Orchid seed sensitivity to nitrate reflects habitat preferences and soil nitrate content

• Orchids are distributed around the world, however, the factors shaping their specific distribution and habitat preferences are largely unknown. Moreover, many orchids are at risk of becoming threatened as landscapes change, sometimes declining without apparent reason. One important factor affecting plant distribution is nutrient levels in the environment. Nitrates can inhibit not only orchid growth and persistence, but also seed germination.
• We used in vitro axenic cultures to exactly determine the germination sensitivity of seven orchid species to nitrates and correlated this with soil properties of the natural sites and with the species’ habitat preferences.
• We found high variation in response to nitrate between species. Orchids from oligotrophic habitats were highly sensitive, while orchids from more eutrophic habitats were almost insensitive. Sensitivity to nitrate was also associated with soil parameters that indicated a higher nitrification rate.
• Our results indicate that nitrate can affect orchid distribution via direct inhibition of seed germination. Nitrate levels in soils are increasing rapidly due to intensification of agricultural processes and concurrent soil pollution, and we propose this increase could cause a decline in some orchid species.

     

Surprisingly high orchid diversity in travertine and forest areas in the Huanglong valley, China, and implications for conservation

The presence of such a large number of terrestrial orchid species in a small area (ca. 1 km²) of the Huanglong valley in southwestern China is uncommon for this country. Studying the relationship between the distribution patterns of these orchid species and their microenvironments may help us understand this uncommon phenomenon. We established 662 1 m × 1 m plots, measured the cover of each species and found that there were 33 orchid species distributed mainly in two different habitats, i.e. travertine areas and forest. In the travertine areas, 30 orchid species were found; the six most common ones being Cypripedium bardolphianum, Cypripedium flavum, Cypripedium tibeticum, Galearis diantha, Ponerorchis chusua and Phaius delavayi. However in the forested habitat, we found 21 orchid species; the most common ones being Tipularia szechuanica and Goodyera repens. Travertine areas had a higher number of orchid species as well as higher numbers of orchid species per plot as compared to forest. Light availability seems critical to the performance and distribution of orchid species. Stream flow through the travertine area during the orchids growing season appears to be an important factor in shaping and maintaining stable microenvironments favorable to the growth and reproduction of orchids. The results presented in this study suggest that some orchid species in the travertine area might be threatened if the travertine stream flows were to change or be disrupted.     

Genetic inference of epiphytic orchid colonization; it may only take one

Colonization of vacant habitat is a fundamental ecological process that affects the ability of species to persist and undergo range modifications in continually shifting landscapes. Thus, understanding factors that affect and limit colonization has important ecological and conservation implications. Epiphytic orchids are increasingly threatened by various factors, including anthropogenic habitat disturbance. As cleared areas (e.g. pastures) are recolonized by suitable host trees, the establishment and genetic composition of epiphytic orchid populations are likely a function of their colonization patterns. We used genetic analyses to infer the prevailing colonization pattern of the epiphytic orchid, Brassavola nodosa. Samples from three populations (i.e. individuals within a tree) from each of five pastures in the dry forest of Costa Rica were genotyped with neutral nuclear and chloroplast markers. Spatial autocorrelation and hierarchical genetic structure analyses were used to assess the relatedness of individuals within populations, among populations within pastures and among populations in different pastures. The results showed significant relatedness within populations (mean r = 0.166) and significant but lower relatedness among populations within a pasture (mean r = 0.058). Our data suggest that colonization of available habitats is by few individuals with subsequent population expansion resulting from in situ reproduction, and that individuals within a tree are not a random sample of the regional seed pool. Furthermore, populations within a pasture were likely colonized by seeds produced by founders of a neighbouring population within that pasture. These results have important ramifications for understanding conservation measures needed for this species and other epiphytic orchids.     

Niche analysis of orchids of serpentine and non-serpentine areas: Implications for conservation

Orchids are known for their species richness, intriguing ecology, rarity and the fact that they grow in almost all terrestrial ecosystems. Although numerous studies about their ecology have been carried out concerning calcareous areas, little is known about orchids that occur in serpentine habitats. The aim of this study was to investigate the ecological preferences of orchids in serpentine and non-serpentine areas on the model of the Valjevo Mountain Range (W Serbia). Niche analysis of orchids was performed using outlying mean index analysis. Data concerning geographical coordinates, altitude, habitat type, inclination, bedrock type, light regime, soil moisture, acidity, nitrogen and temperature were used as explanatory variables. Data of 33 orchid taxa from 407 localities were analysed. The most important gradients that govern orchid distribution were geological bedrock, light regime and temperature. The results have shown that only Anacamptis morio and Gymnadenia conopsea have statistically significantly larger populations on serpentine compared with non-serpentine bedrocks. This study highlights the importance of serpentine habitats as orchid habitats, bearing in mind the occurrence of rare species and species which were found exclusively in serpentine habitats.     

In vitro propagation of Paphiopedilum orchids

Paphiopedilum is one of the most popular and rare orchid genera. Members of the genus are sold and exhibited as pot plants and cut flowers. Wild populations of Paphiopedilum are under the threat of extinction due to over-collection and loss of suitable habitats. A reduction in their commercial value through large-scale propagation in vitro is an option to reduce pressure from illegal collection, to attempt to meet commercial needs and to re-establish threatened species back into the wild. Although they are commercially propagated via asymbiotic seed germination, Paphiopedilum are considered to be difficult to propagate in vitro, especially by plant regeneration from tissue culture. This review aims to cover the most important aspects and to provide an up-to-date research progress on in vitro propagation of Paphiopedilum and to emphasize the importance of further improving tissue culture protocols for ex vitro-derived explants.     

Fungal biodiversity in aquatic habitats

Fungal biodiversity in freshwater, brackish and marine habitats was estimated based on reports in the literature. The taxonomic groups treated were those with species commonly found on submerged substrates in aquatic habitats: Ascomycetes (exclusive of yeasts), Basidiomycetes, Chytridiomycetes, and the non-fungal Saprolegniales in the Class Oomycetes. Based on presence/absence data for a large number and variety of aquatic habitats, about 3,000 fungal species and 138 saprolegnialean species have been reported from aquatic habitats. The greatest number of taxa comprise the Ascomycetes, including mitosporic taxa, and Chytridiomycetes. Taxa of Basidiomycetes are, for the most part, excluded from aquatic habitats. The greatest biodiversity for all groups occurs in temperate areas, followed by Asian tropical areas. This pattern may be an artifact of the location of most of the sampling effort. The least sampled geographic areas include Africa, Australia, China, South America and boreal and tropical regions worldwide. Some species overlap occurs among terrestrial and freshwater taxa but little species overlap occurs among freshwater and marine taxa. We predict that many species remain to be discovered in aquatic habitats given the few taxonomic specialists studying these fungi, the few substrate types studied intensively, and the vast geographical area not yet sampled.     

Symbiotic seed germination of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid

The rapid loss of native orchid habitat throughout ecologically important areas (e.g., Florida) has prompted researchers to develop appropriate plans for the propagation and reintroduction of many native orchid species. Ideally, symbiotic orchid seed germination methods are utilized in the production of orchid seedlings to be used in plant reintroduction programs. In the current study we (1) describe an efficient symbiotic seed germination protocol to germinate seeds of the rare sub-tropical terrestrial orchid Habenaria macroceratitis; (2) discuss the in vitro fungal specificity demonstrated by this species; and (3) describe the effects of three photoperiods (0/24 h, 16/8 h, 24/0 h L/D) on in vitro symbiotic seed germination of H. macroceratitis. Six fungal mycobionts were isolated from both vegetative and flowering plants of H. macroceratitis from two geographically distinct sites. Symbiotic seed germination percent was highest (65.7%) and protocorm development was most advanced (Stage 2) when seeds were cultured with fungal mycobiont Hmac-310. Seeds of H. macroceratitis demonstrated a degree of specificity toward fungal mycobionts isolated from plants originating from the same site where seed was collected. Continual darkness (0/24 h L/D) inhibited initial seed germination (Stage 1; 17.1%), but stimulated subsequent protocorm development (Stage 2; 53.5%). These findings will aid in developing an efficient symbiotic seed germination protocol for the conservation of this rare Florida terrestrial orchid, and may prove useful in the conservation of other sub-tropical terrestrial orchid species.     

Dust seed production and dispersal in Swedish Pyroleae species

Dust seeds are the smallest seeds in angiosperms weighing just about a few micrograms. These seeds are characteristic of most orchids, and several studies have been performed on seed features, fecundity and dispersal of orchid dust seeds. In this study we examine seed features, seed production and seed dispersal in another plant group with dust seeds, the Pyroleae (Monotropoideae, Ericaceae), focusing on six species: Pyrola chlorantha, P. minor, P. rotundifolia, Chimaphila umbellata, Moneses uniflora and Orthilia secunda. Seed production per capsule among these species was in the range between ca 1000 and 7800, and seed production per capsule bearing shoot was in the range between ca 7000 and 60 000. Combining our results with published information on pollen‐ovule ratios suggests that these Pyroleae species have a generally efficient pollination system. The most fecund species was P. minor, the only species among the investigated that is probably largely self‐pollinating. The investigated Pyroleae species have a seed production comparable to the less fecund orchid species. We studied seed dispersal in the field in one of the species, P. chlorantha. Despite the extremely small and potentially buoyant seeds, the vast majority of seeds are deposited close to the seed source, within a few meters. Further studies on the recruitment ecology of the investigated Pyroleae species are currently under way.     

Sea level rise may increase extinction risk of a saltmarsh ontogenetic habitat specialist

Specialist species are more vulnerable to environmental change than generalist species. For species with ontogenetic niche shifts, specialization may occur at a particular life stage making those stages more susceptible to environmental change. In the salt marshes in the northeast U.S., accelerated sea level rise is shifting vegetation patterns from flood‐intolerant species such as Spartina patens to the flood‐tolerant Spartina alterniflora. We tested the potential impact of this change on the coffee bean snail, Melampus bidentatus, a numerically dominant benthic invertebrate with an ontogenetic niche shift. From a survey of eight marshes throughout the northeast U.S., small snails were found primarily in S. patens habitats, and large snails were found primarily in stunted S. alterniflora habitats. When transplanted into stunted S. alterniflora, small snails suffered significantly higher mortality relative to those in S. patens habitats; adult snail survivorship was similar between habitats. Because other habitats were not interchangeable with S. patens for young snails, these results suggest that Melampus is an ontogenetic specialist where young snails are habitat specialists and adult snails are habitat generalists. Temperature was significantly higher and relative humidity significantly lower in stunted S. alterniflora than in S. patens. These data suggest that thermal and desiccation stress restricted young snails to S. patens habitat, which has high stem density and a layer of thatch that protects snails from environmental stress. Other authors predict that if salt marshes in the northeast U.S. are unable to migrate landward, sea level rise will eliminate S. patens habitats. We suggest that if a salt marsh loses its S. patens habitats, it will also lose its coffee bean snails. Our results demonstrate the need to consider individual life stages when determining a species’ vulnerability to global change.     

Studies of Mycorrhizal Fungi of Chinese Orchids and Their Role in Orchid Conservation in China—A Review

China has over 1,200 species of native orchids in nearly 173 genera. About one fourth of native species are of horticultural merit. Some species are of Chinese medicinal value. In fact, the demand on orchid species with high Chinese medicinal values such as Gastrodia elata, Dendrobium offcinale, along with demands on species of cultural importance, such as those in the genus of Cymbidium, is a major factor causing wild populations to diminish and in some cases, drive wild populations to the brink of extinction. These market demands have also driven studies on the role of mycorrhizal fungi in orchid seed germination, seedling and adult growth, and reproduction. Most of these mycorrhizal studies of Chinese orchids, however, are published in Chinese, some in medical journals, and thus overlooked by the mainstream orchid mycorrhizal publications. Yet some of these studies contained interesting discoveries on the nature of the mycorrhizal relationships between orchids and fungi. We present a review of some of these neglected publications. The most important discovery comes from the mycorrhizal studies on G. elata, in which the researchers concluded that those fungi species required to stimulate seed germination are different from those that facilitate the growth of G. elata beyond seedling stages. In addition, presence of the mycorrhizal fungi associated with vegetative growth of post-seedling G. elata hindered the germination of seeds. These phenomena were unreported prior to these studies. Furthermore, orchid mycorrhizal studies in China differ from the mainstream orchid studies in that many epiphytic species (in the genus of Dendrobium, as medicinal herbs) were investigated as well as terrestrial orchids (mostly in the genus Cymbidium, as traditional horticultural species). The different responses between epiphytic and terrestrial orchid seeds to fungi derived from roots suggest that epiphytic orchids may have a more general mycorrhizal relationship with fungi than do terrestrial orchid species during the seed germination stage. To date, orchid mycorrhizal research in China has had a strongly commercial purpose. We suggest that this continuing research on orchid mycorrhizal relationships are a solid foundation for further research that includes more rare and endangered taxa, and more in-situ studies to assist conservation and restoration of the endangered orchids. Knowledge on the identities and roles of mycorrhizal fungi of orchids holds one of the keys to successful restoration and sustainable use of Chinese orchids.     

Endophytic fungi from Pecteilis susannae (L.) Rafin (Orchidaceae), a threatened terrestrial orchid in Thailand

Eight endophytic fungi were isolated from roots of the threatened terrestrial orchid, Pecteilis susannae (L.) Rafin. Phylogenetic analysis based on an alignment of internal transcribed spacer regions of nuclear rDNA indicated that seven isolates belonged to the genus Epulorhiza and one to Fusarium. All fungal isolates were cultured with orchid seeds collected from three field sites near Doi Suthep-Pui National Park, Chiang Mai, Thailand. Seed germination and protocorm development were evaluated up to 70 days after sowing. Percent symbiotic seed germination was highest (86.2%) when seeds were cultured with Epulorhiza (CMU-Aug 013). The protocorm development was the most advanced up to stage 2, continued embryo enlargement, or rupture of the testa, and the highest percentage was 17.8% when seeds were cultured with Epulorhiza (CMU-Aug 007). Without fungi, seed germination and protocorm development were 62.1% and 11.1%, respectively. The dependency of P. susannae on fungal symbionts for early seedling development is yet to be determined. Optimizing seed germination and seedling fitness will assist the conservation of this threatened orchid in Thailand.     

Characterization of microsatellite loci in the endangered grand spider orchid Caladenia huegelii (Orchidaceae)

The orchid genus Caladenia is species rich with many threatened and endangered taxa. We report on the isolation and characterization of microsatellite loci in the rare Caladenia huegelii for molecular evaluation of this and the closely related C. thinicola as part of the development of conservation initiatives. Eight di‐ and trinucleotide loci were screened using 30 samples from each species. All loci were highly variable, with similar levels of heterozygosity and number of alleles across both species. These markers will be highly informative for population studies in both species.     

A narrowly endemic photosynthetic orchid is non‐specific in its mycorrhizal associations

Mycorrhizal association is a common characteristic in a majority of land plants, and the survival and distribution of a species can depend on the distribution of suitable fungi in its habitat. Orchidaceae is one of the most species‐rich angiosperm families, and all orchids are fully dependent on fungi for their seed germination and some also for subsequent growth and survival. Given this obligate dependence, at least in the early growth stages, elucidating the patterns of orchid–mycorrhizal relationships is critical to orchid biology, ecology and conservation. To assess whether rarity of an orchid is determined by its specificity towards its fungal hosts, we studied the spatial and temporal variability in the host fungi associated with one of the rarest North American terrestrial orchids, Piperia yadonii. The fungal internal transcribed spacer region was amplified and sequenced by sampling roots from eight populations of P. yadonii distributed across two habitats, Pinus radiata forest and maritime chaparral, in California. Across populations and sampling years, 26 operational taxonomic units representing three fungal families, the Ceratobasidiaceae, Sebacinaceae and Tulasnellaceae, were identified. Fungi belonging to the Sebacinaceae were documented in orchid roots only at P. radiata forest sites, while those from the Ceratobasidiaceae and Tulasnellaceae occurred in both habitats. Our results indicate that orchid rarity can be unrelated to the breadth of mycorrhizal associations. Our data also show that the dominance of various fungal families in mycorrhizal plants can be influenced by habitat preferences of mycorrhizal partners.     

Taxonomic and functional characterisation of fungi from the Sebacina vermifera complex from common and rare orchids in the genus Caladenia

The terrestrial orchid genus Caladenia contains many species which are threatened with extinction. They have highly specific associations with Sebacina vermifera and closely related fungi, and conservation of these terrestrial orchids, in part, relies on symbiotic propagation to produce plants for reintroduction and ex situ conservation collections. However, little is known of the diversity of mycorrhizal fungi associating with natural populations. Here, restriction fragment polymorphism analysis, internal transcribed spacer and nuclear large subunit sequencing and symbiotic seed germination were used to investigate the taxonomic and functional diversity of fungal isolates from single populations of six endangered Caladenia species and one common species across the same biogeographic range. Fifty-nine fungal isolates were collected for investigation including ten isolates from the six endangered species Caladenia audasii, Caladenia amoena, Caladenia sp. aff. fragrantissima (Central Victoria), Caladenia sp. aff. patersonii, Caladenia rosella and Caladenia orientalis and 49 isolates from six populations of the common species Caladenia tentaculata. While the common species associated with three distinct S. vermifera-like taxa, the six endangered species were restricted to one of these fungal taxa. No direct relationship between the taxonomic identity of the fungi and their ability to stimulate seed germination was observed; however, the majority of the fungi isolated from the Caladenia species were capable of germinating seed in vitro, indicating their mycorrhizal status and potential for symbiotic propagation in conservation programmes.     

Seed morphology in the tribe Chloraeeae (Orchidaceae): combining traditional and geometric morphometrics

Previous work on orchid seeds has shown that characters associated with the seed coat may be useful for classification and phylogeny at a suprageneric level. The seed morphology of several species of the tribe Chloraeeae was analysed using traditional morphometrics, and the seed shape was studied, for the first time, using tools of geometric morphometrics. Seed characters were evaluated by their discriminative power and the information they may provide in a phylogenetic context. By contrast with previous findings, seed shape resulted in a continuum among the taxa studied, and in only a few cases could genera or groups of species be discriminated on the basis of shape. However, seed size, expressed as centroid size, was a variable character and informative at a phylogenetic level. Traditional measures of seed coat, mainly those of seed coat cells, were also helpful for the discrimination of genera and species, agreeing with previous statements about their utility in taxonomy and phylogeny. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 160 , 171–183.     

Sexual reproduction is light-limited as marsh grasses colonize maritime forest

Climate change is driving abiotic shifts that can threaten the conservation of foundation species and the habitats they support. Directional range shifting is one mechanism of escape, but requires the successful colonization of habitats where interspecific interactions may differ from those to which a species has adapted. For plants with multiple reproductive strategies, these range-edge interactions may alter the investment or allocation toward a given reproductive strategy. In this study, we quantified sexual reproduction of the clonal marsh grass Spartina patens across an inland colonization front into maritime forest being driven by sea-level rise. We find that flowering is variable across S. patens meadows, but consistently reduced in low light conditions like those of the forest understory. Observational surveys of S. patens flowering at four sites in the Delmarva Peninsula agreed with the results of two experimental manipulations of light availability (shading experiment in S. patens-dominated marsh and a forest dieback manipulation). These three approaches pinpointed light limitation as a principal control on S. patens flowering capacity, suggesting that light competition with taller upland species can suppress S. patens flowering along its upland migration front. Consequently, all propagation in shaded conditions must occur clonally or via seeds from the marsh, a reproductive restriction that could limit the potential for local adaptation and reduce genetic diversity. Future research is needed to determine whether the lack of flowering is the result of a trade-off between sexual and clonal reproduction or results from insufficient photosynthetic products needed to achieve either reproductive method.     

Analysis of fungal diversity in <Emphasis Type="Italic">Orchis tridentata</Emphasis> Scopoli

We have assessed the identities of fungi associated with Orchis tridentata, an endangered orchid species growing in open woodland and poor grassland of Central and Southern Europe. Fungal diversity in ten O. tridentata adult individuals collected in two protected areas of Central Italy was analysed by means of morphological and molecular methods. Sequencing of the cloned ITS fungal inserts corresponding to the dominant PCR products obtained from amplification of total root DNA with ITS1F and ITS4 primers revealed a variety of fungal species occurring in O. tridentata roots. Among them, members of the basidiomycete families Ceratobasidiaceae, Tulasnellaceae and Hymenogastraceae were recovered, together with ascomycetes belonging to Leptodontidium and Terfezia. The implications of these results in the understanding of O. tridentata biology and for the conservation of this threatened orchid species are discussed.     

兰科石斛属植物菌根真菌研究进展

石斛属(Dendrobium)隶属于兰科(Orchidaceae)树兰亚科(Epidendroideae)石斛兰族(Dendrobiinae),是兰科最大的属之一,终生附生于树上或岩石上。石斛属很多种类具有很高的药用价值与观赏价值。由于人为过度采挖和野生生境的破坏,使得野生石斛资源濒临灭绝。石斛属植物为典型的兰科菌根植物,在自然条件下需要与真菌共生,才能完成生活史。菌根真菌对于石斛属植物的种子萌发和植株生长具有重要的作用。对石斛属植物菌根的形成、菌根真菌的作用、菌根真菌多样性及菌根技术在石斛属植物中的应用做了评述,并对今后的研究内容和重点提出了一些思路。     

Prey use by dingoes in a contested landscape: Ecosystem service provider or biodiversity threat?

In Australia, dingoes (Canis lupus dingo) have been implicated in the decline and extinction of a number of vertebrate species. The lowland Wet Tropics of Queensland, Australia is a biologically rich area with many species of rainforest‐restricted vertebrates that could be threatened by dingoes; however, the ecological impacts of dingoes in this region are poorly understood. We determined the potential threat posed by dingoes to native vertebrates in the lowland Wet Tropics using dingo scat/stomach content and stable isotope analyses of hair from dingoes and potential prey species. Common mammals dominated dingo diets. We found no evidence of predation on threatened taxa or rainforest specialists within our study areas. The most significant prey species were northern brown bandicoots (Isoodon macrourus), canefield rats (Rattus sordidus), and agile wallabies (Macropus agilis). All are common species associated with relatively open grass/woodland habitats. Stable isotope analysis suggested that prey species sourced their nutrients primarily from open habitats and that prey choice, as identified by scat/stomach analysis alone, was a poor indicator of primary foraging habitats. In general, we find that prey use by dingoes in the lowland Wet Tropics does not pose a major threat to native and/or threatened fauna, including rainforest specialists. In fact, our results suggest that dingo predation on “pest” species may represent an important ecological service that outweighs potential biodiversity threats. A more targeted approach to managing wild canids is needed if the ecosystem services they provide in these contested landscapes are to be maintained, while simultaneously avoiding negative conservation or economic impacts.     

The role of alien plants in the natural coastal vegetation in central-northern Spain

Alien plants in coastal habitats and their influence on natural vegetation are studied. After 5 years working on this subject in the Basque Country and surrounding areas, a number of results from the coastal ecosystems are presented. These ecosystems are one of the most threatened and affected by the invasion of alien plants, especially shore dunes, saltmarshes and cliffs. These kinds of habitats, especially the dunes, experience significant pressure from human activities which favours the expansion of some of these species: Arctotheca calendula, Sporobolus indicus and Oenothera spp. The presence and abundance of these invasive plants and others such as Baccharis halimifolia, Cortaderia selloana, Spartina patens and Carpobrotus edulis in the plant communities in an area between the French border and the western part of the region of Cantabria have been studied. The degree of invasion of each plant in each syntaxonomic unit has been analysed.