How distinct are arbuscular mycorrhizal fungal communities associating with grapevines?

Grapevines form associations with arbuscular mycorrhizal (AM) fungi. These root-dwelling fungi have the potential to contribute to crop vigor, productivity, pathogen protection, and nutrient content in grapes. In this study the arbuscular mycorrhizal fungal communities of grapevines and the surrounding interrow and native vegetation are compared. We found over 40 different taxa associating with both vines and interrow vegetation, but these communities differed based on host plant identity. These differences were apparent even after accounting for differences in soil chemical properties and differences in host plant diversity between vinerows and interrows, indicating that Vitis preferentially interacts with a subset of the viticultural fungal community. Since AM fungal communities play a major role in grapevine health, our results suggest that host identity and the diversity of AM fungal hosts in a vineyard can have strong effects on arbuscular mycorrhizal fungi community structure. In this paper, we used high throughput sequencing of the large subunit rDNA to analyze the diversity of AM fungi growing in a vineyard.     

Global diversity and distribution of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi form associations with most land plants and can control carbon, nitrogen, and phosphorus cycling between above- and belowground components of ecosystems. Current estimates of AM fungal distributions are mainly inferred from the individual distributions of plant biomes, and climatic factors. However, dispersal limitation, local environmental conditions,and interactions among AM fungal taxa may also determine local diversity and global distributions. We assessed the relative importance of these potential controls by collecting 14,961 DNA sequences from 111 published studies and testing for relationships between AM fungal community composition and geography, environment, and plant biomes. Our results indicated that the global species richness of AM fungi was up to six times higher than previously estimated, largely owing to high beta diversity among sampling sites. Geographic distance, soil temperature and moisture, and plant community type were each significantly related to AM fungal community structure, but explained only a small amount of the observed variance. AM fungal species also tended to be phylogenetically clustered within sites, further suggesting that habitat filtering or dispersal limitation is a driver of AM fungal community assembly. Therefore, predicted shifts in climate and plant species distributions under global change may alter AM fungal communities.     

Solubilization of insoluble inorganic zinc compounds by ericoid mycorrhizal fungi derived from heavy metal polluted sites

Ericoid mycorrhizal fungi increase the ability of their host plants to colonize soils polluted with toxic metals, although the underlying mechanisms are not clearly understood. Two mycorrhizal strains of Oidiodendron maius isolated from contaminated soil were previously shown to tolerate high concentrations of toxic metals. We investigated further the biological mechanisms that may explain metal tolerance, focussing on the interactions between insoluble metal species and extracellular fungal metabolites. In particular, we demonstrate that fungal strains derived from polluted and unpolluted soils mobilize insoluble inorganic zinc compounds to different extents. Strains from polluted soils showed in fact little ability to solubilize Zn from both ZnO and Zn₃(PO₄)₂, whereas strains from unpolluted soils showed a higher solubilization potential. This different behaviour was confirmed when the solubilization abilities of a wider range of fungal strains (25 isolates) was examined. Induction of organic acids (malate and citrate) by the metal compounds was at least in part responsible for metal solubilization. Our results suggest that ericoid mycorrhizal strains from polluted and unpolluted soils may interact differently with metal compounds. We speculate that this may reflect specific strategies to maintain homeostasis of essential metals under different soil conditions.     

Arbuscular mycorrhiza and soil nitrogen cycling

Nitrogen is a major nutrient that frequently limits primary productivity in terrestrial ecosystems. Therefore, the physiological responses of plants to soil nitrogen (N) availability have been extensively investigated, and the study of the soil N-cycle has become an important component of ecosystem ecology and biogeochemistry. The bulk of the literature in these areas has, however, overlooked the fact that most plants form mycorrhizal associations, and that nutrient uptake is therefore mediated by mycorrhizal fungi. It is well established that ecto- and ericoid mycorrhizas influence N nutrition of plants, but roles of arbuscular mycorrhizas in N nutrition are less well established; perhaps even more importantly, current conceptual models ignore possible influences of arbuscular mycorrhizal (AM) fungi on N-cycling processes. We review evidence for the interaction between the AM symbiosis with microbes and processes involved in soil N-cycling. We show that to date investigations have rather poorly addressed such interactions and discuss possible reasons for this. We outline mechanisms that could potentially operate with regards to AM fungal – N-cycling interactions, discuss experimental designs aimed at studying these, and conclude by pointing out priorities for future research.     

A colorimetric method for estimating vesicular-arbuscular mycorrhizal infection in roots

A colorimetric method measuring the conversion of fungal chitin to glucosamine has been used to estimate the intensity of vesicular-arbuscular mycorrhizal infection in roots. The technique has been used successfully with four plant genera and with four different endophytes.     

Solubilization of toxic metal minerals and metal tolerance of mycorrhizal fungi

This work investigates the ability of ericoid mycorrhizal (ErM) and ectomycorrhizal (EcM) fungi to solubilize different toxic metal (Cd, Cu, Pb, Zn)-containing minerals. Minerals were incorporated into solidified agar media and solubilization assessed by measuring clearing of the agar after fungal growth. Measurement of radial growth and biomass dry weight provided indications of metal tolerance: accumulated metal in the biomass was measured by atomic absorption spectrophotometry. Metal tolerance and solubilizing ability varied widely between different mineral and fungal species, and strains derived from sites of differing degrees of metal pollution. Zinc phosphate exhibited the least toxicity and was the easiest to solubilize by the majority of tested fungal isolates. Solubilization of toxic metal minerals was connected with both the pH of the medium and growth and tolerance of fungi and it seems that acidification of the medium was the main mechanism of mineral dissolution for most of the mycorrhizal fungi studied. A very strong lethal effect was observed for ectomycorrhizal isolates (>60% of strains) in the presence of Pb phosphate, carbonate, sulphide and tetraoxide. In contrast, ericoid mycorrhizal isolates were able to grow on Pb-mineral-amended media. A significant proportion of ericoid mycorrhizal cultures (70-90%) solubilized Cd and Cu phosphates and cuprite. None of the ericoid mycorrhizal and ectomycorrhizal fungi were able to produce a clear zone in Pb mineral-containing agar. However, many fungi were able to accumulate mobilized Pb in their mycelia. Differences in toxic metal mineral tolerance, mineral solubilization and metal uptake between populations isolated from metal-polluted and uncontaminated sites were related to the toxic metal which was the main pollutant in the original contaminated environment. In general, metal-tolerant fungi grew and solubilized toxic metal minerals better than non-tolerant isolates.     

Plant pathogen protection by arbuscular mycorrhizas: A role for fungal diversity?

Arbuscular mycorrhizal (AM) fungi can confer protection to host plants against some root pathogens, and several mechanisms for these phenomena have been proposed. If AM fungal taxa vary in the ways that they limit the negative effects of pathogens on host plants, additive and/or synergistic interactions among members of diverse AM fungal assemblages and communities may result in a greater pathogen protection than is currently predicted. However, in a review of the literature on interactions between AM and pathogenic fungi, we found few examples that compared the effectiveness of single- and multi-species AM fungal assemblages. Here, we briefly recount the generally recognized mechanisms of pathogen protection by AM fungi and present evidence, where appropriate, for functional diversity among AM fungal taxa with regard to these mechanisms. We propose that functional complementarity of AM fungal taxa in interactions with pathogens could mimic, or even be the cause of, previously observed relationships between AM fungal diversity and plant productivity.     

Root colonisation by arbuscular mycorrhizal, fine endophytic and dark septate fungi across a pH gradient in acid beech forests

Colonisation by root endophytes can be beneficial to plants growing on acid, nutrient-poor soils. Arbuscular mycorrhizal (AM) fungi can supply herbs with nutrients and may give protection against aluminium toxicity. Two other root colonising fungi, fine endophytes (FE) and dark septate fungi (DSE), are less well known but are potentially of benefit to their host plant. AM fungi are the most prevalent symbionts in herbs at neutral to acidic soil pH. At extremely low pH, fungal growth can be limited and AM colonisation is usually rare. Fine and dark septate endophytes, on the other hand, have been observed more often under these conditions. In order to relate endophyte colonisation to a gradient in soil pH, we investigated root colonisation by AM, FE and DSE in Maianthemum bifolium, Galium odoratum, Mercurialis perennis and Stellaria nemorum, from a range of acidic beech forests. With decreasing pH, colonisation by AM decreased, whereas the other two endophytes increased. AM and FE colonisation were inversely correlated in Maianthemum bifolium. We compared changes in root colonisation with those in chemical composition of soil and leaf samples and found a positive correlation between leaf magnesium concentrations and the presence of DSE in Galium odoratum. Aluminium concentration in Maianthemum bifolium tended to be lower when FE colonisation was high, suggesting a possible role for the fungi in plant protection against Al. We suggest that FE and DSE may replace AM fungi in herbaceous vegetation at extremely low pH, counteracting some of the negative effects of high soil acidity on plants.     

Arbuscular mycorrhizal fungi in tree-based intercropping systems: A review of their abundance and diversity

Arbuscular mycorrhizal (AM) fungi are important components of agroecosystems as they form symbiotic associations with the majority of agricultural crops. The mycorrhizal association is normally mutualistic and can provide a number of benefits to the host plant including increased nutrient uptake, improved water relations, and protection from pathogens. However, conventional agricultural practices have been shown to have a negative impact on the abundance and diversity of AM fungi. The use of more diverse and sustainable land use practices such as tree-based intercropping can have the potential to reduce the negative impact of agricultural practices on AM fungi. This paper reviews the literature to investigate the effect of temperate and tropical tree-based intercropping systems on the abundance and diversity of AM fungi. Evidence from these studies suggests that tree-based intercropping systems support a more abundant and diverse AM fungal community compared to conventionally managed systems. However, there are studies that observed zero or negative significant effects on the AM fungal community as a result of incorporating trees into agricultural systems. The variable effect of tree-based intercropping systems on AM fungi observed may be a function of the different cultivation techniques, climatic variation, or diverse tree-crop combinations used within the different tree-based intercropping systems. To further our understanding of AM fungal dynamics in tree-based intercropping systems, future research should focus on the influence of tree species with varying mycorrhizal associations and the functional role of common mycelial networks in these systems, while utilizing applicable molecular techniques.     

Molecular characterization and glomalin production of arbuscular mycorrhizal fungi colonizing a heavy metal polluted ash disposal island, downtown Venice

In this work the arbuscular mycorrhizal (AM) fungal communities colonizing a polluted ash dump island, downtown Venice, were studied by using a multimodal approach. The island, Sacca San Biagio, was covered with a thick layer of municipal solid waste residues produced by an incinerator operating from 1973, to 1984. Such residues contained high levels of heavy metals (Cu, Pb and Zn). We characterized the AMF communities present in soils on Sacca San Biagio island by using molecular methods. Nine AM fungal sequence types were detected in the roots of three plant species, representative of the dominant flora, by using partial SSU ribosomal RNA genes. The most abundant sequence types corresponded to Glomus intraradices/Glomus fasciculatum, and to Glo18, a sequence detected so far only in planta. Two sequences were new to science. Glomalin-related soil protein (GRSP), extracted from rhizosphere soil of dominant plant species, ranged from 1.6 to 2.3 mg g⁻¹. The occurrence of an active AM fungal community able to live in such harsh environment was evinced by the correlation between mycorrhizal colonization and GRSP content.     

Relationship between soil enzyme activities, nutrient cycling and soil fungal communities in a northern hardwood forest

Soil fungi are highly diverse and act as the primary agents of nutrient cycling in forests. These fungal communities are often dominated by mycorrhizal fungi that form mutually beneficial relationships with plant roots and some mycorrhizal fungi produce extracellular and cell-bound enzymes that catalyze the hydrolysis of nitrogen (N)- and phosphorus (P)- containing compounds in soil organic matter. Here we investigated whether the community structure of different types of mycorrhizal fungi (arbuscular and ectomycorrhizal fungi) is correlated with soil chemistry and enzyme activity in a northern hardwood forest and whether these correlations change over the growing season. We quantified these relationships in an experimental paired plot study where white-tailed deer (access or excluded 4.5 yrs) treatment was crossed with garlic mustard (presence or removal 1 yr). We collected soil samples early and late in the growing season and analyzed them for soil chemistry, extracellular enzyme activity and molecular analysis of both arbuscular mycorrhizal (AM) and ectomycorrhizal/saprotrophic fungal communities using terminal restriction fragment length polymorphism (TRFLP). AM fungal communities did not change seasonally but were positively correlated with the activities of urease and leucine aminopeptidase (LAP), enzymes involved in N cycling. The density of garlic mustard was correlated with the presence of specific AM fungal species, while deer exclusion or access had no effect on either fungal community after 4.5 yrs. Ectomycorrhizal/saprotrophic fungal communities changed seasonally and were positively correlated with most soil enzymes, including enzymes involved in carbon (C), N and P cycling, but only during late summer sampling. Our results suggest that fine scale temporal and spatial changes in soil fungal communities may affect soil nutrient and carbon cycling. Although AM fungi are not generally considered capable of producing extracellular enzymes, the correlation between some AM taxa and the activity of N acquisition enzymes suggests that these fungi may play a role in forest understory N cycling.     

Rapid uptake of N-ammonium and glycine-C, N by arbuscular and ericoid mycorrhizal plants native to a Northern California coastal pygmy forest

While it is well established that plants are able to acquire nitrogen in inorganic form, there is less information on their ability to ‘short circuit’ the N cycle, compete with microbes, and acquire nitrogen in organic form. Mycorrhizal fungi, known to enhance nutrient uptake by plants, may play a role in organic N uptake, particularly ericoid mycorrhizas. We asked the question—Can mycorrhizal fungi increase the ability of plants to take up organic N, compared to inorganic N? Here, we report on the abilities of three plant species, ericoid mycorrhizal Rhododendron macrophyllum and Vaccinium ovatum and arbuscular mycorrhizal Cupressus goveniana ssp. pigmaea, to acquire C and/or N from an organic and an inorganic N source. All three species are native to a California coastal pygmy forest growing in acidic, low-fertility, highly organic soils. In a pot study, glycine-α¹³C, ¹⁵N and ¹⁵N-ammonium were applied to pygmy forest soil for 17 or 44 h. Ericoid mycorrhizal species did not demonstrate a preference for either inorganic or organic sources of N while Cupressus acquired more NH₄-N than glycine-N. For all species, glycine-N uptake did not increase after 17 h suggesting glycine uptake and glycine immobilization occurred rapidly. Both glycine-N and glycine-C were recovered in shoots and in roots suggesting that all species acquired some N in organic form. Regression analyses of glycine-N and glycine-C recovery in root tissue indicate that much of the glycine was taken up intact and that the minimum proportion of glycine-N recovered in organic form was 85% (Cupressus) and 70% (Rhododendron). Regressions were non-significant for Vaccinium. For all species, glycine-N remained predominantly in roots while glycine-C was transferred to shoots. In contrast, NH₄-N remained in roots of ericoid plants but was transferred to shoots of arbuscular mycorrhizal Cupressus. Since net N mineralization rates in pygmy forest soils are low, our results suggest that organic N may be an important N source for plants in this temperate coniferous ecosystem regardless of mycorrhizal type. Acquisition of amino acid C by these species also may partially offset the carbon cost to plants of hosting mycorrhizal fungi.     

Isolation and characterization of fungal inhibitors from Epichloë festucae

A series of studies was conducted to test the antifungal activity of clavicipitaceous endophytes and to identify potential fungal inhibitors in this symbiotic infection. A diverse group of endophytes was screened for antifungal activity using organic extracts from liquid fermentation cultures. Fungal inhibitors were purified from fermentation cultures of Epichlo? festucae using a bioassay-directed extraction with Cryphonectria parasitica as the test organism. Compounds shown to have antifungal activity were subsequently identified using NMR and GC-MS. Extracts from a wide range of fungal isolates had various degrees of antifungal activity, but the greatest antifungal activity was observed in E. festucae and Neotyphodium tembladerae. Three types of inhibitors were isolated from a batch culture of E. festucae, including several indole derivatives, a sesquiterpene, and a diacetamide. Among the indole derivatives, indole-3-acetic acid and indole-3-ethanol were identified as the major indoles. These compounds were previously reported in endophytic fungi, and this study suggests a role in host disease resistance against other pathogens. The diversity in fungal inhibitors produced by this endophyte also suggests that fungal inhibitors may act additively or synergistically to reduce colonization of endophyte-infected hosts by potential fungal competitors.     

Distribution of roots and arbuscular mycorrhizal associations in tropical forest types of Xishuangbanna,southwest China

Root distribution and mycorrhizal associations were compared in primary, secondary and limestone forests in Xishuangbanna, southwest China. Soil cores to a depth of 20 cm were collected at random points from four 50 m² quadrats in each forest type. Arbuscular mycorrhizal (AM) associations were the only form of mycorrhiza found in all forest types. The primary forest was characterized by high root mass, root lengths and AM colonization levels higher than other forest types. In contrast, secondary forests had greater AM fungal spore numbers and specific root length, indicating that plant species in secondary forests achieved a greater degree of soil exploration with less biomass allocation to roots. Root density, AM colonization and AM fungal spore numbers decreased with soil depth in all forest types. Although the correlation between AM colonization levels and spore numbers was insignificant when all forest types were considered together, significant relationships emerged when each forest type was considered individually. AM colonization and spore numbers were correlated with several root variables.     

Extracellular enzyme activity in the mycorrhizospheres of a boreal fire chronosequence

Saprotrophic microbes are typically credited with producing extracellular enzymes that recycle organic matter, though roots and mycorrhizal fungi also can contribute and may compete with the saprotrophs. We examined extracellular enzyme activity associated with the mycorrhizospheres of arbuscular mycorrhizal, ectomycorrhizal, dual-colonized (arbuscular and ectomycorrhizal), and ericoid mycorrhizal plants in a fire chronosequence in Alaska. Bulk soil and soil from beneath host plants were gathered in July 2004 and assayed for five enzymes that target organic C, P, and N substrates. Compared to bulk soil, activities of the C-targeting enzymes β-1,4-glucosidase and peroxidase were lower in arbuscular mycorrhizospheres and ericoid mycorrhizospheres, respectively. Moreover, extracellular enzyme activity varied among mycorrhizosphere types. Specifically, N-targeting leucine aminopeptidase was highest in arbuscular mycorrhizospheres, followed by ericoid and ectomycorrhizal/dual-colonized mycorrhizospheres; β-1,4-glucosidase had the reverse pattern. In addition, enzymatic stoichiometry suggested that extracellular enzyme producers invested more in C-acquisition than in N-acquisition in recent fire scars compared to mature forests. These data extend previous findings that roots and mycorrhizal fungi compete with saprotrophs by showing that the strength of this effect varies by mycorrhizal host. As a result the community composition of mycorrhizal host plants might mediate enzymatic activity in boreal soils.     

Different mycorrhizal fungal strains determine plant community response to nitrogen and water availability

Most research on the mycorrhizal positive–negative responsiveness continuum (or “mutualism–parasitism continuum”) has focused on individual plant species growing at different levels of P availability. Here, we explore this continuum in an experimental plant community inoculated with three arbuscular mycorrhizal (AM) fungal strains (both single and mixed) growing under four resource availability scenarios. These scenarios are a factorial combination of two levels of water and N availability. Each AM fungal strain had a different origin: an arid ecosystem, a farmland, and a mine. We hypothesized that the response of the plant community to mycorrhizal inoculum would depend on the associated AM fungal strain and would be negatively related with increased nitrogen and water availability. Our results showed that mixed‐strain AM fungal inoculation had more positive effects along a wider range of water and N availability scenarios than single‐strain inoculation. In contrast, mycorrhizal growth response of plants inoculated with a single AM fungal strain shifted from positive to neutral and negative depending on resource availability. Adaptation of each strain to its local conditions might confer different properties to the mycorrhizal symbiosis. Therefore, we conclude that AM fungal origin and environmental limiting resources are crucial factors to predict plant community mycorrhizal growth response in changing ecosystems.     

Time-course of heavy metal uptake in maize and clover as affected by root density and different mycorrhizal inoculation regimes

A pot experiment was conducted to test the effect of three microbial regimes on the time course of heavy metal uptake in clover and maize from an industrially polluted soil. The three treatments included: (1) an intact flora of bacteria and fungi, including indigenous arbuscular mycorrhizal (AM) fungi together with soil microfauna; (2) the indigenous bacterial/fungal flora except AM fungi, reintroduced into sterilized soil; or (3) the same bacterial/fungal flora plus an AM fungus. For the final harvest, two pot sizes were included to assess the effect of root density. Plant uptake of P and heavy metals varied according to plant species, harvest time and soil treatment. For both plant species, shoot concentration of Zn, Cd and Cu decreased and Ni increased with plant age. Plants growing in sterilized soil with reintroduced AM fungi generally grew better, but contained higher concentrations of heavy metals than those colonized by indigenous AM fungi. Plants with mycorrhiza frequently contained more P, Zn, Cd, Cu and Pb in roots and shoots compared to nonmycorrhizal plants. Elevated root/shoot concentration ratios of P and metals indicate a sequestration of metal phosphates in mycorrhizal roots. Mycorrhizal performance was influenced by root density. At low root densities, metal concentrations in mycorrhizal plants were reduced, whereas it had no effect at high root densities when the entire soil volume was efficiently exploited by roots. We conclude that root density data are essential for interpretations of the influence of AM on metal uptake in plants.     

Effect of fungal endophytes on morphological characteristics,nutrients content and longevity of plane trees (Platanus orientalis L.)

The plane tree is one of the most frequently used street trees in urban landscape in Iran and some Mediterranean countries and some age-old ones are found in these countries. Endophytes are microorganisms which associate with healthy plant tissues, persisting without causing diseases symptoms in the host and normally benefitting them through improving plant ability to withstand environmental stresses. In this contribution, the hypothesis of fungal endophytes existence in the plane trees as well as their relationship with morphological characteristics of trees and nutrient elements of leaves were studied in the central part of Iran. The experiment was performed according to an unbalanced completely randomized design. Approximate age and height of plane trees were recorded and then the greenness and visual appearance were evaluated. Healthy shoots and bark pieces were collected and surface sterilized then placed in petri dishes containing PDA. The results showed that endophytes frequency were significantly greater in age-old trees (60.04%) than young ones (39.96%). Endophytes existence had a positive correlation with iron (Fe) and potassium (K) concentrations of the leaves, and trees height and circumference. Also, there was a positive correlation between the presence of fungal endophytes and improved visual appearance of plane trees. The results suggest that fungal endophytes association enhanced nutrient elements assimilation in trees contributing at least partly to elevated survival of age-old plane trees.     

Local diversity of native arbuscular mycorrhizal symbionts differentially affects growth and nutrition of three crop plant species

Intact whole native AMF communities occurring across a 100-m-long field were used for the evaluation of plant performance, as determined by the actual fungal species colonizing host roots. The soil from distinct plots within a “hot spot” field was collected to set up 54 experimental units where three different plant species were grown, in order to test whether the whole native AMF communities were able to differentially affect plant growth, to assess the genetic identity of the AMF actually colonizing the tested plants and to analyse their community composition in the different hosts. Molecular analyses revealed that plant growth and nutrition of the crop plants were differentially affected by the diverse native arbuscular mycorrhizal communities colonizing the roots of the three plants, whose performance varied depending on the identity of plant hosts and fungal symbionts, more than on a rich and diversified AMF community. Such results, improving our understanding of AMF distribution at the local scale, represent a starting point allowing the selection, isolation and characterization of the most efficient AMF assemblages to be used as inoculants in sustainable food production systems.     

Changes in soil organic carbon,total nitrogen,and abundance of arbuscular mycorrhizal fungi along a large-scale aridity gradient

Changes in soil organic carbon, total nitrogen, pH, and the abundance of arbuscular mycorrhizal fungi are examined along a large-scale aridity gradient from southeast to northwest in China. Soil organic carbon and total nitrogen decreased but pH increased with increased aridity. Aboveground plant biomass, spore abundance, and colonization of roots by arbuscular mycorrhizal fungi also declined as the aridity increased. Soil organic carbon and total nitrogen were positively correlated with aboveground plant biomass, and arbuscular mycorrhizal fungal spore number and root colonization were positively correlated with soil organic carbon, total nitrogen, and aboveground plant biomass but were negatively correlated with soil pH. A structural equation model suggested that aridity affected soil organic carbon and total nitrogen by limiting aboveground plant biomass. Aridity exerted a large direct effect and smaller indirect effects (via changes in aboveground plant biomass) on the abundance of arbuscular mycorrhizal fungi. Soil pH also directly influenced arbuscular mycorrhizal fungal abundance. These results suggest that aboveground plant biomass could be a key factor driving the changes of soil organic carbon, total nitrogen, and arbuscular mycorrhizal fungal abundance along this aridity gradient in China.