Common Mycorrhizal Networks Research Articles

Regulation of plants' phosphate uptake in common mycorrhizal networks: Role of intraradical fungal phosphate transporters

ABSTRACTWe have recently identified two genes coding for inorganic phosphate transporters (Pht) in sorghum (Sorghum bicolor) and flax (Linum usitatissimum) that were induced in roots colonized by arbuscular mycorrhizal (AM) fungi. Mycorrhizal acquisition of inorganic phosphorus (Pi) was strongly affected by the combination of plant and AM fungal species, but the expression level of these genes coding for AM-inducible Pi transporters did not explain differences in plant phosphorus acquisition where flax and sorghum are sharing a common mycorrhizal network. In the present study, we investigated the possible role of fungal Pi transporters in the regulation of mycorrhizal Pi acquisition by measuring their expression in roots of flax and sorghum. One Pi transporter of Rhizophagus irregularis (RiPT5) showed a positive correlation with mycorrhizal Pi acquisition of sorghum. This indicates that a possible involvement in the regulation of mycorrhizal Pi acquisition. In general, expression of AMF Pi transporters was more related to mycorrhizal Pi acquisition of sorghum than of flax, indicating plant species-specific differences in the regulation of mycorrhizal Pi acquisition.

Common mycorrhizal networks and their effect on the bargaining power of the fungal partner in the arbuscular mycorrhizal symbiosis

ABSTRACTArbuscular mycorrhizal (AM) fungi form mutualistic interactions with the majority of land plants, including some of the most important crop species. The fungus takes up nutrients from the soil, and transfers these nutrients to the mycorrhizal interface in the root, where these nutrients are exchanged against carbon from the host. AM fungi form extensive hyphal networks in the soil and connect with their network multiple host plants. These common mycorrhizal networks (CMNs) play a critical role in the long-distance transport of nutrients through soil ecosystems and allow the exchange of signals between the interconnected plants. CMNs affect the survival, fitness, and competitiveness of the fungal and plant species that interact via these networks, but how the resource transport within these CMNs is controlled is largely unknown. We discuss the significance of CMNs for plant communities and for the bargaining power of the fungal partner in the AM symbiosis.

Faster acquisition of symbiotic partner by common mycorrhizal networks in early plant life stage

AbstractArbuscular mycorrhizal (AM) fungi usually improve plant performance yet our knowledge about their effects on seed germination and early plant establishment is very limited. We performed a factorial greenhouse experiment where the seeds from four low Arctic co‐occurring mycorrhizal herbs (Antennaria dioica, Campanula rotundifolia, Sibbaldia procumbens, and Solidago virgaurea) were germinated alone or in the vicinity of an adult Sibbaldia plant with or without AM fungi; given either as spores or being present in a common mycorrhizal network (CMN). Three different AM fungal species were examined to assess species‐specific differences in symbiont acquisition rate. Of the four plant species investigated, the presence of AM fungi affected seed germination only in Campanula and this effect was dependent on whether the AM fungi were present in the soil as spores or as a CMN. Overall, after germination, developing seedlings showed AM fungal colonization in their roots as soon as 2 d after cotyledon emergence. Our results show that CMN may provide germinating seedlings faster acquisition of the AM fungal partner in comparison to acquisition from spores. Furthermore, there were AM species‐specific differences in the symbiont acquisition rate highlighting the importance of species identity in AM interactions. These findings suggest that while AM fungi may not play a fundamental role during seed germination, plant community composition may be affected by the species‐specific AM fungal effects on seedling establishment and CMN acquisition.

Effects of Common Mycorrhizal Network on Plant Carbohydrates and Soil Properties in Trifoliate Orange-White Clover Association.

Common mycorrhizal network (CMN) allows nutrients and signals to pass between two or more plants. In this study, trifoliate orange (Poncirus trifoliata) and white clover (Trifolium repens) were planted in a two-compartmented rootbox, separated by a 37–μm nylon mesh and then inoculated with an arbuscular mycorrhizal fungus (AMF), Diversispora spurca. Inoculation with D. spurca resulted in formation of a CMN between trifoliate orange and white clover, whilst the best AM colonization occurred in the donor trifoliate orange–receptor white clover association. In the trifoliate orange–white clover association, the mycorrhizal colonization of receptor plant by extraradical hyphae originated from the donor plant significantly increased shoot and root fresh weight and chlorophyll concentration of the receptor plant. Enzymatic activity of soil β-glucoside hydrolase, protease, acid and neutral phosphatase, water-stable aggregate percentage at 2–4 and 0.5–1 mm size, and mean weight diameter in the rhizosphere of the receptor plant also increased. The hyphae of CMN released more easily-extractable glomalin-related soil protein and total glomalin-related soil protein into the receptor rhizosphere, which represented a significantly positive correlation with aggregate stability. AMF inoculation exhibited diverse changes in leaf and root sucrose concentration in the donor plant, and AM colonization by CMN conferred a significant increase of root glucose in the receptor plant. These results suggested that CMN formed in the trifoliate orange–white clover association, and root AM colonization by CMN promoted plant growth, root glucose accumulation, and rhizospheric soil properties in the receptor plant.

Plant identity and density can influence arbuscular mycorrhizal fungi colonization, plant growth, and reproduction investment in coculture

Reciprocal effects between arbuscular mycorrhizal fungi (AMF) and plant communities are essential to study the complexity of interactions in a grassland ecosystem. Here, we investigated the effects of plant community density and composition on AMF colonization, plant growth, and reproduction investment. We developed an experimental system with three compartments, each containing either three or six Medicago truncatula Gaertn. plants, or three M. truncatula plants associated with three Silene vulgaris (Moench) Garcke plants. All three compartments shared the same common mycorrhizal network built either by Rhizophagus irregularis MUCL 43194, by Rhizophagus clarus MUCL 46238, or by both AMF in association grown in a central compartment on Plantago lanceolata L. Our results demonstrate an absence of effect of plant density but a positive influence of mixed cultures on AMF root colonization compared with monocultures. This higher AMF development resulted in a positive feedback on shoot biomass and number of flowers and fruits produced by M. truncatula. Although both fungal strains were present in root systems, co-inoculation did not generate a synergistic effect on plant development. These results highlight the importance of plant associations on AMF dynamics, which requires further investigation at the community scale to improve our understanding of the intricate AMF – host plant relationships.

The potential of Dark Septate Endophytes to form root symbioses with ectomycorrhizal and ericoid mycorrhizal middle European forest plants.

The unresolved ecophysiological significance of Dark Septate Endophytes (DSE) may be in part due to existence of morphologically indistinguishable cryptic species in the most common Phialocephala fortinii s. l.—Acephala applanata species complex (PAC). We inoculated three middle European forest plants (European blueberry, Norway spruce and silver birch) with 16 strains of eight PAC cryptic species and other DSE and ectomycorrhizal/ericoid mycorrhizal fungi and focused on intraradical structures possibly representing interfaces for plant-fungus nutrient transfer and on host growth response. The PAC species Acephala applanata simultaneously formed structures resembling ericoid mycorrhiza (ErM) and DSE microsclerotia in blueberry. A. macrosclerotiorum, a close relative to PAC, formed ectomycorrhizae with spruce but not with birch, and structures resembling ErM in blueberry. Phialocephala glacialis, another close relative to PAC, formed structures resembling ErM in blueberry. In blueberry, six PAC strains significantly decreased dry shoot biomass compared to ErM control. In birch, one A. macrosclerotiorum strain increased root biomass and the other shoot biomass in comparison with non-inoculated control. The dual mycorrhizal ability of A. macrosclerotiorum suggested that it may form mycorrhizal links between Ericaceae and Pinaceae. However, we were unable to detect this species in Ericaceae roots growing in a forest with presence of A. macrosclerotiorum ectomycorrhizae. Nevertheless, the diversity of Ericaceae mycobionts was high (380 OTUs) with individual sites often dominated by hitherto unreported helotialean and chaetothyrialean/verrucarialean species; in contrast, typical ErM fungi were either absent or low in abundance. Some DSE apparently have a potential to form mycorrhizae with typical middle European forest plants. However, except A. applanata, the tested representatives of all hitherto described PAC cryptic species formed typical DSE colonization without specific structures necessary for mycorrhizal nutrient transport. A. macrosclerotiorum forms ectomycorrhiza with conifers but not with broadleaves and probably does not form common mycorrhizal networks between conifers with Ericaceae.

Ectomycorrhizal fungal communities of Coccoloba uvifera (L.) L. mature trees and seedlings in the neotropical coastal forests of Guadeloupe (Lesser Antilles).

We studied belowground and aboveground diversity and distribution of ectomycorrhizal (EM) fungal species colonizing Coccoloba uvifera (L.) L. (seagrape) mature trees and seedlings naturally regenerating in four littoral forests of the Guadeloupe island (Lesser Antilles). We collected 546 sporocarps, 49 sclerotia, and morphotyped 26,722 root tips from mature trees and seedlings. Seven EM fungal species only were recovered among sporocarps (Cantharellus cinnabarinus, Amanita arenicola, Russula cremeolilacina, Inocybe littoralis, Inocybe xerophytica, Melanogaster sp., and Scleroderma bermudense) and one EM fungal species from sclerotia (Cenococcum geophilum). After internal transcribed spacer (ITS) sequencing, the EM root tips fell into 15 EM fungal taxa including 14 basidiomycetes and 1 ascomycete identified. Sporocarp survey only weakly reflected belowground assessment of the EM fungal community, although 5 fruiting species were found on roots. Seagrape seedlings and mature trees had very similar communities of EM fungi, dominated by S. bermudense, R. cremeolilacina, and two Thelephoraceae: shared species represented 93 % of the taxonomic EM fungal diversity and 74 % of the sampled EM root tips. Furthermore, some significant differences were observed between the frequencies of EM fungal taxa on mature trees and seedlings. The EM fungal community composition also varied between the four investigated sites. We discuss the reasons for such a species-poor community and the possible role of common mycorrhizal networks linking seagrape seedlings and mature trees in regeneration of coastal forests.

Plant phosphorus acquisition in a common mycorrhizal network: regulation of phosphate transporter genes of the Pht1 family in sorghum and flax.

In a preceding microcosm study, we found huge differences in phosphorus (P) acquisition in sorghum (Sorghum bicolor) and flax (Linum usitatissimum) sharing a common mycorrhizal network (CMN). Is the transcriptional regulation of arbuscular mycorrhizal (AM)-induced inorganic orthophosphate (Pi) transporters responsible for these differences? We characterized and analyzed the expression of Pi transporters of the Pht1 family in both plant species, and identified two new AM-inducible Pi transporters in flax. Mycorrhizal Pi acquisition was strongly affected by the combination of plant and AM fungal species. A corresponding change in the expression of two AM-inducible Pht1 transporters was noticed in both plants (SbPT9, SbPT10, LuPT5 and LuPT8), but the effect was very weak. Overall, the expression level of these genes did not explain why flax took up more Pi from the CMN than did sorghum. The post-transcriptional regulation of the transporters and their biochemical properties may be more important for their function than the fine-tuning of their gene expression.

Fungal nutrient allocation in common mycorrhizal networks is regulated by the carbon source strength of individual host plants.

Common mycorrhizal networks (CMNs) of arbuscular mycorrhizal (AM) fungi in the soil simultaneously provide multiple host plants with nutrients, but the mechanisms by which the nutrient transport to individual host plants within one CMN is controlled are unknown. Using radioactive and stable isotopes, we followed the transport of phosphorus (P) and nitrogen (N) in the CMNs of two fungal species to plants that differed in their carbon (C) source strength, and correlated the transport to the expression of mycorrhiza-inducible plant P (MtPt4) and ammonium (1723.m00046) transporters in mycorrhizal roots. AM fungi discriminated between host plants that shared a CMN and preferentially allocated nutrients to high-quality (nonshaded) hosts. However, the fungus also supplied low-quality (shaded) hosts with nutrients and maintained a high colonization rate in these plants. Fungal P transport was correlated to the expression of MtPt4. The expression of the putative ammonium transporter 1723.m00046 was dependent on the fungal nutrient supply and was induced when the CMN had access to N. Biological market theory has emerged as a tool with which the strategic investment of competing partners in trading networks can be studied. Our work demonstrates how fungal partners are able to retain bargaining power, despite being obligately dependent on their hosts.

Hijacking common mycorrhizal networks for herbivore-induced defence signal transfer between tomato plants

Common mycorrhizal networks (CMNs) link multiple plants together. We hypothesized that CMNs can serve as an underground conduit for transferring herbivore-induced defence signals. We established CMN between two tomato plants in pots with mycorrhizal fungus Funneliformis mosseae, challenged a ‘donor' plant with caterpillar Spodoptera litura, and investigated defence responses and insect resistance in neighbouring CMN-connected ‘receiver' plants. After CMN establishment caterpillar infestation on ‘donor' plant led to increased insect resistance and activities of putative defensive enzymes, induction of defence-related genes and activation of jasmonate (JA) pathway in the ‘receiver' plant. However, use of a JA biosynthesis defective mutant spr2 as ‘donor' plants resulted in no induction of defence responses and no change in insect resistance in ‘receiver' plants, suggesting that JA signalling is required for CMN-mediated interplant communication. These results indicate that plants are able to hijack CMNs for herbivore-induced defence signal transfer and interplant defence communication.

丛枝菌根真菌在土壤氮素循环中的作用

作为植物需求量最大的营养元素，氮素是陆地生态系统初级生产力的主要限制因子。丛枝菌根真菌能与地球上80%以上的陆生植物形成菌根共生体，帮助宿主植物吸收土壤中的P、N等矿质养分。目前，丛枝菌根真菌与氮素循环相关研究侧重于真菌对氮素的吸收形态以及共生体中氮的传输代谢机制，却忽略了丛枝菌根真菌在固氮过程、矿化与吸收过程、硝化过程、反硝化过程以及氮素淋洗过程等土壤氮素循环过程中所起到的潜在作用，并且越来越多的证据也表明丛枝菌根真菌是影响土壤氮素循环过程的重要因子。总结了丛枝菌根真菌可利用的氮素形态及真菌的氮代谢转运相关基因的研究现状；重点分析了丛枝菌根真菌在调控土壤氮素循环过程中的潜在作用以及在生态系统中的重要生态学意义，同时提出了丛枝菌根真菌在土壤氮素循环过程中一些需要深入研究的问题。

N2-fixing trees and the transfer of fixed-N for sustainable agroforestry: a review

Many tropical areas lack soil nitrogen (N), an essential nutrient for plant growth and the production of food. Commercial N fertilisers are expensive, with only a fraction of this nutrient reaching the plant, which limits efficiency and potentially increases water contamination. Dinitrogen (N2)-fixing trees are a promising alternative to sustainably fertilise crops. The use of N2-fixing trees in tropical agriculture has garnered attention from researchers, development organisations, governments, and farmers in recent years as a revival of pro-environmental practices. Dinitrogen (N2)-fixing trees can establish in N-deficient soils, replace N lost in harvest and provide an as-of-yet not fully realised benefit to ecosystem services. High N2-fixation rates, upwards of 92 %, have been measured in some N2-fixing trees, using the 15N natural abundance method. The recovery of this fixed-N by associated perennial crops is of particular interest in tropical agroforestry systems. Here, we review N transfer pathways from trees to perennial crops in agroforestry. We focus on Theobroma cacao and Coffea arabica. We also draw on agroforestry systems with herbaceous alleys. We identify three pathways of N transfer from N2-fixers to non-N2-fixing crops: (1) decomposition and mineralisation of organic compounds, e.g., litter, prunings, roots, and nodules, (2) root-to-root direct transfer via exudation, and (3) common mycorrhizal networks. Both 15N natural abundance and 15N enrichment techniques have been used to study N transfer. However, various factors limit the accuracy of estimates within agroforestry systems. Under field conditions, the major limits are (1) improper reference selection and (2) unrepresentative sampling of the receiver plant and/or donor N source. We highlight key findings and provide recommendations to tackle these obstacles.

土壌共生菌がつくりだす 森林の多様性： 大規模移入操作実験と最先端分子同定技術の融合

土壌共生菌がつくりだす 森林の多様性： 大規模移入操作実験と最先端分子同定技術の融合

Common arbuscular mycorrhizal networks amplify competition for phosphorus between seedlings and established plants

Common mycorrhizal networks (CMNs) influence competition between plants, but reports regarding their precise effect are conflicting. We studied CMN effects on phosphorus (P) uptake and growth of seedlings as influenced by various disruptions of network components. Tomato (Solanum lycopersicon) seedlings grew into established networks of Rhizophagus irregularis and cucumber (Cucumis sativus) in two experiments. One experiment studied seedling uptake of (32)P in the network in response to cutting of cucumber shoots; the other analysed seedling uptake of P and nitrogen (N) in the presence of intact or severed arbuscular mycorrhizal fungus networks and at two soil P concentrations. Pre-established and intact networks suppressed growth of tomato seedlings. Cutting of cucumber shoots mitigated P deficiency symptoms of seedlings, which obtained access to P in the extraradical mycelium and thereby showed improved growth. Solitary seedlings growing in a network patch that had been severed from the CMN also grew much better than seedlings of the corresponding CMN. Interspecific and size-asymmetric competition between plants may be amplified rather than relaxed by CMNs that transfer P to large plants providing most carbon and render small plants P deficient. It is likely that grazing or senescence of the large plants will alleviate the network-induced suppression of seedling growth.

Mycorrhiza for all: an under‐earth revolution

Mycorrhiza for all: an under‐earth revolution

Arbuscular-mycorrhizal networks inhibit Eucalyptus tetrodonta seedlings in rain forest soil microcosms.

Eucalyptus tetrodonta, a co-dominant tree species of tropical, northern Australian savannas, does not invade adjacent monsoon rain forest unless the forest is burnt intensely. Such facilitation by fire of seedling establishment is known as the "ashbed effect." Because the ashbed effect might involve disruption of common mycorrhizal networks, we hypothesized that in the absence of fire, intact rain forest arbuscular mycorrhizal (AM) networks inhibit E. tetrodonta seedlings. Although arbuscular mycorrhizas predominate in the rain forest, common tree species of the northern Australian savannas (including adult E. tetrodonta) host ectomycorrhizas. To test our hypothesis, we grew E. tetrodonta and Ceiba pentandra (an AM-responsive species used to confirm treatments) separately in microcosms of ambient or methyl-bromide fumigated rain forest soil with or without severing potential mycorrhizal fungus connections to an AM nurse plant, Litsea glutinosa. As expected, C. pentandra formed mycorrhizas in all treatments but had the most root colonization and grew fastest in ambient soil. E. tetrodonta seedlings also formed AM in all treatments, but severing hyphae in fumigated soil produced the least colonization and the best growth. Three of ten E. tetrodonta seedlings in ambient soil with intact network hyphae died. Because foliar chlorosis was symptomatic of iron deficiency, after 130 days we began to fertilize half the E. tetrodonta seedlings in ambient soil with an iron solution. Iron fertilization completely remedied chlorosis and stimulated leaf growth. Our microcosm results suggest that in intact rain forest, common AM networks mediate belowground competition and AM fungi may exacerbate iron deficiency, thereby enhancing resistance to E. tetrodonta invasion. Common AM networks–previously unrecognized as contributors to the ashbed effect–probably help to maintain the rain forest–savanna boundary.

Common mycorrhizal networks amplify size inequality in Andropogon gerardii monocultures

Arbuscular mycorrhizal fungi can interconnect plant root systems through hyphal common mycorrhizal networks, which may influence the distribution of limiting mineral nutrients among interconnected individuals, potentially affecting competition and consequent size inequality. Using a microcosm model system, we investigated whether the members of Andropogon gerardii monocultures compete via common mycorrhizal networks. We grew A. gerardii seedlings with isolated root systems in individual, adjacent containers while preventing, disrupting or allowing common mycorrhizal networks among them. Fertile soil was placed within the containers, which were embedded within infertile sand. We assessed mycorrhizas, leaf tissue mineral nutrient concentrations, size hierarchies and the growth of nearest neighbors. Plants interconnected by common mycorrhizal networks had 8% greater colonized root length, 12% higher phosphorus and 35% higher manganese concentrations than plants severed from common mycorrhizal networks. Interconnected plants were, on average, 15% larger and had 32% greater size inequality, as reflected by Gini coefficients, than those with severed connections. Only with intact common mycorrhizal networks were whole-plant dry weights negatively associated with those of their neighbors. In the absence of root system overlap, common mycorrhizal networks likely promote asymmetric competition below ground, thereby exaggerating size inequality within A. gerardii populations.

Using common mycorrhizal networks for controlled inoculation of Quercus spp. with Tuber melanosporum: the nurse plant method

The high cost and restricted availability of black truffle spore inoculum for controlled mycorrhiza formation of host trees produced for truffle orchards worldwide encourage the search for more efficient and sustainable inoculation methods that can be applied globally. In this study, we evaluated the potential of the nurse plant method for the controlled inoculation of Quercus cerris and Quercus robur with Tuber melanosporum by mycorrhizal networks in pot cultures. Pine bark compost, adjusted to pH 7.8 by liming, was used as substrate for all assays. Initially, Q. robur seedlings were inoculated with truffle spores and cultured for 12months. After this period, the plants presenting 74% mycorrhizal fine roots were transferred to larger containers. Nurse plants were used for two treatments of two different nursling species: five sterilized acorns or five 45-day-old, axenically grown Q. robur or Q. cerris seedlings, planted in containers around the nurse plant. After 6months, colonized nursling plant root tips showed that mycorrhiza formation by T. melanosporum was higher than 45% in the seedlings tested, with the most successful nursling combination being Q. cerris seedlings, reaching 81% colonization. Bulk identification of T. melanosporum mycorrhizae was based on morphological and anatomical features and confirmed by sequencing of the internal transcribed spacer region of the ribosomal DNA of selected root tips. Our results show that the nurse plant method yields attractive rates of mycorrhiza formation by the Périgord black truffle and suggest that establishing and maintaining common mycorrhizal networks in pot cultures enables sustained use of the initial spore inoculum.

Tracking the carbon source of arbuscular mycorrhizal fungi colonizing C3 and C4 plants using carbon isotope ratios (δ13C)

Arbuscular mycorrhizal fungi (AMF) may colonize several plant species simultaneously, thus receiving their carbon from different plants. In previous work, we have used microcosms with flax (a C3 plant) and sorghum (a C4 plant), connected to a common mycorrhizal network, in order to track the carbon source of AMF, making use of the distinct 13C/12C isotope compositions of C3 and C4 plants (Walder et al., 2012). Here we compare three methods for analysing the stable carbon isotope composition of AMF. Bulk carbon isotope analysis of washed extraradical mycelium is possible, but has the drawback of potential contamination from non-mycorrhizal sources. Bulk carbon isotope analysis of isolated AMF spores yields more reliable results but is rather tedious. We explain, in some detail, a more refined analysis based on the extraction of lipids from soil, followed by analysis of an AMF biomarker, the fatty acid C16:1ω5.

Fungal superhighways: do common mycorrhizal networks enhance below ground communication?

In many natural communities communication between plants and other organisms below ground drives community dynamics. This communication is primarily through the release and detection of infochemicals, which must traverse the soil matrix to be effective. In this opinion article, we propose the Network Enhanced Bioactive Zone (NEBaZ) model, which posits that common mycorrhizal networks (CMNs) increase the bioactive zones of infochemicals by serving as superhighways directly connecting plants below ground. Here we argue that infochemical transport via CMNs allows for systemic defense signaling across plant populations and directed allelochemical delivery to target plants. Plant-animal interactions may also be facilitated by CMNs, suggesting that these fungal networks may be crucial components of many natural ecosystems.