Orchid biology: from Linnaeus via Darwin to the 21st century

Orchidaceae are the largest family of flowering plants, with at least 24 000 species, and perhaps better than any other family of flowering plants, orchids represent the extreme specializations that are possible. As a result, they have long fascinated luminaries of the botanical world including Linnaeus and Darwin, but the size of the family has historically been an impediment to their study. Specifically, the lack of detailed information about relationships within the family made it difficult to formulate explicit evolutionary hypotheses for such a large group, but the advent of molecular systematics has revolutionized our understanding of the orchids. Their complex life histories make orchids particularly vulnerable to environmental change, and as result many are now threatened with extinction. In this Special Issue we present a series of 20 papers on orchid biology ranging from phylogenetics, floral evolutionary development, taxonomy, mycorrhizal associations, pollination biology, population genetics and conservation.     

Reproductive biology and conservation genetics of Goodyera procera (Orchidaceae)

Goodyera procera is an endangered terrestrial orchid in Hong Kong. Information on its reproductive biology and pattern of genetic variation is needed to develop efficient conservation strategies. Pollination experiments showed that the species is self-compatible, but dependent on pollinators for fruit set. Bagged plants produced no fruits. Artificial pollinations resulted in 92% fruit set through selfing, 94% with geitonogamous pollination, and 95% following xenogamous pollination. Fruit set in the open-pollinated control was 75% at the same sites. Allozyme electrophoresis and random amplified polymorphic DNA (RAPD) were used to evaluate genetic variation and structure of 15 populations of Goodyera procera. Despite its outbreeding system, allozyme data revealed low variation both at the population (P = 21.78%, A = 1.22, and H = 0.073) and species (P = 33%, A = 1.33, and H = 0.15) levels, in comparison with other animal-pollinated outbreeding plant species. However, RAPD variation was relatively high (P = 55.13% and H = 0.18 at the population level, and P = 97.03% and H = 0.29 at the species level). G_ST estimates indicated high levels of genetic differentiation among populations (G_ST = 0.52 and I = 0.909 ± 0.049 based on allozyme data, and G_ST = 0.39 and I = 0.859 ± 0.038 based on RAPD data), much above the average for outcrossing species, suggesting that gene flow was limited in this species. Based on these data, suitable strategies were developed for the genetic conservation and management of the species.     

Orchid conservation in China from 2000 to 2020: Achievements and perspectives

We review achievements in the conservation of orchid diversity in China over the last 21 years. We provide updated information on orchid biodiversity and suggestions for orchid conservation in China. We outline national policies of biodiversity conservation, especially of orchid conservation, which provide general guidelines for orchid conservation in China. There are now approximately 1708 known species of Orchidaceae in 181 genera in China, including five new genera and 365 new species described over the last 21 years. The assessment of risk of extinction of all 1502 known native orchid species in China in 2013 indicated that 653 species were identified as threatened, 132 species were treated as data-deficient, and four species endemic to China were classified as extinct. Approximately 1100 species (ca. 65%) are protected in national nature reserves, and another ~66 species in provincial nature reserves. About 800 native orchid species have living collections in major botanical gardens. The pollination biology of 74 native orchid species and the genetic diversity and spatial genetic structure of 29 orchid species have been investigated at a local scale and/or across species distributions. The mycorrhizal fungal community composition has been investigated in many genera, such as Bletilla, Coelogyne, Cymbidium, Cypripedium, and Dendrobium. Approximately 292 species will be included in the list of national key protected wild plants this year. Two major tasks for near future include in situ conservation and monitoring population dynamics of endangered species.     

Floral biology and histochemical analysis of Vanilla edwallii Hoehne (Orchidaceae: Vanilloideae): an orchid pollinated by Epicharis (Apidae: Centridini)

The genus Vanilla is the most diverse in Vanilloideae, with ca 90 species distributed among tropical regions. Despite their economic importance, studies on pollination of Vanilla are very scarce and data on pollinators of species endemic to Brazil are lacking. Based on fieldwork and laboratory investigations, the floral biology of V. edwallii was studied. The pollinators and pollination process were recorded at the Serra do Japi reserve, state of São Paulo, southeastern Brazil, and the presence of floral reward was also investigated. Vanilla edwallii blooms in summer. The lateral inflorescences produce up to four pale green flowers. The white labellum is united to the base of the column forming a mentum. In the studied population V. edwallii is pollinated by Epicharis (Hoplepicharis) affinis, where the males exhibit a territorial behavior, defending flowers from other possible flower visitors. The pollen is deposited on the scutellum of bees when they abandon the flower. The mentum region is dry, suggesting no nectar production. The only secretory structures are osmophores dispersed on the inner surface of the lip responsible for production of a sweet fragrance, which together with color and morphology of flowers is related to bee attraction. The labellum is rich in mucilaginous cells, while the mucilaginous substance is retained inside the cells. The histochemical analysis also detected the presence of phenolic compounds and starch concentrated mainly at the adaxial surface of the lip and around the vascular bundles.     

Fruit set, nectar reward, and rarity in the Orchidaceae

A review of comparative levels of reproductive success among nectariferous and nectarless orchids worldwide was compiled from a comprehensive survey of fruit set from 117 orchid species in the literature and from our own field studies. It confirms the hypothesis that nectariferous orchids are more successful in setting fruit than are nectarless species. Overall fruit set figures for nectarless and nectariferous orchids were 19.5 and 49.3% for North America, 27.7 and 63.1% for Europe, 41.4 and 74.4% for the temperate southern hemisphere, and 11.5 and 24.9% for the tropics, demonstrating that the dichotomy is consistent across all geographical areas. On average, the provision of nectar doubles the probability of fruit set in both temperate and tropical areas, but tropical orchids are remarkable in that all (whether nectarless or nectariferous, or terrestrial or epiphytic) display low fruit productivity (<50%). Fruiting failure in the tropics may be balanced by higher productivity per capsule, since tropical orchid fruits contain on average 150 times more seeds than temperate ones. Hybridization occurs more frequently among nectarless orchids in Britain and Europe than among nectariferous ones, and there is a significant positive association between orchid rarity and lack of nectar reward in the British Isles. Sexual reproduction in the Orchidaceae is predominantly pollinator dependent, but this can sometimes be successfully circumvented by asexual seed production (agamospermy) or, more frequently, by automatic self-pollination (autogamy). The proportion of highly successful nectarless orchids from all geographic areas is very low and comparable with that of orchids offering rewards other than nectar (～14% of species in each case) emphasizing that high reproductive success is only associated with nectar reward (53% of species). It is suggested that the evolution of nectar production within the family has been the most frequent means of escaping the reproductive limitations of low pollinator visitation frequencies.     

Reproductive biology of Trichocentrum pumilum: an orchid pollinated by oil-collecting bees

The reproductive biology, reward production and pollination mechanism of Trichocentrum pumilum were studied in a gallery forest in the interior of the State of São Paulo, southeast Brazil. The floral visitors and pollination mechanism were recorded, and experimental pollinations were carried out in order to determine the breeding system of this species. Trichocentrum pumilum blooms in spring. Each paniculate inflorescence bears an average of 85 flowers that present a central yellow callus and finger‐like trichomes on the lateral lobes of the lip. A lipoidal substance is produced and stored among these trichomes. In the studied population, T. pumilum is exclusively visited and pollinated by two bee species (Tetrapedia diversipes and Lophopedia nigrispinis). Pollinaria are deposited on mouthparts of bees during collection of the lipoidal substance from the lateral lobes of the labellum. Trichocentrum pumilum is self‐incompatible and pollinator‐limited. Natural fruit set was low (9%, compared to 45% in experimentally cross‐pollinated flowers). Potentially viable seed exceed 97% in fruits obtained through cross‐pollination and in natural conditions (open pollination).     

Genetic Diversity and Structure in Fragmented Populations of the Tropical Orchid Myrmecophila christinae var christinae1

In recent years the coastal dune vegetation of the state of Yucatan, Mexico has become highly fragmented due to clearing for development. We evaluated patterns of genetic differentiation and genetic variability among orchid populations (Myrmecophila christinae var christinae) in eight habitat fragments along a west‐to‐east disturbance gradient in which sites located on the western end began experiencing fragmentation earlier than those in the east. Leaf samples from adult individuals and juvenile plants were collected from all eight populations, and analyzed using starch‐gel electrophoresis in a lithium buffer system. Per population estimates of genetic diversity, proportion of polymorphic loci at 95 percent, mean number of alleles per locus, allelic richness, and population structure were calculated, as well as estimate comparisons between generations. Genetic diversity at the loci analyzed did not show significant differences among the eight populations. Based on the results, the model of isolation by distance does not fit the M. christinae populations under study; in some cases, populations close to one another apparently experienced very little genetic exchange. Thus, we propose that so far, fragmentation has not led to significant genetic differences between populations subject to different historical backgrounds of disturbance (i.e., time since onset of disturbance), as well as between generations within each of the M. christinae study populations. Nevertheless, the genetic characteristics of some remnant populations might change over time due to a future decrease in the opportunities for genetic exchange with other populations.     

明党参的濒危机制及其保护对策的研究

本文对明党参的资源状况、种群生态、种子结构、植株结构实率与环境的关系及传粉生物学等方面进行了研究。从种子形态及分子水平对明党参的遗传多样性和遗传结构进行了分析。研究结果表明,明党参种子后熟阶段与梅雨气候的不协调是导致种子萌发率低及更新种群少的主要原因之一;人类活动破坏了明党参的适宜生境和种群数量,使得种群间基因流减小,进而导致严重的遗传漂变及种群间的分化。由此提出了对明常参就地和迁地保护的有效措施。     

Biosystematics of the genus Piperia Rydb. (Orchidaceae)

Piperia is a North American segregate of Habenaria consisting of 5 taxa (4 species and 1 subspecies). The genus is defined and differentiated from other North American genera of the Habenaria alliance by its morphology and pollination mechanism. All taxa have n = 21 chromosomes. Thin-layer chromatography of fresh leaf extracts and a morphological assessment revealed discrete, but variable, taxa. All species are partially interfertile; however, a high degree of sympatry suggests these species are reproductively isolated. One new combination is proposed: P. elongata subsp. michaelii.     

Reproductive biology of Cyrtopodium punctatum in situ: implications for conservation of an endangered Florida orchid

Cyrtopodium punctatum (Linnaeus) Lindley is an endangered epiphytic orchid restricted in the USA to southern Florida. This species has been extensively collected from the wild since the early 1900s, and today only a few plants remain in protected areas. As part of a conservation plan, a reproductive biology study was conducted to better understand the ecology of this species in Florida. Cyrtopodium punctatum relies on a deceit pollination system using aromatic compounds to attract pollinators. Nine aromatic compounds were identified as components of the fragrance of C. punctatum inflorescences, including two compounds that are known to be Euglossine bee attractants. However, this group of bees is not native to Florida. Of the four bee species observed to visit C. punctatum flowers in the present study, carpenter bees ( Xylocopa spp.) are likely to be the main pollinators. Pollination experiments demonstrated that C. puntatum is self-compatible, but requires a pollinator and thus does not exhibit spontaneous autogamy. In addition, the rates of fruit set were significantly higher for flowers that were outcrossed (xenogamy) than for those that were self-crossed. Thus, the species has evolved a degree of incompatibility. Examples of natural pollination and fruit set were observed during the present study (2007–2008), but the rates of reproduction were modest as a consequence of the low plant numbers and possible changes in insect densities as a result of anthropogenic influences.     

Composition of Orchid Scents Attracting Euglossine Bees

Fragrance composition of flowers from 101 plant species, especially orchids, were analyzed. Several compounds, including allo-aromadendrene, β-bourbonene, α-copaene, α-cubebene, 1,2-dimethoxybenzene, 1,3,5-trimethoxybenzene, epoxygeranyl acetate, 7,11-epoxymegastigma-5(6)-en-9-one, two γ-lactones, germacradienol, germacrene D, humulene, methoxy-phenyl-ethyl acetate, myrcene epoxide, sabinene, styrene, and undecatriene were detected for the first time in orchids. Fragrance composition of flowers pollinated by male euglossine bee species (perfume flowers) of four plant families are compared and contrasted with those of orchid species with other pollination systems. Melittophilous, but highly specialized orchids, produce fragrances rich in different sesquiterpenes and other rare compounds. In the species that are exclusively pollinated by fragrance-collecting male euglossine bees, the fragrances are primary attractants that serve both as an attractant and as a reward. The unusually intensely smelling flowers mostly produce esters and monoterpenes. The fragrances of euglossophilous flowers of the three plant families investigated are composed of nearly the same sets of chemical compounds, suggesting convergent evolution. Typically, euglossophilous plant species produce large amounts of few fragrant substances while melittophilous species often produce rare compounds in small amounts.     

广西雅长林区野生兰科植物资源现状与保护策略

广西雅长林区野生兰科植物资源丰富,约有52属156种,其中雅长保护区几乎每座山头都有兰科植物的分布,但其种类与居群基株数量等分布不均匀,且不连续,呈现出破碎化现象,其中面积较大的局部密集分布区有16处.文中根据野生兰科植物的资源现状及生物生态学特性,针对目前兰科植物保育存在的问题,分析人畜干扰及自然条件变化等不利因素,从兰科植物维持机制角度出发,提出雅长野生兰科植物的保护应以就地保护为主、迁地保育为辅的保育措施与策略:保护区应加快建设的步伐,构建野生兰科植物种质资源基因库,同时对密集分布区在人畜干扰大的区域,优先拉设铁丝网围护,并派专人巡护,重点保护,确保野生兰科植物及基因库安全.     

Microbiota of the Orchid Rhizoplane

Six bacterial strains isolated from the underground roots of the terrestrial orchid Calanthe vestitavar. rubro-oculatawere found to belong to the genera Arthrobacter, Bacillus, Mycobacterium, and Pseudomonas.Strains isolated from the aerial roots of the epiphytic orchid Dendrobium moschatumwere classified into the genera Bacillus, Curtobacterium, Flavobacterium, Nocardia, Pseudomonas, Rhodococcus, and Xanthomonas.The rhizoplane of the terrestrial orchid was also populated by cyanobacteria of the genera Nostocand Oscillatoria, whereas that of the epiphytic orchid was populated by one genus, Nostoc.In orchids occupying different econiches, the spectra of the bacterial genera revealed differed. The microbial complex of the terrestrial orchid rhizoplane differed from that of the surrounding soil.     

Diurnal butterfly pollination in the orchid Habenaria radiata

Habenaria is one of the largest orchid genera, with its main centers of diversity in Brazil, eastern Asia and central and southern Africa. In spite of its diversity, the pollinators of few Habenaria spp. have been studied, while most published reports indicate that Habenaria spp. are mainly pollinated by moths. Here we report that the diurnal skipper Parnara guttata can be at least one of the pollinators of Habenaria radiata. Skippers have generally been thought of as nectar thieves, transferring few pollen grains. However, orchids transmit their pollen grains in large groups within pollinia. Such innovation can probably render the skipper a pollinator of H. radiata.     

Visitation rate of pollinators and nectar robbers to the flowers and inflorescences of Tabebuia aurea (Bignoniaceae): effects of floral display size and habitat fragmentation

Large floral displays favour pollinator attraction and the import and export of pollen. However, large floral displays also have negative effects, such as increased geitonogamy, pollen discounting and nectar/pollen robber attraction. The size of the floral display can be measured at different scales (e.g. the flower, inflorescence or entire plant) and variations in one of these scales may affect the behaviour of flower visitors in different ways. Moreover, the fragmentation of natural forests may affect flower visitation rates and flower visitor behaviour. In the present study, video recordings of the inflorescences of a tree species (Tabebuia aurea) from the tropical savannah of central Brazil were used to examine the effect of floral display size at the inflorescence and tree scales on the visitation rate of pollinators and nectar robbers to the inflorescence, the number of flowers approached per visit, the number of visits per flower of potential pollinators and nectar robbers, and the interaction of these variables with the degree of landscape disturbance. Nectar production was quantified with respect to flower age. Although large bees are responsible for most of the pollination, a great diversity of flower insects visit the inflorescences of T. aurea. Other bee and hummingbird species are highly active nectar robbers. Increases in inflorescence size increase the visitation rate of pollinators to inflorescences, whereas increases in the number of inflorescences on the tree decrease visitation rates to inflorescences and flowers. This effect has been strongly correlated with urban environments in which trees with the largest floral displays are observed. Pollinating bees (and nectar robbers) visit few flowers per inflorescence and concentrate visits to a fraction of available flowers, generating an overdispersed distribution of the number of visits per inflorescence and per flower. This behaviour reflects preferential visits to young flowers (including flower buds) with a greater nectar supply.     

Exine Micromorphology of Orchidinae (Orchidoideae,Orchidaceae): Phylogenetic Constraints or Ecological Influences?

BACKGROUND AND AIMS: Pollen characters have been widely used in defining evolutionary trends in orchids. In recent years, information on pollination biology and phylogenetic patterns within Orchidinae has become available. Hence, the aim of the presented work is to re-evaluate exine micromorphology of Orchidinae in light of recent phylogenetic studies and to test whether pollen micromorphology strictly depends on phylogenetic relationships among species or whether it is influenced by the marked differences in pollination ecology also reported among closely related species. METHODS: Pollen sculpturing of 45 species of Orchidinae and related taxa was investigated using scanning electron microscopy. To cover potential intraspecific variation, several accessions of the same species were examined. KEY RESULTS: Orchidinae show remarkable variation in exine sculpturing, with a different level of variation within species groups. In some genera, such as Serapias (rugulate) and Ophrys (psilate to verrucate), intrageneric uniformity corresponds well to a common pollination strategy and close relationships among species. However, little exine variability (psilate-scabrate and scabrate-rugulate) was also found in the genus Anacamptis in spite of striking differences in floral architecture and pollination strategies. A larger variety of exine conditions was found in genera Dactylorhiza (psilate, psilate-scabrate and reticulate) and Orchis s.s. (psilate, reticulate, perforate-rugulate and baculate) where no unequivocal correspondence can be found to either phylogenetic patterns or pollination strategies. CONCLUSIONS: Changes in pollen characteristics do not consistently reflect shifts in pollination strategy. A unique trend of exine evolution within Orchidinae is difficult to trace. However, the clades comprising Anacamptis, Neotinea, Ophrys and Serapias show psilate to rugulate or scabrate pollen, while that of the clade comprising Chamorchis, Dactylorhiza, Gymnadenia, Orchis s.s., Platanthera, Pseudorchis and Traunsteinera ranges from psilate to reticulate. Comparison of the data with exine micromorphology from members of the tribe Orchidieae and related tribes suggests a possible general trend from reticulate to psilate.