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Species delimitation is one of the most fundamental processes in biology. Biodiversity undertakings, for instance, require explicit species concepts and criteria for species delimitation in order to be relevant and translatable. However, a perfect species concept does not exist for Fungi. Here, we review the species concepts commonly used in Basidiomycota, the second largest phylum of Fungi that contains some of the best known species of mushrooms, rusts, smuts, and jelly fungi. In general, best practice is to delimitate species, publish new taxa, and conduct taxonomic revisions based on as many independent lines of evidence as possible, that is, by applying a so-called unifying (or integrative) conceptual framework. However, the types of data used vary considerably from group to group. For this reason we discuss the different classes of Basidiomycota, and for each provide: (i) a general introduction with difficulties faced in species recognition, (ii) species concepts and methods for species delimitation, and (iii) community recommendations and conclusions. [ABSTRACT FROM AUTHOR]

Besides the plant-fungus symbiosis in arbuscular mycorrhizal (AM) and ectomycorrhizal (EM) plants, many endorhizal and rhizosphere bacteria (Root Associated Bacteria, or RAB) also enhance plant fitness, diversity, and coexistence among plants via bi- or tripartite interactions with plant hosts and mycorrhizal fungi. Assuming that bacterial associations are just as important for the obligate mycorrhizal plant family Orchidaceae, surprisingly little is known about the RAB associated with orchids. Herein, we first present the current, underwhelming state of RAB research including their interactions with fungi and the influence of holobionts on plant fitness. We then delineate the need for novel investigations specifically in orchid RAB ecology, and sketch out questions and hypotheses which, when addressed, will advance plant-microbial ecology. We specifically discuss the potential effects of beneficial RAB on orchids as: (1) Plant Growth Promoting Rhizobacteria (PGPR), (2) Mycorrhization Helper Bacteria (MHB), and (3) constituents of an orchid holobiont. We further posit that a hologenomic view should be considered as a framework for addressing co-evolution of the plant host, their obligate Orchid Mycorrhizal Fungi (OMF), and orchid RAB. We conclude by discussing implications of the suggested research for conservation of orchids, their microbial partners, and their collective habitats. [ABSTRACT FROM AUTHOR]

We describe an extraordinary new species in the genus Thismia from the Terengganu State of Malaysia in the Malay Peninsula. The new species, which we name Thismia sitimeriamiae, is distinct from all other Thismia species known to science, most notably in its unique mitre configuration formed by the inner tepals and its floral surface morphology characterised by conspicuous orange trichomes. We discuss our findings in the context of underestimated species diversity in the genus Thismia and implications for their conservation. We recommend assigning T. sitimeriamiae the conservation status as Critically Endangered (CR) according to IUCN criteria. [ABSTRACT FROM AUTHOR]

Orchid distribution and population dynamics are influenced by a variety of ecological factors and the formation of holobionts, which play key roles in colonization and ecological community construction. Seed germination, seedling establishment, reproduction, and survival of orchid species are strongly dependent on orchid mycorrhizal fungi (OMF), with mycorrhizal cheating increasingly observed in photosynthetic orchids. Therefore, changes in the composition and abundance of OMF can have profound effects on orchid distribution and fitness. Network analysis is an important tool for the study of interactions between plants, microbes, and the environment, because of the insights that it can provide into the interactions and coexistence patterns among species. Here, we provide a comprehensive overview, systematically describing the current research status of the effects of OMF on orchid distribution and dynamics, phylogenetic signals in orchid–OMF interactions, and OMF networks. We argue that orchid–OMF associations exhibit complementary and specific effects that are highly adapted to their environment. Such specificity of associations may affect the niche breadth of orchid species and act as a stabilizing force in plant–microbe coevolution. We postulate that network analysis is required to elucidate the functions of fungal partners beyond their effects on germination and growth. Such studies may lend insight into the microbial ecology of orchids and provide a scientific basis for the protection of orchids under natural conditions in an efficient and cost-effective manner. [ABSTRACT FROM AUTHOR]

Background and Aims An arbuscular mycorrhiza is a mutualistic symbiosis with plants as carbon providers for fungi. However, achlorophyllous arbuscular mycorrhizal species are known to obtain carbon from fungi, i.e. they are mycoheterotrophic. These species all have the Paris type of arbuscular mycorrhiza. Recently, two chlorophyllous Paris -type species proved to be partially mycoheterotrophic. In this study, we explore the frequency of this condition and its association with Paris -type arbuscular mycorrhiza. Methods We searched for evidence of mycoheterotrophy in all currently published 13C, 2H and 15N stable isotope abundance patterns suited for calculations of enrichment factors, i.e. isotopic differences between neighbouring Paris - and Arum -type species. We found suitable data for 135 plant species classified into the two arbuscular mycorrhizal morphotypes. Key Results About half of the chlorophyllous Paris -type species tested were significantly enriched in 13C and often also enriched in 2H and 15N, compared with co-occurring Arum -type species. Based on a two-source linear mixing model, the carbon gain from the fungal source ranged between 7 and 93 % with ferns > horsetails > seed plants. The seed plants represented 13 families, many without a previous record of mycoheterotrophy. The 13C-enriched chlorophyllous Paris -type species were exclusively herbaceous perennials, with a majority of them thriving on shady forest ground. Conclusions Significant carbon acquisition from fungi appears quite common and widespread among Paris -type species, this arbuscular mycorrhizal morphotype probably being a pre-condition for developing varying degrees of mycoheterotrophy. [ABSTRACT FROM AUTHOR]

Understanding features that fostered the persistence of Equisetum–Earth's oldest extant vascular plant genus–since Mesozoic times and through episodes of significant global environmental change, is of current interest in view of modern challenges to plant survival. In addition to known structural and physiological adaptations, we hypothesized that microscopy and shotgun metagenomic sequencing might reveal eukaryotic microorganisms such as fungi that may aid Equisetum survival. Here, we report evidence for several lineages of eukaryotic microbes associated with giant Equisetum xylochaetum, which dominates vegetation in saline streambeds of remote valleys in the hyper-arid Atacama Desert, Chile. Plant material was collected and field-preserved at two comparatively low-disturbance sites; DNA extracted in Chile using low-shear methods was later sequenced, 18S and 28S rDNA taxonomic marker sequences were selected for SILVAngs classification, allowing comparisons to eukaryotic microorganisms previously inferred for earlier-diverging plant lineages. SEM, fluorescence microscopy, and/or LM of toluidine blue-stained sections of roots indicated protists, epiphytic and endophytic fungi, and cortical nematodes. Eukaryotic genera inferred from 18S rDNA at >100X mean sequencing depth included the ciliate Engelmanniella, hyphal chytrid Monoblepharella, predatory ascomycete Cephaliophora, a salpingoecid choanoflagellate, and an annelid worm. 23S rDNA sequences indicated ascomycete Capnodiales fungi at one site and four types of Pezizomycotina fungi at the other. No evidence for vesicular-arbuscular mycorrhizal fungi was found, but we hypothesized that Equisetum may benefit from other types of fungal associations, some possibly inherited from ancestral plant lineages. [ABSTRACT FROM AUTHOR]

Photosynthesis with highly photoreactive chlorophyll (Chl) provides energy for plant growth but with simultaneous risk of photooxidative damage and photoprotection costs. Although the leafless orchid Cymbidium macrorhizon mostly depends on mycorrhizal fungi for carbon, it accumulates Chl particularly during fruiting and may not be fully mycoheterotrophic. In fact, stable isotopic analysis suggested that the fruiting C. macrorhizon specimens obtain a significant proportion of its carbon demands through photosynthesis. However, actual photosynthetic characteristics of this leafless orchid are unknown. To reveal the functionality of photosynthetic electron transport in C. macrorhizon , we compared its photosynthetic properties with those of its relative mixotrophic orchid Cymbidium goeringii and the model plant Arabidopsis thaliana. Compared with C. goeringii and A. thaliana , maximum photochemical efficiency of PSII was substantially low in C. macrorhizon. Chl fluorescence induction kinetics revealed that the electron transport capacity of PSII was limited in C. macrorhizon. Chl fluorescence analysis at 77 K suggested partial energetic disconnection of the light-harvesting antenna from the PSII reaction center in C. macrorhizon. Despite its low PSII photochemical efficiency, C. macrorhizon showed photosynthetic electron transport activity both in the field and under laboratory conditions. Cymbidium macrorhizon developed strong nonphotochemical quenching in response to increased light intensity as did C. goeringii , suggesting the functionality of photoprotective systems in this orchid. Moreover, C. macrorhizon fruit developed stomata on the pericarp and showed net O2-evolving activity. Our data demonstrate that C. macrorhizon can perform photosynthetic electron transport in the pericarp, although its contribution to net carbon acquisition may be limited. [ABSTRACT FROM AUTHOR]

Partial mycoheterotrophy, the ability of plants to obtain carbon from fungi throughout their life cycle in combination with photosynthesis, appears to be more common within the Plant Kingdom than previously anticipated. Recent studies using stable isotope analyses have indicated that isotope signatures in partially mycoheterotrophic plants vary widely among species, but the relative contributions of family- or species-specific characteristics and the identity of the fungal symbionts to the observed differences remain unclear. Here, we investigated in detail mycorrhizal communities and isotopic signatures in four co-occurring terrestrial orchids (Platanthera chlorantha, Epipactis helleborine, E. neglecta and the mycoheterotrophic Neottia nidus-avis). All investigated species were mycorrhizal generalists (i.e., associated with a large number of fungi simultaneously), but mycorrhizal communities differed significantly between species. Mycorrhizal communities associating with the two Epipactis species consisted of a wide range of fungi belonging to different families, whereas P. chlorantha and N. nidus-avis associated mainly with Ceratobasidiaceae and Sebacinaceae species, respectively. Isotopic signatures differed significantly between both Epipactis species, with E. helleborine showing near autotrophic behavior and E. neglecta showing significant enrichment in both carbon and nitrogen. No significant differences in photosynthesis and stomatal conductance were observed between the two partially mycoheterotrophic orchids, despite significant differences in isotopic signatures. Our results demonstrate that partially mycoheterotrophic orchids of the genus Epipactis formed mycorrhizas with a wide diversity of fungi from different fungal families, but variation in mycorrhizal community composition was not related to isotope signatures and thus transfer of C and N to the plant. We conclude that the observed differences in isotope signatures between E. helleborine and E. neglecta cannot solely be explained by differences in mycorrhizal communities, but most likely reflect a combination of inherent physiological differences and differences in mycorrhizal communities. [ABSTRACT FROM AUTHOR]

We studied mycorrhizal symbioses of Cephalanthera falcata (Orchidaceae) in a population that included giant individuals, some of which were over 50 cm in height and had more than 10 shoots, as well as in two nearby habitats without giant individuals. For C. falcata adults in the habitat with giant individuals, a fungal lineage in the ectomycorrhiza‐forming clade of Sebacinales was detected as the dominant mycobiont. Furthermore, higher 13C enrichment, which is an indicator of mycoheterotrophy, was found in these C. falcata individuals compared to those in the two nearby habitats. These results indicate that the growth of giant C. falcata individuals may be a result of the increased ability of Sebacinales fungi to transfer carbon to the orchids. Furthermore, we attempted to discern the effect of mycobiont taxa on seedling development in the three habitats. Contrary to our expectations, Sebacinales fungi were not found to significantly impact seedling growth as this orchid was predominantly colonized by Thelephoraceae fungi at the seedling stage. These results suggest that seedling growth is mostly enhanced by symbiosis with Thelephoraceae fungi in this orchid, and that the growth of giant individuals due to colonization by Sebacinales fungi may occur subsequently. [ABSTRACT FROM AUTHOR]

The endophytic microbiota can establish mutualistic or commensalistic interactions within the host plant tissues. We investigated the bacterial endophytic microbiota in three species of Mediterranean orchids (Neottia ovata , Serapias vomeracea , and Spiranthes spiralis) by metabarcoding of the 16S rRNA gene. We examined whether the different orchid species and organs, both underground and aboveground, influenced the endophytic bacterial communities. A total of 1,930 operational taxonomic units (OTUs) were obtained, mainly Proteobacteria and Actinobacteria, whose distribution model indicated that the plant organ was the main determinant of the bacterial community structure. The co-occurrence network was not modular, suggesting a relative homogeneity of the microbiota between both plant species and organs. Moreover, the decrease in species richness and diversity in the aerial vegetative organs may indicate a filtering effect by the host plant. We identified four hub OTUs, three of them already reported as plant-associated taxa (Pseudoxanthomonas, Rhizobium , and Mitsuaria), whereas Thermus was an unusual member of the plant microbiota. Core microbiota analysis revealed a selective and systemic ascent of bacterial communities from the vegetative to the reproductive organs. The core microbiota was also maintained in the S. spiralis seeds, suggesting a potential vertical transfer of the microbiota. Surprisingly, some S. spiralis seed samples displayed a very rich endophytic microbiota, with a large number of OTUs shared with the roots, a situation that may lead to a putative restoring process of the root-associated microbiota in the progeny. Our results indicate that the bacterial community has adapted to colonize the orchid organs selectively and systemically, suggesting an active involvement in the orchid holobiont. [ABSTRACT FROM AUTHOR]

Marchantia polymorpha is a common liverwort whose use as a model plant in physiological and evolutionary processes is increasing in recent years. As far as plant-microorganism interactions are concerned, there are still few studies conducted with M. polymorpha. Specifically, in the interaction of M. polymorpha with arbuscular mycorrhizal fungi (AMF), it has been described how AMF colonize the M. polymorpha tissues, without knowing more about the interaction. In this study, M. polymorpha is inoculated with different AMF formulations, analyzing the direct effect on M. polymorpha's growth and the nutritional content, along with stress responses. Moreover, expression levels of defense genes in M. polymorpha were analyzed. The results obtained showed how M. polymorpha-AMF interaction is detrimental to plant under in vitro conditions. A reduction in its growth and viability of its tissues was observed, in addition to an increase only in nutritional content of those elements related to plant defenses, together with the reactive oxygen species (ROS) content. Rhizophagus fasciculatus is only present in the formulation that causes major damage to the plant, including symptoms of tissue damage, and that mostly colonizes the plant. It suggests its possible role as a plant pathogen, due to the inability of M. polymorpha to defend it against the AMF by the route of salicylic acid (SA). [ABSTRACT FROM AUTHOR]

The vast majority of terrestrial plants exchange nutrients with fungal partners forming different mycorrhizal types. The minority of plants considered as non‐mycorrhizal, however, are not necessarily free of any fungi, but are frequently colonized by elusive fungal endophytes, such as dark septate endophytes (DSE) or fine root endophytes (FRE). While a functional role of FRE in the improvement of nutrient gain was recently elucidated, the function of DSE is still in discussion and was here addressed for 36 plant species belonging to the families Equisetaceae, Cypereaceae and Caryophyllaceae.Molecular and microscopic staining approaches were conducted to verify the presence of DSE in the investigated species. Stable isotope natural abundances of the elements carbon, nitrogen, hydrogen and oxygen and total nitrogen concentrations were analysed for the respective species of the target plant families and accompanying mycorrhizal and non‐mycorrhizal (Brassicaceae) plant species.Staining approaches confirmed the presence of DSE in all investigated species within the families Equisetaceae, Cyperaceae and Caryophyllaceae. A co‐colonization with Paris‐type arbuscular mycorrhiza (AM) was occasionally found by staining and molecular approaches in species of the Equisetaceae. Species of the Equisetaceae, Cyperaceae and Caryophyllaceae were significantly 15N‐enriched in comparison to accompanying plants. In addition, a significant 13C and 2H enrichment and increased total nitrogen concentrations were found for representatives of the Equisetaceae.The 15N enrichment found here for representatives of Equisetaceae, Cyperaceae and Caryophyllaceae provides evidence for a functional role of the ubiquitous DSE fungi. DSE fungi obviously provide access to 15N‐enriched soil organic compounds probably in exchange for organic carbon compounds from plant photosynthesis. As indicated by additional 13C and 2H enrichments, representatives of the Equisetaceae apparently gain simultaneously organic carbon compounds from their AM fungi of the Paris‐morphotype. Thus, species of the Equisetaceae have to be considered as partially, or in case of the achlorophyllous fertile Equisetum arvense, as fully mycoheterotrophic at least in some stages of their life cycle.So far, mostly underappreciated fungi classified as DSE are suggested to occupy an ecologically relevant role similar to mycorrhizae and the occurrence of simultaneous functions of DSE and AM fungi in Equisetaceae is proposed. A free Plain Language Summary can be found within the Supporting Information of this article. [ABSTRACT FROM AUTHOR]

Summary: Despite the ecological significance of ericoid mycorrhizal fungi, little is known about the abiotic and biotic factors driving their diversity and community composition.To determine the relative importance of abiotic and biotic filtering in structuring ericoid mycorrhizal fungal communities, we established 156 sampling plots in two highly contrasting environments but dominated by the same Ericaceae plant species: waterlogged bogs and dry heathlands. Plots were located across 25 bogs and 27 dry heathlands in seven European countries covering a gradient in nitrogen deposition and phosphorus availability. Putatively ericoid mycorrhizal fungal communities in the roots of 10 different Ericaceae species were characterized using high‐throughput amplicon sequencing.Variation in ericoid mycorrhizal fungal communities was attributed to both habitat and soil variables on the one hand and host plant identity on the other. Communities differed significantly between bogs and heathlands and, in a given habitat, communities differed significantly among host plant species. Fungal richness was negatively related to nitrogen deposition in bogs and phosphorus availability in bogs and heathlands.Our results demonstrate that both abiotic and biotic filtering shapes ericoid mycorrhizal fungal communities and advocate an environmental policy minimizing excess nutrient input in these nutrient‐poor ecosystems to avoid loss of ericoid mycorrhizal fungal taxa. [ABSTRACT FROM AUTHOR]

Many adult orchids, especially photoautotrophic species, associate with a diverse range of mycorrhizal fungi, but little is known about the temporal changes that might occur in the diversity and functioning of orchid mycorrhiza during vegetative and reproductive plant growth. Temporal variations in the spectrum of mycorrhizal fungi and in stable isotope natural abundance were investigated in adult plants of Anacamptis morio, a wintergreen meadow orchid. Anacamptis morio associated with mycorrhizal fungi belonging to Tulasnella, Ceratobasidium and a clade of Pezizaceae (Ascomycetes). When a complete growing season was investigated, multivariate analyses indicated significant differences in the mycorrhizal fungal community. Among fungi identified from manually isolated pelotons, Tulasnella was more common in autumn and winter, the pezizacean clade was very frequent in spring, and Ceratobasidium was more frequent in summer. By contrast, relatively small variations were found in carbon (C) and nitrogen (N) stable isotope natural abundance, A. morio samples showing similar (15)N enrichment and (13)C depletion at the different sampling times. These observations suggest that, irrespective of differences in the seasonal environmental conditions, the plant phenological stages and the associated fungi, the isotopic content in mycorrhizal A. morio remains fairly constant over time., (© 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.)

Background and Aims The geographical distribution of plant species is linked fundamentally not only to environmental variables, but also to key traits that affect the dispersal, establishment and evolutionary potential of a species. One of the key plant traits that can be expected to affect standing genetic variation, speed of adaptation and the capacity to colonize and establish in new habitats, and therefore niche breadth and range size, is the plant mating system. However, the precise role of the mating system in shaping range size and niche breadth of plant species remains unclear, and different studies have provided contrasting results. In this study, we tested the hypothesis that range size and niche breadth differed with mating system in the orchid genus Epipactis. Methods We modelled the ecological niches of 14 Epipactis species in Europe using occurrence records and environmental satellite data in Maxent. Niche breadth and niche overlap in both geographic and environmental space were calculated from the resulting habitat suitability maps using ENMTools, and geographic range was estimated using α-hull range definition. Habitat suitability, environmental variable contributions and niche metrics were compared among species with different mating systems. Key Results We did not detect significant differences in niche breadth, occurrence probability or geographical range between autogamous and allogamous Epipactis species, although autogamous species demonstrated notably low variation in niche parameters. We also found no significant differences in niche overlap between species with the same mating system or different mating systems. For all Epipactis species, occurrence was strongly associated with land cover, particularly broad-leafed and coniferous forests, and with limestone bedrock. Conclusions These results suggest that the mating system does not necessarily contribute to niche breadth and differentiation, and that other factors (e.g. mycorrhizal specificity) may be more important drivers of range size and niche breadth in Epipactis and orchids in general. [ABSTRACT FROM AUTHOR]

Mixotrophic plants obtain carbon by their own photosynthetic activity and from their root-associated mycorrhizal fungi. Mixotrophy is deemed a pre-adaptation for evolution of mycoheterotrophic nutrition, where plants fully depend on fungi and lose their photosynthetic activity. The aim of this study was to clarify mycorrhizal dependency and heterotrophy level in various phenotypes of mixotrophic Pyrola japonica (Ericaceae), encompassing green individuals, rare achlorophyllous variants (albinos) and a form with minute leaves, P. japonica f. subaphylla. These three phenotypes were collected in two Japanese forests. Phylogenetic analysis of both plants and mycorrhizal fungi was conducted based on DNA barcoding. Enrichment in 13C among organs (leaves, stems and roots) of the phenotypes with reference plants and fungal fruitbodies were compared by measuring stable carbon isotopic ratio. All plants were placed in the same clade, with f. subaphylla as a separate subclade. Leaf 13C abundances of albinos were congruent with a fully mycoheterotrophic nutrition, suggesting that green P. japonica leaves are 36.8% heterotrophic, while rhizomes are 74.0% heterotrophic. There were no significant differences in δ13C values among organs in both albino P. japonica and P. japonica f. subaphylla, suggesting full and high mycoheterotrophic nutrition, respectively. Among 55 molecular operational taxonomic units (OTUs) detected as symbionts, the genus Russula was the most abundant in each phenotype and its dominance was significantly higher in albino P. japonica and P. japonica f. subaphylla. Russula spp. detected in P. japonica f. subaphylla showed higher dissimilarity with other phenotypes. These results suggest that P. japonica sensu lato is prone to evolve mycoheterotrophic variants, in a process that changes its mycorrhizal preferences, especially towards the genus Russula for which this species has a marked preference. [ABSTRACT FROM AUTHOR]

The activities of soil resident enzymes and fungal communities are critical for soil health and ecosystem functionality. However, their dynamics in response to variations in sowing techniques are poorly understood. This study aimed to investigate the effects of wide‐ridge and narrow‐furrow (WR&NF), film‐mulching ridge (FMR), and conventional drilling (CD) practices on the activities of soil‐residing enzymes and fungal communities in a dryland wheat field on the Loess Plateau. Photometrically, unbuffered, and colorimetric methods were employed to determine the activities of alkaline phosphatases, urease, and sucrase enzymes, respectively. Soil fungal diversity and community composition was measured via the Illumina Miseq 2500 sequencing method. The results revealed that WR&NF and FMR practices significantly enhanced the soil activities of alkaline phosphatases, urease, and sucrase, while improving soil fungal communities including Chytridiomycota phylum and Phoma (Ascomycota), Olpidium (Chytridiomycota), and Trichosporon (Basidiomycota) genera. Both the enzymatic activities and fungal community compositions of the soils under study varied from rhizosphere to non‐rhizosphere, as well as at different soil depths. Remarkably, several soil fungal genera, including Cystofilobasidium (Basidiomycota), Triparticalcar (Chytridiomycota), and Guehomyces (Basidiomycota) were more abundant in the subsoil than topsoil. In general, we observed that ridge‐furrow and film‐mulching practices enhanced the enzyme activities and improved fungal communities of soils. Therefore, the wide‐ridge and narrow‐furrow sowing technique is recommended as an environmentally compatible practice to benefit the arid wheat (Triticum aestivum L.) fields of the Loess Plateau. [ABSTRACT FROM AUTHOR]

Data for stable C and N isotope natural abundances of arbuscular mycorrhizal (AM) fungi are currently sparse, as fungal material is difficult to access for analysis. So far, isotope analyses have been limited to lipid compounds associated with fungal membranes or storage structures (biomarkers), fungal spores and soil hyphae. However, it remains unclear whether any of these components are an ideal substitute for intraradical AM hyphae as the functional nutrient trading organ. Thus, we isolated intraradical hyphae of the AM fungus Rhizophagus irregularis from roots of the grass Festuca ovina and the legume Medicago sativa via an enzymatic and a mechanical approach. In addition, extraradical hyphae were isolated from a sand-soil mix associated with each plant. All three approaches revealed comparable isotope signatures of R. irregularis hyphae. The hyphae were 13C- and 15N-enriched relative to leaves and roots irrespective of the plant partner, while they were enriched only in 15N compared with soil. The 13C enrichment of AM hyphae implies a plant carbohydrate source, whereby the enrichment was likely reduced by an additional plant lipid source. The 15N enrichment indicates the potential of AM fungi to gain nitrogen from an organic source. Our isotope signatures of the investigated AM fungus support recent findings for mycoheterotrophic plants which are suggested to mirror the associated AM fungi isotope composition. Stable isotope natural abundances of intraradical AM hyphae as the functional trading organ for bi-directional carbon-for-mineral nutrient exchanges complement data on spores and membrane biomarkers. [ABSTRACT FROM AUTHOR]

Numerous studies of terrestrial orchids have demonstrated widespread partial mycoheterotrophy, particularly the possibility of obtaining organic matter from surrounding trees through a common fungal network. Fungi are also widespread in epiphytic orchid roots, but there have been no attempts to determine if epiphytes accept organic matter from the living stems of their phorophytes. We hypothesise that such transfer does not exist because epiphytes and phorophytes harbour different fungal communities. To test this hypothesis, we tagged three short Randia sp. trees with 13C-enriched CO2 and examined 13C transfer from the phorophyte into the epiphytic orchids Grosourdya appendiculata, Dendrobium oligophyllum and Gastrochilus sp. in Cat Tien National Park, (South Vietnam, Cat Tien National Park, plot size approx. 1 ha). The coincidence of fungal sequences in the orchid roots and in the branches on which they grew was also examined. We did not detect 13C label moving from phorophytes to epiphytes. Using Illumina sequencing, 162 fungal operational taxonomic units (OTUs) were detected. The fungal communities were significantly different between the roots of epiphytes and branches of phorophytes, although no strict fungal specificity at the species level was found in either epiphytes or phorophytes. [ABSTRACT FROM AUTHOR]

Background and Aims In orchid conservation, quantifying the specificity of mycorrhizal associations, and establishing which orchid species use the same fungal taxa, is important for sourcing suitable fungi for symbiotic propagation and selecting sites for conservation translocation. For Caladenia subgenus Calonema (Orchidaceae), which contains 58 threatened species, we ask the following questions. (1) How many taxa of Serendipita mycorrhizal fungi do threatened species of Caladenia associate with? (2) Do threatened Caladenia share orchid mycorrhizal fungi with common Caladenia ? (3) How geographically widespread are mycorrhizal fungi associated with Caladenia ? Methods Fungi were isolated from 127 Caladenia species followed by DNA sequencing of the internal transcibed spacer (ITS) sequence locus. We used a 4.1–6 % sequence divergence cut-off range to delimit Serendipita operational taxonomic units (OTUs). We conducted trials testing the ability of fungal isolates to support germination and plant growth. A total of 597 Serendipita isolates from Caladenia , collected from across the Australian continent, were used to estimate the geographic range of OTUs. Key Results Across the genus, Caladenia associated with ten OTUs of Serendipita (Serendipitaceae) mycorrhizal fungi. Specificity was high, with 19 of the 23 threatened Caladenia species sampled in detail associating solely with OTU A, which supported plants from germination to adulthood. The majority of populations of Caladenia associated with one OTU per site. Fungal sharing was extensive, with 62 of the 79 Caladenia sampled in subgenus Calonema associating with OTU A. Most Serendipita OTUs were geographically widespread. Conclusions Mycorrhizal fungi can be isolated from related common species to propagate threatened Caladenia. Because of high specificity of most Caladenia species, only small numbers of OTUs typically need to be considered for conservation translocation. When selecting translocation sites, the geographic range of the fungi is not a limiting factor, and using related Caladenia species to infer the presence of suitable fungal OTUs may be feasible. [ABSTRACT FROM AUTHOR]

Orchids form endomycorrhizal associations with fungi mainly belonging to basidiomycetes. The molecular events taking place in orchid mycorrhiza are poorly understood, although the cellular changes necessary to accommodate the fungus and to control nutrient exchanges imply a modulation of gene expression. Here, we used proteomics and transcriptomics to identify changes in the steady-state levels of proteins and transcripts in the roots of the green terrestrial orchid Oeceoclades maculata. When mycorrhizal and non-mycorrhizal roots from the same individuals were compared, 94 proteins showed differential accumulation using the label-free protein quantitation approach, 86 using isobaric tagging and 60 using 2D-differential electrophoresis. After de novo assembly of transcriptomic data, 11,179 plant transcripts were found to be differentially expressed, and 2175 were successfully annotated. The annotated plant transcripts allowed the identification of up- and down-regulated metabolic pathways. Overall, proteomics and transcriptomics revealed, in mycorrhizal roots, increased levels of transcription factors and nutrient transporters, as well as ethylene-related proteins. The expression pattern of proteins and transcripts involved in plant defense responses suggested that plant defense was reduced in O. maculata mycorrhizal roots sampled in nature. These results expand our current knowledge towards a better understanding of the orchid mycorrhizal symbiosis in adult plants under natural conditions. [ABSTRACT FROM AUTHOR]

Background Given the exceptional diversity of orchids (26 000+ species), improving strategies for the conservation of orchids will benefit a vast number of taxa. Furthermore, with rapidly increasing numbers of endangered orchids and low success rates in orchid conservation translocation programmes worldwide, it is evident that our progress in understanding the biology of orchids is not yet translating into widespread effective conservation. Scope We highlight unusual aspects of the reproductive biology of orchids that can have important consequences for conservation programmes, such as specialization of pollination systems, low fruit set but high seed production, and the potential for long-distance seed dispersal. Further, we discuss the importance of their reliance on mycorrhizal fungi for germination, including quantifying the incidence of specialized versus generalized mycorrhizal associations in orchids. In light of leading conservation theory and the biology of orchids, we provide recommendations for improving population management and translocation programmes. Conclusions Major gains in orchid conservation can be achieved by incorporating knowledge of ecological interactions, for both generalist and specialist species. For example, habitat management can be tailored to maintain pollinator populations and conservation translocation sites selected based on confirmed availability of pollinators. Similarly, use of efficacious mycorrhizal fungi in propagation will increase the value of ex situ collections and likely increase the success of conservation translocations. Given the low genetic differentiation between populations of many orchids, experimental genetic mixing is an option to increase fitness of small populations, although caution is needed where cytotypes or floral ecotypes are present. Combining demographic data and field experiments will provide knowledge to enhance management and translocation success. Finally, high per-fruit fecundity means that orchids offer powerful but overlooked opportunities to propagate plants for experiments aimed at improving conservation outcomes. Given the predictions of ongoing environmental change, experimental approaches also offer effective ways to build more resilient populations. [ABSTRACT FROM AUTHOR]

Orchid mycorrhizal fungi (OMF) are critical for seed germination and maintaining natural populations of orchids, yet the degree of specificity of most orchids to their mycorrhizal associates remains unknown. Many orchids are at risk of extinction, whether generalists or specialists, but orchid species of narrow fungal specificity are arguably under increased threat due to their requirement for specific fungal symbionts. This study characterises the fungi associated with Aerangis ellisii, a lithophytic orchid from a site in the Central Highlands of Madagascar. Culturable OMF isolated from spontaneous protocorms of this species from the wild were used for seed germination. In vitro germination and seedling development of A. ellisii were achieved with fungi derived from A. ellisii and an isolate from a different Aerangis species 30 km away. The significance of these findings and their importance to conservation strategies for this species and other Aerangis spp. is discussed. These results have important implications for the conservation of A. ellisii populations in Madagascar. [ABSTRACT FROM AUTHOR]

Aims: Arbuscule-producing fine root endophytes (FRE) (previously incorrectly Glomus tenue) were recently placed within subphylum Mucoromycotina; the first report of arbuscules outside subphylum Glomeromycotina. Here, we aimed to estimate nutrient concentrations in plant and fungal structures of FRE and to test the utility of cryo-scanning electron microscopy (cryoSEM) for studying these fungi. Methods: We used replicated cryoSEM and X-ray microanalysis of heavily colonized roots of Trifolium subterraneum. Results: Intercellular hyphae and hyphae in developed arbuscules were consistently very thin; 1.35 ± 0.03 μm and 0.99 ± 0.03 μm in diameter, respectively (mean ± SE). Several intercellular hyphae were often adjacent to each other forming "hyphal ropes." Developed arbuscules showed higher phosphorus concentrations than senesced arbuscules and non-colonized structures. Senesced arbuscules showed greatly elevated concentrations of calcium and magnesium. Conclusion: While uniformly thin hyphae and hyphal ropes are distinct features of FRE, the morphology of fully developed arbuscules, elevated phosphorus in fungal structures, and accumulation of calcium with loss of structural integrity in senesced arbuscules are similar to glomeromycotinian fungi. Thus, we provide evidence that FRE may respond to similar host-plant signals or that the host plant may employ a similar mechanism of association with FRE and AMF. [ABSTRACT FROM AUTHOR]

This article is a Commentary on Suetsugu et al. (2020), 227: 1519–1529. [ABSTRACT FROM AUTHOR]

Background: Achlorophyllous orchids are mycoheterotrophic plants, which lack photosynthetic ability and associate with fungi to acquire carbon from different environmental sources. In tropical latitudes, achlorophyllous forest orchids show a preference to establish mycorrhizal relationships with saprotrophic fungi. However, a few of them have been recently found to associate with ectomycorrhizal fungi and there is still much to be learned about the identity of fungi associated with tropical orchids. The present study focused on mycorrhizal diversity in the achlorophyllous orchid C. inverta, an endangered species, which is endemic to southern China. The aim of this work was to identify the main mycorrhizal partners of C. inverta in different plant life stages, by means of morphological and molecular methods. Results: Microscopy showed that the roots of analysed C. inverta samples were extensively colonized by fungal hyphae forming pelotons in root cortical cells. Fungal ITS regions were amplified by polymerase chain reaction, from DNA extracted from fungal mycelia isolated from orchid root samples, as well as from total root DNA. Molecular sequencing and phylogenetic analyses showed that the investigated orchid primarily associated with ectomycorrhizal fungi belonging to a narrow clade within the family Ceratobasidiaceae, which was previously detected in a few fully mycoheterotrophic orchids and was also found to show ectomycorrhizal capability on trees and shrubs. Russulaceae fungal symbionts, showing high similarity with members of the ectomycorrhizal genus Russula, were also identified from the roots of C. inverta, at young seedling stage. Ascomycetous fungi including Chaetomium, Diaporthe, Leptodontidium, and Phomopsis genera, and zygomycetes in the genus Mortierella were obtained from orchid root isolated strains with unclear functional role. Conclusions: This study represents the first assessment of root fungal diversity in the rare, cryptic and narrowly distributed Chinese orchid C. inverta. Our results provide new insights on the spectrum of orchid-fungus symbiosis suggesting an unprecedented mixed association between the studied achlorophyllous forest orchid and ectomycorrhizal fungi belonging to Ceratobasidiaceae and Russulaceae. Ceratobasidioid fungi as dominant associates in the roots of C. inverta represent a new record of the rare association between the identified fungal group and fully mycoheterotrophic orchids in nature. [ABSTRACT FROM AUTHOR]

Summary: The root‐associated habit has evolved on numerous occasions in different fungal lineages, suggesting a strong evolutionary pressure for saprotrophic fungi to switch to symbiotic associations with plants. Species within the ubiquitous, saprotrophic genus Mycena are frequently major components in molecular studies of root‐associated fungal communities, suggesting that an evaluation of their trophic status is warranted. Here, we report on interactions between a range of Mycena species and the plant Betula pendula.In all, 17 Mycena species were inoculated onto B. pendula seedlings. Physical interactions between hyphae and fine roots were examined using differential staining and fluorescence microscopy. Physiological interactions were investigated using 14C and 32P to show potential transfer between symbionts.All Mycena species associated closely with fine roots, showing hyphal penetration into the roots, which in some cases were intracellular. Seven species formed mantle‐like structures around root tips, but none formed a Hartig net. Mycena pura and Mycena galopus both enhanced seedling growth, with M. pura showing significant transfer of 32P to the seedlings.Our results support the view that several Mycena species can associate closely with plant roots and some may potentially occupy a transitional state between saprotrophy and biotrophy. [ABSTRACT FROM AUTHOR]

Fully mycoheterotrophic plants are thought to obtain carbon exclusively from their root‐associated fungal partners. The general enrichment of these plants in the heavy isotopes 13C and 15N suggests that fungi are the main nutrient source for these plants. Yet, the majority of studies have targeted mycoheterotrophic plants associated with ectomycorrhizal, orchid mycorrhizal and saprotrophic fungi, while mycoheterotrophic plants living on arbuscular mycorrhizal fungi remain understudied.Here, we sampled 13 species of arbuscular mycorrhizal fully mycoheterotrophic plants from five families and co‐occurring autotrophic reference plants growing in forests of tropical South America, tropical South East Asia and temperate Australasia. We measured stable isotope natural abundances (δ13C, δ15N, δ2H and δ18O), determined total nitrogen concentrations and used high‐throughput DNA sequencing to characterize the arbuscular mycorrhizal fungal communities associated with the sampled mycoheterotrophic plants.We observed a general enrichment in 13C and 15N isotopes across mycoheterotrophic plant families and geographic regions. We confirm cases where no 15N enrichment is present, but we show that in general arbuscular mycoheterotrophic plants are enriched in 15N. Moreover, we demonstrate for the first time that these plants are significantly enriched in 2H but not in 18O in relation to their autotrophic references. The fungal communities targeted by the mycoheterotrophs mainly consist of Glomeraceae and show strong association with the isotopic signatures and geographic origin of the plants.Synthesis. Our findings enlarge the limited knowledge on the multi‐element stable isotopic signatures of mycoheterotrophic plants living on arbuscular mycorrhizal fungi. We show that these plants are enriched in 13C and 2H as expected due to their mycoheterotrophic nutrition, and that in general they are also enriched in 15N, despite some exceptions. Variation in stable isotope signatures is likely influenced by plant taxonomy, geography and fungal community composition. [ABSTRACT FROM AUTHOR]

Discreet heterotrophs: green plants that receive fungal carbon through Paris-type arbuscular mycorrhiza Keywords: arbuscular mycorrhiza (AM); Arum maculatum; Arum-type; mycoheterotrophy; Paris quadrifolia; Paris-type; stable isotope natural abundance EN arbuscular mycorrhiza (AM) Arum maculatum Arum-type mycoheterotrophy Paris quadrifolia Paris-type stable isotope natural abundance 960 966 7 04/20/20 20200515 NES 200515 Arbuscular mycorrhiza (AM) represents a symbiosis between plants and Glomeromycotina fungi, and it is distributed throughout the plant kingdom and all terrestrial ecosystems. Arbuscules presumably are the site of mineral nutrient fluxes from fungus to plant, and together with intercellular hyphae are also involved in photosynthate transfer from plant to fungus (Smith & Smith, [49]; Dickson I et al. i , [13]; Wipf I et al. i , [56]). In recent years a steadily increasing number of such species have been identified as "partial mycoheterotrophs", because their stable isotope composition values lie between nonmycoheterotrophic neighboring plants and full mycoheterotrophs (Hynson I et al. i , [25], [27]; Gebauer I et al. i , [20]). [Extracted from the article]

C |$_{4}$| photosynthesis is a complex trait that sustains fast growth and high productivity in tropical and subtropical conditions and evolved repeatedly in flowering plants. One of the major C |$_{4}$| lineages is Andropogoneae, a group of |$\sim $| 1200 grass species that includes some of the world's most important crops and species dominating tropical and some temperate grasslands. Previous efforts to understand C |$_{4}$| evolution in the group have compared a few model C |$_{4}$| plants to distantly related C |$_{3}$| species so that changes directly responsible for the transition to C |$_{4}$| could not be distinguished from those that preceded or followed it. In this study, we analyze the genomes of 66 grass species, capturing the earliest diversification within Andropogoneae as well as their C |$_{3}$| relatives. Phylogenomics combined with molecular dating and analyses of protein evolution show that many changes linked to the evolution of C |$_{4}$| photosynthesis in Andropogoneae happened in the Early Miocene, between 21 and 18 Ma, after the split from its C |$_{3}$| sister lineage, and before the diversification of the group. This initial burst of changes was followed by an extended period of modifications to leaf anatomy and biochemistry during the diversification of Andropogoneae, so that a single C |$_{4}$| origin gave birth to a diversity of C |$_{4}$| phenotypes during 18 million years of speciation events and migration across geographic and ecological spaces. Our comprehensive approach and broad sampling of the diversity in the group reveals that one key transition can lead to a plethora of phenotypes following sustained adaptation of the ancestral state. [Adaptive evolution; complex traits; herbarium genomics; Jansenelleae; leaf anatomy; Poaceae; phylogenomics.] [ABSTRACT FROM AUTHOR]

All orchids and pyroloids are mycoheterotrophic at least in the early stage. Many species are predisposed to mycoheterotrophic nutrition even in the adult stage, due to the initial mycoheterotrophy during germination. Although other green plants, such as gentian species, also produce numerous minute seeds, whose germination may depend on fungal associations to meet C demands, physiological evidence for partial mycoheterotrophy in the adult stage is lacking for most candidate taxa. Here, we compared the natural abundances of 13C and 15N isotopes in the AM-associated gentian species Pterygocalyx volubilis growing in high-light-intensity habitats with those of co-occurring autotrophic C3 and C4 plants and AM fungal spores. We found that P. volubilis was significantly enriched in 13C compared with the surrounding C3 plants, which suggests the transfer of some C from the surrounding autotrophic plants through shared AM networks. In addition, the intermediate δ15N values of P. volubilis, between those of autotrophic plants and AM fungal spores, provide further evidence for partial mycoheterotrophy in P. volubilis. Although it is often considered that light deficiency selects partial mycoheterotrophy, we show that partial mycoheterotrophy in AM-forming plants can evolve even under light-saturated conditions. The fact that there have been relatively few descriptions of partial mycoheterotrophy in AM plants may not necessarily reflect the rarity of such associations. In conclusion, partial mycoheterotrophy in AM plants may be more common than hitherto believed. [ABSTRACT FROM AUTHOR]

Several species of the family Orchidaceae have limited distribution and small populations, in which the probability of extinction is much higher than in broadly distributed species. As orchid seeds are very small and lack energy reserves, they require association with compatible mycorrhizal fungi that provide nutrients and carbon needed for germination and early development. Therefore, identification of putative orchid mycorrhizal partners is essential for the design of conservation strategies for these species. We investigated the mycorrhizal associations of two endangered rare species of orchids, Bipinnula apinnula and B. volckmannii, and compared our results with the mycorrhizal associations of two broadly distributed species of Bipinnula. We isolated mycorrhizal fungi and directly amplified DNA from colonized root segments and found an extremely low diversity of mycorrhizal fungi. The rarefaction curves of rare species populations saturated at a single OTU, showing a highly specialized association and considerably lower diversity than the other two species of Bipinnula. Nevertheless, symbiotic germination experiments with fungal strains of this specific fungal OTU only reached the stage of seed coat rupture in both orchid species, without formation of protocorms. Our results suggest that seeds may require additional treatment for successful germination and that fungal partners isolated from adult plants may not be the same that support the germination of seeds. Further studies are needed to improve germination of thiese orchid species and to contribute to their conservation. [ABSTRACT FROM AUTHOR]

Aim: Interactions with mycorrhizal fungi are increasingly recognized as an important factor underlying the distribution and abundance of orchid species. However, the geographical distribution of orchid mycorrhizal fungi (OMF) and how their communities vary over large geographical areas are less well understood. Because climatic and environmental similarity may decrease with geographical distance or because some OMF have limited dispersal capabilities, similarities in orchid mycorrhizal communities can be expected to decrease with increasing distances separating orchid populations. However, up till now empirical evidence is largely lacking. Location: Eurasia. Taxa: Gymnadenia conopsea (L.) R. Brown and Epipactis helleborine (L.) Crantz. Methods: High‐throughput sequencing was used to perform a cross‐continental comparison of OMF that associate with two widespread Eurasian terrestrial orchids, Epipactis helleborine and Gymnadenia conopsea. Both phylogenetic and nonphylogenetic measures of community dissimilarity and their components were calculated and related to geographical distances using Mantel tests. Results: Our results showed that in both orchid species similarity in mycorrhizal communities decreased significantly with geographical distance. Decomposing the contribution of spatial turnover and nestedness to overall dissimilarity showed that the observed dissimilarity was mainly the result of species replacement between regions, and not of species loss. Similarly, a strong relationship was observed between phylogenetic community dissimilarity and geographical distance. Decomposing PCD values into a nonphylogenetic and phylogenetic component showed that orchid populations located closely next to each other were likely to contain the same operational taxonomic units (OTUs), but that the non‐shared taxa came from different phylogenetic clades. Species indicator analyses showed that the majority of OMF OTUs were restricted to particular geographical areas. However, some OTUs occurred in both continents, indicating that some fungi have very wide distributions. Main conclusions: Overall, these results demonstrate that orchid mycorrhizal communities differ substantially across large geographical areas, but that the distribution of orchids is not necessarily restricted by the distribution of particular OMF. Hence, widespread orchid species can be considered mycorrhizal generalists that are flexible in the OMF with which they associate across large geographical areas. [ABSTRACT FROM AUTHOR]

Background: Orchids maintain a symbiotic relationship with mycorrhizal fungi in the lifecycle. Previous reports indicated that diverse mycobionts may have different roles during orchid growth and development. Although various mycorrhizal fungi have been isolated from Dendrobium roots and protocorms, little is known about their specific effects on seed germination and seedling growth. To understand the specific role of isolated fungal strains (i.e., Tulasnella and Sebacina), we used symbiotic culture to compare the effect of 6 fungal strains on seed germination and seedling growth of Dendrobium officinale, an important Chinese medicinal orchid. Results: In symbiotic germination tests, 6 fungal strains (4 Tulasnella strains and 2 Sebacina strains) promoted seed germination with different efficiencies. Seeds inoculated with Tulasnella strains S6 and S7 conferred higher germination percentage and faster protocorm development than other fungal strains. In symbiotic cultures, seedlings inoculated with Sebacina strain S3 had optimal fresh and dry matter yield. Also, Tulasnella strains S6 and S7 promoted seedling growth with good fresh and dry matter yield. Sebacina strain S2 inoculation greatly enhanced root and tiller production and the content of total crude polysaccharides, although seedlings were smaller with less fresh and dry matter yield than other seedlings. Conclusions: Tulasnella and Sebacina strains could promote seed germination and seedling growth of D. officinale with different efficiencies. Our results suggest a non-specific mycorrhizal association and development-dependent preference. Our data provide the basic knowledge for use of different fungal strains in conservation and/or production practices of D. officinale. [ABSTRACT FROM AUTHOR]

Lycopods are tracheophytes in the Kingdom Plantae and represent one of the oldest lineages of living vascular plants. Symbiotic interactions between these plants with fungi and bacteria, including fine root endophytes in Endogonales, have been hypothesized to have helped early diverging plant lineages colonize land. However, attempts to study the lycopod rhizobiome in its natural environment are still limited. In this study, we used Illumina amplicon sequencing to characterize fungal and bacterial diversity in nine Lycopodiaceae (club moss) species collected in New Zealand. This was done with generic fungal ITS rDNA primers, as well as Endogonales- and arbuscular mycorrhizal fungi (AMF)-selective primer sets targeting the 18S rDNA, and generic bacterial primers targeting the V4 region of the 16S rDNA. We found that the Lycopodiaceae rhizobiome was comprised of an unexpected high frequency of Basidiomycota and Ascomycota coincident with a low abundance of Endogonales and Glomerales. The distribution and abundance of Endogonales varied with host lycopod, and included a novel taxon as well as a single operational taxonomic unit (OTU) that was detected across all plant species. The Lycopodiaceae species with the greatest number and also most unique OTUs was Phlegmariurus varius , while the plant species that shared the most fungal OTUs were Lycopodiella fastigiatum and Lycopodium scariosum. The bacterial OTU distribution was generally not consistent with fungal OTU distribution. For example, community dissimilarity analysis revealed strong concordance between the evolutionary histories of host plants with the fungal community but not with the bacterial community, indicating that Lycopodiaceae have evolved specific relationships with their fungal symbionts. Notably, nearly 16% of the ITS rDNA fungal diversity detected in the Lycopodiaceae rhizobiome remained poorly classified, indicating there is much plant-associated fungal diversity left to describe in New Zealand. [ABSTRACT FROM AUTHOR]

Some mixotrophic plants from temperate forests use the mycorrhizal fungi colonizing their roots as a carbon source to supplement their photosynthesis. These fungi are also mycorrhizal on surrounding trees, from which they transfer carbon to mixotrophic plants. These plants are thus reputed difficult to transplant, even when their protection requires it. Here, we take profit of a successful ex situ pot cultivation over 1 to 3 years of the mixotrophic orchid Epipacis helleborine to investigate its mycorrhizal and nutrition status. Firstly, compared with surrounding autotrophic plants, it did not display the higher N content and higher isotopic (13C and 15N) abundance that normally feature mixotrophic orchids because they incorporate N-, 13C-, and 15N-rich fungal biomass. Second, fungal barcoding by next-generation sequencing revealed that the proportion of ectomycorrhizal fungi (expressed as percentage of the total number of either reads or operational taxonomic units) was unusually low compared with E. helleborine growing in situ: instead, we found a high percentage of rhizoctonias, the usual mycorrhizal partners of autotrophic orchids. Altogether, this supports autotrophic survival. Added to the recently published evidence that plastid genomes of mixotrophic orchids have intact photosynthetic genes, this suggests that at least some of them have abilities for autotrophy. This adds to the ecological plasticity of mixotrophic plants, and may allow some reversion to autotrophy in their evolution. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

There is little known about the function of rice hexokinases (HXK s) in planta. We characterized hxk5-1 , a Tos17 mutant of OsHXK5 that is up-regulated in maturing pollen, a stage when starch accumulates. Progeny analysis of self-pollinated heterozygotes of hxk5-1 and reciprocal crosses between the wild-type and heterozygotes revealed that loss of HXK5 causes male sterility. Homozygous hxk5-1 , produced via anther culture, and additional homozygous hxk5-2 , hxk5-3 and hxk5-4 lines created by CRISPR/Cas9 confirmed the male-sterile phenotype. In vitro pollen germination ability and in vivo pollen tube growth rate were significantly reduced in the hxk5 mutant pollen. Biochemical analysis of anthers with the mutant pollen revealed significantly reduced hexokinase activity and starch content, although they were sufficient to produce some viable seed. However, the mutant pollen was unable to compete successfully against wild-type pollen. Expression of the catalytically inactive OsHXK5-G113D did not rescue the hxk5 male-sterile phenotype, indicating that its catalytic function was responsible for pollen fertility, rather than its role in sugar sensing and signaling. Our results demonstrate that OsHXK5 contributes to a large portion of the hexokinase activity necessary for the starch utilization pathway during pollen germination and tube growth, as well as for starch biosynthesis during pollen maturation. [ABSTRACT FROM AUTHOR]

Urban expansion threatens biodiversity worldwide, therefore urban spaces need to be amenable to biodiversity conservation. On trees in urban environments, natural colonisation and successful translocation of epiphytic orchids are necessary to enhance urban biodiversity, and depend on the availability of compatible orchid mycorrhizal fungi (OMF). However, the extent of OMF presence and distribution, as well as niche requirements for the OMF, remain poorly studied. To identify and quantify OMF on urban trees as well as assess their suitability for native epiphytic orchids, we conducted high-throughput sequencing on tree bark and orchid root samples. OMF were detected at 60% of the study sites on 16% of 270 bark samples (from stem, fork, and branch microsites within each tree). OMF presence and richness on bark samples were related to multiple biophysical factors; in general, humus presence and precipitation levels were positively predictive of OMF presence and richness. We found Ceratobasidiaceae- and Serendipitaceae-associated OMF both on bark and within roots. Orchid species also showed differing mycorrhizal specificity. Sites associated with fungal genera Ceratobasidium, Rhizoctonia, and Serendipita were considered suitable habitats for seven orchid species. The results suggest that urban trees support OMF and are therefore suitable for native orchid species; however, OMF availability are largely constrained by biophysical factors. To maximise the likelihood of translocation success and consequent natural establishment, we propose that (micro)sites are screened for compatible OMF prior to any intervention. [ABSTRACT FROM AUTHOR]

Copyright of Anbar Journal of Agricultural Sciences is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

For germination and establishment, orchids depend on carbon (C) and nutrients supplied by mycorrhizal fungi. As adults, the majority of orchids then appear to become autotrophic. To compare the proportional C and nitrogen (N) gain from fungi in mycoheterotrophic seedlings and in adults, here we examined in the field C and N stable isotope compositions in seedlings and adults of orchids associated with ectomycorrhizal and saprotrophic fungi. Using a new highly sensitive approach, we measured the isotope compositions of seedlings and adults of four orchid species belonging to different functional groups: fully and partially mycoheterotrophic orchids associated with narrow or broad sets of ectomycorrhizal fungi, and two adult putatively autotrophic orchids associated exclusively with saprotrophic fungi. Seedlings of orchids associated with ectomycorrhizal fungi were enriched in (13) C and (15) N similarly to fully mycoheterotrophic adults. Seedlings of saprotroph-associated orchids were also enriched in (13) C and (15) N, but unexpectedly their enrichment was significantly lower, making them hardly distinguishable from their respective adult stages and neighbouring autotrophic plants. We conclude that partial mycoheterotrophy among saprotroph-associated orchids cannot be identified unequivocally based on C and N isotope compositions alone. Thus, partial mycoheterotrophy may be much more widely distributed among orchids than hitherto assumed., (© 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.)

The fern and lycophyte flora of Japan comprising 721 native taxa (including subspecies and varieties) plus 371 interspecific hybrids was reassessed using a nearly comprehensively sampled distribution map at 10 km resolution vouchered by 216,687 specimens, up-to-date cytotaxonomic information covering 74% of the taxa, and an rbcL sequence dataset covering 97.9% of the taxa. Spatial distribution of species richness and phylogenetic diversity was visualized. Apomixis was observed in 11.0% of the native taxa whose reproductive modes are known. The number of sexually reproducing polyploid taxa (n = 199) is less than sexual diploids (n = 241), and 30 of them are evidently allopolyploid, in contrast with the low number of possible autopolyploids (n = 4). Apomictic taxa were found to have smaller latitudinal ranges than sexual taxa or taxa with multiple reproductive modes. A morphological character dataset in Lucid format is provided for taxonomic identification of the native taxa. [ABSTRACT FROM AUTHOR]

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The Chinese occurrences of the marattioid fern genus Christensenia have been considered as requiring protection because of its extreme rarity and very small population size. Here, we explored different biological aspects to enable protection of these rare ferns, well known as Mesozoic living fossils. Firstly, we documented the cytology of the Chinese occurrences for the first time. This is the second tetraploid record of Christensenia worth for further studies to confirm its taxonomic status. Secondly, we obtained the first complete plastid genome of this genus, which confirmed the proposed conservatism of the plastid genome structure in marattioid ferns. By comparing the chloroplast genome with other marattioids, we identified several candidate regions to develop highly variable markers to investigate the intra-species diversity of marattioid ferns. Thirdly, phylogenetic analyses of rbcL sequences implied that there are at least two distinct species of Christensenia. Finally, we re-assessed the conservation status of Christensenia in the context of its local and global distribution by assessing specimen information extracted from publications and digitized voucher information. This assessment confirmed the need to obtain more accurate information about the distribution of this genus to assess the status incorporating the disjunct distribution from southern China and India in the North towards the Solomon Islands in the South. [ABSTRACT FROM AUTHOR]

Heterotrophic plants are evolutionary experiments in genomic, morphological, and physiological change. Yet, genomic sampling gaps exist among independently derived heterotrophic lineages, leaving unanswered questions about the process of genome modification. Here, we have sequenced complete plastid genomes for all species of the leafless orchid genus Hexalectris , including multiple individuals for most, and leafy relatives Basiphyllaea and Bletia. Our objectives are to determine the number of independent losses of photosynthesis and to test hypotheses on the process of genome degradation as a result of relaxed selection. We demonstrate four to five independent losses of photosynthesis in Hexalectris based on degradation of the photosynthetic apparatus, with all but two species displaying evidence of losses, and variation in gene loss extending below the species level. Degradation in the atp complex is advanced in Hexalectris warnockii , whereas only minimal degradation (i.e. physical loss) has occurred among some "housekeeping" genes. We find genomic rearrangements, shifts in Inverted Repeat boundaries including complete loss in one accession of H. arizonica , and correlations among substitutional and genomic attributes. Our unprecedented finding of multiple, independent transitions to a fully mycoheterotrophic lifestyle in a single genus reveals that the number of such transitions among land plants is likely underestimated. This study underscores the importance of dense taxon sampling, which is highly informative for advancing models of genome evolution in heterotrophs. Mycoheterotrophs such as Hexalectris provide forward-genetic opportunities to study the consequences of radical genome evolution beyond what is possible with mutational studies in model organisms alone. [ABSTRACT FROM AUTHOR]

Aim: Mycoheterotrophy is a mode of life where plants cheat the mycorrhizal symbiosis, receiving carbon via their fungal partners. Despite being widespread, mycoheterotrophic plants are locally rare, hampering the understanding of their global environmental drivers. Here, we explore global environmental preferences of mycoheterotrophy, and investigate environmental drivers of differential habitat preferences of mycoheterotrophic plants associated with arbuscular (AM) and ectomycorrhizal (EM) fungi. Location: Global. Time period: Current. Major taxa studied: Mycoheterotrophic flowering plants. Methods: We compiled the largest global dataset of epiparasitic mycoheterotrophic plant species occurrences and examined which environmental factors, including soil type, climate, vegetation type and distribution patterns of mycorrhizal autotrophic plants, relate to occurrence patterns of mycoheterotrophic plant species associated with AM and EM fungi. Results: Mycoheterotrophic plant species avoid cold and highly seasonal climates and show a strong preference for forests. AM‐associated mycoheterotrophs are predominantly found in broadleaved tropical evergreen forests whereas EM‐associated mycoheterotrophs occur in temperate regions, mostly in broadleaved deciduous and evergreen needleleaved forests. The abundance of AM and EM autotrophic plants was a weaker predictor for mycoheterotrophs occurrences than forest type. Temperature and precipitation variables – but not edaphic factors – were the best predictors explaining the distribution patterns of mycoheterotrophs after accounting for the effects of forest type. For individual lineages, major differences in environmental preferences (often related to edaphic factors) occurred that were significantly associated with plant evolutionary relationships, indicating that these cheater plants have limited adaptive capabilities. Main conclusions: The strong global geographical segregation of AM and EM mycoheterotrophs does not reflect the abundance of their potential autotrophic hosts, but seems to be driven by differential climate and habitat preferences. Our results highlight the non‐trivial nature of mycorrhizal interactions, and indicate that identity of the partners is not enough to understand the underlying mechanisms promoting plant–fungal interactions in mycoheterotrophic plants. [ABSTRACT FROM AUTHOR]