Characters evolution of Encyclia (Laeliinae-Orchidaceae) reveals a complex pattern not phylogenetically determined: insights from macro- and micromorphology.

Encyclia is the second-largest genus in the neotropical subtribe Laeliinae (Orchidaceae) and has more than 150 species, which are characterized by fairly consistent flower morphology. Its taxonomy and species boundaries, however, seem to be still under debate. In the present study, we first examined the lip micromorphology of 61 species of Encyclia sensu stricto. We correlated our results with external flower morphology and phylogenetic analyses performed on a combined dataset that included both nuclear (ITS, Xdh, PhyC) and plastid markers (ycf1, rpl32, and trnL-trnF). Phylogenetic reconstruction showed that Encyclia sensu stricto species form a coherent, monophyletic group. However, it is difficult to determine the relationships between the different groups within one larger clade. The groups all form distinct lineages that evolved from a common ancestor. The UPGMA cluster analysis for the seven qualitative micromorphological features clearly divides the genus into two main groups, the larger of which is further subdivided into two subgroups. None of these, however, overlap with any of the phylogeographic units distinguished in previously published papers or in presented article. It is worth noting that the groups resulting from the UPGMA analysis cannot be defined by macromorphological features. The pattern of similarities between species, taking into account both macro- and micromorphological features, is eminently mosaic in nature, and only a multifaceted approach can explain this enigmatic group.

Schiedeella bajaverapacensis (Orchidaceae, Spiranthinae), a New Orchid Species from Guatemala.

Guatemala is recognized for its diverse and rich flora and fauna. It is estimated that over 1200 orchid species, classified in 223 genera, are known to occur in this rather small, yet megadiverse country. While studying the diversity of this plant group in the department of Baja Verapaz, we found individuals that clearly belonged to the genus Schiedeella, but whose features did not fit any previously known species. At that time, nine terrestrial taxon representatives were known to occur in Guatemala. We conducted the morphological analysis in accordance with the standard procedures of classical taxonomy. For phylogenetic reconstruction, 59 sequences of the ITS region and 48 of the trnL-trnF marker were applied. The topology of trees was obtained based on the Bayesian inference. Schiedeella bajaverapacensis was described and illustrated based on morphological evidence, and its taxonomic position was confirmed by phylogenetic analyses. The new entity is the 10th Schiedeella representative known from Guatemala.

Active Compounds with Medicinal Potential Found in Maxillariinae Benth. (Orchidaceae Juss.) Representatives-A Review.

Orchids are widely used in traditional medicine for the treatment of a whole range of different health conditions, and representatives of the Neotropical subtribe Maxillariinae are not an exception. They are utilized, for instance, for their spasmolytic and anti-inflammatory activities. In this work, we analyze the literature concerning the chemical composition of the plant extracts and secretions of this subtribe's representatives published between 1991 and 2022. Maxillariinae is one of the biggest taxa within the orchid family; however, to date, only 19 species have been investigated in this regard and, as we report, they produce 62 semiochemicals of medical potential. The presented review is the first summary of biologically active compounds found in Maxillariinae.

Floral attractants in the black orchid Brasiliorchis schunkeana (Orchidaceae, Maxillariinae): clues for presumed sapromyophily and potential antimicrobial activity.

BACKGROUND: Orchids have evolved various strategies that aim to ensure their reproduction success. These may include the production of rewards for pollinators, or on the contrary, deception. Specific sets of features such as flower morphology, color, nectar, and odor presence (or lack thereof) are considered to determine suitability for pollination by different groups of animals. Stingless bees are thought to be the primary pollinators of the orchids of the Neotropical subtribe Maxillariinae. However, almost black flowered Brasiliorchis schunkeana at first glance presents floral adaptations that may suggest another pollination syndrome-sapromyophily. RESULTS: A few traces of secretion were noticed on the glabrous lip callus and lip apex built by conical to villiform papillae (SEM analysis). Histochemical studies revealed huge amounts of lipids in the epidermis, subepidermis, and some parenchyma cells (SBB test) with various stages of lipids accumulation between cells. Further TEM analysis showed a heterogeneous (lipoid and phenolic) nature of secretion. The dense osmiophilic cytoplasm contained organelles (RER, free ribosomes, dictyosomes, plastids with plastoglobuli, nucleus) and vesicles migrating to plasmalemma. The vesicles, osmiophilic globules, and flocculent material were visible in periplasmic space. The central vacuole possessed osmiophilic phenolic content and flocculent material. GC-MS analysis revealed in floral extract the presence of 7,9-di-tert-butyl-1-oxaspiro(4,5)deca-6,9-diene-2,8-dione (77.06%) and 2,5-di-tert-butyl-1,4-benzoquinone (16.65%). Both compounds are known for their biological activity. CONCLUSIONS: The juxtaposition of results led us to the conclusion that the labellar tissue produces lipoid and phenolic material, which is responsible for the glossiness and rotten herring scent. This type of secretion could be classified as a phenolic resin. The chemical analysis revealed the presence of five semiochemicals that are known to be attractants for some Diptera, which together with the rest of the results constitutes a strong premise that representatives of this order could be potential pollinators of B. schunkeana. Field observations however are still needed to confirm this pollination syndrome.

Ornithophily in the subtribe Maxillariinae (Orchidaceae) proven with a case study of Ornithidium fulgens in Guatemala.

Ornithophily has been long speculated to occur in the subtribe Maxillariinae (Orchidaceae), relying either solely on micromorphological analyses or scarce field observations of undefined species. In Guatemala we were able to observe regular visits of the azure-crowned hummingbirds feeding on flowers of Ornithidium fulgens. These observations have led us to investigation of floral attractants by means of scanning and transmission microscopy, histochemical and chemical analyses (GC-MS). Conducted investigation revealed that the epidermis of basal protuberance of column-foot has features proving the secretory activity and that secreted nectar is sucrose-dominant. Slight secretion on the middle part of the lip is puzzling. The presence of other potential pollinators has not been reported. Based on the results of this study, we confirmed that the flowers of O. fulgens meet all criteria of ornithophily and thus that the hypothesis about bird pollination in the subtribe Maxillariinae is proven. The presented results confirm that the previously described floral features predicting the bird pollination in this group are justified. This strengthens the theory about floral adaptations to different pollinators and gives valid reasons to consider species with flowers with a certain set of traits as ornithophilous, even in the absence of the pollination observation.

Maxillariaanacatalinaportillae (Orchidaceae, Maxillariinae), a new remarkable species from Ecuador.

Neotropical genus Maxillaria Ruiz & Pav. belongs to one of the most diverse and species-rich groups of orchids. Several of its representatives are popular, horticultural plants with large and showy flowers, often nicely fragranced. It is not uncommon that some distinctly colored individuals are introduced to the commercial market under names of similar, more or less related species, as informal varieties or color forms, largely causing confusion. While investigating the diversity of Maxillaria in Ecuador, we have encountered plants that were commercially referred to as M.sanderianaxanthina. In the course of conducted morphological and micromorphological analyses, we concluded that it is a new, separate species and hereby, we describe it as M.anacatalinaportillae.

Crepidium sect. Crepidium (Orchidaceae, Malaxidinae)-Chemical and Morphological Study of Flower Structures in the Context of Pollination Processes.

Crepidium is a large genus of mainly pantropical orchids. The lips of its flowers are upwardly directed and do not serve as landing platforms for pollinators. This role is assumed by the dorsal sepal and/or gynostemium. Information about the pollination and floral morphology of this genus is scarce. To date, no papers have been published on these topics. Field observations have revealed that the flowers are visited by small flies, midges, fruit flies, other small dipterans, ants, spiders, and mites. Preliminary observations revealed at least two forms of small liquid droplets secreted on the lip surface of Crepidium species: simple secretions from epidermal cells, and cell sap released upon the rupturing of raphide-producing cells. Further research revealed that this was the first time liquid secretion was recorded in this genus. Floral secretions were subjected to sequential organic solvent extraction and gas chromatography-mass spectrometry (GC-MS). Floral parts were investigated by means of scanning (SEM) and transmission electron microscopy (TEM), and histochemical tests. The presence of liquid droplets on the lip of Crepidium, the presence of a food reward, and the sequence of raphide development are reported here for the first time.

Labellum structure of Bulbophyllum echinolabium J.J. Sm. (section Lepidorhiza Schltr., Bulbophyllinae Schltr., Orchidaceae Juss.).

This micromorphological, chemical and ultrastructural study is a continuation of research conducted on the section Lepidorhiza. The Bulbophyllum echinolabium flowers comprised features that characterize a sapromyophilous syndrome, having large, glistening parts that emit an intense scent of rotten meat. The secretory activity was described in the hypochile (nectary in longitudinal groove and in the prickles) and the epichile (putative osmophore). The ultrastructural studies revealed a dense cytoplasm in the epidermis and subepidermal tissue with large nuclei and numerous mitochondria, the profiles of SER and RER, and dictyosomes. Large amounts of heterogeneous residues of secreted material (possibly phenolic) were present on the cuticle surface, similar to the unusual prominent periplasmic space with flocculent secretory material. The chemical analysis (GC/MS) of the scent profile of lips comprised carbohydrates and their derivatives (29.55% of all compounds), amino acids (1.66%), lipids (8.04%) and other organic compounds (60.73%). A great number of identified compounds are Diptera attractants (mainly Milichiidae, Tephritidae, Drosophilidae, Muscidae, Sarcophagidae, Tachinidae). The examination of visual and olfactory features indicates correlation between colour of flowers and the type of olfactory mimicry, where a dark colour labellum emits strong smell of rotten waste.