Arbuscular Mycorrhizal Fungal Networks Research Articles

Thresholds of Nitrogen and Phosphorus Input Substantially Alter Arbuscular Mycorrhizal Fungal Communities and Wheat Yield in Dryland Farmland.

Arbuscular mycorrhizal (AM) fungi are essential for preserving the multifunctionality of ecosystems. The nitrogen (N)/phosphorus (P) threshold that causes notable variations in the AM fungus community of the soil and plant productivity is still unclear. Herein, a long-term (18 years) field experiment with five N and five P fertilizer levels was conducted to investigate the change patterns of soil AM fungus, multifunctionality, and wheat yield. High-N and -P fertilizer inputs did not considerably increase the wheat yield. In the AM fungal network, a statistically significant positive correlation was observed between ecosystem multifunctionality and the biodiversity of two primary ecological clusters (N: Module #0 and P: Module #3). Furthermore, fertilizer input thresholds for N (92-160 kg ha-1) and P (78-100 kg ha-1) significantly altered the AM fungal community, soil characteristics, and plant productivity. Our study provided a basis for reduced N and P fertilizer application and sustainable agricultural development from the aspect of soil AM fungi.

Cd contamination determined assembly processes and network stability of AM fungal communities in an urban green space ecosystem

The effects of cadmium (Cd) contamination on the assembly mechanism and co-occurrence patterns of arbuscular mycorrhizal (AM) fungal communities remain unclear, especially in urban green spaces. This study sequenced AM fungal communities in greenbelt soils in Zhengzhou (China). The effects of Cd contamination on the AM fungal diversity, community assembly processes, and co-occurrence patterns were explored. We found that (1) an increase in Cd contamination changed the community composition, which resulted in a significant improvement in the diversity of specialists of AM fungi and a significant decrease in the diversity of generalists. (2) Deterministic processes dominated the community assembly of specialists and stochastic processes dominated the community assembly of generalists. (3) Specialists played a more important role than generalists in maintaining the stability of AM fungal networks under Cd contamination. Overall, Cd contamination affected the ecological processes of AM fungi in urban green space ecosystems. However, the effects on the assembly processes and network stability of different AM fungi taxa (specialists and generalists) differed significantly. The present study provides deeper insight into the effect of Cd contamination on the ecological processes of AMF and is helpful in further exploring the ecological risk of Cd contamination in urban green spaces.

Arbuscular mycorrhiza can be disadvantageous for weedy annuals in competition with paired perennial plants

Arbuscular mycorrhizal (AM) fungi can support the establishment of mycotrophic plants in new environments. However, the role of mycorrhizal symbiosis in interactions between perennial and weedy annual plants is not well understood. In our current study, we examine how widespread generalist AM fungi and soil disturbance, including disturbance of AM fungal networks (CMNs), affect the performance of two late-successional perennial plants of Central Europe, Senecio jacobaea and Crepis biennis, co-occurring with weedy annual forbs, Conyza canadensis and Erigeron annuus. Although presence of weedy annual E. annuus or C. canadensis did not affect the performance of the paired perennials, AM fungi supported perennial C. biennis in competition with weedy annual E. annuus. However, this AM-aided underpinning was independent of disturbance of CMNs. Conversely, although AM fungi benefited perennial S. jacobaea, this did not affect its competitive abilities when grown with weedy annual C. canadensis. Similarly, soil disturbance, independent of AM fungal presence, improved plant tissue P and biomass production of S. jacobaea, but not its competitive abilities. Our results show AM fungi may be advantageous for perennial plants growing in competition with weedy annual plants. Therefore, maintaining healthy soils containing an abundance of AM fungi, may encourage late successional perennial plants, potentially limiting establishment of weedy annual plant species.

Effects of Conventional and Organic Agriculture on Soil Arbuscular Mycorrhizal Fungal Community in Low-Quality Farmland.

Arbuscular mycorrhizal (AM) fungi have promising applications in low-quality farmlands all over the world, but research on their responses to conventional and organic farming systems in low-quality soil is limited. We hypothesized that the colonization activity and community diversity of AM fungi in conventional farming systems may not be lower than in organic farming on low-quality farmlands where beneficial symbiosis is required. We collected soil and maize root samples from medium to low fertility farmlands with conventional or organic farming systems in western Jilin Province, China. The colonization percentage and intensity, taxonomic and phylogenetic diversity, community composition of soil AM fungi, and soil factors were detected and compared between the two farming systems. The colonization intensity and operational taxonomic unit (OTU) taxonomic diversity on conventional farms were higher than on organic farms. Glomus was the most common genus on conventional farms, whereas Paraglomus and Glomus were the most common on organic farms. We also found a simpler AM fungal network structure with lower OTU phylogenetic diversity on conventional farms. Our findings suggested that though the conventional farming system resulted in different compositions and simpler structures of soil AM fungal community, there are potential diverse OTU resources currently present on conventional farms. This work has potential impacts on understanding the influence of different farming systems on soil AM fungi in low-quality farmlands and the development of efficient mycorrhizal inoculant production.

Impacts of biochar-based fertilization on soil arbuscular mycorrhizal fungal community structure in a karst mountainous area.

The application of biochar-based fertilizer can improve soil properties in part by stimulating microbial activity and growth. Karst ecosystems, which make up large areas of Southwest China, are prone to degradation. Understanding the response of arbuscular mycorrhizal fungal (AMF) community structure to biochar-based fertilizer application is of great significance to karst soil restoration. A field experiment was conducted in a typical karst soil (calcareous sandy loam) in Southwest China. A high-throughput sequencing approach was used to investigate the effect of biochar-based fertilization on AMF community structure in the karst soil. With the control (CK), compost with NPK fertilizer (MF), biochar (B), a lower amount of biochar with compost and NPK fertilizer (B1MF), biochar with compost and NPK fertilizer (BMF), and a higher amount of biochar with compost and NPK fertilizer (B4MF), the field trials were set up for 24 months. Soil amendments increased soil nutrient content and AMF diversity. The composition and structure of the AMF community varied among the treatments. AMF community composition was significantly impacted by soil chemical properties such as TC (total carbon), TN (total nitrogen), TP (total phosphorus), and AP (available phosphorus). Furthermore, network analysis showed that biochar-based fertilization increased the scale and complexity of the microbial co-occurrence network. Biochar-based fertilization enabled more keystone species (such as order Diversisporales and Glomerales) in the soil AMF network to participate in soil carbon resource management and soil nutrient cycling, indicating that biochar-based fertilizer is beneficial for the restoration of degraded karst soils.

A coumarin exudation pathway mitigates arbuscular mycorrhizal incompatibility in Arabidopsis thaliana.

Overexpression of genes involved in coumarin production and secretion can mitigate mycorrhizal incompatibility in nonhost Arabidopsis plants. The coumarin scopoletin, in particular, stimulates pre-penetration development and metabolism in mycorrhizal fungi. Although most plants can benefit from mutualistic associations with arbuscular mycorrhizal (AM) fungi, nonhost plant species such as the model Arabidopsis thaliana have acquired incompatibility. The transcriptional response of Arabidopsis to colonization by host-supported AM fungi switches from initial AM recognition to defense activation and plant growth antagonism. However, detailed functional information on incompatibility in nonhost-AM fungus interactions is largely missing. We studied interactions between host-sustained AM fungal networks of Rhizophagus irregularis and 18 Arabidopsis genotypes affected in nonhost penetration resistance, coumarin production and secretion, and defense (salicylic acid, jasmonic acid, and ethylene) and growth hormones (auxin, brassinosteroid, cytokinin, and gibberellin). We demonstrated that root-secreted coumarins can mitigate incompatibility by stimulating fungal metabolism and promoting initial steps of AM colonization. Moreover, we provide evidence that major molecular defenses in Arabidopsis do not operate as primary mechanisms of AM incompatibility nor of growth antagonism. Our study reveals that, although incompatible, nonhost plants can harbor hidden tools that promote initial steps of AM colonization. Moreover, it uncovered the coumarin scopoletin as a novel signal in the pre-penetration dialogue, with possible implications for the chemical communication in plant-mycorrhizal fungi associations.

Impacts of Urbanization Undermine Nestedness of the Plant-Arbuscular Mycorrhizal Fungal Network.

Cities are prone to ecological problems, yet the impacts of rapid global urbanization on the feedback between above- and belowground subsystems remain largely unknown. We sampled the roots of 8 common herbaceous plants within the Fifth Ring (urban areas) and in Jiufeng National Forest Park (rural areas) in Beijing (China) to assess the impacts of urbanization on the network of plant-arbuscular mycorrhizal (AM) fungal associations. Using Illumina MiSeq sequencing, 81 AM fungal OTUs were identified in 78 herb root samples. The Shannon, Simpson, and Pielou indices of root AM fungi in urban areas were significantly higher than those in rural areas. In this study, a significantly nested mycorrhizal association network was observed in rural areas (NODF = 64.68), whereas a non-nested pattern was observed in urban areas (NODF = 55.50). The competition index C-score (0.0769) of AM fungi in urban areas was slightly lower than that in rural areas (0.1431), and the species specialization (d’) of 8 host plants and fungal dissimilarity among 8 host plants in urban areas were significantly lower than those in rural areas. Convergent associations among hosts may be an important factor influencing this non-nested pattern of the plant-AM fungi network in urban areas. Generalists, rather than specialists, were enhanced during the establishment of mycorrhizal associations in urban areas. Our results suggest that reduced selectivity of host plants, and generalist promotion and specialist reduction of AM fungi during urbanization may contribute to the non-nested network of plant-AM fungal associations.

Soil fungal networks moderate density-dependent survival and growth of seedlings.

Pathogenic and mutualistic fungi have contrasting effects on seedling establishment, but it remains unclear whether density-dependent survival and growth are regulated by access to different types of mycorrhizal fungal networks supported by neighbouring adult trees. Here, we conducted an extensive field survey to test how mycorrhizal and pathogenic fungal colonization of arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) seedlings in a subtropical forest respond to density of neighbouring adult trees. In addition, we undertook a hyphal exclusion experiment to explicitly test the role of soil fungal networks in driving density-dependent effects on seedling growth and survival. Conspecific adult density was a strong predictor for the relative abundance of putative pathogens, which was greater in roots of AM than of ECM seedlings, while mycorrhizal fungal abundance and colonization were not consistently affected by conspecific adult density. Both ECM and AM fungal networks counteracted conspecific density-dependent mortality, but ECM fungi were more effective at weakening the negative effects of high seedling density than AM fungi. Our findings reveal a critical role of common fungal networks in mitigating negative density-dependent effects of pathogenic fungi on seedling establishment, which provides mechanistic insights into how soil fungal diversity shapes plant community structure in subtropical forests.

Seasonal variation of diversity and co-occurrence patterns of arbuscular mycorrhizal fungal communities in mixed broadleaf-conifer forests

Arbuscular mycorrhizal (AM) fungi are an important determinant of plant performance. The temporal distribution patterns of AM fungal communities, important groups in forest soil, are affected by various environmental factors. However, the effect of seasonal variation on AM fungal communities in forest soil is still not fully understood. To improve knowledge regarding the seasonal dynamics of AM fungal communities in forest soil, the Juglans mandshurica rhizosphere soil in a 30-year-old natural J. mandshurica and Larix gmelinii mixed forest in northeastern China was sampled and studied during different seasons. AM fungal community composition and structure in the rhizosphere soil were determined by using the Illumina MiSeq sequencing platform. The results showed that forest type significantly affected soil pH and the microbial biomass carbon (Cmic), microbial biomass nitrogen (Nmic), C, N, P, NH4+-N and NO3−-N contents. Similarly, soil moisture significantly decreased with seasonal progression in all forest types. Glomus and Diversispora were the dominant genera. The Shannon, Sobs and Chao1 indices of the mixed forest were higher in July than in May and September. The NO3−-N and water-soluble phenol (WSPh) contents were the most important factors driving the shift in the soil AM fungal community across the seasonal gradient. Network analysis revealed that AM fungal network structure was significantly different between seasons, and the number of links between species significantly increased with time. We observed a clear tendency for mixed forest management to significantly affect AM fungal community structure and composition but not diversity. Furthermore, the co-occurrence network and keystone taxa among AM fungi can change in response to environmental filtering due to mixed management in different seasons.

Dead Rhizophagus irregularis biomass mysteriously stimulates plant growth.

Arbuscular mycorrhizal (AM) fungi establish symbiotic associations with many plant species, transferring significant amounts of soil nutrients such as phosphorus to plants and receiving photosynthetically fixed carbon in return. Functioning of AM symbiosis is thus based on interaction between two living partners. The importance of dead AM fungal biomass (necromass) in ecosystem processes remains unclear. Here, we applied either living biomass or necromass (0.0004 potting substrate weight percent) of monoxenically produced AM fungus (Rhizophagus irregularis) into previously sterilized potting substrate planted with Andropogon gerardii. Plant biomass production significantly improved in both treatments as compared to non-amended controls. Living AM fungus, in contrast to the necromass, specifically improved plant acquisition of nutrients normally supplied to the plants by AM fungal networks, such as phosphorus and zinc. There was, however, no difference between the two amendment treatments with respect to plant uptake of other nutrients such as nitrogen and/or magnesium, indicating that the effect on plants of the AM fungal necromass was not primarily nutritional. Plant growth stimulation by the necromass could thus be either due to AM fungal metabolites directly affecting the plants, indirectly due to changes in soil/root microbiomes or due to physicochemical modifications of the potting substrate. In the necromass, we identified several potentially bioactive molecules. We also provide experimental evidence for significant differences in underground microbiomes depending on the amendment with living or dead AM fungal biomass. This research thus provides the first glimpse into possible mechanisms responsible for observed plant growth stimulation by the AM fungal necromass.

Interactions between arbuscular mycorrhizal fungi and non-host Carex capillacea.

A topic of confusion over the interactions between arbuscular mycorrhizal (AM) fungi and plants is the mycorrhizal status of some plant families such as Cyperaceae, which is generally considered to be non-mycorrhizal. Here, we conducted experiments to explore how the abiotic environmental conditions and AM network influence the interactions between AM fungi and Carex capillacea. We grew Carex capillacea alone or together with a mycorrhizal host species Medicago sativa in the presence or absence of AM fungi (soil inoculum from Mount Segrila and Rhizophagus intraradices from the Chinese Bank of the Glomeromycota, BGC). Plants were grown in a growth chamber and at two elevational sites of Mount Segrila, respectively. The results indicate that mycorrhizal host plants ensured the presence of an active AM fungal network whether under growth chamber or alpine conditions. The AM fungal network significantly depressed the growth of C. capillacea, especially when native inocula were used and the plants grew under alpine site conditions, although root colonization of C. capillacea increased in most cases. Moreover, the colonization level of C. capillacea was much higher (≤ 30%) when growing under alpine conditions compared with growth chamber conditions (< 8.5%). Up to 20% root colonization by Rhizophagus intraradices was observed in monocultures under alpine conditions. A significant negative relationship was found between shoot phosphorus concentrations in M. sativa and shoot dry mass of C. capillacea. These results indicate that growing conditions, AM network, and inoculum source are all important factors affecting the susceptibility of C. capillacea to AM fungi, and growing conditions might be a key driver of the interactions between AM fungi and C. capillacea.

Phylogenetic, structural, and functional characterization of AMT3;1, an ammonium transporter induced by mycorrhization among model grasses.

In the arbuscular mycorrhizal (AM) symbiosis, plants satisfy part of their nitrogen (N) requirement through the AM pathway. In sorghum, the ammonium transporters (AMT) AMT3;1, and to a lesser extent AMT4, are induced in cells containing developing arbuscules. Here, we have characterized orthologs of AMT3;1 and AMT4 in four other grasses in addition to sorghum. AMT3;1 and AMT4 orthologous genes are induced in AM roots, suggesting that in the common ancestor of these five plant species, both AMT3;1 and AMT4 were already present and upregulated upon AM colonization. An artificial microRNA approach was successfully used to downregulate either AMT3;1 or AMT4 in rice. Mycorrhizal root colonization and hyphal length density of knockdown plants were not affected at that time, indicating that the manipulation did not modify the establishment of the AM symbiosis and the interaction between both partners. However, expression of the fungal phosphate transporter FmPT was significantly reduced in knockdown plants, indicating a reduction of the nutrient fluxes from the AM fungus to the plant. The AMT3;1 knockdown plants (but not the AMT4 knockdown plants) were significantly less stimulated in growth by AM fungal colonization, and uptake of both 15N and 33P from the AM fungal network was reduced. This confirms that N and phosphorus nutrition through the mycorrhizal pathway are closely linked. But most importantly, it indicates that AMT3;1 is the prime plant transporter involved in the mycorrhizal ammonium transfer and that its function during uptake of N cannot be performed by AMT4.

Constraining the role of early land plants in Palaeozoic weathering and global cooling.

How the colonization of terrestrial environments by early land plants over 400 Ma influenced rock weathering, the biogeochemical cycling of carbon and phosphorus, and climate in the Palaeozoic is uncertain. Here we show experimentally that mineral weathering by liverworts—an extant lineage of early land plants—partnering arbuscular mycorrhizal (AM) fungi, like those in 410 Ma-old early land plant fossils, amplified calcium weathering from basalt grains threefold to sevenfold, relative to plant-free controls. Phosphate weathering by mycorrhizal liverworts was amplified 9–13-fold over plant-free controls, compared with fivefold to sevenfold amplification by liverworts lacking fungal symbionts. Etching and trenching of phyllosilicate minerals increased with AM fungal network size and atmospheric CO2 concentration. Integration of grain-scale weathering rates over the depths of liverwort rhizoids and mycelia (0.1 m), or tree roots and mycelia (0.75 m), indicate early land plants with shallow anchorage systems were probably at least 10-fold less effective at enhancing the total weathering flux than later-evolving trees. This work challenges the suggestion that early land plants significantly enhanced total weathering and land-to-ocean fluxes of calcium and phosphorus, which have been proposed as a trigger for transient dramatic atmospheric CO2 sequestration and glaciations in the Ordovician.

Arbuscular mycorrhizal fungi reduce growth and infect roots of the non‐host plant Arabidopsis thaliana

The arbuscular mycorrhizal (AM) symbiosis is widespread throughout the plant kingdom and important for plant nutrition and ecosystem functioning. Nonetheless, most terrestrial ecosystems also contain a considerable number of non-mycorrhizal plants. The interaction of such non-host plants with AM fungi (AMF) is still poorly understood. Here, in three complementary experiments, we investigated whether the non-mycorrhizal plant Arabidopsis thaliana, the model organism for plant molecular biology and genetics, interacts with AMF. We grew A. thaliana alone or together with a mycorrhizal host species (either Trifolium pratense or Lolium multiflorum) in the presence or absence of the AMF Rhizophagus irregularis. Plants were grown in a dual-compartment system with a hyphal mesh separating roots of A. thaliana from roots of the host species, avoiding direct root competition. The host plants in the system ensured the presence of an active AM fungal network. AM fungal networks caused growth depressions in A. thaliana of more than 50% which were not observed in the absence of host plants. Microscopy analyses revealed that R. irregularis supported by a host plant was capable of infecting A. thaliana root tissues (up to 43% of root length colonized), but no arbuscules were observed. The results reveal high susceptibility of A. thaliana to R. irregularis, suggesting that A. thaliana is a suitable model plant to study non-host/AMF interactions and the biological basis of AM incompatibility.

Absence of carbon transfer between Medicago truncatula plants linked by a mycorrhizal network, demonstrated in an experimental microcosm

Carbon transfer between plants via a common extraradical network of arbuscular mycorrhizal (AM) fungal hyphae has been investigated abundantly, but the results remain equivocal. We studied the transfer of carbon through this fungal network, from a Medicago truncatula donor plant to a receiver (1) M. truncatula plant growing under decreased light conditions and (2) M. truncatula seedling. Autotrophic plants were grown in bicompartmented Petri plates, with their root systems physically separated, but linked by the extraradical network of Glomus intraradices. A control Myc-/Nod- M. truncatula plant was inserted in the same compartment as the receiver plant. Following labeling of the donor plant with 13CO2, 13C was recovered in the donor plant shoots and roots, in the extraradical mycelium and in the receiver plant roots. Fatty acid analysis of the receiver's roots further demonstrated 13C enrichment in the fungal-specific lipids, while almost no label was detected in the plant-specific compounds. We conclude that carbon was transferred from the donor to the receiver plant via the AM fungal network, but that the transferred carbon remained within the intraradical AM fungal structures of the receiver's root and was not transferred to the receiver's plant tissues.

Common mycorrhizal networks provide a potential pathway for the transfer of hydraulically lifted water between plants

Plant roots may be linked by shared or common mycorrhizal networks (CMNs) that constitute pathways for the transfer of resources among plants. The potential for water transfer by such networks was examined by manipulating CMNs independently of plant roots in order to isolate the role(s) of ectomycorrhizal (EM) and arbuscular mycorrhizal fungal (AMF) networks in the plant water balance during drought (soil water potential -5.9 MPa). Fluorescent tracer dyes and deuterium-enriched water were used to follow the pathways of water transfer from coastal live oak seedlings (Quercus agrifolia Nee; colonized by EM and AMF) conducting hydraulic lift (HL) into the roots of water-stressed seedlings connected only by EM (Q. agrifolia) or AMF networks (Q. agrifolia, Eriogonum fasciculatum Benth., Salvia mellifera Greene, Keckiella antirrhinoides Benth). When connected to donor plants by hyphal linkages, deuterium was detected in the transpiration flux of receiver oak plants, and dye-labelled extraradical hyphae, rhizomorphs, mantles, and Hartig nets were observed in receiver EM oak roots, and in AMF hyphae of Salvia. Hyphal labelling was scarce in Eriogonum and Keckiella since these species are less dependent on AMF. The observed patterns of dye distribution also indicated that only a small percentage of mycorrhizal roots and extraradical hyphae were involved with water transfer among plants. Our results suggest that the movement of water by CMNs is potentially important to plant survival during drought, and that the functional ecophysiological traits of individual mycorrhizal fungi may be a component of this mechanism.

The fungus does not transfer carbon to or between roots in an arbuscular mycorrhizal symbiosis.

• Carbon transfer from fungus to plant in the arbuscular mycorrhizal (AM) symbiosis has been reported, but its significance and even its existence have been called into question and the issue remains controversial. We investigated carbon movement from fungus to plant and from one mycorrhizal root system to another via a common AM fungal network in monoxenic cultures to avoid limitations of some previous studies. • 13 C and 14 C labeled substrates were supplied to functioning in vitro AM mycorrhizas between Ri T-DNA transformed carrot (Daucus carota) roots and Glomus intraradices to follow carbon movement into and between host and fungal metabolite pools. • Fungal triacylglycerol and trehalose were labeled when permeant substrates were supplied to the extraradical mycelium (ERM), but host-specific compounds in the roots did not become labeled. When labeled glucose was provided to a donor root system, label moved to recipient roots via a common AM fungal network but remained in fungal compounds. • We conclude that carbon flow in the AM symbiosis is normally unidirectional from plant to fungus and that while carbon is translocated by the fungus from one metabolically active root system to another, it remains within the intraradical mycelium (IRM).