Wetland dicots and monocots differ in colonization by arbuscular mycorrhizal fungi and dark septate endophytes

As an initial step towards evaluating whether mycorrhizas influence composition and diversity in calcareous fen plant communities, we surveyed root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytic fungi (DSE) in 67 plant species in three different fens in central New York State (USA). We found colonization by AMF and DSE in most plant species at all three sites, with the type and extent of colonization differing between monocots and dicots. On average, AMF colonization was higher in dicots (58±3%, mean±SE) than in monocots (13±4%) but DSE colonization followed the opposite trend (24±3% in monocots and 9±1% in dicots). In sedges and cattails, two monocot families that are often abundant in fens and other wetlands, AMF colonization was usually very low (<10%) in five species and completely absent in seven others. However, DSE colonization in these species was frequently observed. Responses of wetland plants to AMF and DSE are poorly understood, but in the fen communities surveyed, dicots appear to be in a better position to respond to AMF than many of these more abundant monocots (e.g., sedges and cattails). In contrast, these monocots may be more likely to respond to DSE. Future work directed towards understanding the response of these wetland plants to AMF and DSE should provide insight into the roles these fungal symbionts play in influencing diversity in fen plant communities.     

Root fungal symbionts interact with mammalian herbivory, soil nutrient availability and specific habitat conditions

Herbivory, competition and soil fertility interactively shape plant communities and exhibit an important role in modifying conditions for host-dependent fungal symbionts. However, field studies on the combined impacts of natural herbivory, competition and soil fertility on root fungal symbionts are rare. We asked how mammalian herbivory, fertilization, liming and plant–plant competition affect the root colonization of arbuscular mycorrhizal fungi (AMF) and dark septate endophytic (DSE) fungi of the dicot herb, Solidago virgaurea. The 2-year full-factorial experiment was conducted in two contrasting habitats: non-acidic and acidic mountain tundra. We found that herbivory increased arbuscular colonization (i.e. the site of resource exchange) at fertile non-acidic sites, where vegetation was rich in species having AMF symbionts, whereas at infertile acidic sites, where plants having AMF symbiont are scarce, the response was the opposite. Herbivory of the host plant negatively affected DSE hyphal and sclerotial colonization in unfertilized plots, possibly due to reduced carbon flow from the host plant while there was no effect of herbivory in fertilized plots. DSE colonization was highest in unfertilized exclosures where soil nutrient concentrations were also lowest. Liming had a negative effect on DSE hyphal colonization, and its effect also interacted with herbivory and the habitat. Biomass removal of the neighboring plants did not affect the root colonization percent of either arbuscules or DSE. Our results show that the impacts of aboveground mammalian herbivory, soil nutrient availability and specific habitat conditions on belowground root fungal symbionts are highly dependent on each other. Arbuscule response to herbivory appeared to be regulated by specific habitat conditions possibly caused by differences in the AMF availability in the soil while DSE response was associated with availability of host-derived carbon. Our result of the relationship between herbivory and soil nutrients suggests an important role of DSE in ecosystem processes.     

丛枝菌根真菌与深色有隔内生真菌生态修复功能与作用

生态修复是目前全球关注的热点问题,如何增加植被的覆盖度及生态修复效率是目前研究的重点。丛枝菌根真菌(arbuscular mycorrhiza fungi,AMF)和深色有隔内生真菌(dark septate endophyte,DSE)均是自然界植物根际分布广泛的一类内生真菌,均能与植物形成菌根共生体,具有一定的促进植物生长、抵抗逆境及修复污染土壤等功能与作用,在生态修复中具有广泛的应用潜力。本文综述了AMF及DSE两种微生物的功能、作用及其在生态修复应用中的研究进展,并进一步对AMF和DSE在生态修复中存在的问题和前景进行展望。     

The dark side is not fastidious--dark septate endophytic fungi of native and invasive plants of semiarid sandy areas

Dark septate endophytic (DSE) fungi represent a frequent root-colonizing fungal group common in environments with strong abiotic stress, such as (semi)arid ecosystems. This work aimed to study the DSE fungi colonizing the plants of semiarid sandy grasslands with wood steppe patches on the Great Hungarian Plain. As we may assume that fungi colonizing both invasive and native species are generalists, root associated fungi (RAF) were isolated from eight native and three invasive plant species. The nrDNA sequences of the isolates were used for identification. To confirm that the fungi were endophytes an artificial inoculation system was used to test the isolates: we considered a fungus as DSE if it colonized the roots without causing a negative effect on the plant and formed microsclerotia in the roots. According to the analyses of the ITS sequence of nrDNA the 296 isolates clustered into 41 groups. We found that 14 of these 41 groups were DSE, representing approximately 60% of the isolates. The main DSE groups were generalist and showed no specificity to area or season and colonized both native and invasive species, demonstrating that exotic plants are capable of using the root endophytic fungi of the invaded areas. The DSE community of the region shows high similarity to those found in arid grasslands of North America. Taking into account a previous hypothesis about the common root colonizers of those grasslands and our results reported here, we hypothesize that plants of (semi)arid grasslands share common dominant members of the DSE fungal community on a global scale.     

Coexistence of arbuscular mycorrhizae and dark septate endophytic fungi in an undisturbed and a disturbed site of an arid ecosystem

Effect of disturbance on root colonization and vertical distribution of arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) was investigated at two adjacent sites of Lal Suhanra Biosphere Reserve, Pakistan. Disturbance clearly affected AMF and DSE colonization, vertical distribution of AMF and plant community structure. Mean colonization of AMF and DSE was slightly less at the disturbed site. Average spore densities, diversity and richness of AMF and DSE were higher at the undisturbed site. A study of the vertical distribution of AMF associated with the five plant species most common to each study site indicated that beside AMF and DSE colonization disturbance may affect AMF species composition. Correlation of AMF with DSE is also discussed.     

Fungal root endophyte associations of plants endemic to the Pamir Alay Mountains of Central Asia

The fungal root endophyte associations of 16 species from 12 families of plants endemic to the Pamir Alay Mountains of Central Asia are presented. The plants and soil samples were collected in Zeravshan and Hissar ranges within the central Pamir Alay mountain system. Colonization by arbuscular mycorrhizal fungi (AMF) was found in 15 plant species; in 8 species it was of the Arum type and in 4 of the Paris type, while 3 taxa revealed intermediate arbuscular mycorrhiza (AM) morphology. AMF colonization was found to be absent only in Matthiola integrifolia, the representative of the Brassicaceae family. The AM status and morphology are reported for the first time for all the species analyzed and for the genera Asyneuma, Clementsia, and Eremostachys. Mycelia of dark septate endophytes (DSE) accompanied the AMF colonization in ten plant species. The frequency of DSE occurrence in the roots was low in all the plants, with the exception of Spiraea baldschuanica. However, in the case of both low and higher occurrence, the percentage of DSE root colonization was low. Moreover, the sporangia of Olpidium spp. were sporadically found inside the root epidermal cells of three plant species. Seven AMF species (Glomeromycota) found in the trap cultures established with soils surrounding roots of the plants being studied were reported for the first time from this region of Asia. Our results provide information that might well be of use to the conservation and restoration programmes of these valuable plant species. The potential application of beneficial root-inhabiting fungi in active plant protection projects of rare, endemic and endangered plants is discussed.     

Colonization Characteristics and Composition of Dark Septate Endophytes (DSE) in a Lead and Zinc Slag Heap in Southwest China

The colonization characteristics and diversity of dark septate endophytes (DSE) associated with plants growing in a lead and zinc slag heap with a more than 250-year history in southwestern China were assessed. A total of 117 plant samples belonging to 29 species and 19 families were examined, and 97% (28/29) were colonized by dark septate fungal hyphae or microsclerotia. DSE colonization intensity varied greatly across plant species and also across individuals of the same species, showing a hyphal colonization range of 0.0–35.7%. In total, 38 melanized root-associated fungi with various morphological characteristics were isolated and 82% (31/38) of fungi were closely related to well-known DSE fungi, such as Exophiala spp., Phialophora (= Cadophora) spp., Phialocephala spp., and Leptodontidium spp. Among these, Phialophora and Exophiala were the dominant genera. Our results showed that DSE dominantly colonized the roots of the pioneer plants growing in such an ancient Pb-Zn slag heap. By contributing to metal tolerance and nutrient acquisition, DSE may be integral to the function of metal mine ecosystems.     

Medicinal plants as hosts of arbuscular mycorrhizal fungi and dark septate endophytes

Arbuscular mycorrhizal and dark septate endophyte associations of 31 medicinal plant species collected from the Garden of Medicinal Plants of the Faculty of Pharmacy, Jagiellonian University, Collegium Medicum in Kraków were investigated. Arbuscular mycorrhiza (AM) was found in 30 species; 23 were of the Arum-type, 5—Paris and 2 taxa revealed intermediate morphology. Many plants were strongly colonized by arbuscular mycorrhizal fungi (AMF). The mycelium of dark septate endophytes (DSE) was observed in 21 taxa. However, the percentage of root colonization by these fungi was low. Spores of 15 species of AMF (Glomeromycota) were found in the rhizosphere of the investigated plants. Our results are the first detailed report of both AMF and DSE associations of these plant species. The use of AMF and DSE during the process of medicinal plant cultivation for pharmaceutical purposes is discussed.     

Spatial variability of arbuscular mycorrhizal fungal spores in two natural plant communities

Geostatistical techniques were used to assess the spatial patterns of spores of arbuscular mycorrhizal fungi (AMF) in soils from two contrasting plant communities: a salt marsh containing only arbuscular mycorrhizal and non-mycorrhizal plants in a distinct clumped distribution pattern and a maquis with different types of mycorrhiza where most plants were relatively randomly distributed. Also evaluated was the relationship between the spatial distribution of spores and AM plant distribution and soil properties. A nested sampling scheme was applied in both sites with sample cores taken from nested grids. Spores of AMF and soil characteristics (organic matter and moisture) were quantified in each core, and core sample location was related to plant location. Semivariograms for spore density indicated strong spatial autocorrelation and a patchy distribution within both sites for all AM fungal genera found. However, the patch size differed between the two plant communities and AM fungal genera. In the salt marsh, AM fungal spore distribution was correlated with distance to AM plants and projected stand area of AM plants. In maquis, spatial AM fungal spore distribution was correlated with organic matter. These results suggest that spore distribution of AMF varied between the two plant communities according to plant distribution and soil properties.     

The co-existence between DSE and AMF symbionts affects plant P pools through P mineralization and solubilization processes

The capacity of dark septate endophytes (DSE; Phialocephala turiciensis, Acephala applanata, P. glacialis and Phaeomollisia piceae) to solubilize inorganic phosphate (P) and to mineralize the organic form was studied. We analysed the effect of DSE strains on P uptake by Trifolium repens in the presence or absence of arbuscular mycorrhizal fungi (AMF). Phosphatases were observed both in the absence of the host plant and the organic resource, showing that the P mineralization process is not induced by the enzyme substrate or the host. DSE were more efficient at mineralizing organic P. Independently of the presence of AMF, DSE increased the pool of P in the soil, with significant differences being found in P levels among the different DSE. In contrast, plant P uptake was increased by AMF. The P content of plants increased with the co-inoculation of AMF and P. turiciensis or P. piceae. We hypothesize a close relationship between DSE and AMF in relation to P availability and uptake in plants. Whereas DSE increase the pool of P in the rhizosphere, AMF are responsible for P transfer to the host, with co-colonization of plants by DSE and AMF showing a synergistic outcome.     

Assessment of root-associated fungal communities colonizing two species of tropical grasses reveals incongruence to fungal communities of North American native grasses

This study characterized the root-associated fungal (RAF) communities inhabiting Eustachys petraea and Panicum maximum, two tropical and weedy grass species on an island off the coast of Honduras, Central America. Media-based analyses revealed that fungi colonizing these grasses exhibited similar morphotypes (albeit in different proportions). Conversely, molecular-based analyses suggested that the community of fungal OTUs were dissimilar, with several OTUs commonly present on only one plant and no AMF sequences present in either plant species. When compared to various datasets of RAF obtained from the North American mainland (with similar methods), the Honduran grasses harbored a different set of fungal OTUs. Interestingly, some cosmopolitan taxonomic clades commonly encountered on the mainland were also isolated in Honduras, though based on ITS rDNA the fungi were only related distantly to those on the mainland. Additional work is necessary to determine how the RAF communities change over geographic distance and whether invasive grasses are constitutionally set up to accept relationships with a different suite of RAF species.     

Cooccurring Gentiana verna and Gentiana acaulis and their neighboring plants in two Swiss upper montane meadows harbor distinct arbuscular mycorrhizal fungal communities

The community composition of arbuscular mycorrhizal fungi (AMF) was analyzed in roots of Gentiana verna, Gentiana acaulis, and accompanying plant species from two species-rich Swiss alpine meadows located in the same area. The aim of the study was to elucidate the impact of host preference or host specificity on the AMF community in the roots. The roots were analyzed by nested PCR, restriction fragment length polymorphism screening, and sequencing of ribosomal DNA small-subunit and internal transcribed spacer regions. The AMF sequences were analyzed phylogenetically and used to define monophyletic sequence types. The AMF community composition was strongly influenced by the host plant species, but compositions did not significantly differ between the two sites. Detailed analyses of the two cooccurring gentian species G. verna and G. acaulis, as well as of neighboring Trifolium spp., revealed that their AMF communities differed significantly. All three host plant taxa harbored AMF communities comprising multiple phylotypes from different fungal lineages. A frequent fungal phylotype from Glomus group B was almost exclusively found in Trifolium spp., suggesting some degree of host preference for this fungus in this habitat. In conclusion, the results indicate that within a relatively small area with similar soil and climatic conditions, the host plant species can have a major influence on the AMF communities within the roots. No evidence was found for a narrowing of the mycosymbiont spectrum in the two green gentians, in contrast to previous findings with their achlorophyllous relatives.     

AMF和DSE组合菌剂促生防线虫病效应

本试验旨在探究丛枝菌根真菌（AMF）和暗隔内生真菌（DSE）组合菌剂对南方根结线虫Meloidogyne incognita发育、侵染、黄瓜根结线虫病以及黄瓜生长发育的影响,为进一步探索AMF和DSE协同发挥生理生态效应的作用机制奠定基础。试验对“津优35号”黄瓜接种南方根结线虫、Phoma leveillei（DSE）、Funneliformis mosseae（AMF）、Glomus versiforme（AMF）、Acaulospora laevis（AMF）和/或Scutellospora aurigloba（AMF）。结果表明,供试AMF与DSE 能促进黄瓜生长、提高产量和增强抗病性。AMF+DSE组合处理的菌根和DSE的定殖数量、株高、茎粗、地上部和根系干重、单株产量等显著优于单接种AMF或单接种DSE处理。以F. mosseae + P. leveillei组合抑制南方根结线虫的发育、降低线虫繁殖数量、根内定殖数量、发病率和根结指数的效果最好。     

Contrasting impacts of defoliation on root colonization by arbuscular mycorrhizal and dark septate endophytic fungi of Medicago sativa

Individual plants typically interact with multiple mutualists and enemies simultaneously. Plant roots encounter both arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungi, while the leaves are exposed to herbivores. AMF are usually beneficial symbionts, while the functional role of DSE is largely unknown. Leaf herbivory may have a negative effect on root symbiotic fungi due to decreased carbon availability. However, evidence for this is ambiguous and no inoculation-based experiment on joint effects of herbivory on AM and DSE has been done to date. We investigated how artificial defoliation impacts root colonization by AM (Glomus intraradices) and DSE (Phialocephala fortinii) fungi and growth of Medicago sativa host in a factorial laboratory experiment. Defoliation affected fungi differentially, causing a decrease in arbuscular colonization and a slight increase in DSE-type colonization. However, the presence of one fungal species had no effect on colonization by the other or on plant growth. Defoliation reduced plant biomass, with this effect independent of the fungal treatments. Inoculation by either fungal species reduced root/shoot ratios, with this effect independent of the defoliation treatments. These results suggest AM colonization is limited by host carbon availability, while DSE may benefit from root dieback or exudation associated with defoliation. Reductions in root allocation associated with fungal inoculation combined with a lack of effect of fungi on plant biomass suggest DSE and AMF may be functional equivalent to the plant within this study. Combined, our results indicate different controls of colonization, but no apparent functional consequences between AM and DSE association in plant roots in this experimental setup.     

Arbuscular mycorrhizal fungal communities along a pedo-hydrological gradient in a Central Amazonian terra firme forest

Little attention has been paid to plant mutualistic interactions in the Amazon rainforest, and the general pattern of occurrence and diversity of arbuscular mycorrhizal fungi (AMF) in these ecosystems is largely unknown. This study investigated AMF communities through their spores in soil in a ‘terra firme forest’ in Central Amazonia. The contribution played by abiotic factors and plant host species identity in regulating the composition, abundance and diversity of such communities along a topographic gradient with different soils and hydrology was also evaluated. Forty-one spore morphotypes were observed with species belonging to the genera Glomus and Acaulospora, representing 44 % of the total taxa. Soil texture and moisture, together with host identity, were predominant factors responsible for shaping AMF communities along the pedo-hydrological gradient. However, the variability within AMF communities was largely associated with shifts in the relative abundance of spores rather than changes in species composition, confirming that common AMF species are widely distributed in plant communities and all plants recruited into the forest are likely to be exposed to the dominant sporulating AMF species.     

Effects of Alliaria petiolata (garlic mustard; Brassicaceae) on mycorrhizal colonization and community structure in three herbaceous plants in a mixed deciduous forest

Herbaceous plant species are important components of forest ecosystems, and their persistence in forests may be affected by invasive plant species that reduce mycorrhizal colonization of plant roots. I examined the effect of the invasive plant Alliaria petiolata on arbuscular mycorrhizal fungi (AMF) colonizing the roots of three forest plant species. AMF root colonization and community structure was examined from plants that were growing either in the absence or presence of Alliaria under natural forest conditions. AMF root colonization varied among the plant species but was not significantly affected by Alliaria. With molecular methods, ～12 different taxa of AMF could be distinguished among the root samples, and these taxa belonged to the genera Acaulospora and Glomus, with Glomus dominating AMF communities. There were significant differences between the community of AMF colonizing roots of Maianthemum racemosum and Trillium grandiflorum, but only AMF communities of Maianthemum roots were significantly affected by Alliaria. Indicator species analysis found that an Acaulospora species type was a significant indicator of Maianthemum plants grown in the absence of Alliaria. These results suggest invasive plants like Alliaria may selectively suppress AMF fungi, and this suppression can affect AMF communities colonizing the roots of some native plant species.     

Cooccurring Gentiana verna and Gentiana acaulis and Their Neighboring Plants in Two Swiss Upper Montane Meadows Harbor Distinct Arbuscular Mycorrhizal Fungal Communities

The community composition of arbuscular mycorrhizal fungi (AMF) was analyzed in roots of Gentiana verna, Gentiana acaulis, and accompanying plant species from two species-rich Swiss alpine meadows located in the same area. The aim of the study was to elucidate the impact of host preference or host specificity on the AMF community in the roots. The roots were analyzed by nested PCR, restriction fragment length polymorphism screening, and sequencing of ribosomal DNA small-subunit and internal transcribed spacer regions. The AMF sequences were analyzed phylogenetically and used to define monophyletic sequence types. The AMF community composition was strongly influenced by the host plant species, but compositions did not significantly differ between the two sites. Detailed analyses of the two cooccurring gentian species G. verna and G. acaulis, as well as of neighboring Trifolium spp., revealed that their AMF communities differed significantly. All three host plant taxa harbored AMF communities comprising multiple phylotypes from different fungal lineages. A frequent fungal phylotype from Glomus group B was almost exclusively found in Trifolium spp., suggesting some degree of host preference for this fungus in this habitat. In conclusion, the results indicate that within a relatively small area with similar soil and climatic conditions, the host plant species can have a major influence on the AMF communities within the roots. No evidence was found for a narrowing of the mycosymbiont spectrum in the two green gentians, in contrast to previous findings with their achlorophyllous relatives.     

A survey of the mycorrhization of Southeast Asian ferns and lycophytes

The colonisation of land by plants may not have been possible without mycorrhizae, which supply the majority of land plants with nutrients, water and other benefits. In this sense, the mycorrhization of basal groups of land plants such as ferns and lycophytes is of particular interest, yet only about 9% of fern and lycophyte species have been sampled for their mycorrhization status, and no community‐level analyses exist for tropical fern communities. In the present study, we screened 170 specimens of ferns and lycophytes from Malaysia and Sulawesi (Indonesia), representing 126 species, and report the mycorrhization status for 109 species and 19 genera for the first time. Mycorrhizal colonisations were detected in 96 (56.5%) of the specimens, 85 of which corresponded to arbuscular mycorrhizae (AMF), three to dark‐septate endophytes (DSE) and four to mixed colonisations (AMF + DSE). DSE colonisations were lower than in comparable samples of ferns from the Andes, suggesting a geographical or taxonomic pattern in this type of colonisation. Epiphytes had significantly lower levels of colonisation (26.1%) than terrestrial plants (70.7%), probably due to the difficulty of establishment of mycorrhizal fungi in the canopy habitat.     

Fungal root symbionts and their relationship with fine root proportion in native plants from the Bolivian Andean highlands above 3,700?m elevation

Here, we examined the colonization by fungal root symbionts in the cultivated Andean grain Chenopodium quinoa and in 12 species that dominate plant communities in the Bolivian Altiplano above 3,700 m elevation and explore for the possible relationships between fungal colonization and fine root proportion. The 12 most abundant species in the study area were consistently colonized by AMF and DSE. In contrast, the annual Andean grain C. quinoa showed negligible or absence of mycorrhizal fungi colonizing roots. On the other hand, C. quinoa, Junelia seriphioides and Chersodoma jodopappa were infected to a varying degree by the root pathogen Olpidium sp. We observed no relationship between AMF and DSE colonization and proportion of fine roots in the root system, but instead, the ratio between DSE and AMF colonization (ratio DSE/AMF) negatively related with proportion of fine roots. Our findings support the hypothesis regarding the importance of DSE at high altitudes and suggest a functional relationship between the rate of DSE/AMF and proportion of fine roots. The colonization by the root pathogen Olpidium sp. in C. quinoa deserves further study since this Andean grain is increasingly important for the local economy in these marginal areas.     

Molecular biodiversity of arbuscular mycorrhizal fungi in trace metal-polluted soils

We assessed the indigenous arbuscular mycorrhizal fungi (AMF) community structure from the roots and associated soil of Plantago major (plantain) plants growing on sites polluted with trace metals (TM) and on unpolluted sites. Uncontaminated and TM-contaminated sites containing As, Cd, Cu, Pb, Sn and Zn were selected based on a survey of the TM concentration in soils of community gardens in the City of Montréal. Total genomic DNA was extracted directly from these samples. PCR followed by denaturing gradient gel electrophoresis (PCR-DGGE), augmented by cloning and sequencing, as well as direct sequencing techniques, was all used to investigate AMF community structure. We found a decreased diversity of native AMF (assessed by the number of AMF ribotypes) in soils and plant roots harvested from TM-polluted soils compared with unpolluted soils. We also found that community structure was modified by TM contamination. Various species of Glomus, Scutellospora aurigloba and S. calospora were the most abundant ribotypes detected in unpolluted soil; ribotypes of G. etunicatum, G. irregulare/G. intraradices and G. viscosum were found in both polluted and unpolluted soils, while ribotypes of G. mosseae and Glomus spp. (B9 and B13) were dominant in TM-polluted soils. The predominance of G. mosseae in metal-polluted sites suggests the tolerance of this species to TM stress, as well as its potential use for phytoremediation. These data are relevant for our understanding of how AMF microbial communities respond to natural environments that contain a broad variety of toxic inorganic compounds and will substantially expand our knowledge of AMF ecology and biodiversity.