Aerial and terrestrial root habits influence the composition of the cell walls of Vanilla phaeantha (Orchidaceae).

In response to the restrictions imposed by their epiphytic habit, orchids have developed structural traits that allow greater efficiency in water uptake and use, such as a complex adventitious root system with velamen. The composition of cell wall of this specialized epidermis can be altered according to the substrate to which it is fixed, influencing wall permeability, absorption, and storage of water in roots. The current study aimed to evaluate the cell wall composition of adventitious roots of Vanilla phaeantha (Orchidaceae) that grow attached to the phorophyte, fixed in the soil, or hung free. Immunocytochemical analyses were used to determine the protein, hemicellulose, and pectin composition of the cell walls of aerial and terrestrial roots. We observed that pectins are present in the different tissues of the aerial roots, while in the terrestrial roots, they are concentrated in the cortical parenchyma. The deposition of xyloglucans, extensins, and arabinogalactans was greater in the epidermis of the free side of the roots attached to the phorophyte. The strong labeling of pectins in aerial roots may be related to the influx of water and nutrients, which are generally scarce in this environment. The arrangement of hemicelluloses and proteins with the pectins may be associated with increased cell rigidity and sustainability, a feature of interest for the aerial roots. In summary, the habit of roots can interfere with the non-cellulosic composition of the cell walls of V. phaeantha, possibly related to changes in cell functionality.

Orchid seeds are not always short lived in a conventional seed bank!

BACKGROUND AND AIMS: Orchid seeds are reputed to be short lived in dry, cold storage conditions, potentially limiting the use of conventional seed banks for long-term ex situ conservation. This work explores whether Cattleya seeds are long lived or not during conventional storage (predried to ~12 % relative humidity, then stored at -18 °C). METHODS: We explored the possible interaction of factors influencing seed lifespan in eight species of the genus Cattleya using physiological (germination and vigour), biochemical (gas chromatography), biophysical (differential scanning calorimetry) and morphometric methods. Seeds were desiccated to ~3 % moisture content and stored at -18 °C for more than a decade, and seed quality was measured via three in vitro germination techniques. Tetrazolium staining was also used to monitor seed viability during storage. The morphometric and germination data were subjected to ANOVA and cluster analysis, and seed lifespan was subjected to probit analysis. KEY RESULTS: Seeds of all Cattleya species were found to be desiccation tolerant, with predicted storage lifespans (P50y) of ~30 years for six species and much longer for two species. Cluster analysis showed that the three species with the longest-lived seeds had smaller (9-11 %) airspaces around the embryo. The post-storage germination method impacted the quality assessment; seeds equilibrated at room temperature for 24 h or in 10 % sucrose solution had improved germination, particularly for the seeds with the smallest embryos. Chromatography revealed that the seeds of all eight species were rich in linoleic acid, and differential scanning calorimetry identified a peak that might be auxiliary to selecting long-lived seeds. CONCLUSIONS: These findings show that not all orchids produce seeds that are short lived, and our trait analyses might help to strengthen prediction of seed longevity in diverse orchid species.

Micropropagation of Guarianthe skinneri (Bateman) Dressler & W. E. Higging in Temporary Immersion Bioreactors.

Guarianthe skinneri (Bateman) Dressler & W. E. Higgins is an orchid valued for its ornamental characteristics. However, it is an orchid classified as threatened with extinction due to the illegal extraction from its natural habitat. In addition, its propagation through seed germination is very low, as is the case with most members of the family Orchidaceae. Its asexual propagation through pseudobulb separation is slow and produces a few propagules. For this reason, in vitro propagation techniques are an alternative to increase the number of plants obtained and thus be able to recover this valuable plant genetic resource. Temporary immersion systems (TIS) offer the advantage of mass-propagating plants for different purposes. This chapter describes a large-scale micropropagation protocol for Guarianthe skinneri using temporary immersion bioreactors (TIB).

Micropropagation of Encyclia cordigera (Kunth) Dressler in Ebb-and-Flow Bioreactor.

The use of new technologies for micropropagation such as temporary immersion systems (TISs) is important, because it reduces costs by 40% lowering labor, agar and containers. TISs are containers designed for large-scale, semiautomatic production of plants in a liquid medium, which has been used in propagation of commercial orchids. This tool has high potential for application in micropropagation of medicinal and endangered orchids for conservation and commercial purposes. In this chapter, we describe a detailed protocol for propagation and development of Encyclia cordigera to be used in research projects for small-scale production. This protocol comprises all steps from explant preparation to the establishment orchids plantlets.

Orchid Micropropagation Using Temporary Immersion Systems: A Review.

Temporary immersion systems (TIS) have been used for orchid micropropagation. The main advantage of TIS use for micropropagation is that the explant is periodically immersed in nutrient media, and then, the nutrient solution is drained, which allows the explant tissue to stay in air. The current review resumes the application of TIS in orchid propagation. Fifty-three papers are discussed considering: explant, culture media, TIS bioreactor type, frequency and immersion time, and the TIS effects in acclimatization phase.

Complexity of the Male Perfume of Eulaema nigrita from Forest and Woody Physiognomies of the Brazilian Savanna: Is There a Relationship with Body Size and Wing Wear?

Male orchid bees collect volatile and semi-volatile compounds from the environment for storage and accumulation in specialized hind legs. Later, these compounds form a perfume blend used during courtship to lure conspecific females for mating. Male orchid bees perfume has been suggested to play an important role as a sexual signaling trait involved in pre-mating isolation of species, functioning as an indicator of male genetic quality. Eulaema nigrita Lepeletier (Apidae: Euglossini) is a common species in both forested and woody savanna (Cerrado stricto sensu) physiognomies of the Brazilian savanna biome. By identifying the chemical composition of male E. nigrita perfume, we tested for differences in the bouquet chemical profile in populations from remnants of seasonal semideciduous forest and woody savanna. In addition, we assessed the relation between perfume complexity and morphological traits associated with size and age of males. Our analysis showed a low effect of physiognomies on differences in the perfume chemical profile of sampled males. Nevertheless, we observed significant differences in the chemical profile of individuals from two seasonal semideciduous forest remnants, which suggests an environmental effect in individual bouquet. Wing wear measurements were positively related to perfume complexity, consistent with the premise that perfumes from older individuals are indicators of survival capacity in male orchid bees.

Euglossini bee diversity is driven by forest cover in coastal Amazon.

Habitat loss and fragmentation are drivers of biodiversity loss, such as Euglossini bees in continental regions. Knowledge about these effects on this group of pollinators in coastal regions is still incipient and needs to be further investigated. This study aimed to evaluate the effects of landscape structure on the abundance, richness, diversity and species composition of Euglossini bees on the coast of the Brazilian Amazon. We mapped the surrounding landscape around 48 sampling points in the east of the island of Marajó, Pará, Brazil where we collected bees using chemical baits. We used Generalized Linear Models (GLMs) to evaluate the effects of landscape structure (composition and configuration) on the abundance, richness, diversity and composition of Euglossini bees. We collected a total of 1017 males belonging to four genera and 22 species. Forest cover (%) and landscape heterogeneity were the best predictors of the bee community. Increased forest cover positively affected the abundance, richness and diversity of bees at a local scale. On the other hand, abundance, richness and diversity decreased with increasing landscape heterogeneity, also at a local scale. The hypothesis that the amount of habitat favors Euglossini communities was corroborated by our results. Based on our conclusions, landscapes with greater forest cover can effectively contribute to the conservation of these bees and their pollination services along the Amazon coast.

An elevational gradient in floral traits and pollinator assemblages in the Neotropical species Costus guanaiensis var. tarmicus in Peru.

Different populations of plant species can adapt to their local pollinators and diverge in floral traits accordingly. Floral traits are subject to pollinator-driven natural selection to enhance plant reproductive success. Studies on temperate plant systems have shown pollinator-driven selection results in floral trait variation along elevational gradients, but studies in tropical systems are lacking. We analyzed floral traits and pollinator assemblages in the Neotropical bee-pollinated taxon Costus guanaiensis var. tarmicus across four sites along a steep elevational gradient in Peru. We found variations in floral traits of size, color, and reward, and in the pollinator assemblage along the elevational gradient. We examined our results considering two hypotheses, (1) local adaptation to different bee assemblages, and (2) the early stages of an evolutionary shift to a new pollinator functional group (hummingbirds). We found some evidence consistent with the adaptation of C. guanaiensis var. tarmicus to the local bee fauna along the studied elevational gradient. Corolla width across sites was associated with bee thorax width of the local most frequent pollinator. However, we could not rule out the possibility of the beginning of a bee-to-hummingbird pollination shift in the highest-studied site. Our study is one of the few geographic-scale analyses of floral trait and pollinator assemblage variation in tropical plant species. Our results broaden our understanding of plant-pollinator interactions beyond temperate systems by showing substantial intraspecific divergence in both floral traits and pollinator assemblages across geographic space in a tropical plant species.

Diferentes poblaciones de una especie de planta pueden adaptarse a sus polinizadores locales y, en consecuencia, divergir en los rasgos florales. Los rasgos florales están sujetos a la selección natural impulsada por los polinizadores para mejorar el éxito reproductivo de la planta. Los estudios en sistemas de plantas de zonas templadas han demostrado que la selección impulsada por los polinizadores da como resultado una variación de los rasgos florales a lo largo de gradientes de altitud, pero faltan estudios en sistemas de plantas de zonas tropicales. Analizamos las características florales y los ensamblajes de polinizadores en el taxón Neotropical polinizado por abejas Costus guanaiensis var. tarmicus en cuatro sitios a lo largo de un empinado gradiente altitudinal en Perú. Encontramos variación en los rasgos florales de tamaño, color y recompensa, y en los ensamblajes de polinizadores a lo largo del gradiente altitudinal. Examinamos nuestros resultados considerando dos hipótesis, (1) la adaptación local a diferentes ensamblajes de abejas polinizadoras y (2) las primeras etapas de un desplazamiento evolutivo a un nuevo grupo funcional polinizador (colibríes). Encontramos evidencia que respalda la adaptación de C. guanaiensis var. tarmicus a la fauna de abejas local a lo largo del gradiente altitudinal estudiado. El ancho de la corola de la flor a lo largo de los sitios de estudio se asoció con el ancho del tórax de la especie de abeja local más frecuentemente registrada en las flores. Sin embargo, no pudimos descartar la posibilidad del comienzo de un desplazamiento de polinización de abeja a colibrí en el sitio más alto estudiado. Nuestro estudio es uno de los pocos análisis a escala geográfica de variación de rasgos florales y ensamblaje de polinizadores en una especie tropical. Nuestros resultados amplían la comprensión de las interacciones planta­polinizador más allá de los sistemas templados al mostrar una sustancial divergencia intraespecífica tanto en los rasgos florales como en los ensamblajes de polinizadores en una especie tropical.

In Vitro Polyploidization of Brassolaeliocattleya Hybrid Orchid.

The Cattleya (Orchidaceae-Laeliinae subtribe) intergeneric hybrids, such as Brassolaeliocattleya (Blc.), have great ornamental value, due to their compact-size, with large and high color diversity of flowers. Artificial induction of polyploidy brings agronomic, ornamental and genetic benefits to plants. Polyploidization efficiency depends on factors, such as the type of antimitotic, polyploidization method, concentrations, exposure times and type of explant. This study aimed to develop a protocol to polyploidize Blc. orchids, by testing two types of explants (seeds and protocorms), concentrations and exposure times to colchicine. The effects of colchicine on the in vitro development of explants were also investigated. The responses of explants to colchicine depended on the concentrations, exposure time and the interaction of these factors. Flow cytometric analysis evidenced high endopolyploidy and allowed the separation of polyploidized (4C, 8C and 16C peaks) from non-polyploidized (only 2C and 4C peaks) plants. The highest percentage of polyploid plants was regenerated from protocorms (16.4%) treated with colchicine instead of seeds (3.2%). Protocorms treated with colchicine at 500-750 µM for 18 h resulted in the best percentage of polyploidization. Additionally, in vitro natural polyploidization using protocorms was reported (11.5%). Cytological analyses allowed an estimation of the number of chromosomes of the parents (≡70), polyploidized (≡140) and non-polyploidized progeny (≡70).

High species richness and turnover of vascular epiphytes is associated with water availability along the elevation gradient of Volcán Maderas, Nicaragua.

Research that has been conducted documenting species richness patterns on tropical mountains has resulted in conflicting observations: monotonic declines with increasing elevation, monotonic increases with increasing elevation, and a mid-elevation "bulge." Currently, it is unclear if these differences are due to environmental differences among study areas, the taxonomic groups or ecological groups (e.g., growth form) sampled, or the scale of study along elevation gradients. Because of the difficulty in sampling and identifying canopy-dwelling plants, the number of inventories quantifying tropical epiphytes is relatively limited and recent. In this study, we provide a detailed qualitative and quantitative assessment of the vascular epiphyte flora and its spatial distribution on Volcán Maderas, Isla de Ometepe, Nicaragua, including weather and environmental measurements along the entire elevation gradient of the volcano. We sampled epiphytes in five distinct forest types associated with increasing elevation as follows: dry forest, humid forest, wet forest, cloud forest, and elfin forest. Five weather stations were placed along the elevation gradient for us to relate observed patterns to environmental conditions. A mid-elevation peak in species richness was detected for all vascular epiphytes at approximately 1000 m in elevation (cloud forest), yet epiphyte abundance increased with increasing elevation. In total we identified 206 taxa of vascular epiphytes belonging to 26 families and 73 genera. The most species-rich family was the Orchidaceae with 55 species for the entire elevation gradient, followed by Bromeliaceae (29 species), Araceae (23), Polypodiaceae (25), Dryopteridaceae (16), and Piperaceae (11), with all other families represented by fewer than 10 species each. We found that richness patterns differ phylogenetically across epiphyte groups, possibly due to different adaptive strategies, and species for the most part appear to be narrowly distributed within specific habitat zones along the elevation gradient. Variables associated with moisture, precipitation, humidity, mist, or cloud cover are key to understanding the observed patterns.

Structural plasticity in roots of the hemiepiphyte Vanilla phaeantha Rchb.f. (Orchidaceae): a relationship between environment and function.

The aerial environment appears to structurally modify roots, which frequently show specializations for absorbing water and nutrients. Among those specializations are the velamen, a multiseriate epidermis generally composed of dead mature cells, and greater degrees of lignification in the endodermis, exodermis, and pith. Vanilla phaeantha is a hemiepiphyte used here as a model of study to determine which root characteristics demonstrate the most plasticity in response to aerial and terrestrial environments. It produces roots growing under three conditions: (1) aerial and free, growing from the highest branches towards the ground; (2) aerial roots attached to the phorophyte; and (3) terrestrial. Samples taken 3 cm from the apices were used to prepare histological slides. The tissues and other anatomical structures were measured and histochemically characterized. The most plastic characteristics were the external periclinal thicknesses of the exodermis and the total area occupied by the aerenchyma lacunae. The free roots were the longest, did not evidence root hairs, and had the largest number of the aerenchyma lacunae; they also evidenced greater thicknesses of the exodermis in contact with the epidermis walls that helped maintain their shapes. Terrestrial roots had root hairs around the entire circumference and intense infestations of mycorrhiza, indicating their involvement in nutrient acquisition. The adhering roots evidenced free regions similar to those of aerial roots, as well as adhering regions showed characteristics similar to terrestrial roots (with root hairs and mycorrhiza infestations).

Diversity of Root-Associated Fungi of the Terrestrial Orchids Gavilea lutea and Chloraea collicensis in a Temperate Forest Soil of South-Central Chile.

The diversity of orchid mycorrhizal fungi (OMF) and other beneficial root-associated fungi in temperate forests has scarcely been examined. This study aimed to analyze the diversity of mycorrhizal and rhizosphere-associated fungal communities in the terrestrial orchids Gavilea lutea and Chloraea collicensis growing in high-orchid-population-density areas in the piedmont of the Andes Cordillera with native forest (Nothofagus-Araucaria) and Coastal Cordillera with an exotic plantation (Pinus-Eucalyptus) in south-central Chile. We focused on rhizosphere-inhabiting and peloton-associated OMF in a native forest (Andes Cordillera) and a mixed forest (Coastal Cordillera). The native terrestrial orchids G. lutea and C. collicensis were localized, mycorrhizal root segments were taken to isolate peloton-associated OMF, and rhizosphere soil was taken to perform the metabarcoding approach. The results revealed that Basidiomycota and Ascomycota were the main rhizosphere-inhabiting fungal phyla, showing significant differences in the composition of fungal communities in both sites. Sebacina was the most-abundant OMF genera in the rhizosphere of G. lutea growing in the native forest soil. In contrast, Thanatephorus was the most abundant mycorrhizal taxa growing in the rhizosphere of orchids from the Coastal Cordillera. Besides, other OMF genera such as Inocybe, Tomentella, and Mycena were detected. The diversity of OMF in pelotons differed, being mainly related to Ceratobasidium sp. and Tulasnella sp. These results provide evidence of differences in OMF from pelotons and the rhizosphere soil in G. lutea growing in the Andes Cordillera and a selection of microbial communities in the rhizosphere of C. collicensis in the Coastal Cordillera. This raises questions about the efficiency of propagation strategies based only on mycorrhizal fungi obtained by culture-dependent methods, especially in orchids that depend on non-culturable taxa for seed germination and plantlet development.

Validation of the traditional medicinal use of a Mexican endemic orchid (Prosthechea karwinskii) through UPLC-ESI-qTOF-MS/MS characterization of its bioactive compounds.

Ethnopharmacological relevance: The orchid Prosthechea karwinskii is a medicinal orchid in Oaxaca, Mexico, used to treat diabetes, cough, wounds, and burns, prevent miscarriage and assist in labor. Each part of the plant (leaves, pseudobulbs, or flowers) is used by healers for certain treatment conditions, indicating that each part has different biocompounds with specific pharmacological activity. Aim of the study: To characterize the biocompounds in extracts from leaves, pseudobulbs, and flowers of P. karwinskii and evaluate their ROS inhibition capacity to associate it with medicinal uses. Materials and methods: The compounds present in extracts from leaves, pseudobulbs, and flowers of P. karwinskii were identified by UPLC-ESI-qTOF-MS/MS. The chemical differentiation of each extract was tested by principal component analysis (PCA) using compound intensity values. For each extract, total phenol and flavonoid contents were quantified. Their antioxidant capacity was evaluated ex vivo by inhibition of ROS with DCFH-DA and in vitro with DPPH radical. Results: Based on the PCA, it was observed that some compounds were completely separated from others according to the correlation that they presented. The compounds common to all three plant parts were quinic, malic, succinic, azelaic, and pinellic acids. Among the compounds identified, two were exclusive to leaves, four to pseudobulbs, and ten to flowers. Some of the identified compounds have well-known antioxidant activity. The leaves had the highest content of total phenols and flavonoids, and the highest in vitro and ex vivo antioxidant capacity. A strong correlation was observed between phenol and flavonoid contents, and antioxidant capacity ex vivo and in vitro. Conclusions: It was found that the bioactive compounds and antioxidant capacity of each part of the plant were associated with its traditional medicinal use. A pharmacological potential was also found in P. karwinskii for further biological studies because of the type of compounds it contained.

Mesoamerican Cypripedium: Mycorrhizal Contributions to Promote Their Conservation as Critically Endangered Species.

In the valuable orchid genus Cypripedium, the section Irapeana consists of a distinctive group of Mesoamerican species that is formed by Cypripedium dickinsonianum Hágsater, C. irapeanum Lex., and C. molle Lindl. All lady slipper orchids exhibit different distributions and abundances. Data analysis that used herbarium accessions and field investigations indicated that the habitats of these three species have been dramatically reduced. Prospecting for suitable habitats based on climatic, vegetation, and soil parameters allows us to predict potential distributions. Conservation strategies, such as ex situ propagation by asymbiotic and symbiotic approaches, have indicated that the culture media used are a determining factor for seedling development. Mycorrhizal isolates play a main role in the compatibility and further development of germinated seeds. The fungi isolated from adult plants belong to two different families, which makes it possible that widely distributed C. irapeanum populations will be fungal-specific as well as restricted for C. molle. Root mycorrhization patterns occur high on the secondary roots. In contrast with other species of the genus, in situ germination can occur over a short period of two months, but we have documented periods as long as ten years. Cypripedium is a highly problematic genus for ex situ conservation because the germination requirements and cultures are poorly documented, and there is great urgency for in situ conservation to develop strategies for identifying hotspot habitats and actualize the protection status to avoid extinction of this genus.

Advances and Perspectives for Polyploidy Breeding in Orchids.

The orchid market is a dynamic horticultural business in which novelty and beauty command high prices. The two main interests are the development of flowers, from the miniature to the large and showy, and their fragrance. Overall organ size might be modified by doubling the chromosome number, which can be accomplished by careful study of meiotic chromosome disjunction in hybrids or species. Meiosis is the process in which diploid (2n) pollen mother cells recombine their DNA sequences and then undergo two rounds of division to give rise to four haploid (n) cells. Thus, by interfering in chromosome segregation, one can induce the development of diploid recombinant cells, called unreduced gametes. These unreduced gametes may be used for breeding polyploid progenies with enhanced fertility and large flower size. This review provides an overview of developments in orchid polyploidy breeding placed in the large context of meiotic chromosome segregation in the model plants Arabidopsis thaliana and Brassica napus to facilitate molecular translational research and horticultural innovation.

Descripción morfológica y caracterización molecular de los hongos asociados a la raíz de Masdevallia coccinea Linden ex Lindl / Morphological description and molecular characterization of fungi associated with the root of Masdevallia coccinea Linden ex Lindl

RESUMEN Masdevallia coccinea es una orquídea llamativa, endémica de Colombia, empleada en la generación de híbridos ornamentales con más de una década, clasificada como una especie en peligro de extinción. Entre las técnicas usadas para la propagación in vitro y ex situ de especies de la familia Orchidaceae, se estudia la simbiosis micorrízica, debido a que esta familia micoheterótrofa depende de una correlación con estos hongos para subsistir en las primeras etapas de desarrollo en estado silvestre. Con el objetivo de caracterizar e identificar los hongos asociados a las raíces de M. coccinea, se realizó un estudio histológico en raíces y, a partir de micropreparados, se caracterizó morfológicamente micro, macroscópica y molecularmente diez aislamientos. Se identificó a M. coccinea como una orquídea que presenta diferentes patrones de colonización micorrízicos y con posibles efectos endófitos de los géneros Aspergillus, Scopulariopsis, Trichoderma, Ilyonectria y del orden Xylariales en condiciones ex situ.

ABSTRACT Masdevallia coccinea is a striking orchid, endemic to Colombia, used in the generation of ornamental hybrids, with more than a decade classified as an endangered species. Among the techniques used for in vitro and ex situ propagation of species of the Orchidaceae family, mycorrhizal symbiosis is studied, because this mycoheterotrophic family depends on a correlation with these fungi to survive in the early stages of development in the wild. In order to characterize and identify the fungi associated with the roots of M. coccinea, a histological study was carried out on roots and 10 isolates were morphologically, macroscopically and molecularly characterized from micropreparations. M. coccinea is identified as an orchid with different mycorrhizal colonization patterns and with possible endophytic effects of the genera Aspergillus, Scopulariopsis, Trichoderma, Ilyonectria and the order Xylariales under ex situ conditions.

Endogenous Hormone Profile and Sugars Display Differential Distribution in Leaves and Pseudobulbs of Laelia anceps Plants Induced and Non-Induced to Flowering by Exogenous Gibberellic Acid.

A profile of endogenous hormones and sugars in leaves and pseudobulbs of Laelia anceps subsp. anceps (Orchidaceae) plants induced and non-induced to flowering by the effect of different doses of exogenous gibberellic acid (GA3), considering the current and back growth structures (CGS and BGS), were investigated. A factorial experiment with five doses of GA3 and two growth structures was designed. Adult plants with undifferentiated vegetative buds were selected and sprayed with doses of 0, 400, 600, 800, and 1000 mg GA3 L-1. The main results showed a strong interaction between GA3 dose and growth structures, which promoted the highest kinetin (KIN) concentration in CGS. Exogenous GA3 increased endogenous GA3 in leaves and pseudobulbs induced (I-Leaf and I-PSB) and non-induced (NI-Leaf and NI-PSB) to flowering. For sugar concentration, the 400 mg L-1 GA3 dose promotes significant interaction with the CGS in NI-PSB. In general, the hormone profile revealed opposite balances of endogenous hormone concentrations for KIN, zeatin (ZEA), trans-zeatin (T-ZEA), indoleacetic acid (IAA), indole-3-butyric acid (IBA) and GA3, not only for growth structures but also for vegetative organs analyzed, depending on whether the plants were induced or not induced to flowering, with the highest concentration of endogenous hormones in pseudobulbs. Likewise, different sugar concentration balances were observed. These balances of both endogenous hormones and sugars are likely to be involved in the flowering of L. anceps.

Germination and initial development in vitro, and acclimatization of Oeceoclades maculata (Orchidaceae), aiming at its reintroduction in natural environments

Oeceoclades maculata is a terrestrial orchid species that has potential for commercial purposes. Taking that into consideration, the present investigation aimed at studying its germination and initial development in vitro as well as its acclimatization. The influence of Murashige and Skoog (MS), Knudson C (KC), and Vacin and Went (VW) media in the presence and absence of 0.3% activated charcoal on in vitro germination and protocorm development were investigated. The effects of different concentrations of BA in combination with 0.5 mg L-1 NAA on seedling multiplication and growth were evaluated. The possibility of using dark-grown stem segments for micropropagation and acclimatization under laboratory and field conditions was also assessed. The results indicated that the most adequate media for germination were full-strength MS enriched with activated charcoal or KC supplemented with 1.5 mg L-1 BA in combination with 0.5 mg L-1 NAA. In terms of protocorm development, KC supplemented with 1.5 mg L-1 BA alone or in combination with 0.5 mg L-1 NAA provided the best results. The addition of 1.5 mg L-1 BA in combination with 0.5 mg L-1 NAA to KC medium favored the best results for seedling multiplication and development. The use of dark-grown stem segments is a viable alternative for the micropropagation of O. maculata. Regarding acclimatization, 100% survival of plants was observed during the initial phases and under field conditions average survival was 53.33%.(AU)

Are chromosome number and genome size associated with habit and environmental niche variables? Insights from the Neotropical orchids.

BACKGROUND AND AIMS: The entangled relationship of chromosome number and genome size with species distribution has been the subject of study for almost a century, but remains an open question due to previous ecological and phylogenetic knowledge constraints. To better address this subject, we used the clade Maxillariinae, a widely distributed and karyotypically known orchid group, as a model system to infer such relationships in a robust methodological framework. METHODS: Based on the literature and new data, we gathered the chromosome number and genome size for 93 and 64 species, respectively. We built a phylogenetic hypothesis and assessed the best macroevolutionary model for both genomic traits. Additionally, we collected together ecological data (preferences for bioclimatic variables, elevation and habit) used as explanatory variables in multivariate phylogenetic models explaining genomic traits. Finally, the impact of polyploidy was estimated by running the analyses with and without polyploids in the sample. KEY RESULTS: The association between genomic and ecological data varied depending on whether polyploids were considered or not. Without polyploids, chromosome number failed to present consistent associations with ecological variables. With polyploids, there was a tendency to waive epiphytism and colonize new habitats outside humid forests. The genome size showed association with ecological variables: without polyploids, genome increase was associated with flexible habits, with higher elevation and with drier summers; with polyploids, genome size increase was associated with colonizing drier environments. CONCLUSIONS: The chromosome number and genome size variations, essential but neglected traits in the ecological niche, are shaped in the Maxillariinae by both neutral and adaptive evolution. Both genomic traits are partially correlated to bioclimatic variables and elevation, even when controlling for phylogenetic constraints. While polyploidy was associated with shifts in the environmental niche, the genome size emerges as a central trait in orchid evolution by the association between small genome size and epiphytism, a key innovation to Neotropical orchid diversification.

Floral Scent Chemistry and Pollinators of a Sexually Dimorphic Neotropical Orchid.

Catasetum is a speciose Neotropical orchid genus of which male and female flowers emit scents acting both as attractant and reward for their exclusive pollinators, male orchid bees (Euglossini: Apidae). In Catasetum, it is well known that flowers display a remarkably morphological sexual dimorphism. However, it remains poorly investigated whether this is also true for floral scents. Here, we investigated the pollination ecology and floral scent traits (chemistry and total emission) of C. maranhense, a species endemic to the Brazilian N/NE region. Males of Euglossa securigera are the only pollinators of C. maranhense. The floral scent of C. maranhense is composed of 29 volatile compounds, with eucalyptol, indole, (E)-Methyl p-methoxycinnamate, and (Z)-Methyl p-methoxycinnamate accounting for more than 80% of the scent bouquet. No sexual dimorphism was detected in any of the traits investigated. We discuss the ecological and evolutionary significance of our findings to Catasetum species and other unisexual perfume plants.