BACKGROUND: Hybridization associated with polyploidy studies is rare in the tropics. The genus Zygopetalum (Orchidaceae) was investigated here as a case study of Neotropical plants. In the rocky highlands of the Ibitipoca State Park (ISP), southeast Brazil, individuals with intermediate colors and forms between the species Z. maculatum and Z. triste were commonly identified.
RESULTS: Chromosomal analysis and DNA quantity showed a uniform population. Regardless of the aspects related to the color and shape of floral structures, all individuals showed 2n = 96 chromosomes and an average of 14.05 pg of DNA. Irregularities in meiosis associated with chromosome number and C value suggest the occurrence of polyploidy. The genetic distance estimated using ISSR molecular markers revealed the existence of genetic variability not related to morphological clusters. Morphometric measurements of the flower pieces revealed that Z. maculatum shows higher variation than Z. triste although lacking a defined circumscription.
CONCLUSIONS: The observed variation can be explained by the polyploid and phenotypic plasticity resulting from the interaction of the genotypes with the heterogeneous environments observed in this habitat.

The Orchidaceae, which is one of the most interesting families of angiosperms, contains a large number of rare species. Despite their acknowledged importance, little attention has been paid to the study of orchids distributed in northern territories. In this study, we determined the syntaxonomical diversity and ecological parameters of orchid habitats in two of Europe\'s largest protected areas, the Pechoro-Ilychsky Reserve and the Yugyd Va National Park (northeastern European Russia), and then compared our findings to those in other parts of orchid distribution ranges. For this purpose, we studied 345 descriptions of plant communities (releves) containing species from Orchidaceae and defined habitat parameters using Ellenberg indicator values with the community weight mean approach, nonmetric multidimensional scaling (NMS), and relative niche width. We found that orchids were distributed in eight habitat types and 97 plant associations. The largest number of orchid species is found in forest communities. Half of the orchid species under study occur in the mires and rock habitats with open vegetation. Several orchids consistently occur in areas disturbed by human activity. In addition, our study indicates that the main drivers of orchid distribution across the vegetation types are light and soil nitrogen. Our analysis of the ecological parameters of orchid habitats indicates that some orchid species can be classified as habitat specialists that are confined to a relatively narrow ecological niche in the Urals (e.g., Goodyera repens, Cypripedium guttatum and Dactylorhiza maculata). Several other species (e.g. Neottia cordata and Dactylorhiza fuchsia) grow under diverse ecological parameters.

BACKGROUND: The flowers of some species of orchids produce nectar as a reward for pollination, the process of transferring pollen from flower to flower. Epipactis albensis is an obligatory autogamous species, does not require the presence of insects for pollination, nevertheless, it has not lost the ability to produce nectar, the chemical composition of which we examined by gas chromatography-mass spectrometry (GC-MS) method for identification of potential insect attractants.
RESULTS: During five years of field research, we did not observe any true pollinating insects visiting the flowers of this species, only accidental insects as ants and aphids. As a result of our studies, we find that this self-pollinating orchid produces in nectar inter alia aliphatic saturated and unsaturated aldehydes such as nonanal (pelargonal) and 2-pentenal as well as aromatic ones (i.e., syringaldehyde, hyacinthin). The nectar is low in alkenes, which may explain the absence of pollinating insects. Moreover, vanillin and eugenol derivatives, well-known as important scent compounds were also identified, but the list of chemical compounds is much poorer compared with a closely related species, insect-pollinating E. helleborine.
CONCLUSIONS: Autogamy is a reproductive mechanism employed by many flowering plants, including the orchid genus Epipactis, as an adaptation to growing in habitats where pollinating insects are rarely observed due to the lack of nectar-producing plants they feed on. The production of numerous chemical attractants by self-pollinated E. albensis confirms the evolutionary secondary process, i.e., transition from ancestral insect-pollinating species to obligatory autogamous.

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.

The phylogeny of the genus Paphiopedilum based on the plastome is consistent with morphological analysis. However, to date, none of the analyzed nuclear markers has confirmed this. Topology incongruence among the trees of different nuclear markers concerns entire sections of the subgenus Paphiopedilum. The low-copy nuclear protein-coding gene PHYC was obtained for 22 species representing all sections and subgenera of Paphiopedilum. The nuclear-based phylogeny is supported by morphological characteristics and plastid data analysis. We assumed that an incongruence in nuclear gene trees is caused by ancestral homoploid hybridization. We present a model for inferring the phylogeny of the species despite the incongruence of the different tree topologies. Our analysis, based on six low-copy nuclear genes, is congruent with plastome phylogeny and has been confirmed by phylogenetic network analysis.

The South Korean government is seeking information on the environmental conservation value (ECV) of Cypripedium japonicum, a plant on the first-level priority endangered species list of the Korea Ministry of Environment, as well as on the Red List of the International Union for Conservation of Nature. This research paper aims to assess the ECV by employing contingent valuation (CV) to elicit people\'s willingness to pay (WTP) for its conservation. To achieve this aim, in May 2020, a CV survey of 1,000 interviewees in South Korea was carried out employing a closed-ended question. The average household WTP is estimated to be KRW 3,770 (USD 3.07) per annum and secures statistical significance. From a national point of view, using information on the national population, this value comes to KRW 76.72 billion (USD 62.42 million) per year. Although the cost of conserving the species has not yet been accurately estimated, the ECV seems to be larger than the cost of conservation. Thus, it is socially desirable to conserve Cypripedium japonicum, and the conservation should be carried out in a stable and continuous manner.

Developing a monitoring system and a conservation strategy against the negative impact of global change on threatened plant species, is nowadays the challenge for conservation experts. The Plant Micro-Reserve (PMR) approach became a highly effective approach in protecting plant species, since mild active management of vegetation plots and protection of plant populations takes place. The PMR has greatly evolved since its initial concept of managing a large network of PMRs, to having fewer protected areas subject to intensive scientific monitoring (e.g. Intensive Monitoring PMR; IM-PMR). This study further improved the IM-PMR approach by focusing on the threatened plant species of Ophrys kotschyi in Cyprus. The proposed IM-PMR enhances the available knowledge on the biology, physiology and ecology of the targeted plant species, through implementing an intensive monitoring system and assessing its genetic diversity. Within the framework of IM-PMR, the population size of O. kotschyi recorded statistically significant differentiation during the monitoring period, most likely due to the vegetative dormancy of the species. The subpopulation size and dormancy in O. kotschyi was correlated with precipitation and air temperature for specific months. In addition, the different local climatic conditions and the species dormancy between years seem to influence the flowers production among individuals, in the four monitoring years. Nevertheless, the low natural fecundity compared to the artificially pollinated plants and the absent correlation with any of the climatic parameters, might be closely related to the lack of pollinators at this site. The genetic diversity (HT = 0.456) is higher compared to other endemic and short-lived perennial species, while the genetic differentiation among the subpopulations of O. kotschyi showed significant substructure (FSTFIS=0.5 = 0.097*). The subpopulation within IM-PMR showed relatively lower genetic diversity among the largest subpopulations of O. kotschyi, and the highest percentage of linked loci. Such observations support the non-random association of different loci in this subpopulation, and the ineffective pollen flow within this single subpopulation. The improvement of the original PMR approach in the current IM-PMR proposal denotes that different ecological aspects are taken into account towards gaining a holistic knowledge on a target species. The IM-PMR approach as implemented for O. kotschyi, could lead to the development of an integrated conservation approach for rare, threatened, or endangered species.

The geographic distribution of species depends on their relationships with climate and on the biotic interactions of the species. Ecological Niche Models (ENMs) mainly consider climatic variables only and may tend to overestimate these distributions, especially for species strongly restricted by biotic interactions. We identified the preference of Laelia speciosa for different host tree species and include this information in an ENM. The effect of habitat loss and climate change on the distribution of these species was also estimated. Although L. speciosa was recorded as epiphyte at six tree species, 96% of the individuals were registered at one single species (Quercus deserticola), which indicated a strong biotic interaction. We included the distribution of this host tree as a biotic variable in the ENM of L. speciosa. The contemporary distribution of L. speciosa is 52,892 km2, which represent 4% of Mexican territory and only 0.6% of the distribution falls within protected areas. Habitat loss rate for L. speciosa during the study period was 0.6% per year. Projections for 2050 and 2070 under optimistic and pessimistic climate change scenarios indicated a severe reduction in its distribution. Climaticaly suitable areas will also shift upwards (200-400 m higher). When estimating the distribution of a species, including its interactions can improve the performance of the ENMs, allowing for more  accurate estimates of the actual distribution of the species, which in turn allows for better conservation strategies.

Understanding how environmental pollutants influence plant occurrence, growth, and development is key for effective management plans and potential bioremediation. Rare plants, such as orchids, may occur in modified habitats and on soils containing heavy metals, yet their ecological and physiological responses to heavy metals is poorly understood. We investigated the influence of heavy metal pollution on orchid growth rates and interactions with soil fungal mutualists by comparing a large population of the orchid Epipactis helleborine (L.) Crantz subsp. tremolsii (Pau) E. Klein that grows on mine tailings in south-west Sardinia (Italy) with a population that grows on non-contaminated soils in central Sardinia. Soils of the contaminated site had high levels of heavy metals and low organic matter and nutritive elements content. We performed a morphological analysis on twenty individuals that have been subjected to measurement of bioaccumulation and translocation of heavy metals. Fungi associated with the roots of plants from the contaminated and uncontaminated site were grown and identified by DNA barcoding approach. Plants from the contaminated site were smaller than the ones growing in the uncontaminated site and were found to be able to tolerate heavy metals from the soil and to accumulate and translocate them into their organs. Fungi belonging to the genus Ilyonectria (Ascomycota) were found both in contaminated and uncontaminated sites, while an unidentified fungus was isolated from roots in the contaminated site only. These results are discussed in terms of orchids\' tolerance to heavy metals and its physiological and ecological mechanisms. The role of contaminated habitats in harbouring orchids and peculiar taxa is also discussed.

Members of the Orchidaceae, one of the largest families of flowering plants, evolved the crassulacean acid metabolism (CAM) photosynthesis strategy. It is thought that CAM triggers adaptive radiation into new niche spaces, yet very little is known about its origin and diversification on different continents. Here, we assess the prevalence of CAM in Dendrobium, which is one of the largest genera of flowering plants and found in a wide range of environments, from the high altitudes of the Himalayas to relatively arid habitats in Australia. Based on phylogenetic time trees, we estimated that CAM, as determined by δ 13C values less negative than -20.0‰, evolved independently at least eight times in Dendrobium. The oldest lineage appeared in the Asian clade during the middle Miocene, indicating the origin of CAM was associated with a pronounced climatic cooling that followed a period of aridity. Divergence of the four CAM lineages in the Asian clade appeared to be earlier than divergence of those in the Australasian clade. However, CAM species in the Asian clade are much less diverse (25.6%) than those in the Australasian clade (57.9%). These findings shed new light on CAM evolutionary history and the aridity levels of the paleoclimate on different continents.