. [ABSTRACT FROM AUTHOR]

The SPME-GC-MS analysis of the aroma components of Gymnadenia conopsea subsp. conopsea, subsp. densiflora, var. alpina and Gymnadenia odoratissima var. odoratissima, var. idae were reported. The main components of in total 78 found in G. conopsea subsp. conopsea were elemicin, cis-9-hexadecenal, hexadecanal, isoelemicin and (Z)-11-hexadecen-1-ol acetate; in subsp. densiflora benzyl benzoate, eugenol and trans-isoeugenol; in var. alpina benzyl benzoate, methyleugenol and elemicin. In the scent of G. odoratissima var. odoratissima were found 2-phenylethyl acetate, eugenol and pentadecane, in var. idae mainly C15–C21 alkanes and C16, C18 carbonic acids and some isoprenoid-derivatives. As all tested Gymnadenia-taxa are allogamous, the differences in scent composition may play a role in pollinator attraction. [ABSTRACT FROM AUTHOR]

Pollinator sharing between close relatives can be costly and can promote pollination niche partitioning and floral divergence. This should be reflected by a higher species divergence in sympatry than in allopatry. We tested this hypothesis in two orchid congeners with overlapping distributions and flowering times. We characterized floral traits and pollination niches and quantified pollen limitation in 15 pure and mixed populations, and we measured phenotypic selection on floral traits and performed controlled crosses in one mixed site. Most floral traits differed between species, yet pollinator sharing was extensive. Only the timing of scent emission diverged more in mixed sites than in pure sites, and this was not mirrored by the timing of pollinator visitation. We did not detect divergent selection on floral traits. Seed production was pollen limited in most populations but not more severely in mixed sites than in pure sites. Interspecific crosses produced the same or a higher proportion of viable seeds than intraspecific crosses. The two orchid species attract the same pollinator species despite showing divergent floral traits. However, this does not promote character displacement, implying a low cost of pollinator sharing. Our results highlight the importance of characterizing both traits and ecological niches in character displacement studies. [ABSTRACT FROM AUTHOR]

Abstract Closely related species often differ in traits that influence reproductive success, suggesting that divergent selection on such traits contribute to the maintenance of species boundaries. Gymnadenia conopsea ss. and Gymnadenia densiflora are two closely related, perennial orchid species that differ in (a) floral traits important for pollination, including flowering phenology, floral display, and spur length, and (b) dominant pollinators. If plant–pollinator interactions contribute to the maintenance of trait differences between these two taxa, we expect current divergent selection on flowering phenology and floral morphology between the two species. We quantified phenotypic selection via female fitness in one year on flowering start, three floral display traits (plant height, number of flowers, and corolla size) and spur length, in six populations of G. conopsea s.s. and in four populations of G. densiflora. There was indication of divergent selection on flowering start in the expected direction, with selection for earlier flowering in two populations of the early‐flowering G. conopsea s.s. and for later flowering in one population of the late‐flowering G. densiflora. No divergent selection on floral morphology was detected, and there was no significant stabilizing selection on any trait in the two species. The results suggest ongoing adaptive differentiation of flowering phenology, strengthening this premating reproductive barrier between the two species. Synthesis: This study is among the first to test whether divergent selection on floral traits contribute to the maintenance of species differences between closely related plants. Phenological isolation confers a substantial potential for reproductive isolation, and divergent selection on flowering time can thus greatly influence reproductive isolation and adaptive differentiation.

Circumscriptions of both the genus Gymnadenia and the 11–27 species that it contains are highly controversial. These Eurasian terrestrial orchids are nectar-rewarding and pollinated primarily by Lepidoptera. Opinions expressed on the number of species occurring in the British Isles range from one to four, though there exists broad agreement that at least three recognizable, ecologically differentiated taxa are widespread. Here, we use a large-scale morphometric survey of these supposedly 'cryptic' taxa to determine whether phenotypic differentiation exists alongside their documented genotypic and ecological differentiation, seeking the most diagnostic morphological characters. Ten Gymnadenia plants were measured in each of 29 populations that encompassed the entire taxonomic range and geographic distribution of the genus within the British Isles. Results were subjected to detailed multivariate and univariate analyses, and interpreted in the context of molecular phylogenies, including a bespoke Europe-wide nrITS phylogeny that includes 17 plants sampled across Britain and Ireland. Floral micromorphology was investigated through scanning electron microscopy. Ecotypes occurring respectively in calcareous grasslands (G. conopsea s.s.), acid heaths (G. borealis) and calcareous to neutral marshes (G. densiflora) are subtly but reliably distinct, both morphologically and molecularly, though locally their morphological distinctiveness is somewhat weakened by the occurrence of infrequent populations in dune slacks and chalk downs that are intermediate in phenotype between the grassland and marsh ecotypes ('subsp. cf. friesica'). A polythetic taxonomic key emphasizes the characters and character-state ranges that are demonstrably the most diagnostic. Given that they are reliably genetically differentiable, and individuals can be distinguished with ∼85% confidence using morphological characters such as flower dimensions, depth of flower colour, and leaf number and size, we uphold our previous arguments that the three ecotypes merit species-level recognition. In the continued absence of field sequencing devices, identification is best attempted through quantitative examination of morphology focused at the population level, rejecting records of isolated plants. Despite their strong morphological similarity, genus-wide molecular data show that the British species are not each other's closest relatives. Gymnadenia borealis remains one of only three orchid species putatively endemic to the British Isles. [ABSTRACT FROM AUTHOR]

Recent phylogenetic trees of the phenotypically diverse Eurasian orchid genus Gymnadenia s.l., based on Sanger sequencing of nrITS and next-generation RAD-seq data, largely agree on species boundaries but disagree radically regarding relationships among those species. Moreover, both contrasting topologies receive support from a recent transcriptome study. But these genetic studies are not matched by a taxonomically comprehensive analysis of phenotype. Population-level in situ morphometric analyses of 41 characters for 150 plants spanning seven putative species are used to determine overall phenetic similarities of the species, thereby allowing both phenotypic species circumscription and comparison of estimated phenotypic and genotypic disparities. The three widely recognized short-spurred lineages prove highly morphometrically divergent from each other and from the long-spurred species, which differ from each other only subtly. Phenetic disparity correlates strongly with genetic disparity for RAD-seq data but not with that for nrITS data. The RAD-seq topology is appealing because it implies a fractal evolutionary pattern, but it also requires that later-divergent, recently evolved species are far more geographically widespread than their putative antecedents. Current knowledge of this intensively studied genus allows stronger conclusions on circumscription of taxa than on determining their evolutionary relationships; the clade is unequivocally a single genus that, on present evidence, consists of approximately 12 species. I emphasize the importance of rigorously describing and analysing phenotype, of tree rooting, and of critically appraising reconstructed phylogenies, irrespective of the robustness of the data and the determination with which they are analysed. [ABSTRACT FROM AUTHOR]

Closely related species often differ in traits that influence reproductive success, suggesting that divergent selection on such traits contribute to the maintenance of species boundaries. Gymnadenia conopsea ss. and Gymnadenia densiflora are two closely related, perennial orchid species that differ in (a) floral traits important for pollination, including flowering phenology, floral display, and spur length, and (b) dominant pollinators. If plant–pollinator interactions contribute to the maintenance of trait differences between these two taxa, we expect current divergent selection on flowering phenology and floral morphology between the two species. We quantified phenotypic selection via female fitness in one year on flowering start, three floral display traits (plant height, number of flowers, and corolla size) and spur length, in six populations of G. conopsea s.s. and in four populations of G. densiflora. There was indication of divergent selection on flowering start in the expected direction, with selection for earlier flowering in two populations of the early‐flowering G. conopsea s.s. and for later flowering in one population of the late‐flowering G. densiflora. No divergent selection on floral morphology was detected, and there was no significant stabilizing selection on any trait in the two species. The results suggest ongoing adaptive differentiation of flowering phenology, strengthening this premating reproductive barrier between the two species. Synthesis: This study is among the first to test whether divergent selection on floral traits contribute to the maintenance of species differences between closely related plants. Phenological isolation confers a substantial potential for reproductive isolation, and divergent selection on flowering time can thus greatly influence reproductive isolation and adaptive differentiation. [ABSTRACT FROM AUTHOR]

The orchids (Orchidaceae) constitute one of the largest and most diverse families of flowering plants. They have evolved a great variety of adaptations to achieve pollination by a diverse group of pollinators. Many orchids reward their pollinators, typically with nectar, but the family is also well-known for employing deceptive pollination strategies in which there is no reward for the pollinator, in the most extreme case by mimicking sexual signals of pollinators. In the European flora, two examples of these different pollination strategies are the sexually deceptive genus Ophrys and the rewarding genus Gymnadenia, which differ in their level of pollinator specialization; Ophrys is typically pollinated by pseudo-copulation of males of a single insect species, whilst Gymnadenia attracts a broad range of floral visitors. Here, we present and describe the annotated floral transcriptome of Ophrys iricolor, an Andrena-pollinated representative of the genus Ophrys that is widespread throughout the Aegean. Furthermore, we present additional floral transcriptomes of both sexually deceptive and rewarding orchids, specifically the deceptive Ophrys insectifera, Ophrys aymoninii, and an updated floral transcriptome of Ophrys sphegodes, as well as the floral transcriptomes of the rewarding orchids Gymnadenia conopsea, Gymnadenia densiflora, Gymnadenia odoratissima, and Gymnadenia rhellicani (syn. Nigritella rhellicani). Comparisons of these novel floral transcriptomes reveal few annotation differences between deceptive and rewarding orchids. Since together, these transcriptomes provide a representative sample of the genus-wide taxonomic diversity within Ophrys and Gymnadenia (Orchidoideae: Orchidinae), we employ a phylogenomic approach to address open questions of phylogenetic relationships within the genera. Specifically, this includes the controversial placement of O. insectifera within the Ophrys phylogeny and the placement of “Nigritella”-type morphologies within the phylogeny of Gymnadenia. Whereas in Gymnadenia, several conflicting topologies are supported by a similar number of gene trees, a majority of Ophrys gene topologies clearly supports a placement of O. insectifera as sister to a clade containing O. sphegodes.

Aim: Interactions with mycorrhizal fungi are increasingly recognized as an important factor underlying the distribution and abundance of orchid species. However, the geographical distribution of orchid mycorrhizal fungi (OMF) and how their communities vary over large geographical areas are less well understood. Because climatic and environmental similarity may decrease with geographical distance or because some OMF have limited dispersal capabilities, similarities in orchid mycorrhizal communities can be expected to decrease with increasing distances separating orchid populations. However, up till now empirical evidence is largely lacking. Location: Eurasia. Taxa: Gymnadenia conopsea (L.) R. Brown and Epipactis helleborine (L.) Crantz. Methods: High‐throughput sequencing was used to perform a cross‐continental comparison of OMF that associate with two widespread Eurasian terrestrial orchids, Epipactis helleborine and Gymnadenia conopsea. Both phylogenetic and nonphylogenetic measures of community dissimilarity and their components were calculated and related to geographical distances using Mantel tests. Results: Our results showed that in both orchid species similarity in mycorrhizal communities decreased significantly with geographical distance. Decomposing the contribution of spatial turnover and nestedness to overall dissimilarity showed that the observed dissimilarity was mainly the result of species replacement between regions, and not of species loss. Similarly, a strong relationship was observed between phylogenetic community dissimilarity and geographical distance. Decomposing PCD values into a nonphylogenetic and phylogenetic component showed that orchid populations located closely next to each other were likely to contain the same operational taxonomic units (OTUs), but that the non‐shared taxa came from different phylogenetic clades. Species indicator analyses showed that the majority of OMF OTUs were restricted to particular geographical areas. However, some OTUs occurred in both continents, indicating that some fungi have very wide distributions. Main conclusions: Overall, these results demonstrate that orchid mycorrhizal communities differ substantially across large geographical areas, but that the distribution of orchids is not necessarily restricted by the distribution of particular OMF. Hence, widespread orchid species can be considered mycorrhizal generalists that are flexible in the OMF with which they associate across large geographical areas. [ABSTRACT FROM AUTHOR]

The orchids (Orchidaceae) constitute one of the largest and most diverse families of flowering plants. They have evolved a great variety of adaptations to achieve pollination by a diverse group of pollinators. Many orchids reward their pollinators, typically with nectar, but the family is also well-known for employing deceptive pollination strategies in which there is no reward for the pollinator, in the most extreme case by mimicking sexual signals of pollinators. In the European flora, two examples of these different pollination strategies are the sexually deceptive genus Ophrys and the rewarding genus Gymnadenia , which differ in their level of pollinator specialization; Ophrys is typically pollinated by pseudo-copulation of males of a single insect species, whilst Gymnadenia attracts a broad range of floral visitors. Here, we present and describe the annotated floral transcriptome of Ophrys iricolor , an Andrena- pollinated representative of the genus Ophrys that is widespread throughout the Aegean. Furthermore, we present additional floral transcriptomes of both sexually deceptive and rewarding orchids, specifically the deceptive Ophrys insectifera , Ophrys aymoninii , and an updated floral transcriptome of Ophrys sphegodes, as well as the floral transcriptomes of the rewarding orchids Gymnadenia conopsea, Gymnadenia densiflora, Gymnadenia odoratissima , and Gymnadenia rhellicani (syn. Nigritella rhellicani). Comparisons of these novel floral transcriptomes reveal few annotation differences between deceptive and rewarding orchids. Since together, these transcriptomes provide a representative sample of the genus-wide taxonomic diversity within Ophrys and Gymnadenia (Orchidoideae: Orchidinae), we employ a phylogenomic approach to address open questions of phylogenetic relationships within the genera. Specifically, this includes the controversial placement of O. insectifera within the Ophrys phylogeny and the placement of " Nigritella "-type morphologies within the phylogeny of Gymnadenia. Whereas in Gymnadenia , several conflicting topologies are supported by a similar number of gene trees, a majority of Ophrys gene topologies clearly supports a placement of O. insectifera as sister to a clade containing O. sphegodes. [ABSTRACT FROM AUTHOR]

• RADseq supports a sister relationship of the orchid genera Gymnadenia and Nigritella. • Restricted gene flow between the two genera may explain discordant tree topologies. • An origin of Gymnigritella runei from extant G. conopsea and N. nigra is supported. • The separation of G. densiflora from G. conopsea s.str. is supported. • Affiliation of Nigritella polyploid taxa into three distinct groups is supported. The orchid genus Nigritella is closely related to Gymnadenia and has from time to time been merged with the latter. Although Nigritella is morphologically distinct, it has been suggested that the separating characters are easily modifiable and subject to rapid evolutionary change. So far, molecular phylogenetic studies have either given support for the inclusion of Nigritella in Gymnadenia , or for their separation as different genera. To resolve this issue, we analysed data obtained from Restriction-site associated DNA sequencing, RADseq, which provides a large number of SNPs distributed across the entire genome. To analyse samples of different ploidies, we take an analytical approach of building a reduced genomic reference based on de novo RADseq loci reconstructed from diploid accessions only, which we further use to map and call variants across both diploid and polyploid accessions. We found that Nigritella is distinct from Gymnadenia forming a well-supported separate clade, and that genetic diversity within Gymnadenia is high. Within Gymnadenia , taxa characterized by an ITS-E ribotype (G. conopsea s.str. (early flowering) and G. odoratissima), are divergent from taxa characterized by ITS-L ribotype (G. frivaldii, G. densiflora and late flowering G. conopsea). Gymnigritella runei is confirmed to have an allopolyploid origin from diploid Gymnadenia conopsea and tetraploid N. nigra ssp. nigra on the basis of RADseq data. Within Nigritella the aggregation of polyploid members into three clear-cut groups as suggested by allozyme and nuclear microsatellite data was further supported. [ABSTRACT FROM AUTHOR]

Seeds of 21 taxa of the genera Dactylorhiza (incl. Coeloglossum), Gymnadenia (incl. Nigritella) and Pseudorchis were examined by light microscope and SEM. Qualitative and quantitative characters were analyzed. In Dactylorhiza, the seeds are fusiform, but some populations of D. viridis show clavate seeds. According to the ornamentation of the periclinal walls, 3 types of seeds are recognized in this genus. The considerable variation in the seed coat ornamentation pattern in the taxa belonging to majalis, maculata, and praetermissa groups of the genus Dactylorhiza is congruent with the genetic processes that occurred during the history of this genus. Gymnadenia shows clavate seeds with stout and straight to slightly curved anticlinal walls, although these are straight to wavy in some taxa considered previously as Nigritella. These taxa also show low values of seed length. Pseudorchis has fusiform seeds without ornamentation in the periclinal walls and fine anticlinal walls. Our study supports the inclusion of the former genus Coeloglossum in Dactylorhiza. [ABSTRACT FROM AUTHOR]

Floral signaling, especially through floral scent, is often highly complex, and little is known about the molecular mechanisms and evolutionary causes of this complexity. In this study, we focused on the evolution of 'floral scent genes' and the associated changes in their functions in three closely related orchid species of the genus Gymnadenia. We developed a benchmark repertoire of 2,571 expressed sequence tags (ESTs) in Gymnadenia odoratissima. For the functional characterization and evolutionary analysis, we focused on eugenol synthase, as eugenol is a widespread and important scent compound. We obtained complete coding complementary DNAs (cDNAs) of two copies of putative eugenol synthase genes in each of the three species. The proteins encoded by these cDNAs were characterized by expression and testing for activity in Escherichia coli. While G. odoratissima and Gymnadenia conopsea enzymes were found to catalyze the formation of eugenol only, the Gymnadenia densiflora proteins synthesize eugenol, as well as a smaller amount of isoeugenol. Finally, we showed that the eugenol and isoeugenol producing gene copies of G. densiflora are evolutionarily derived from the ancestral genes of the other species producing only eugenol. The evolutionary switch from production of one to two compounds evolved under relaxed purifying selection. In conclusion, our study shows the molecular bases of eugenol and isoeugenol production and suggests that an evolutionary transition in a single gene can lead to an increased complexity in floral scent emitted by plants. [ABSTRACT FROM AUTHOR]

Background and Aims Patterns of ploidy variation among and within populations can provide valuable insights into the evolutionary mechanisms shaping the dynamics of plant systems showing ploidy diversity. Whereas data on majority ploidies are, by definition, often sufficiently extensive, much less is known about the incidence and evolutionary role of minority cytotypes. Methods Ploidy and proportions of endoreplicated genome were determined using DAPI (4',6-diamidino-2-phenylindole) flow cytometry in 6150 Gymnadenia plants (fragrant orchids) collected from 141 populations in 17 European countries. All widely recognized European species, and several taxa of less certain taxonomic status were sampled within Gymnadenia conopsea sensu lato. Key Results Most Gymnadenia populations were taxonomically and/or ploidy heterogeneous. Two majority (2x and 4x) and three minority (3x, 5x and 6x) cytotypes were identified. Evolution largely proceeded at the diploid level, whereas tetraploids were much more geographically and taxonomically restricted. Although minority ploidies constituted <2 % of the individuals sampled, they were found in 35 % of populations across the entire area investigated. The amount of nuclear DNA, together with the level of progressively partial endoreplication, separated all Gymnadenia species currently widely recognized in Europe. Conclusions Despite their low frequency, minority cytotypes substantially increase intraspecific and intrapopulation ploidy diversity estimates for fragrant orchids. The cytogenetic structure of Gymnadenia populations is remarkably dynamic and shaped by multiple evolutionary mechanisms, including both the ongoing production of unreduced gametes and heteroploid hybridization. Overall, it is likely that the level of ploidy heterogeneity experienced by most plant species/populations is currently underestimated; intensive sampling is necessary to obtain a holistic picture. [ABSTRACT FROM AUTHOR]

On the basis of the material that is in the Herbarium of "Alexandru Ioan Cuza" University of Iaşi [I], the paper presents the taxonomic and chorologic revision of 5 genera of Orchidaceae family. There were revised 276 herbarium sheets with specimens collected from different parts from Romania during 1894-2005. In the collection of the herbarium are: 133 specimens of Gymnadenia conopsea, 1 specimen of Gymnadenia odoratissima, 9 specimens of Listera cordata, 33 specimens of Listera ovata, 7 specimens of Nigritella nigra subsp. nigra, 12 specimens of Nigritella nigra subsp. rubra, 61 specimens of Platanthera bifolia, 5 specimens of Platanthera chlorantha, 15 specimens of Pseudorchis albida. Also, it was analyzed the dynamics of species entry into the herbarium collection: the most part of the herbarium material was collected in period 1961-1980. [ABSTRACT FROM AUTHOR]

Thoroughly sampled molecular phylogenies of the dominantly European orchid subtribe Orchidinae were used to identify a pair and a triplet of recently diverged species in which: (1) divergence involved substantial changes in floral morphology, particularly in the labellar lobes and spur; and (2) the polarity of those changes could be inferred phylogenetically. Floral ontogeny in the selected species was documented in detail through macromorphological, light microscopic, and scanning electron microscopic study of a wide range of ontogenetic stages. All study species showed differentiation of perianth segments earlier than the gynostemium. Unsurprisingly, component parts of the basic floral organs (gymnostemial auricles and rostellum, labellar lateral lobes, and spur) were initiated relatively late, the spur and ovary continuing to expand beyond anthesis. The predominant evolutionary pattern identified in the two case studies was paedomorphosis via progenesis (earlier offset of growth); this credibly explained the reduction in spur size and lateral lobing of the labellum in Gymnadenia odoratissima and, especially, G. austriaca relative to G. conopsea. Loss of resupination in G. austriaca was best viewed as the deletion of a formerly terminal ontogenetic stage. Radical reduction of the spur of Dactylorhiza viridis relative to D. fuchsii was also attributed to progenesis, although the long, narrow outline and relatively short central lobe of its labellum were attributed to increased growth of the lateral lobes (i.e. hypermorphosis resulting in peramorphosis). Microscopic study of epidermal cell types on the labellum and spur suggested a degree of decoupling of micromorphological from macromorphological transitions, although both were subject to heterochronic shifts. Each of the two case studies was consistent with, but not proof of, saltational macroevolution operating via functional changes in one or more key developmental genes. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 157, 429–454. [ABSTRACT FROM AUTHOR]

Previous research has shown that the presence of a reward on average doubles reproductive success in orchids by correspondingly increasing the frequency of pollinator visitations. We examined whether such reward-induced increases have a downstream effect on the behaviour of ambush predators concealed in orchid inflorescences, extending observations begun at Downe by Charles Darwin. Specifically, we studied three orchid-rich sites in southern England, in order to compare the occurrence of crab spiders on three coexisting species of terrestrial orchids: the nectariferous Gymnadenia conopsea versus the nectar-less Dactylorhiza fuchsii and Anacamptis pyramidalis. No significant difference was observed between rewarding and non-rewarding inflorescences at Risborough, whereas at Aston Clinton the nectar-less species supported significantly more crab spiders, albeit mainly in relatively short grass. Comparison of the two non-rewarding species present at Downe approximated a significant preference by the spiders for Anacamptis, which more closely resembles the rewarding Gymnadenia. The presence of a floral reward does not result in a higher frequency of crab spiders. We speculate that concealment quality of the inflorescence, the nature of the dominant pollinator(s), and/or ease of movement of spiders between inflorescences may be more important than frequency of visits by potential prey, particularly when no other form of concealment is readily available. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society, 2007, 92, 763–771. [ABSTRACT FROM AUTHOR]

Although originally ascribed to the genus Gymnadenia R. Br. (Orchidinae: Orchidaceae), the Balkan endemic orchid (2. frivaldii Hampe ex Griseb. has since been more frequently assigned to Pseudorchis Séguier (syn. Leucorchis E. Mey., Bicchia Parl.). Molecular phylogenetic analysis using the ITS region of rDNA reveals a large disparity between the two genera and demonstrates that frivaldii is embedded well within Gymnadenia s.s. Macromorphological and SEM studies further elucidate the floral and vegetative similarities between G. frivaldii and Pseudorchis, notably the heterochronically reduced gynostemium and small, short-spurred labellum; these similarities represent convergent evolutionary transitions, whereas other characters such as contrasting stigma and tuber morphologies provide stronger phylogenetic signals. The sequence-based phylogeny suggests that G. frivaldii represents one of three cases of independent paedomorphic floral reduction inferred in the genus; simplification has been more severe than in G. odoratissima but less severe than in the closely related Gymnadenia subgenus Nigritella. Alternatively, an effectively instantaneous evolutionary origin through hybridisation with a (most likely diploid) species of subgenus Nigritella remains possible. Reports of rare hybridisation between G. frivaldii and members of subgenus Nigritella are acceptably well documented, whereas reports of hybridisation with several other more phylogenetically distant orchid species (including the often sympatric Pseudorchis albida) are considered less secure. [ABSTRACT FROM AUTHOR]

Patterns and levels of genetic diversity mayhave significant influence on the long termpersistence of local populations and revealingsuch information is important in protectingrare species. In this study we investigated thegenetic pattern in five microsatellite lociwithin five Swedish populations of the rareorchid species Gymnadenia odoratissima. Thegeographic distribution of G. odoratissima isrestricted to Europe and in Scandinavia it isonly found in three provinces in southernSweden; Östergötland,Västergötland and on the island ofGotland.Compared with the more widespread congener G.conopsea our results indicate lower levels ofgenetic variation within and higher degrees ofgenetic differentiation among populations of G. odoratissima (HEL = 0.6–0.8 in G. conopseaand 0.3–0.7 in G. odoratissima; FST over allpopulations = 0.06 in G. conopsea and 0.19 in G. odoratissima). Also, we found a cleardistinction among mainland and islandpopulations of G. odoratissima wherepopulations on the island of Gotland seem toexhibit higher levels of gene flow andintragenetic variation, probably as a result ofa larger number of existing populations.Future conservation of this species shouldfocus on facilitation on colonisation events,especially on the mainland, and preservation ofthe genetically more variable Gotlandpopulations. [ABSTRACT FROM AUTHOR]

Ten species of orchid plants belonging to the genera Orchis (7), Dactylorhiza (2), and Gymnadenia (1) were analyzed by enzyme electrophoresis. Each species can be identified by a combination of enzyme bands different from those of all other species examined. The electrophoretic data were used for the construction of phenetic and phylogenetic trees with the help of computer programs. The trees were almost identical regardless which method was used. Our results differ considerably from a classification based on morphological evidence. The electrophoretic data indicate that the genus Orchis is not a monophyletic group. [ABSTRACT FROM AUTHOR]