Using lipid analysis and hyphal length to quantify AM and saprotrophic fungal abundance along a soil chronosequence

We evaluate the use of signature fatty acids and direct hyphal counts as tools to detect and quantify arbuscular mycorrhizal (AM) and saprotrophic fungal (SF) biomass in three Hawaiian soils along a natural soil fertility gradient. Phospholipids16:1ω5c and 18:2ω6,9c were used as an index of AM and saprotrophic fungal biomass, respectively. Both phospholipid analysis and hyphal length indicated that the biomass of AMF was greatest at the highest fertility site, and lowest where phosphorus limits plant growth. Saprotrophic fungal biomass did not vary. Hyphal length counts appeared to under-estimate SF abundance, while the phospholipid AMF:SF ratio was in line with expectations. This study indicates that phospholipids may be a valuable and reliable tool for studying the abundance, distribution, and interactions between AM and saprotrophic fungi in soil.     

Spatial distribution of arbuscular mycorrhizal fungi, glomalin and soil enzymes under the canopy of Astragalus adsurgens Pall. in the Mu Us sandland, China

To measure and manage plant growth in arid and semi-arid sandlands, improved understanding of the spatial patterns of desert soil resources and the role of arbuscular mycorrhizal (AM) fungi is needed. Spatial patterns of AM fungi, glomalin and soil enzyme activities were investigated in five plots located in the Mu Us sandland, northwestern China. Soils to 50 cm depth in the rhizosphere of Astragalus adsurgens Pall. were sampled. The study demonstrated that A. adsurgens Pall. could form strong symbiotic relationships with AM fungi. Arbuscular mycorrhizal fungal status and distributions were significantly different among the five studied plots. Correlation coefficient analysis demonstrated that spore density was significantly and positively correlated with soil organic carbon (SOC), soil acid phosphatase and to two Bradford-reactive soil protein (BRSP) fractions (P < 0.01). Colonization of arbuscules and vesicles were positively correlated with protease activity. The BRSP fractions were also significantly and positively correlated to edaphic factors (e.g. SOC, available nitrogen, and Olsen phosphorus) and soil enzymes (e.g. soil urease and acid phosphatase). The means of total BRSP and easily extractable BRSP were 0.95 mg g⁻¹ and 0.5 mg g⁻¹ in all data, respectively. The levels of BRSP in the desert soil were little lower than those in native and arable soils, but the ratios of BRSP to SOC were much higher than farmland soils. The results of this study support the conclusion that glomalin could be an appropriate index related to the level of soil fertility, especially in desert soil. Moreover, AM fungal colonizations and glomalin might be useful to monitor desertification and soil degradation.     

Independent roles of ectomycorrhizal and saprotrophic communities in soil organic matter decomposition

The relative roles of ectomycorrhizal (ECM) and saprotrophic communities in controlling the decomposition of soil organic matter remain unclear. We tested the hypothesis that ECM community structure and activity influences the breakdown of nutrient-rich biopolymers in soils, while saprotrophic communities primarily regulate the breakdown of carbon-rich biopolymers. To test this hypothesis, we used high-throughput techniques to measure ECM and saprotrophic community structure, soil resource availability, and extracellular enzyme activity in whole soils and on ECM root tips in a coastal pine forest. We found that ECM and saprotroph richness did not show spatial structure and did not co-vary with any soil resource. However, species richness of ECM fungi explained variation in the activity of enzymes targeting recalcitrant N sources (protease and peroxidase) in bulk soil. Activity of carbohydrate- and organic P- targeting enzymes (e.g. cellobiohydrolase, β-glucosidase, α-glucosidase, hemicellulases, N-acetyl-glucosaminidase, and acid phosphatase) was correlated with saprotroph community structure and soil resource abundance (total soil C, N, and moisture), both of which varied along the soil profile. These observations suggest independent roles of ECM fungi and saprotrophic fungi in the cycling of N-rich, C-rich, and P-rich molecules through soil organic matter. Enzymatic activity on ECM root tips taken from the same soil cores used for bulk enzyme analysis did not correlate with the activity of any enzyme measured in the bulk soil, suggesting that ECM contributions to larger-scale soil C and nutrient cycling may occur primarily via extramatrical hyphae outside the rhizosphere.     

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.     

Changes in soil aggregation and glomalin-related soil protein content as affected by the arbuscular mycorrhizal fungal species Glomus mosseae and Glomus intraradices

Arbuscular mycorrhizal (AM) fungi are key organisms of the soil/plant system, influencing soil fertility and plant nutrition, and contributing to soil aggregation and soil structure stability by the combined action of extraradical hyphae and of an insoluble, hydrophobic proteinaceous substance named glomalin-related soil protein (GRSP). Since the GRSP extraction procedures have recently revealed problems related to co-extracting substances, the relationship between GRSP and AM fungi still remains to be verified. In this work the hypothesis that GRSP concentration is positively correlated with the occurrence of AM fungi was tested by using Medicago sativa plants inoculated with different isolates of Glomus mosseae and Glomus intraradices in a microcosm experiment. Our results show that (i) mycorrhizal establishment produced an increase in GRSP concentration - compared to initial values - in contrast with non-mycorrhizal plants, which did not produce any change; (ii) aggregate stability, evaluated as mean weight diameter (MWD) of macroaggregates of 1-2 mm diameter, was significantly higher in mycorrhizal soils compared to non-mycorrhizal soil; (iii) GRSP concentration and soil aggregate stability were positively correlated with mycorrhizal root volume and weakly correlated with total root volume; (iv) MWD values of soil aggregates were positively correlated with values of total hyphal length and hyphal density of the AM fungi utilized.The different ability of AM fungal isolates to affect GRSP concentration and to form extensive and dense mycelial networks, which may directly affect soil aggregates stability by hyphal enmeshment of soil particles, suggests the possibility of selecting the most efficient isolates to be utilized for soil quality improvement and land restoration programs.     

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.     

降水变化和氮沉降对荒漠草原土壤丛枝菌根真菌群落结构的影响

降水变化和氮沉降是影响植物、微生物和土壤环境变化的两个重要方面。尽管丛枝菌根（AM）真菌在陆地生态系统中起着至关重要的作用，但人们对降水变化和氮添加如何交互影响AM真菌群落仍知之甚少。本研究以短花针茅荒漠草原为研究对象，采用裂区设计，主区为自然降雨(CK)、增雨30%(W)和减雨30%(R)三个水分梯度，副区为0(N0)，30(N30)，50(N50)和100(N100) kg?hm-2?a-1 四个氮素梯度共12个处理，通过高通量测序分析了土壤中AM真菌群落的多样性和组成。结果发现，水分处理对土壤AM真菌的Alpha多样性有促进作用，氮素处理抑制了土壤AM真菌的Alpha多样性，水分增加和氮素添加的交互作用促进了AM真菌的Alpha多样性增加，并改变了土壤AM真菌群落组成。水分和氮素刺激了各功能型植物生物量的增加，氮添加使多年生杂草和半灌木、小半灌木生物量显著增加，多年生禾草生物量显著减少。此外，多型孢子菌科的相对丰度与一二年生植物和半灌木、小半灌木生物量呈显著正相关，一二年生植物和半灌木、小半灌木生物量在氮添加和增雨处理下增加。本研究证明了AM真菌群落在短期气候变化下的稳定性。此外，AM真菌在科水平上的丰度与各生活型植物地上生物量的相关性证明了地上和地下生态系统的连通性。     

Fraxinus-Glomus-Pisolithus symbiosis: Plant growth and soil aggregation effects

It has been established that arbuscular mycorrhizal (AM) fungi are involved in the conservation of soil structure. However, the effect of ectomycorrhizal (EM) fungi alone or in interaction with AM fungi in soil structure has been much less studied. This experiment evaluated EM and AM fungi effects on soil aggregation and plant growth. Ash plants (Fraxinus uhdei) were grown in pots, and were inoculated with Glomus intraradices and Pisolithus tinctorius separately but also in combination. Our results showed that F. uhdei established a symbiotic association with EM and AM fungi, and that these organisms, when interacting, showed synergistic and additive effects on plant growth compared to singly inoculated treatments. EM and AM fungi prompted changes in root morphology and increased water-stable aggregates. AM fungi affect mainly small-sized macroaggregates, while EM and EM-AM fungi interaction mainly affected aggregates bigger than 0.5 mm diameter. These results suggest that ectomyccorrhizal as well as arbuscular mycorrhizal fungi should be considered in restoration programs with Fraxinus plants.     

Dynamics of arbuscular mycorrhizal fungi and glomalin in the rhizosphere of Artemisia ordosica Krasch. in Mu Us sandland, China

To understand the ecological significance of arbuscular mycorrhizal (AM) associations in semi-arid and arid lands, the temporal and spatial dynamics of AM fungi and glomalin were surveyed in Mu Us sandland, northwest China. Soil samples in the rhizosphere of Artemisia ordosica Krasch. were collected in May, July and October 2007, respectively. Arbuscular, hyphal and total root infection and spore density of AM fungi peaked in summer. The mean contents of total Bradford-reactive soil proteins (T-BRSPs, TG) and easily extractable Bradford-reactive soil proteins (EE-BRSPs, EEG) reached maximal values in spring. Spore density and two BRSPs fractions were the highest in the 0-10 cm soil layer, but the ratios of two BRSPs fractions to soil organic carbon (SOC) were the highest in the 30-50 cm soil layer. Hyphal infection was negatively correlated with soil enzymatic activity (soil urease and acid phosphatase) (P < 0.05). Arbuscular infection was negatively correlated with soil acid phosphatase (P < 0.01). Spore density was positively correlated with edaphic factors (soil available N, Olsen P, and SOC) and soil enzymatic activity (soil acid and alkaline phosphatase) (P < 0.01). Two BRSPs fractions were positively correlated with edaphic factors (soil available N and SOC) and soil enzymatic activity (soil urease, acid and alkaline phosphatase) (P < 0.01). TG was positively correlated with soil Olsen P (P < 0.05). We concluded that the dynamics of AM fungi and glomalin have highly temporal and depth patterns, and influenced by nutrient availability and enzymatic activity in Mu Us sandland, and suggest that glomalin are useful indicators for evaluating soil quality and function of desert ecosystem on the basis of its relationship to AM fungal community, soil nutrient dynamics and carbon cycle.     

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.     

Interactions between phosphorus availability and an AM fungus (Glomus intraradices) and their effects on soil microbial respiration, biomass and enzyme activities in a calcareous soil

The interactions between soil P availability and mycorrhizal fungi could potentially impact the activity of soil microorganisms and enzymes involved in nutrient turnover and cycling, and subsequent plant growth. However, much remains to be known of the possible interactions among phosphorus availability and mycorrhizal fungi in the rhizosphere of berseem clover (Trifolium alexandrinum L.) grown in calcareous soils deficient in available P. The primary purpose of this study was to look at the interaction between P availability and an arbuscular mycorrhizal (AM) fungus (Glomus intraradices) on the growth of berseem clover and on soil microbial activity associated with plant growth. Berseem clover was grown in P unfertilized soil (−P) and P fertilized soil (+P), inoculated (+M) and non-inoculated (−M) with the mycorrhizal fungus for 70 days under greenhouse conditions. We found an increased biomass production of shoot and root for AM fungus-inoculated berseem relative to uninoculated berseem grown at low P levels. AM fungus inoculation led to an improvement of P and N uptake. Soil respiration (SR) responded positively to P addition, but negatively to AM fungus inoculation, suggesting that P limitation may be responsible for stimulating effects on microbial activity by P fertilization. Results showed decreases in microbial respiration and biomass C in mycorrhizal treatments, implying that reduced availability of C may account for the suppressive effects of AM fungus inoculation on microbial activity. However, both AM fungus inoculation and P fertilization affected neither substrate-induced respiration (SIR) nor microbial metabolic quotients (qCO₂). So, both P and C availability may concurrently limit the microbial activity in these calcareous P-fixing soils. On the contrary, the activities of alkaline phosphatase (ALP) and acid phosphatase (ACP) enzymes responded negatively to P addition, but positively to AM fungus inoculation, indicating that AM fungus may only contribute to plant P nutrition without a significant contribution from the total microbial activity in the rhizosphere. Therefore, the contrasting effects of P and AM fungus on the soil microbial activity and biomass C and enzymes may have a positive or negative feedback to C dynamics and decomposition, and subsequently to nutrient cycling in these calcareous soils. In conclusion, soil microbial activity depended on the addition of P and/or the presence of AM fungus, which could affect either P or C availability.     

Suppression of other soil microorganisms by mycelium of arbuscular mycorrhizal fungi in root-free soil

The influence of mycelium of two arbuscular mycorrhizal (AM) fungi, Glomus intraradices and Glomus mosseae, on other soil microorganisms, was examined in root-free soil with and without organic substrate amendment in terms of cellulose. The AM fungi were grown in symbiosis with cucumber in a compartmented growth system, which allowed AM fungal external mycelium to grow into root-free compartments. The fungicide Benomyl was applied to the root-free compartments to create an alternative non-mycorrhizal control treatment. Whole cell biomarker fatty acids were employed to quantify different groups of soil microorganisms including the two AM fungi. Abundance of most microbial groups were reduced by external mycelium of both AM fungi, though differential effects on the microbial community composition were observed between the two AM fungi as revealed from principal component analysis. Inhibition of other soil microorganisms was more pronounced in root-free soil with mycelium of G. mosseae than with mycelium of G. intraradices. In general, cellulose increased the amount of biomarker fatty acids of most groups of soil microorganisms, but cellulose did not affect the influence of AM fungi on other soil microorganisms. Benomyl suppressed growth of the external mycelium of the two AM fungi and had limited non-target effects on other microbial groups. In conclusion, our results show differential effects of external mycelium of AM fungi on other soil microbial communities, though both AM fungi included in the study overall inhibited most microbial groups as examined using whole cell biomarker fatty acids.     

Phylogenetic structure of arbuscular mycorrhizal community shifts in response to increasing soil fertility

Understanding the underlying mechanisms driving responses of belowground communities to increasing soil fertility will facilitate predictions of ecosystem responses to anthropogenic eutrophication of terrestrial systems. We studied the impact of fertilization of an alpine meadow on arbuscular mycorrhizal (AM) fungi, a group of root-associated microorganisms that are important in maintaining sustainable ecosystems. Species and phylogenetic composition of AM fungal communities in soils were compared across a soil fertility gradient generated by 8 years of combined nitrogen and phosphorus fertilization. Phylogenetic patterns were used to infer the ecological processes structuring the fungal communities. We identified 37 AM fungal virtual taxa, mostly in the genus Glomus. High fertilizer treatments caused a dramatic loss of Glomus species, but a significant increase in genus richness and a shift towards dominance of the lineage of Diversispora. AM fungal communities were phylogenetically clustered in unfertilized soil, random in the low fertilizer treatment and over-dispersed in the high fertilizer treatments, suggesting that the primary ecological process structuring communities shifted from environmental filtering (selection by host plants and fungal niches) to a stochastic process and finally to competitive exclusion across the fertilization gradient. Our findings elucidate the community shifts associated with increased soil fertility, and suggest that high fertilizer inputs may change the dominant ecological processes responsible for the assembly of AM fungal communities towards increased competition as photosynthate from host plants becomes an increasingly limited resource.     

Woodland trees modulate soil resources and conserve fungal diversity in fragmented landscapes

Resource islands around woody plants are thought to define the structure and function of many semiarid and arid ecosystems, but their role in patterning of soil microbial communities remains largely unexamined in dry environments. This study examined soil resource distribution and associated fungal communities in two Allocasuarina luehmannii (buloke) remnants of semiarid north-western Victoria, Australia. These savannah-like woodlands are listed as endangered due to extensive clearing for agriculture. We used the DNA-based profiling technique T-RFLP and ordination-based statistical methods to compare fungal community compositions in surface soils from two remnants (located 1.6 km apart) and three sampling positions (beneath individual buloke canopies; grassy inter-canopy areas; and adjoining cleared paddocks). Resource island formation beneath buloke trees was clearly evident in soil physicochemical properties (e.g. threefold concentrations of total carbon and nitrogen in canopy versus non-canopy soils). This heterogeneity of resources was moderately correlated with soil fungal community compositions, which were distinct for each sampling position. We argue that fungal composition patterns reflected multiple roles of fungi in dryland ecosystems, namely: responses of saprotrophic fungi to tree organic matter inputs; specificity of ectomycorrhizal fungi to tree rooting zones; and fungal involvement in biological soil crusts that variably covered non-canopy soils. Our data did not indicate that buloke canopy areas were particular hotspots of soil fungal diversity, but that they increased landscape-level diversity by supporting a distinct suite of fungi. In addition, we provide evidence of phylogenetic differentiation of soil fungal communities between our two remnants, which adds to growing evidence of fungal genetic structure at localised scales. These findings highlight the importance of remnant trees in conserving both soil resources and microbial genetic diversity. In addition, evidence of differentiation of soil fungal phylogenetics between nearby but isolated remnants suggests that conserving soil fungal diversity requires conservation of host habitats over their entire (remaining) range, and indicates previously unseen consequences of tree loss from extensively cleared landscapes.     

Fermentation of sugar beet waste by Aspergillus niger facilitates growth and P uptake of external mycelium of mixed populations of arbuscular mycorrhizal fungi

Sugar beet waste has potential value as a soil amendment and this work studied whether fermentation of the waste by Aspergillus niger would influence the growth and P uptake of arbuscular mycorrhizal (AM) fungi. Plants were grown in compartmentalised growth units, each with a root compartment (RC) and two lateral root-free compartments (RFC). One RFC contained untreated soil while the other RFC contained soil, which was uniformly mixed with sugar beet waste, either untreated (SB) or degraded by A. niger (ASB) in a rock phosphate (RP)-supplied medium. The soil in each pair of RFC was labelled with ³³P and ³²P in order to measure P uptake by the AM fungal mycelium, of which length density was also measured. Whole cell fatty acid (WCFA) signatures were used as biomarkers of the AM fungal mycelium and other soil microorganisms. The amount of biomarkers of saprotrophic fungi and both Gram-positive and Gram-negative bacteria was higher in SB than in ASB treatments. Whilst ASB increased growth and activity of AM mycelium, SB had the opposite effect. Moreover, shoot P content was increased by the addition of ASB, and by inoculation with AM fungi. Modification of soil microbial structure and production of exudates by A. niger, as a consequence of fermentation process of sugar beet waste, could possibly explain the increase of AM growth in ASB treatments. On the other hand, the highest P uptake was a result of the solubilisation of rock phosphate by A. niger during the fermentation.     

Effects of soil application of olive mill wastewaters on the structure and function of the community of arbuscular mycorrhizal fungi

Recycling of olive mill wastewaters (OMW) into agricultural soils is a controversial issue since benefits to soil fertility should counterbalance potential short-term toxicity effects. We investigated the short-term effects of OMW on the soil-plant system, regarding the diversity, structure and root colonization capacity of arbuscular mycorrhizal (AM) fungi and the respective growth response of Vicia faba L, commonly used as green manure in olive-tree plantations. A compartmentalized pot system was used that allowed the establishment of an AM fungal community in one compartment (feeder) and the application of three OMW dose levels in an adjacent second compartment (receiver). At 0, 10, and 30 days after OMW treatment (DAT), V. faba pre-germinated seeds were seeded in the receiver compartment. At harvest, shoot and root dry weights, AM fungal root colonization, soil hyphal length and P availability were recorded in the receiver compartment. In addition, OMW effects on AM fungal diversity in plant roots were studied by DGGE. A transient effect of OMW application was observed; plant growth and AM fungal colonization were initially inhibited, whereas soil hyphal length was stimulated, but in most cases differences were absent when seeding was performed 30 DAT. Similarly, changes induced in the structure of the root AM fungal community were of transient nature. Cloning and sequencing of all the major DGGE bands showed that roots were colonized by Glomus spp. The transient effects of OMW on the structure and function of AM fungi could be attributed to OMW-derived phytoxicity to V. faba plants or to an indirect effect via alteration of soil nutritional status. The high OMW dose significantly increased soil P availability in the presence of AM fungi, suggesting efficient involvement of AM fungi in organic-P minerilization. Overall our results indicate that soil application of OMW would cause transient changes in the AM fungal colonization of V. faba plants, which, would not impair their long-term plant growth promoting ability.     

Effects of mycorrhizal roots and extraradical hyphae on N uptake from vineyard cover crop litter and the soil microbial community

The objectives of this study were to evaluate the contribution of arbuscular mycorrhizal (AM) fungal hyphae to ¹⁵N uptake from vineyard cover crop litter (Medicago polymorpha), and to examine the soil microbial community under the influence of mycorrhizal roots and extraradical hyphae. Mycorrhizal grapevines (Vitis vinifera) were grown in specially designed containers, within which a polyvinyl chloride (PVC) mesh core was inserted. Different sizes of mesh allowed mycorrhizal roots (mycorrhizosphere treatment) or extraradical hyphae (hyphosphere treatment) to access dual labeled ¹⁵N and ¹³C cover crop litter that was placed inside the cores after 4 months of grapevine growth. Mesh cores in the bulk soil treatment, which served as a negative control, had the same mesh size as the hyphosphere treatment, but frequent rotation prevented extraradical hyphae from accessing the litter. Grapevines and soils were harvested 0, 7, 14, and 28 days after addition of the cover crop litter and examined for the presence of ¹⁵N. Soil microbial biomass and the soil microbial community inside the mesh cores were examined using phospholipid fatty acid analysis. ¹⁵N concentrations in grapevines in the hyphosphere treatment were twice that of grapevines in the bulk soil treatment, suggesting that extraradical hyphae extending from mycorrhizal grapevine roots may have a role in nutrient utilization from decomposing vineyard cover crops in the field. Nonetheless, grapevines in the mycorrhizosphere treatment had the highest ¹⁵N concentrations, thus highlighting the importance of a healthy grapevine root system in nutrient uptake. We detected similar peaks in soil microbial biomass in the mycorrhizosphere and hyphosphere treatments after addition of the litter, despite significantly lower microbial biomass in the hyphosphere treatment initially. Our results suggest that although grapevine roots play a dominant role in the uptake of nutrients from a decomposing cover crop, AM hyphae may have a more important role in maintaining soil microbial communities associated with nutrient cycling.     

Impact of soil nitrogen concentration on Glomus spp.-Sinorhizobium interactions as affecting growth, nitrate reductase activity and protein content of Medicago sativa

Our objective was to evaluate how increasing levels of N in the medium (0, 4, 8 and 16 mmol N added kg^-1 soil) affect the interaction between Sinorhizobium and arbuscular mycorrhiza (AM) fungi in the tripartite symbiosis with Medicago sativa. Growth response, nutrient acquisition, protein content, and nitrate reductase (NR) activity were measured both in plant shoots and roots. Results showed that N levels in soil did not affect mycorrhizal colonization but they strongly influenced nodulation, particularly of mycorrhizal plants. Mycorrhizal colonization was required for a proper nodulation when no N was applied to soil. In contrast, the addition of 4 mmol N kg^-1 soil reduced nodulation only in mycorrhizal plants and 8 mmol N added kg^-1 soil allowed nodule formation only in non-mycorrhizal plants. Nodulation was totally inhibited in all treatments with the addition of 16 mmol N added kg^-1 soil. N addition enhanced NR activity in all the treatments, while AM colonization increased the proportion of NR allocated to roots. This effect was more pronounced under the lowest N levels in the medium. The two AM fungal species showed different distribution pattern of enzymatic activities in plant tissues indicating specific physiological traits. Protein content as well as the relative proportion of protein in roots were greatly increased after mycorrhizal colonization. Glomus intraradices-colonized plants had the highest protein content in shoot and root. Mycorrhizal effects on growth, N acquisition and biochemical variables cannot be interpreted as an indirect P-mediated effect since P content was lower in mycorrhizal plants than in those which were P fertilized. Mycorrhizal colonization increased the N content in plants irrespective of the N level, but the effectiveness of AM fungi on plant N acquisition depended on the AM fungus involved, G. intraradices being the most effective, particularly at the highest N rate. N₂ fixation, enhanced by AM colonization, contributed to N acquisition when a moderate N quantity was available in the soil. Nevertheless, under a high N amount the nodulating process and/or fixing capacity by Sinorhizobium was reduced in AM plants. In contrast, the AM fungal mycelium from a particular mycorrhizal fungus may continue to contribute efficiently to the N uptake from the soil even at high N levels. These results demonstrate the particular sensitivity of AM fungal species in terms of their growth and/or function to increasing N amounts in the medium. A selection of AM fungi used to address specific environmental conditions, such as N fertilization regimes comparable to those used in agronomic practices, is required for a better use of N applied to soil.     

Disentangling the contributions of arbuscular mycorrhizal fungi to soil multifunctionality

Soil multifunctionality represents a range of soil processes driven by the interactions between soil abiotic and biotic components. As a group of ubiquitous fungi that form mutualistic symbiotic associations with a vast array of terrestrial plants, arbuscular mycorrhizal (AM) fungi may play a critical role in maintaining soil multifunctionality, but the characteristics of their contributions remain to be unraveled. This mini review aims to disentangle the contributions of AM fungi to soil multifunctionality. We provide a framework of concepts about AM fungi making crucial contributions to maintaining multiple soil functions, including primary productivity, nutrient cycling, water regulation and purification, carbon and climate regulation, habitat for biodiversity, disease and pest control, and pollutant degradation and detoxification, via a variety of pathways, particularly contributing to soil and plant health. This review contends that AM fungi, as a keystone component of soil microbiome, can govern soil multifunctionality, ultimately promoting ecosystem services.     

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.