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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.

Se aportan algunas novedades corológicas y taxonómicas de 16 taxones de orquídeas de la Comunidad Foral de Navarra. Se proponen tres nuevos taxones, uno de ellos a nivel subespecífico y los otros dos hibridógenos.

Four species of Gymnadenia are native to Kosovo: G. conopsea, G. frivaldii, G. nigra, and G. odoratissima. In this study, field expedition data, phytosociological relevés, herbarium specimens along with extensive literature sources were used to analyse vegetation and ecological characteristics, habitat types, distributional patterns as well as provide notes on conservation. Gymnadenia conopsea is distributed throughout the country, from lowlands to the alpine belt. It was recorded in various plant communities. Gymnadenia frivaldii grows in the alpine zone of mountains, close to streams and in wet meadows. Its relevés belong to the class Scheuchzerio-Caricetea fuscae. This species is classified as “Near Threatened” (NT) in Kosovo. Gymnadenia nigra grows in subalpine and alpine grassland on preferably calcareous substrate. It has been found in almost all mountains reaching >2000 m a.s.l., and occurs in different plant communities belonging to the class: Elyno-Seslerietea. Gymnadenia odoratissima was recorded from one locality only in Kosovo, on the massif of Maja e Zezë, Sharri Mts. It was growing in degraded beech forest and meadows on silicate bedrock. Its floristically diverse relevés associate with the class: Mulgedio-Aconitetea. Of the four studied species, G. frivaldii deserves more conservation attention because of its fragile populations.

This study was conducted to investigate the vascular plants which can be an indicator of environmental change in Seseok of Jirisan National Park. The purpose of this study was to find out the basic information for the subalpine species. This study showed that the area had a total 295 taxa including 63 families, 177 genus, 252 species, 2 subspecies, 36 variants 5 forms and Pte-Q was 1.7. Among them, Korea endemic plants were 15 taxa such as Abies koreana. Rare plants were 14 taxa such as Viola diamantiaca. Naturalized plants consisted of 5 taxa such as Rumex crispus. The floristic regional indicator plants found in surveyed area were 72 taxa comprising 3 taxa of grade Ⅴ, 7 taxa of grade Ⅳ and 24 taxa of grade Ⅲ. In summary, The subalpine plants of the survey region consisted of 37 taxa including Abies koreana and Gymnadenia cucullata. As rare plants distributed in subalpine zone respond sensitively to climate change, accuracy distribution range, environmental characteristics of habitat and population dynamics of rare plants should be evaluated quantitatively.

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. ispartof: JOURNAL OF BIOGEOGRAPHY vol:47 issue:2 pages:421-433 status: published

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., We document divergent selection on flowering phenology in two closely related orchids. The results suggest ongoing adaptive differentiation, where selection is strengthening divergence in flowering phenology, an important premating reproductive barrier between the two species.

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.

A Frivaldszky-bibircsvirág a Délkeleti-Kárpátok orchideaflórájának egyik ritkasága, amelynek fajneve híres Balkán kutatónk, Frivaldszky Imre (1799–1870) emlékét őrzi. Hegyvidéki lápokon, vízszivárgós, üde helyeken fordul elő a Balkán-félsziget hegyvidékein (Bulgária, Románia, Görögország, Macedónia, Montenegró, Albánia). A fajra a Kárpátokban először Heuffel János bukkant rá a Szárkő-hegységben. Habár a fajt Simonkai 1874-ben gyűjtötte a Retyezátban (Zenóga- és Bukura-tó), a példánya (BP33967) azonosságában bizonytalan volt, ezért az a halvány bibircsvirág (Gymnadenia albida) alá sorolva szerepel monográfiájában. Azóta több helyről előkerült a Retyezátból, így a Peleaga-havasról, a Zănoaga-, Zănoguţa-, Ana- és Bucura-tavak mellől és a Judele-völgyből, továbbá a Szárkő-hegység egy-két pontjáról is. Az utolsó szerzőnek a faj egy öt évtizeddel korábbi, csupán egy fitocönológiai tabellában dokumentált előfordulását sikerült megerősítenie a Hargita hegységben (Keleti-Kárpátok). Ez a lelőhely a faj areájának északi határán helyezkedik el, ezért növényföldrajzi szempontból igazán fontos adat. A közlemény bemutatja a faj felfedezésének történetét és jelenleg ismert elterjedését és veszélyeztetettségi státuszát a Kárpátokban, továbbá a Hargita hegységi élőhelyét.

The digitate-tubered clade (Dactylorhiza s.l. plus Gymnadenia s.l.) within subtribe Orchidinae is an important element of the North-temperate orchid flora and has become a model system for studying the genetic and epigenetic consequences of organism-wide ploidy change. Here, we integrate morphological phylogenetics with Sanger sequencing of nrITS and the plastid region trnL-F in order to explore phylogenetic relationships and phenotypic character evolution within the clade. The resulting morphological phylogenies are strongly incongruent with the molecular phylogenies, instead reconstructing through parsimony the genus-level boundaries recognised by traditional 20th Century taxonomy. They raise fresh doubts concerning whether Pseudorchis is sister to Platanthera or to Dactylorhiza plus Gymnadenia. Constraining the morphological matrix to the topology derived from ITS sequences increased tree length by 20%, adding considerably to the already exceptional level of phenotypic homoplasy. Both molecular and morphological trees agree that D. viridis and D. iberica are the earliest-diverging species within Dactylorhiza (emphasising the redundancy of the former genus Coeloglossum). Morphology and ITS both suggest that the former genus Nigritella is nested within (and thus part of) Gymnadenia, the Pyrenean endemic 'N.' gabasiana apparently forming a molecular bridge between the two radically contrasting core phenotypes. Comparatively short subtending molecular branches plus widespread (though sporadic) hybridisation indicate that Dactylorhiza and Gymnadenia approximate the minimum level of molecular divergence acceptable in sister genera. They share similar tuber morphologies and base chromosome numbers, and both genera are unusually prone to polyploid speciation. Another prominent feature of multiple speciation events within Gymnadenia is floral paedomorphosis. The 'traditional' morphological and candidate-gene approaches to phylogeny reconstruction are critically appraised.

Floral spurs are regarded as features affecting pollinator behaviour. Anacamptis pyramidalis is regarded as a deceitful, non-rewarding orchid species. In the form fumeauxiana, additional spurs occur on the lateral sepals. In this study we analyse micromorphological and ultrastructural floral features and suggest the mechanism of deception in A. pyramidalis and A. pyramidalis f. fumeauxiana. In f. fumeauxiana, the adaxial surface of the lip, the lip calli, the tips of the lateral sepals, the abaxial and adaxial epidermises of the lip spur, and the lateral sepal spur have a secretory function. Numerous stomata were observed on the abaxial surfaces of spurs and sepals. The characteristic features of the ultrastructure of osmophore cells were noted: dense cytoplasm with numerous profiles of ER, mitochondria, plastids with plastoglobuli and tubular structures, a large nucleus, lipid droplets, and vesicles fusing with the plasmalemma. The similarity of the floral morphology and anatomy, the flowering period, and pollinators of A. pyramidalis, A. pyramidalis f. fumeauxiana and Gymnadeniaconopsea suggest a possible food-deceptive mechanism—imitation of nectar presence produced in the spurs of Gymnadenia.

The fragrant orchid Gymnadenia conopsea s.l. is a controversial taxon with two commonly distinguished species, G. conopsea s.str. and G. densiflora. Despite morphological similarity, differentiation between the taxa has been reported for several characters; however, character variation within taxa has obviated a clear consensus. We assessed ITS sequences, microsatellite variation and chromosome numbers on the European scale (1,420 samples) and conducted morphological analyses for 626 samples from Germany. ITS analysis revealed a 2% nucleotide divergence between the taxa, similar to the divergence between other Gymnadenia species. The ITS sequences of G. densiflora form a well-supported monophyletic group sharing a most recent common ancestor with G. nigra and G. austriaca. Thus, G. conopsea and G. densiflora are not sister species, and a species rank is supported for G. densiflora (Wahlenb.) Dietrich and G. conopsea (L.) R.Br. s.str. This was confirmed by the microsatellite analysis, which revealed a strong genetic differentiation between the taxa because of largely non-overlapping sets of alleles. Chromosome numbers showed that G. conopsea was either diploid or tetraploid, whereas G. densiflora was diploid throughout. Morphologically, the taxa differed significantly in the mean value of a number of diagnostic characters. However, a discriminant analysis showed that the morphological variability is substantial, and on the individual level an unequivocal assignment is not possible as 96% of G. conopsea, but only 77% of G. densiflora could be assigned correctly. Further studies are needed on character variation within and among species and ploidy levels to allow for a better identification of the genetically differentiated but morphologically similar taxa.

Different authors have demonstrated the usefulness of single-copy nuclear genes in unravelling the evolution of polyploid species' complexes. We used the chalcone synthase gene family to analyse relationships among several Dactylorhiza species belonging to the main diploid and tetraploid lineages of this genus and to evaluate relationships between Dactylorhiza and other orchid genera that traditionally have been considered to be closely related. Overall, 59 sequences from 19 taxa were analysed using parsimony and Bayesian inference. In our results, we recover a sister relationship between Dactylorhiza and Gymnadenia. In addition, our results support the idea that genera Coeloglossum and Dactylorhiza should be merged into one taxonomic entity. Finally, we add some information on the origin of allotetraploid Dactylorhiza taxa, such as D. praetermissa and D. elata. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 163, 155–165.

Polyploidy has played a key role in plant evolution and diversification. Despite this, the processes governing reproductive isolation among cytotypes growing in mixed-ploidy populations are still largely unknown. Theoretically, coexistence of diploid and polyploid individuals in sympatric populations is unlikely unless cytotypes are prezygotically isolated through assortative pollination. Here, we investigated the pre-mating barriers involved in the maintenance of three co-occurring cytotypes from the genus Gymnadenia (Orchidaceae): tetraploid and octoploid G. conopsea and tetraploid G. densiflora. We assessed differences in flowering phenology, floral morphology, and visual and olfactory cues, which could lead to assortative mating. Gas chromatography coupled with electroantennographic detection was used to identify scent compounds with physiological activity in the two main pollinators, Deilephila porcellus and Autographa gamma. The importance of olfactory cues was also assessed in the field by analysing the moths’ responses to the olfactory display of the plants, and by following the pollinator’s behaviour on artificial arrays. Our complex approach demonstrated that the coexistence of Gymnadenia cytotypes in mixed-ploidy populations was only partly explained by differences in floral phenology, as cytotypes with overlapping flowering (i.e., octoploid G. conopsea and tetraploid G. densiflora) might freely exchange pollen due to only 1 mm differences in spur lengths and the lack of assortative behaviour of pollinators. While floral colour among the cytotypes was similar, floral scent differed significantly. Though both pollinator species seemed to physiologically detect these differences, and the floral scent alone was sufficient to attract them, pollinators did not use this cue to discriminate the cytotypes in the field. The absence of pre-mating barriers among cytotypes, except partial temporal segregation, suggests the existence of other mechanisms involved in the cytotypes’ coexistence. The genetic differences in ITS sequences among cytotypes were used to discuss the cytotype’s origin.

The tropical Asian Brachycorythis helferi complex has presented serious identification problems, even after the genus was monographed in 1955. In the present study, morphological variation in the complex was summarized by principal components analysis (PCA) performed on data from herbarium specimens and probable misidentifications were detected by cluster analysis combined with information from protologues and type specimens. Species delimitations are revised accordingly, leading to recognition of five species: B. henryi, B. neglecta (sp. nov.), B. helferi, B. laotica and B. acuta. Brachycorythis obovalis and B. menglianensis are reduced to synonyms of B. acuta and B. henryi, respectively. Description of B. neglecta from Burma and Thailand leaves B. henryi with a much narrower distribution than previously recognized (apparently restricted to the provinces of Guizhou and Yunnan in China and the Shan State in Burma). The names Gymnadenia sesamoides (a synonym of Brachycorythis helferi) and B. laotica are lectotypified, and the latter species is reported as new to Thailand. A dichotomous key to the five accepted species is provided, together with brief descriptions. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162, 64–76.

The Asiatic orchid species Neolindleya camtschatica (Cham.) Nevski has been omitted from the majority of relevant taxonomic surveys, including the recent Genera Orchidacearum. In most studies where the species has been included, it has been assigned to the species-rich genera Gymnadenia or Platanthera. A few morphologists recognised a new monotypic genus, Neolindleya Kraenzl., to accommodate the species. More recent molecular phylogenetic studies showed barely sufficient molecular disparity to justify generic separation of this species, but demonstrated clearly that the genus Neolindleya is only distantly related to Gymnadenia. However, the molecular phylogenies show relationships of equal strength of Neolindleya with Galearis (including Amerorchis) on the one hand and Platanthera s.l. on the other. In an attempt to better resolve the phylogenetic placement of Neolindleya, and to more clearly understand its biology and ecology, we have re-examined the morphology of this enigmatic species. Our results, based partly on SEM studies, reinforce the validity of Neolindleya as a genus, indicate a closer relationship with Galearis s.l. than with Platanthera s.l., and strongly suggest that the species became an autogam following ‘accidental’ loss of a functional bursicle. Ecologically, N. camtschatica is an opportunistic species that benefits from anthropogenic habitat disturbance.

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.

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. ADDITIONAL KEYWORDS: Anacamptis ‐ crab spider ‐ Dactylorhiza ‐ floral predator ‐ Gymnadenia ‐ nectar ‐ Orchidaceae ‐ tritrophic interaction.

Although originally ascribed to the genus Gymnadenia R. Br. (Orchidinae: Orchidaceae), the Balkan endemic orchid G. frivaldii Hampe ex Griseb. has since been more frequently assigned to Pseudorchis Seguier (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.

Internal transcribed spacer (ITS nuclear rDNA) data have been obtained from 190 terrestrial orchid species, encompassing all genera and the great majority of the widely recognized species of Orchidinae, a heterogeneous selection of species of Habenariinae, and single species of Satyriinae and Disinae (the latter serving as outgroup). The resulting parsimony-based phylogeny reveals 12 well-resolved clades within the Orchidinae, based on Anacamptis s.l., Serapias, Ophrys, Steveniella–Himantoglossum s.l. (including ‘Comperia’ and ‘Barlia’, most species being 2n = 36), Neotinea s.l., Traunsteinera–Chamorchis, Orchis s.s., Pseudorchis–Amerorchis–Galearis–Neolindleya–Platanthera s.l. (most 2n = 42), Dactylorhiza s.l., Gymnadenia s.l. (most 2n = 40, 80), Ponerorchis s.l.–Hemipilia s.l.–Amitostigma–Neottianthe, and Brachycorythis (most 2n = 42). Relationships are less clearly resolved among these 12 clades, as are those within Habenariinae; the subtribe appears either weakly supported as monophyletic or as paraphyletic under maximum parsimony, and the species-rich genus Habenaria is clearly highly polyphyletic. The triphyly of Orchis as previously delimited is confirmed, and the improved sampling allows further generic transfers to Anacamptis s.l. and Neotinea s.l. In addition, justifications are given for: (1) establishing Steveniella as the basally divergent member of an appreciably expanded Himantoglossum that incorporates the former genera ‘Barlia’ and ‘Comperia’, (2) reuniting ‘Piperia’ with a broadly defined Platanthera as section Piperia, necessitating ten new combinations, (3) broadening Ponerorchis to include Chusua, and Hemipilia to include single ‘orphan’ species of Ponerorchis and Habenaria, and (4) recognizing ‘Gymnadenia’camtschatica as the monotypic Neolindleya camtschatica within the Pseudorchis∼Platanthera clade. Few further generic transfers are likely in Orchidinae s.s., but they are anticipated among habenariid genera, on acquisition of additional morphological and molecular evidence; one probable outcome is expansion of Herminium. Species-level relationships are also satisfactorily resolved within most of the major clades of Orchidinae, with the notable exceptions of Serapias, the derived sections of Ophrys, Himantoglossum s.s., some sections within Dactylorhiza, the former genus ‘Nigritella’ (now tentatively placed within Gymnadenia s.l.), Hemipilia s.l., and possibly Ponerorchis s.s. Relationships among the 12 major clades broadly accord with bona fide records of intergeneric hybridization. Current evidence supports the recently recognized 2n = 36 clade; it also indicates a 2n = 40 clade that is further diagnosed by digitate root-tubers, and is derived relative to the recently recognized clade of exclusively Asian genera (Ponerorchis s.l.–Hemipilia s.l.–Amitostigma–Neottianthe). This in turn appears derived relative to the Afro-Asiatic Brachycorythis group; together, these two clades identify the plesiomorphic chromosome number as 2n = 42. If the African genus Stenogolottis is correctly placed as basally divergent within a monophyletic Habenariinae, the tribe Orchideae and subtribes Orchidinae and Habenariinae could all have originated in Africa, though in contrast the Asiatic focus of the basally divergent members of most major clades of Orchidinae suggests an Asiatic radiation of the subtribe. Morphological characters informally ‘mapped’ across the molecular phylogeny and showing appreciable levels of homoplasy include floral and vegetative pigmentation, flower shape, leaf posture, gynostemium features, and various pollinator attractants. Qualitative comparison of, and reciprocal illumination between, degrees of sequence and morphological divergence suggests a nested set of radiations of progressively decreasing phenotypic magnitude. Brief scenarios, both adaptive and non-adaptive, are outlined for specific evolutionary transitions. Recommendations are made for further species sampling, concentrating on Asian Orchidinae (together with the Afro-Asiatic Brachycorythis group) and both Asian and Southern Hemisphere Habenariinae, and adding plastid sequence data. Taxonomic changes listed are: Anacamptis robusta (T.Stephenson) R.M.Bateman, comb. nov., A. fragrans (Pollini) R.M.Bateman, comb. nov., A. picta (Loiseleur) R.M.Bateman, comb. nov., Neotinea commutata (Todari) R.M.Bateman, comb. nov., N. conica (Willdenow) R.M.Bateman, comb. nov., Platanthera elegans Lindley ssp. maritima (Rydberg) R.M.Bateman, comb. nov., P. elegans Lindley ssp. decurtata (R.Morgan & Glicenstein) R.M.Bateman, comb. nov., P. elongata (Rydberg) R.M.Bateman, comb. nov., P. michaelii (Greene) R.M.Bateman, comb. nov., P. leptopetala (Rydberg) R.M.Bateman, comb. nov., P. transversa (Suksdorf) R.M.Bateman, comb. nov., P. cooperi (S.Watson) R.M.Bateman, comb. nov., P. colemanii (R.Morgan & Glicenstein) R.M.Bateman, comb. nov., P. candida (R.Morgan & Ackerman) R.M.Bateman, comb. nov. and P. yadonii (R.Morgan & Ackerman) R.M.Bateman, comb. nov. © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society, 2003, 142, 1–40.

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.

Gymnadenia conopsea s.l. is a common orchid in central Europe, where early- and late-flowering populations can be distinguished. The early-flowering form is recognized as subspecies conopsea and the late-flowering form as subspecies densiflora. The two subspecies can occur in sympatry, but their flowering periods are separated. We investigated whether early- and late-flowering subspecies are genetically differentiated, whether they diverged once or repeatedly, and we tried to identify potential evolutionary forces involved in the divergence of the two subspecies. We used genetic markers to estimate genetic divergence within and among populations of early- and late-flowering G. conopsea, and to reconstruct their evolutionary history. In addition, we assessed morphological variation between subspecies. Allozyme variation indicated that subspecies conopsea was significantly more variable than ssp. densiflora and that gene flow among populations of ssp. conopsea was higher than among populations of ssp. densiflora. Gene flow between subspecies was low, indicating that the difference in flowering phenology represented an effective barrier to gene flow. A neighbor-joining tree based on allozyme frequencies indicated that early- and late- flowering populations did not diverge repeatedly in sympatry. Levels of cpDNA variation were generally low, even between G. conopsea s.l. and Gymnadenia odoratissima, chosen as an outgroup. Four cpDNA haplotypes were found, which differed only in the number of microsatellite repeats. Their distribution among subspecies of G. conopsea s.l. and G. odoratissima indicates that microsatellite haplotypes have evolved repeatedly, and their occurrence in different taxa thus represents a homoplasy. Floral characters were variable within and among populations and subspecies but did not consistently separate early- from late-flowering populations. A weak separation between subspecies was found in vegetative characters that presumably reflected habitat and competitive differences experienced by early- and late-flowering populations.

Summary Polyploidy is widely recognized as a major mechanism of sympatric speciation in plants, yet little is known about its effects on interactions with other organisms. Mycorrhizal fungi are among the most common plant symbionts and play an important role in plant nutrient supply. It remains to be understood whether mycorrhizal associations of ploidy-variable plants can be ploidy-specific. We examined mycorrhizal associations in three cytotypes (2x, 3x, 4x) of the Gymnadenia conopsea group (Orchidaceae), involving G. conopsea s.s. and G. densiflora, at different spatial scales and during different ontogenetic stages. We analysed: adults from mixed- and single-ploidy populations at a regional scale; closely spaced adults within a mixed-ploidy site; and mycorrhizal seedlings. All Gymnadenia cytotypes associated mainly with saprotrophic Tulasnellaceae (Basidiomycota). Nonetheless, both adults and seedlings of diploids and their autotetraploid derivatives significantly differed in the identity of their mycorrhizal symbionts. Interploidy segregation of mycorrhizal symbionts was most pronounced within a site with closely spaced adults. This study provides the first evidence that polyploidization of a plant species can be associated with a shift in mycorrhizal symbionts. This divergence may contribute to niche partitioning and facilitate establishment and co-existence of different cytotypes.

Gymnadenia conopsea (L.) R. Br. ��� Secteur San Petrone, sentier au N du col de Prato, en direction du Monte Compoli, entre la Bocca di Pruno et la Punta di San Paolo, pr��s du point cot�� 1144, dans une pelouse fraiche �� proximit�� d���une source, 1.7.2010, Delage, A. photographies. Nouvelle station pour cette esp��ce rare en Corse, qui compl��te sa r��partition connue; dans le massif du San Petrone, les stations les plus proches sont situ��es pr��s de Sant���Andrea di Boziu (GAMISANS & al., Candollea 54: 394, 1999) et au Monte tre Pieve (GAMISANS, Candollea 28: 44, 1973); la station d��crite fait le lien entre ces localit��s et celles du massif de Tenda. A. DELAGE, Published as part of Jeanmonod, Daniel & Schl��ssel, Andr��, 2012, Notes et contributions �� la flore de Corse, XXIV, pp. 293-321 in Candollea 67 (2) on page 299, DOI: 10.15553/c2012v672a11, http://zenodo.org/record/5777445

Orchis conopsea Linnaeus, Species Plantarum 2: 942. 1753. “Habitat in Europae pratis montosis.” RCN: 6825. Lectotype (Baumann & al. in Mitteilungsbl. Arbeitskr. Heim. Orchid. Baden-W iirttemberg 2 1: 482, Abb. 18. 1989): [icon] “ Satyrium basilicum mas” in Fuchs, Hist. Stirp.: 711, 712. 1542. Current name: Gymnadenia conopsea (L.) R. Br. (Orchidaceae)., Published as part of Jarvis, Charlie, 2007, pp 705 - 708 in: Order out of Chaos, pp. 705-708 in Order out of Chaos: Linnaean Plant Names and their Types, London :The Linnean Society of London on page 705, DOI: 10.5281/zenodo.250878

Orchis odoratissima Linnaeus, Systema Naturae, ed. 10, 2: 1243. 1759. [“Habitat in Italia, Gallia, Germania, rarissima in Svecia. ”] Sp. PI., ed. 2, 2: 1335 (1763). RCN: 6824. Lectotype (Cribb & Wood in Cafferty & Jarvis in Taxon 48: 48. 1999): Herb. Linn. No. 1054.37 (LINN). Current name: Gymnadenia odoratissima (L.) Rieh. (Orchidaceae)., Published as part of Jarvis, Charlie, 2007, pp 705 - 708 in: Order out of Chaos, pp. 705-708 in Order out of Chaos: Linnaean Plant Names and their Types, London :The Linnean Society of London on page 707, DOI: 10.5281/zenodo.250878

To develop an effective propagation method, a symbiotic culture was attempted in nine species of Japanese terrestrial orchids. Seeds of three species of Goodyera (Goodyera biflora var. macrantha, G. foliosa var. laevis, and G. hachijonesis var. matsumurana), when inoculated non-symbiotically or with fungi from the binucleate Rhizoctonia fungus group among the 20 fungus strains tested germinated with virtually the same TTC activity level as embryos. Those on a symbiotic culture with binucleate Rhizoctonia grew markedly well compared with seeds on a non-symbiotic culture. The low germination rates in six of the species in the TTC test results are attributed to the unsuitability of the test culture conditions. There were numerous effective fungal strains with 5 species other than Cypripedium macranthos var. speciosum, which developed a symbiotic relationship with only one of the fungal isolates tested. Among the 3 species (Aorchis cyclochila, Dactylorhiza aristata, and Gymnadenia camtschatica) the binucleate Rhizoctonia fungal group would be suitable for symbiotic germination and subsequent growth, whereas the 2 species Amitostigma kinoshitae and Ponerorchis graminifolia var. graminifolia. responded better to R. repens

Oecologia, 142 (4), ISSN:0029-8549, ISSN:1432-1939

Using fast-evolving microsatellites, more slowly evolving ITS markers and performing habitat analyses, we demonstrated a drastic genetic divergence and significant habitat differentiation between early- and late-flowering variants of plants morphologically belonging to Gymnadenia conopsea ssp conopsea. The two phenological variants can either be found in separate or in mixed populations. Information from microsatellite markers and ITS sequences indicated the occurrence of an early historical split between the two flowering-time variants, a split that has been maintained until the present time even within sympatric populations. Early-flowering variants were also far more genetically diverse, had more alleles per microsatellite locus and a wider habitat amplitude than late-flowering variants. As a comparison, we included G. odoratissima in the sequencing study. We found G. odoratissima to be most closely related to the early-flowering type. This indicates a more ancient divergence event between the two flowering-time variants within G. conopsea ssp conopsea than between the two different species G. odoratissima and the early-flowering variant of G. conopsea. Possible explanations to the results arrived at and possible mechanisms maintaining the genetic separation are discussed.

Small populations are facing a particular risk of extinction due to a lack of appropriate genetic diversity and associated negative effects, factors dealt with in the discipline of conservation genetics. Many orchid species exhibit characteristics that make them a perfect study object in the scope of conservation genetics. The aim with this thesis was to investigate genetic structure at different levels in two orchid species Gymnadenia conopsea, geographically widespread, although diminishing and G. odoratissima with a long history of being rare. Microsatellite markers, developed in and used in studies of G. conopsea were also used in the study of G. odoratissima. Populations of G. conopsea expressed high levels of genetic variation and a certain amount of gene flow, although investigated mating pattern in a small population indicated non-random mating among individuals, with the majority of pollen exchange between near neighbours, and noticeable levels of geitonogamous pollinations. Further a pronounced year to year variation in flowering frequency among individuals was found. It was also discovered that flowering time variants (early and late) within the species G. conopsea were highly differentiated and seem to have had a more ancient historical separation than the separation between the two different species, G. conopsea and G. odoratissima. Levels of genetic variation in the rare congener, G. odoratissima differed between island and mainland populations where the more numerous island populations expressed larger levels of genetic variation and were less differentiated compared to the few remaining and genetically depauperated mainland populations. Uppsala University Library, Box 510, 75120, Uppsala, Sweden

Gymnadenia R. Br. Handwurz, Nacktdrüse 1. Sporn 11/2-2 mal so lang wie der Fruchtknoten 2. Blätter 0.5-1.5 cm breit; Blüte 5-6 G. conopsea 2*. Blätter 1-2.5 cm breit; Blüte 6-7 G. densiflora 1*. Sporn höchstens so lang wie der Fruchtknoten G. odoratissima Bemerkungen. Bastarde zwischen den Arten sind verbreitet. G. conopsea s.l. wird erst seit kurzer Zeit in die beiden hier erwähnten, miteinander durch Übergänge verbundenen Taxa unterteilt: G. conospea (L.) R.Br. s. str. und G. densiflora Wahlenb. Diese unterscheiden sich neben der Blattbreite vor allem durch die Blütenfarbe (hellviolet - dunkelviolet), durch die Art des Blü- tenstandes (lockerblütig - dichtblütig) und durch die Blütezeit (Mai bis Juni - Juni bis August). Die erste Art wird vor allem durch Tagfalter, die zweite durch Nachtfalter bestäubt. G. conopsea s.l. wird erst seit kurzer Zeit in die beiden hier erwähnten, miteinander durch Übergänge verbundenen Taxa unterteilt: G. conospea (L.) R.Br. s. str. und G. densiflora Wahlenb. Diese unterscheiden sich neben der Blattbreite vor allem durch die Blütenfarbe (hellviolet - dunkelviolet), durch die Art des Blü- tenstandes (lockerblütig - dichtblütig) und durch die Blütezeit (Mai bis Juni - Juni bis August). Die erste Art wird vor allem durch Tagfalter, die zweite durch Nachtfalter bestäubt.

487. Gymnadenia densifloraWahlenb. Dichtblütige Handwurz Blüte. Dunkelviolett bis lila. 6-8. Standort. Kollin bis subalpin. Wechselfeuchte, humusreiche Böden. Riedwiesen, lichte Hangwälder. Literatur. Keine Fundortsangaben, da bis heute nicht unterschieden. Herbar. 2: Kleeweid (1925). 3: Döltschi (1906); Friesenberg (1895). 10: Hönggerberg (1871). Verbreitung. Im Norden selten; im Süden zerstreut; ausserhalb überbauter Zonen. U:4, H/2/2, E: 4, W: I. - Mitteleuropa? Veränderungen. =/>/>. - g, h. - Gefährdung. NT//VU/- Verbreitungsmuster. m., Published as part of Elias Landolt, 2001, Orchidaceae, pp. 232-258 in Flora des Sihltals von der Stadt Zürich bis zum Höhronen, Basel :Fachstelle Naturschutz Kanton Zürich on page 248, DOI: 10.5281/zenodo.248047