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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.

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

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.

Recent molecular phylogenetic studies of the genus Gymnadenia have demonstrated that it contains sibling taxa, i.e., species that are hardly distinguishable according to morphological traits, yet are phylogenetically rather distant and distinctly distinguishable by molecular methods, which is a rare phenomenon for angiosperms. By sequencing the ITS1-5.8S rDNA-ITS2 fragment the presence of G. densiflora was confirmed for Russia. Our data supported a high degree of genetic differentiation between G. conopsea s. str. and G. densiflora, which confirms their taxonomic rank of species. Morphological analysis has shown that the features that are best for the discrimination between these two species in Northwestern Russia are the length of the lower bract, length of the mid-lobe of the lip, and width of leaves. The ecological and phenological differentiation between G. conopsea s. str. and G. densiflora is briefly reviewed. The ITS sequence variation in these species has been analyzed; the molecular genetic differences of the G. conopsea individuals from the eastern part of the distribution area have been discovered for the first time. Different taxonomic interpretations of Gymnadenia phylogenetic tree topology taking into account the presence of sibling species are discussed in general.

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.

Members of the orchid genus Nigritella are widespread in European mountains, but species circumscriptions and evolutionary patterns in the genus are subjects to conflicting opinions. We analyzed a representative material of Nigritella for differentiation at nuclear and plastid marker loci. In agreement with predictions from embryological studies, diploid members of Nigritella are sexual and mostly out-crossing, whereas triploid, tetraploid and pentaploid members are apomicts. The diploid taxa were poorly differentiated in the investigated molecular markers, except for the western N. gabasiana, which was separated in plastid haplotypes. Polyploid Nigritella aggregate into three groups and within each of these groups apomictic polyploids have given rise to new species. Within the N. nigra group, the tetraploid N. nigra subsp. austriaca is somewhat differentiated from the triploid subsp. nigra at nuclear as well as plastid loci. Fusion of an unreduced egg cell from subsp. nigra with a haploid microgamete from Gymnadenia conopsea gave rise to Gymnigritella runei. Within the N. widderi group, N. archiducis-joannis is poorly separated from N. widderi in molecular markers, and the pentaploid N. buschmanniae has evolved by fusion of an unreduced egg cell from N. widderi with a haploid microgamete from a diploid Nigritella. Within the N. miniata group, N. stiriaca is somewhat differentiated from N. miniata at nuclear loci, but no other segregates of N. miniata are supported at species level. Polyploid Nigritella species accumulate genetic diversity by somatic mutations. In the widespread N. nigra subsp. austriaca and N. miniata this diversity is correlated to geography. Although some polyploids may be of recent origins, each polyploid contain genetic markers no longer encountered in diploid members of the genus. According to plastid marker data, Nigritella and Gymnadenia may constitute monophyletic sister genera. (Less)

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.

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.

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 (L.) R. Br., or the fragrant orchid, is one of many plant species negatively influenced by new practices in agriculture and forestry during the last decades. This study describes the level of microsatellite variation within and among 10 Swedish populations of this species. It was not possible to detect strong effects of small population size or fragmentation. In general, the species had high genetic variation within and low genetic divergence among populations, although the correlation between population size and number of alleles was close to significance at the 95% level. Also, a significant isolation by distance effect was observed, indicating the presence of modest restrictions in gene dispersal between the investigated populations.

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.

Spermosporina gymnadeniae (Hyphomycetes) on Gymnadenia sp. (Orchidaceae) collected in the Polish part of the Tatra Mts was described in 2003. The fungus was recently found at new localities in Poland and Slovakia. It was recorded on Gymnadenia conopsea s.l. and Dactylorhiza fuchsii, which are new hosts of this parasite. The fungus is described and additional illustrations are provided.

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.

Reinhammar, L.-G. 1995. Evidence for two distinct species of Pseudorchis (Orchidaceae) in Scandinavia. - Nord. J. Bot. 15: 469–481. Copenhagen. ISSN 0107–055X. Nordic populations of Pseudorchis albida s.l. were studied for variation in morphological characters. Canonical variates analysis (CVA) arrange populations in two discrete groups, corresponding to P. albida S.S. and P. straminea, respectively. In principal component analysis (PCA) of the total material a small overlap could be seen. However, separate PCA:s on members of the two taxa from the same geographic region show perfect separation. Since the two taxa are distinct in morphology as well as in additional characters, e.g. habitat preference in the area of sympatry, they are best treated as separate species. The generic name Pseudorchis is considered to be validly published and is then preferred over the later Leucorchis. A lectotype is selected for Gymnadenia albida f. subalpina.

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

The great botanist Hofmeister (1849) was the first to gave a comprehensive picture of the sexual process based on the study of 30 species belonging to 19 genera of angiosperms. He described pollen germination on the stigma and the possible participation of pollen tube in fertilization, using live materials (Orchis and Gymnadenia). However, he adhered to the opinion that “the male fertilizing element” penetrated into an egg cell by diffusion. Horkel (1836), Schleiden (1837, 1839) and Schacht (1859), who were keen pollination biologists, believed that embryo arose from the tip of a pollen tube. However, in 1856 they admitted that this idea was erroneous.

Ten species of orchid plants belonging to the generaOrchis (7),Dactylorhiza (2), andGymnadenia (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 genusOrchis is not a monophyletic group.

Three new combinations, Dactylorhiza incarnata (L.) Soo ssp. incarnata var. hyphaematodes (Neum.) Lojtnant, Gymnadenia albida (L.) L. C. Rich. ssp. straminea (Fern.) Lojtnant, and Platanthera bifolia (L.) L. C. Rich. ssp. latiflora (Drejer) Lojtnant are made. Some nomenclatural problems of Pseudorchis and Platanthera are discussed and the differences between P. bifolia ssp. bifolia and ssp. latiflora are illustrated by means of photographs, drawings, and biometric data. Drei neue Kombinationen werden vorgeschlagen: Dactylorhiza incarnata (L.) Soo ssp. incarnata var. hyphaematodes (Neum.) Lojtnant, Gymnadenia albida (L.) L. C. Rich. ssp. straminea (Fern.) Lojtnant und Platanthera bifolia (L.) L. C. Rich. ssp. latiflora (Drejer) Lojtnant. Einige nomenklatorische Probleme von Pseudorchis und Platanthera werden diskutiert, und die Verschiedenheiten zwischen P. bifolia ssp. bifolia und ssp. latiflora werden vermittelst Photographien, Zeichnungen und biometrischer Daten illustriert.

Examination of material in spirit, however, shows that the viscidia are quite naked, there being no bursicles as invariably in the genus Orchis, while the viscidia themselves are linear and placed in a parallel position as in Gymnadenia. Vermeulen also could find no bursicles when examining dried material in the Kew Herbarium. It seems, therefore, that the species is correctly placed in the genus Gymnadenia, not in Orchis, and this is borne out by the general structure of the flower. So far as I can see from the material available the tubers are elongate, cylindrical or ovoid and possibly shortly lobed ; I have not seen any that are spherical, as illustrated and described by Reichenbach. Tang and Wang state that the type specimen of G. graminifolia in the Reichenbach Herbarium lacks any tubers so it is not clear from what source Reichenbach obtained his information. EPIGENEIUM Gagnep. In 1853 Lindley (Lindl. & Paxt. Fl. Garden, 1, 136, 155) proposed the name Sarcopodium for a group of orchids. Some of these are now by common consent included in the genus Bulbophyllum, while the remainder have been treated by many authors as a section of Dendrobium. Rolfe and Kraenzlin, however, both considered that this latter part, which contained Lindley's type species (S. amplum (Lindl.) Lindl.) should be maintained as a genus distinct from Dendrobium, a view with which the present writer concurs. In a recent paper in Lloydia (19, 94-98 : 1956) A. D. Hawkes pointed out that the name Sarcopodium Lindl. is a later homonym of Sarcopodium Ehrenb. ex Schlechtend., a genus of Fungi. He therefore proposed the new name Katherinea for the orchid genus, as emended by Kraenzlin and Rolfe, which he also maintained as distinct from Dendrobium, and he made a number of new combinations for the various species which he recognised.