Simulated Nitrogen Deposition Causes a Decline of Intra- and Extraradical Abundance of Arbuscular Mycorrhizal Fungi and Changes in Microbial Community Structure in Northern Hardwood Forests

Increased nitrogen (N) deposition caused by human activities has altered ecosystem functioning and biodiversity. To understand the effects of altered N availability, we measured the abundance of arbuscular mycorrhizal fungi (AMF) and the microbial community in northern hardwood forests exposed to long-term (12 years) simulated N deposition (30 kg N ha⁻¹ y⁻¹) using phospholipid fatty acid (PLFA) analysis and hyphal in-growth bags. Intra- and extraradical AMF biomass and total microbial biomass were significantly decreased by simulated N deposition by 36, 41, and 24%, respectively. Both methods of extraradical AMF biomass estimation (soil PLFA 16:1ω5c and hyphal in-growth bags) showed comparable treatment responses, and extraradical biomass represented the majority of total (intra-plus extraradical) AMF biomass. N deposition also significantly affected the microbial community structure, leading to a 10% decrease in fungal to bacterial biomass ratios. Our observed decline in AMF and total microbial biomass together with changes in microbial community structure could have substantial impacts on the nutrient and carbon cycling within northern hardwood forest ecosystems.     

Lack of arbuscular mycorrhizal colonisation in tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis> L.) plants cultivated in Northern Iran

Soil and roots associated with different tea clones and nearby weeds (Veronica sp., Setaria sp., Salvia sp., Senecio sp. and Tripogon sp.) were sampled for arbuscular mycorrhizal fungi (AMF) in the tea gardens of Northern Iran. Spores were searched for in the soil and AMF colonisation determined microscopically and fatty acid signatures in roots was determined. Root samples from mycorrhizal and non-mycorrhizal clover were used as positive and negative controls. AMF spores were abundant in the tea garden soils; the genera Glomus and Acaulospora dominated. Microscopic observations of stained tea roots showed no sign of AMF. To confirm this, the roots were analysed for fatty acid signature compounds. The average level of PLFA 16:1ω5 as signature molecule for AMF in tea roots was 2 nmol g⁻¹ dry root, while the NLFA 16:1ω5 was not detectable. In mycorrhizal and non-mycorrhizal clover roots, the PLFA 16:1ω5 was 141and 5.74 nmol g⁻¹ dry root, respectively. In roots of weeds in tea plantations, the amount of PLFA 16:1ω5 was in the range 4.9 to 31.1 nmol g⁻¹ dry root. Thus, there was no evidence for AMF association in tea roots and weeds are thought to be the source of the spores in the soils. Finally, no mycorrhizal colonisation was found when tea plant seedlings were inoculated with AMF in pot cultures.     

Does percent root length colonization and soil hyphal length reflect the extent of colonization for all AMF?

Percent root length colonization may not be an appropriate measure of root colonization by arbuscular mycorrhizal fungi (AMF) in all cases. We suggest that AMF will differ in how well percent root length colonization measures the amount of AMF colonization in the root due to differences among AMF in hyphal structure and hyphal aggregation. Although soil hyphal length accounts for hyphal density, we suggest that it does not consider differences in hyphal structure in measurements of external colonization and thus might also misrepresent the true amount of AMF in the soil. To test these suggestions, we measured and compared percent root length colonization and soil hyphal length with root ergosterol and soil ergosterol, respectively, for 21 different species of AMF from three families in a greenhouse experiment. Percent root length colonization predicted intra-radical colonization best for Glomaceae and Acaulosporaceae isolates, while soil hyphal length best represented soil ergosterol for Gigasporaceae isolates. The results show that conventional methods for estimating AMF colonization are not universal for all AMF. Caution is advised when drawing inferences for different groups of AMF.     

Effects of long-term fertilization on AM fungal community structure and Glomalin-related soil protein in the Loess Plateau of China

Arbuscular mycorrhizal fungi (AMF) are crucial for ecosystem functioning, and thus have potential use for sustainable agriculture. In this study, we investigated the impact of organic and mineral fertilizers on the AMF community composition and content of Glomalin-related soil protein (GRSP) in a field experimental station which was established in 1979, in the Loess Plateau of China. Roots and soils were sampled three times during the growing period of winter wheat in 2008. The treatments including: N (inorganic N), NP (inorganic N and P), SNP (straw, inorganic N and P), M (farmyard manure), MNP (farmyard manure, inorganic N and P), and CK (no fertilization). AMF communities of root and soil samples were analyzed using PCR-DGGE, cloning and sequencing techniques; and GRSP content was determined by Bradford assay. Our results indicated that spore density, GRSP, and AMF community varied significantly in soils of long-term fertilization plots at three different wheat growing stages. The effects of wheat growing period on AMF community in roots were much more evident than fertilization regimes. However, the diversity of AMF was low in our study field. Up to five AMF phylotypes appeared in each sample, with the overwhelming dominance of a Glomus-like phylotype affiliated to G. mosseae. GRSP content was correlated positively with organic carbon, total phosphorus, available phosphorus, soil pH, and spore densities, but correlated negatively with soil C/N (P?<?0.05). The results of our study highlight that the richness of AMF in Loess Plateau agricultural region is low, and long-term fertilization, especially amendments with manure and straw, has beneficial effects on accumulation of soil organic carbon, spore density, GRSP content, and AMF diversity. Host phenology, edaphic factors (influenced by long-term fertilization), and habitats interacted to affect the AMF community and agoecosystem functioning. Additionally, soil moisture and pH make a greater contribution than other determined soil parameters to the AMF community dynamics in such a special semi-arid agroecosystem where crops rely greatly on rainfall.     

N-P fertilization did not reduce AMF abundance or diversity but alter AMF composition in an alpine grassland infested by a root hemiparasitic plant

Fertilization has been shown to have suppressive effects on arbuscular mycorrhizal fungi (AMF) and root hemiparasites separately in numerous investigations, but its effects on AMF in the presence of root hemiparasites remain untested. In view of the contrasting nutritional effects of AMF and root hemiparasites on host plants, we tested the hypothesis that fertilization may not show strong suppressive effects on AMF when a plant community was infested by abundant hemiparasitic plants. Plants and soil samples were collected from experimental field plots in Bayanbulak Grassland, where N and P fertilizers had been applied for three continuous years for control against a spreading root hemiparasite, Pedicularis kansuensis. Shoot and root biomass of each plant functional group were determined. Root AMF colonization levels, soil spore abundance, and extraradical hyphae length density were measured for three soil depths (0-10 cm, 10-20 cm, 20-30 cm). Partial 18S rRNA gene sequencing was used to detect AMF diversity and community composition. In addition, we analyzed the relationship between relative abundance of different AMF genera and environmental factors using Spearman's correlation method. In contrast to suppressive effects reported by many previous studies, fertilization showed no significant effects on AMF root colonization or AMF species diversity in the soil. Instead, a marked increase in soil spore abundance and extraradical hyphae length density were observed. However, fertilization altered relative abundance and AMF composition in the soil. Our results support the hypothesis that fertilization does not significantly influence the abundance and diversity of AMF in a plant community infested by P. kansuensis.     

Growth of ectomycorrhizal mycelia and composition of soil microbial communities in oak forest soils along a nitrogen deposition gradient

Deciduous forests may respond differently from coniferous forests to the anthropogenic deposition of nitrogen (N). Since fungi, especially ectomycorrhizal (EM) fungi, are known to be negatively affected by N deposition, the effects of N deposition on the soil microbial community, total fungal biomass and mycelial growth of EM fungi were studied in oak-dominated deciduous forests along a nitrogen deposition gradient in southern Sweden. In-growth mesh bags were used to estimate the production of mycelia by EM fungi in 19 oak stands in the N deposition gradient, and the results were compared with nitrate leaching data obtained previously. Soil samples from 154 oak forest sites were analysed regarding the content of phospholipid fatty acids (PLFAs). Thirty PLFAs associated with microbes were analysed and the PLFA 18:2ω6,9 was used as an indicator to estimate the total fungal biomass. Higher N deposition (20 kg N ha⁻¹ y⁻¹ compared with 10 kg N ha⁻¹ y⁻¹) tended to reduce EM mycelial growth. The total soil fungal biomass was not affected by N deposition or soil pH, while the PLFA 16:1ω5, a biomarker for arbuscular mycorrhizal (AM) fungi, was negatively affected by N deposition, but also positively correlated to soil pH. Other PLFAs positively affected by soil pH were, e.g., i14:0, a15:0, 16:1ω9, a17:0 and 18:1ω7, while some were negatively affected by pH, such as i15:0, 16:1ω7t, 10Me17:0 and cy19:0. In addition, N deposition had an effect on the PLFAs 16:1ω7c and 16:1ω9 (negatively) and cy19:0 (positively). The production of EM mycelia is probably more sensitive to N deposition than total fungal biomass according to the fungal biomarker PLFA 18:2ω6,9. Low amounts of EM mycelia covaried with increased nitrate leaching, suggesting that EM mycelia possibly play an important role in forest soil N retention at increased N input.     

Soil disturbance alters plant community composition and decreases mycorrhizal carbon allocation in a sandy grassland

We have studied how disturbance by ploughing and rotavation affects the carbon (C) flow to arbuscular mycorrhizal (AM) fungi in a dry, semi-natural grassland. AM fungal biomass was estimated using the indicator neutral lipid fatty acid (NLFA) 16:1ω5, and saprotrophic fungal biomass using NLFA 18:2ω6,9. We labeled vegetation plots with ¹³CO₂ and studied the C flow to the signature fatty acids as well as uptake and allocation in plants. We found that AM fungal biomass in roots and soil decreased with disturbance, while saprotrophic fungal biomass in soil was not influenced by disturbance. Rotavation decreased the ¹³C enrichment in NLFA 16:1ω5 in soil, but ¹³C enrichment in the AM fungal indicator NLFA 16:1ω5 in roots or soil was not influenced by any other disturbance. In roots, ¹³C enrichment was consistently higher in NLFA 16:1ω5 than in crude root material. Grasses (mainly Festuca brevipila) decreased as a result of disturbance, while non-mycorrhizal annual forbs increased. This decreases the potential for mycorrhizal C sequestration and may have been the main reason for the reduced mycorrhizal C allocation found in disturbed plots. Disturbance decreased the soil ammonium content but did not change the pH, nitrate or phosphate availability. The overall effect of disturbance on C allocation was that more of the C in AM fungal mycelium was directed to the external phase. Furthermore, the functional identity of the plants seemed to play a minor role in the C cycle as no differences were seen between different groups, although annuals contained less AM fungi than the other groups.     

不同施肥处理对丛枝菌根真菌生态分布的影响

研究了在东北海伦实验站长期定位培肥实验地不同施肥处理下丛枝菌根（Arbuscular Mycorrhizal,AM）真菌生长发育状况（包括侵染率、菌丝量和孢子数）,同时还分析了不同施肥处理下AM真菌群落生态分布和特征.结果表明施肥处理,尤其是磷肥（NP2K）处理显著降低AM真菌侵染玉米根系,而根外菌丝长度和孢子数并无显著变化,这和施肥处理下AM真菌的种群结构发生变化有关.随着土壤肥力的增高,土壤中AM真菌种的丰度和密度都有增加的趋势,而当肥力增高到一定程度后（磷肥和钾肥继续增加到NP2K和NPK2处理后）,土壤中AM真菌种的丰度和密度都有下降的趋势;从AM真菌属在不同肥力处理下出现的频度来看,Glomus属在7个处理中出现的频度最高,在每一个肥力处理中都有分布,Acaullospora属次之,Entrohospora属则只是出现在NK处理下;而Glomus属中出现频度最高的种是Glomus mosseae,其次是Glomus caledonium;再次是Glomus diaphanium,这说明施肥处理会影响到AM真菌种属的分布,进而影响到AM真菌的群落结构和生态分布.     

Fluorescein Diacetate Hydrolysis as a Measure of Fungal Biomass in Soil

The fatty acid methyl esters of lipids extracted from an agricultural soil in the preharvest period of soybean or middle growth cycle from wheat were characterized and quantified by gas-liquid chromatography. The fatty acids 18:2ω6 and 16:1ω5 were used as markers of saprotrophic and arbuscular mycorrhizal fungi. In parallel, biomass estimation through plate counts in selective media for cellulolytic and saprotrophic fungi was also performed all throughout a soybean crop or middle growth cycle of wheat. As an enzymatic method, the fluorescein diacetate (FDA) hydrolytic activity of the samples was determined. Owing to the high relationship exhibited by FDA hydrolysis with organic carbon and total nitrogen content of soil, the enzymatic activity was correlated with the microbial biomass estimated through marker lipids or plate counts. The results obtained point out that FDA hydrolysis may be used as a rapid, cheap, and reliable estimator of fungal biomass. Received: 3 August 2000 / Accepted: 13 October 2000     

Effect of long-term tillage and mineral phosphorus fertilization on arbuscular mycorrhizal fungi in a humid continental zone of Eastern Canada

Aims

Evidence shows that tillage modifies soil properties, especially phosphorus (P) dynamics. Our objective was to disentangle long-term effects of P-fertilization and tillage on arbuscular mycorrhizal fungal (AMF) proliferation and community structure.

Methods

Changes in the community structure of AMF and in the density of their hyphae and spores induced by moldboard plow (MP) or no till (NT), and fertilization with 0, 17.5, or 35 kg?P?ha^?1 were sought in the 0–15 cm and 15–30 cm soil layers after soybean harvest, at a long-term (17 years) experimental site in a humid continental zone of eastern Canada. The relationships among AMF, soil and plant attributes were examined.

Results

The 0–15 cm and 15–30 cm soil layers had different properties under NT, but were similar under MP, after 17 years, and MP increased soil available P levels. Phosphorus fertilization increased P levels in soil and in soybean. Treatment effects on AMF spore and hyphal density at 0–15 cm were greater than that at 15–30 cm, whereas effects on AMF community structure did not change with soil depths. At 0–15 cm, P-fertilization increased AMF spore density and reduced AMF hyphal density, and MP reduced AMF spore density. A total of eight AMF phylotypes were detected. Phosphorus fertilization reduced AMF phylotype richness and Shannon diversity index. Soil P availability increased under MP and hence the influence of P-fertilization treatments on the frequency of AMF phylotype detection varied with tillage system; it declined with P-fertilization under MP, but increased under NT.

Conclusions

Phosphorus fertilization shifts resource partitioning in AMF propagules rather than in their hyphae, and degrades the genetic diversity of AMF in soil; tillage increases soil P availability and hence aggravates the impact of P-fertilization.     

Seasonal changes of arbuscular mycorrhizal fungi as affected by tillage practices and fertilization : Hyphal density and mycorrhizal root colonization

The influence of tillage practices on native arbuscular mycorrhizal fungi (AMF) was studied in two, consecutive years in eastern Canada, in two 11 year-old long-term tillage-fertilizer experimental field soils, a sandy loam and a clay, growing corn in monoculture. The three tillage practices were: 1) conventional tillage (CT; fall plowing plus spring disking), reduced tillage (RT; spring disking) and no-till (NT). The corn crop received either inorganic (N and K) or organic (liquid dairy manure) fertilizers. Mycorrhizal hyphal density was estimated from soil samples obtained in early spring (before disking), at the 12–14 leaf stage, at silking, and at harvest. The percentage of corn root colonization by AMF at the 12–14 leaf stage, at silking and at harvest was also determined. The sandy loam was sampled over two consecutive seasons and the clay soil over one season.Densities of total and metabolically active soil hyphae, and mycorrhizal root colonization were significantly lower in CT soil than in RT and NT soil. Lowest soil hyphal densities were observed in early spring. The levels of intra- and extraradical fungal colonization always increased from spring to silking and decreased thereafter. Spring disking had only a small and transient negative effect on hyphal abundance in soil. Fertilization did not influence mycorrhizal colonization of corn or abundance of soil hyphae in the sandy loam soil, but in the clay soil metabolically active hyphae were more abundant with manure application than with mineral fertilization. In 1992, in both soils different tillage systems had same grain yield, however, in 1993, corn yield was higher in NT compared to CT system.     

Long-Term Phosphorus Fertilization Impacts Soil Fungal and Bacterial Diversity but not AM Fungal Community in Alfalfa

Soil function may be affected by cropping practices impacting the soil microbial community. The effect of different phosphorus (P) fertilization rates (0, 20, or 40 kg P₂O₅ ha⁻¹) on soil microbial diversity was studied in 8-year-old alfalfa monocultures. The hypothesis that P fertilization modifies soil microbial community was tested using denaturing gradient gel electrophoresis and phospholipids fatty acid (PLFA) profiling to describe soil bacteria, fungi, and arbuscular mycorrhizal (AM) fungi diversity. Soil parameters related to fertility (soil phosphate flux, soluble P, moisture, phosphatase and dehydrogenase assays, and carbon and nitrogen content of the light fraction of soil organic matter) were also monitored and related to soil microbial ribotype profiles. Change in soil P fertility with the application of fertilizer had no effect on crop yield in 8 years, but on the year of this study was associated with shifts in the composition of fungal and bacterial communities without affecting their richness, as evidenced by the absence of effect on the average number of ribotypes detected. However, variation in soil P level created by a history of differential fertilization did not significantly influence AM fungi ribotype assemblages nor AM fungi biomass measured with the PLFA 16:1ω5. Fertilization increased P flux and soil soluble P level but reduced soil moisture and soil microbial activity, as revealed by dehydrogenase assay. Results suggest that soil P fertility management could influence soil processes involving soil microorganisms. Seasonal variations were also recorded in microbial activity, soil soluble P level as well as in the abundance of specific bacterial and fungal PLFA indicators of soil microbial biomass.     

The growth of external AM fungal mycelium in sand dunes and in experimental systems

We estimated the biomass and growth of arbuscular mycorrhizal (AM) mycelium in sand dunes using signature fatty acids. Mesh bags and tubes, containing initially mycelium-free sand, were buried in the field near the roots of the dune grass Ammophila arenaria L. AM fungal mycelia were detected at a distance of about 8.5 cm from the roots after 68 days of growth by use of neutral lipid fatty acid (NLFA) 16:1ω5. The average rate of mycelium extension during September and October was estimated as 1.2 mm day⁻¹. The lipid and fatty acid compositions of AM fungal mycelia of isolates and from sand dunes were analysed and showed all to be of a similar composition. Phospholipid fatty acids (PLFAs) can be used as indicators of microbial biomass. The mycelium of G. intraradices growing in glass beads contained 8.3 nmol PLFAs per mg dry biomass, and about 15% of the PLFAs in G. intraradices, G. claroideum and AM fungal mycelium extracted from sand dunes, consisted of the signature PLFA 16:1ω5. We thus suggest a conversion factor of 1.2 nmol PLFA 16:1ω5 per mg dry biomass. Calculations using this conversion factor indicated up to 34 μg dry AM fungal biomass per g sand in the sand dunes, which was less than one tenth of that found in an experimental system with Glomus spp. growing with cucumber as plant associate in agricultural soil. The PLFA results from different systems indicated that the biomass of the AM fungi constitutes a considerable part of the total soil microbial biomass. Calculations based on ATP of AM fungi in an experimental growth system indicated that the biomass of the AM fungi constituted approximately 30% of the total microbial biomass. This revised version was published online in June 2006 with corrections to the Cover Date.     

Determinants of Soil Microbial Communities: Effects of Agricultural Management, Season, and Soil Type on Phospholipid Fatty Acid Profiles

Abstract Phospholipid fatty acid (PLFA) profiles were measured in soils from organic, low-input, and conventional farming systems that are part of the long term Sustainable Agriculture Farming Systems (SAFS) Project. The farming systems differ in whether their source of fertilizer is mineral or organic, and in whether a winter cover crop is grown. Sustained increases in microbial biomass resulting from high organic matter inputs have been observed in the organic and low-input systems. PLFA profiles were compared to ascertain whether previously observed changes in biomass were accompanied by a change in the composition of the microbial community. In addition, the relative importance of environmental variables on PLFA profiles was determined. Redundancy analysis ordination showed that PLFA profiles from organic and conventional systems were significantly different from April to July. On ordination plots, PLFA profiles from the low-input system fell between organic and conventional systems on most sample dates. A group of fatty acids (i14:0, a15:0, 16:1ω7c, 16:1ω5c, 14:0, and 18:2ω6c) was enriched in the organic plots throughout the sampling period, and another group (10Me16:0, 2OH 16:1 and 10Me17:0) was consistently lower in relative abundance in the organic system. In addition, another group (15:0, a17:0, i16:0, 17:0, and 10Me18:0) was enriched over the short term in the organic plots after compost incorporation. The relative importance of various environmental variables in governing the composition of microbial communities could be ranked in the order: soil type > time > specific farming operation (e.g., cover crop incorporation or sidedressing with mineral fertilizer) > management system > spatial variation in the field. Measures of the microbial community and soil properties (including microbial biomass carbon and nitrogen, substrate induced respiration, basal respiration, potentially mineralizable nitrogen, soil nitrate and ammonium, and soil moisture) were seldom associated with the variation in the PLFA profiles. Received: 3 February 1997; Accepted: 7 August 1997     

Increased allocation to external hyphae of arbuscular mycorrhizal fungi under CO2 enrichment

Prunella vulgaris was inoculated with different arbuscular mycorrhizal fungi (AMF) and grown at two concentrations of CO₂ (ambient, 350 μl l⁻¹, and elevated, 600 μl l⁻¹) to test whether a plants response to elevated CO₂ is dependent on the species of AMF colonizing the roots. Using compartments accessible only to AMF hyphae but not to roots, we also tested whether elevated CO₂ affects the growth of external AMF hyphae. Plant biomass was significantly greater at elevated than at ambient CO₂; the biomass of the root system, for example, increased by a factor of 2. The colonization of AMF inside the root remained constant, indicating that the total AMF inside the root system also increased by a factor of 2. The length of external AMF hyphae at elevated CO₂ was up to 5 times that at ambient CO₂, indicating that elevated CO₂ promoted allocation of AMF biomass to the external hyphae. The concentration and content of phosphorus in the stolons differed significantly between ambient and elevated CO₂ but this resulted in either an increase or a decrease, according to which AMF isolate occupied the roots. We hypothesized that an increase in external hyphal growth at elevated CO₂ would result in increased P acquistion by the plant. To test this we supplied phosphorus, in a compartment only accessible to AMF hyphae. Plants did not acquire more phosphorus at elevated CO₂ when phosphorus was added to this compartment. Large increases in AMF hyphal growth could, however, play a significant role in the movement of fixed carbon to the soil and increase soil aggregation. Received: 28 March 1998 / Accepted: 27 August 1998     

Toxicity of diesel fuel to germination,growth and colonization of <Emphasis Type="Italic">Glomus intraradices</Emphasis> in soil and <Emphasis Type="Italic">in vitro</Emphasis> transformed carrot root cultures

Phytoremediation is the use of selected plants to decontaminate polluted environments. Arbuscular mycorrhizal fungi (AMF) may potentially be useful for phytoremediation, but it is not known how petroleum hydrocarbons influence AMF spore germination and hyphal growth. To address this question, germination of spores and germ tube growth of Glomus intraradices Schenck and Smith and Glomus aggregatum Schenck and Smith were assessed in soil contaminated with up to 3% (w/v) of F2 diesel oil or HAGO reference oil. Hyphal growth, colonization and progeny spore production were assessed in vitro using transformed root cultures of Daucus carota and G. intraradices spores in a F2 diesel contaminated medium. In addition, extraradical hyphal growth of G. intraradices colonizing Daucus carota in the presence of F2 diesel was studied. Neither F2 diesel nor HAGO reference oil affected spore germination or germ tube growth in soil. However, in the presence of plant roots, germ tube growth of G. intraradices was reduced and delayed in the presence of F2 diesel and root colonization was not detected. Hyphal growth of pre-colonized carrot roots by G. intraradices was reduced and delayed in F2 contaminated medium compared to controls. F2 diesel did not inhibit spore germination of these AMF species but did reduce colonization, germ tube and hyphal growth. These results suggest that AMF inoculum can be established in petroleum-contaminated sites. However, it may prove beneficial to plant pre-colonized plants to increase the probability of sufficient AMF colonization and growth. The likely mechanism(s) of petroleum toxicity in this plant-microbe system was discussed.     

Seasonality of arbuscular mycorrhizal hyphae and glomalin in a western Montana grassland

In order to more fully understand the basic biology of arbuscular mycorrhizal fungi (AMF), and their role in natural ecosystems, it is necessary to document seasonal changes of various aspects of the life history of these fungi. Due to their unique position at the root-soil interface, AMF have been described as `keystone mutualists' in ecosystems. Despite the importance of AMF in ecosystems, few studies exist that examine the seasonality of external hyphae and their exuded products (e.g. glomalin), the AMF variables directly related to ecosystem function through their contributions to soil aggregation. This study examined seasonal dynamics of several soil variables, with a specific interest in the seasonality of external hyphae and glomalin, a glycoprotein produced by AMF, which is correlated with soil aggregate stability. Here we measured glomalin concentrations and external AMF and non-AMF hyphal length, as well as soil moisture, percent fungal root colonization (AMF and non-AMF), and root length in soil in an intermountain grassland in western Montana over one growing season (13 time points). Of the glomalin pools and hyphal lengths measured, significant seasonal changes occurred for total glomalin (TG; 24.5% change), immunoreactive easily extractable glomalin (IREEG; 53.8% change), and AM hyphal length (107% change). Prior studies on glomalin in natural systems have not considered seasonal effects. The small seasonal change in glomalin pools lends further support to the hypothesis that glomalin is relatively stable in soils, and suggests that one-time sampling may be sufficient to satisfactorily capture this response variable. However, the generality of this observation has yet to be tested in a wider range of ecosystems.     

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

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

Phialophora graminicola, a dark septate fungus, is a beneficial associate of the grass Vulpia ciliata ssp. ambigua

The influence of three organic compounds and bakers' dry yeast on growth of external mycelium and phosphorus uptake of the arbuscular mycorrhizal fungus Glomus intraradices Schenck & Smith (BEG 87) was examined. Two experiments were carried out in compartmentalized growth systems with root-free sand or soil compartments. The sand and soil in the root-free compartments were left untreated or uniformly mixed with one of the following substrates (0.5 mg g⁻¹ soil): bakers' dry yeast, bovine serum albumin, starch or cellulose. Effects of the organic substrates on biomass and hyphal length density of the arbuscular mycorrhizal fungus were examined by using specific fatty acid signatures in combination with direct microscopy. Micro-organisms other than the arbuscular mycorrhizal fungus were measured by fatty acid signatures, and radioactive ³³P labelling of the root-free soil was used to determine arbuscular mycorrhizal hyphal phosphorus uptake. In general, hyphal growth of G. intraradices was enhanced by yeast and bovine serum albumin, whereas the carbon sources, starch and cellulose, depressed fungal growth. By analysing the fatty acid 16:1ω5 from phospholipids (indicating mycelium) and neutral lipids (indicating storage structures) it was shown that increased fungal growth due to yeast was mainly in vegetative hyphae and less in storage structures. Arbuscular mycorrhizal hyphal phosphorus uptake was decreased by cellulose, but unaffected by the other substrates compared with the control. This means that both growth and phosphorus transport by the arbuscular mycorrhizal fungus were decreased under cellulose treatment. However, the composition of the microbial community varied under different substrate conditions indicating a possible interactive component with arbuscular mycorrhizal hyphal growth and phosphorus uptake.     

Effects of long-term NP-fertilization on abundance and diversity of arbuscular mycorrhizal fungi under a maize cropping system

Diversity of arbuscular mycorrhizal fungi (AMF) in 27-year long-term NP-fertilization plots under a maize cropping system in Thailand was studied through spore morphological characterization. The plots received 0–0, 60–60, 120–120 and 180–180 kg N-P₂O₅ ha^–1 year^–1 as ammonium sulfate and triple superphosphate. The plots were sampled monthly for one year, the AMF spores were counted and morphotyped, and taxa were identified after morphotyping and monospecific pot culture. Spore number g^–1 soil, relative spore abundance and Shannon-Wiener indexes were calculated. Sixteen putative taxa were recorded from the field of which nine sporulated on maize roots in pot culture. The long-term fertilization caused decreases in AMF total spore numbers and variation in species diversity depended on sampling time. Effects of fertilization on spore number and also relative spore abundance varied with species and sampling time. Among the nine species sporulating under maize, only Acaulospora sp.1 showed no change (P > 0.003 after Bonferroni correction) in spore number with fertilization in the field; and was therefore classified as an AMF species insensitive to fertilization. Spores of Entrophospora schenckii, Glomus mosseae, Glomus sp.1, Glomus geosporum-like and Scutellospora fulgida, though they decreased in absolute numbers in response to fertilization, showed no change (P > 0.003 after Bonferroni correction) in relative abundance; these species were classified as AMF species slightly sensitive to fertilization. Three unidentified species of Glomus, though they decreased in absolute numbers in response to fertilization, showed decreases (P < 0.003 after Bonferroni correction) in relative abundance; these species were classified as AMF species highly sensitive to fertilization.