Fungal diversity in set-aide agricultural soil investigated using terminal-restriction fragment length polymorphism

As part of the restoration of biodiversity on former agricultural land there has been focused on methods to enhance the rate of transition from agricultural land towards natural grasslands or forest ecosystems. Management practices such as sowing seed mixtures and inoculating soil of later successional stages have been used. The aim of this study was to determine the effects of a managed plant community on the diversity of soil fungi in a newly abandoned agricultural land. A field site was set up consisting of 20 plots where the plant diversity was managed by either sowing 15 plant species, or natural colonization was allowed to occur. The plant mixture contained five species each of grasses, legumes and forbs that all were expected to occur at the site. A subset of the plots (five from each treatment) was inoculated with soil cores from a late successional stage. The plant community composition was subject to a principal component analysis based on the coverage of each species. Five years after abandonment, soil samples were taken from the plots, DNA was extracted and the ITS region of the rDNA gene was amplified using fluorescently labelled fungal specific primers (ITS 1F/ITS 4). The PCR products were digested using HinfI and TaqI and sequenced. Results from both restriction enzymes were combined and a principal component analysis performed on the presence/absence of fragments. Also the fungal diversity expressed as number of restriction fragments were analysed. There was significantly higher fungal species richness in the experimental plots compared to the forest and field soils, but no differences between sown and naturally colonized plots. The different plant treatments did not influence the below ground fungal community composition. Soil water content on the other hand had an impact on the fungal community composition.     

土壤微生物多样性研究的新方法

传统的分离培养和鉴定土壤微生物方法所具有的困难性和局限性 ,是造成难以深入了解土壤微生物生态学特性和多样性组成方面的主要障碍。本文运用分子生物学技术 ,以澳大利亚两种主要森林类型的土壤微生物多样性研究为实例 ,介绍了从土壤中直接提取土壤微生物DNA的方法以及末端限制性酶切片段长度多态性 (T RFLP)分析的基本原理和方法。作者认为 ,用该方法提取的土壤真菌DNA的纯度高 ,完全适合PCR扩增和T RFLP分析的要求。T RFLP已成为国外深入研究土壤微生物多样性的理想方法之一     

RNA- and DNA-based profiling of soil fungal communities in a native Australian eucalypt forest and adjacent Pinus elliotti plantation

Total and active soil fungal communities in a native eucalypt forest and first rotation Pinus elliotti plantation were investigated by direct extraction of DNA and RNA from soil. Terminal restriction fragment length polymorphism (T-RFLP) analysis of internal transcribed spacer (ITS) and 18S rRNA profiles indicated that total and active fungal communities differed significantly in both forest types. This was supported by DGGE profile analysis on an individual plot basis for both forest types and when groups in the canonical analysis were redefined to allow comparison between forest types. Analyses of both ITS and 18S T-RFLP profiles indicated that conversion from native eucalypt forest to P. elliottii plantation may significantly alter total and active soil fungal communities. ITS DGGE (DNA) and 18S (RNA) profiles also suggested differences in fungal communities in the two forest types. No significant separation of the fungal communities in the two forest types was observed, however, when ITS DGGE (RNA) profiles were compared. Overall, the data suggest that conversion from native eucalypt forest to P. elliottii plantation at the Beerburrum State Forest in subtropical Australia has significantly altered soil fungal communities.     

Soil carbon content drives the biogeographical distribution of fungal communities in the black soil zone of northeast China

Black soils (Mollisols) are one of the most important soil resources for maintaining food security in China, and they are mainly distributed in northeast China. A previous comprehensive study revealed the biogeographical distribution patterns of bacterial communities in the black soil zone. In this study, we used the same soil samples and analyzed the 454 pyrosequencing data for the nuclear ribosomal internal transcribed spacer (ITS) region to examine the fungal communities in these black soils. A total of 220,812 fungal ITS sequences were obtained from 26 soil samples that were collected across the black soil zone. These sequences were classified into at least 5 phyla, 20 classes, greater than 70 orders and over 350 genera, suggesting a high fungal diversity across the black soils. The diversity of fungal communities and distribution of several abundant fungal taxa were significantly related to the soil carbon content. Non-metric multidimensional scaling and canonical correspondence analysis plots indicated that the fungal community composition was most strongly affected by the soil carbon content followed by soil pH. This finding differs from the bacterial community results, which indicated that soil pH was the most important edaphic factor in determining the bacterial community composition of these black soils. A variance partitioning analysis indicated that the geographic distance contributed 20% of the fungal community variation and soil environmental factors that were characterized explained approximately 35%. A pairwise analysis revealed that the diversity of the fungal community was relatively higher at lower latitudes, which is similar to the findings for the bacterial communities in the same region and suggests that a latitudinal gradient of microbial community diversity might occur in the black soil zone. By incorporating our previous findings on the bacterial communities, we can conclude that contemporary factors of soil characteristics are more important than historical factor of geographic distance in shaping the microbial community in the black soil zone of northeast China.     

Response of the soil fungal community to multi-factor environmental changes in a temperate forest

Both environmental and climatic changes are known to influence soil microbial biomes in terrestrial ecosystems. However, there are limited data defining the interactive effects of multi-factor environmental disturbances, including N-deposition, precipitation, and air temperature, on soil fungal communities in temperate forests. A 3-year outdoor pot experiment was conducted to examine the temporal shifts of soil fungal communities in a temperate forest following N-addition, precipitation and air temperature changes. The shifts in the structure and composition of soil fungal communities were characterized by denaturing gradient gel electrophoresis and DNA sequencing. N-addition regimen induced significant alterations in the composition of soil fungal communities, and this effect was different at both higher and lower altitudes. The response of the soil fungal community to N-addition was much stronger in precipitation-reduced soils compared to soils experiencing enhanced precipitation. The combined treatment of N-addition and reduced precipitation caused more pronounced changes in the lower altitude versus those in the higher one. Certain fungal species in the subphylum Pezizomycotina and Saccharomycotina distinctively responded to N fertilization and soil water control at both altitudes. Redundancy discrimination analysis showed that changes in environmental factors and soil physicochemical properties explained 43.7% of the total variability in the soil fungal community at this forest ecosystem. Variations in the soil fungal community were significantly related to the altitude, soil temperature, total soil N content (TN) and pH value (P < 0.05). We present evidence for the interactive effects of N-addition, water manipulation and air temperature to reshape soil fungal communities in the temperate forest. Our data could provide new insights into predicting the response of soil micro-ecosystem to climatic changes.     

Diversity of laccase genes from basidiomycetes in a forest soil

Fungal oxidative exo-enzymes lacking substrate specificity play a central role in the cycling of soil organic matter. Due to their broad ecological impact and available knowledge of their gene structure, laccases appeared to be appropriate markers to monitor fungi with this kind of oxidative potential in soils. A degenerate PCR-primer pair Cu1F/Cu2R, specific for basidiomycetes, was designed to assess directly the diversity of laccase genes in soils. PCR amplification of mycelial cultures and fruit-bodies of a wide spectrum of basidiomycetes, covering all functional groups (saprophytes, symbionts, and pathogens), produced multiple DNA fragments around 200 bp. A neighbor-joining tree analysis of the PCR-amplified laccase sequences showed a clear species-specificity, but also revealed that most fungal taxa possess several laccase genes showing a large sequence divergence. This sequence diversity precluded the systematic attribution of amplified laccase of unknown origin to specific taxa. Amplification of laccase sequences from DNA, extracted from a brown (moder) forest soil, showed a specific distribution of laccase genes and of the corresponding fungal species in the various soil horizons (O_h, A_h, B_v). The most organic O_h-horizon displayed the highest gene diversity. Saprophytic fungi appeared to be less widespread through the soil horizons and displayed a higher diversity of laccase genes than the mycorrhizal ones.     

不同土地利用方式对三峡库区消落带土壤细菌和真菌多样性的影响

  目的  明确不同土地利用方式下第四纪古红土细菌和真菌群落特征,为古红土健康评价提供重要的生物性状数据,并为古红土资源的合理利用和科学管理提供指导。  方法  以疏林荒草地、荒草地、林地、耕地第四纪古红土和附近处于同一地层的埋藏第四纪古红土为研究对象,并以埋藏古红土作为对照组,利用高通量测序技术对不同土地利用方式下第四纪古红土细菌和真菌群落的丰富度、多样性和群落组成的变化进行分析,结合古红土理化性状,系统揭示不同土地利用方式下第四纪古红土细菌和真菌的群落特征。  结果  ①不同土地利用方式下第四纪古红土间微生物α 多样性指数存在显著差异,较埋藏古红土,疏林荒草地、荒草地、林地和耕地古红土的细菌丰富度指数和多样性指数以及真菌的丰富度指数均显著增加,耕地的真菌多样性指数显著降低。②埋藏古红土出露地表后不同利用方式下第四纪古红土细菌和真菌优势菌群的相对丰度发生显著变化。较埋藏古红土,其他不同土地利用方式下古红土中变形菌门的相对丰度均显著降低,酸杆菌门、绿弯菌门和芽单胞菌门的相对丰度均显著增加;疏林荒草地、荒草地和林地的子囊菌门的相对丰度显著降低,疏林荒草地和耕地的担子菌门的相对丰度显著增加,林地的被孢霉门的相对丰度显著增加。③古红土细菌和真菌群落的主坐标分析以及层次聚类分析显示,不同土地利用方式下第四纪古红土细菌和真菌群落结构发生变化,其中,荒草地与林地的群落结构最为相近。  结论  埋藏古红土出露地表后不同土地利用方式下第四纪古红土细菌和真菌丰富度、多样性以及群落组成都发生显著变化。研究结果可为开展古红土健康状况评价提供重要的生物性状数据,并为科学地管理与利用古红土资源奠定基础。     

Converting Australian tropical rainforest to native Araucariaceae plantations alters soil fungal communities

Native rainforest tree plantations are increasingly viewed as potentially important for high value timber production and provision of a range of ecological services in tropical and subtropical areas. In order to determine the extent to which conversion of rainforest to native Araucariaceae plantation influences soil fungi, we compared soil fungal communities under native rainforest and 73-74 year-old Araucaria bidwillii, Araucaria cunninghamii and Agathis robusta plantations at Gadgarra State Forest, Queensland, Australia. Following direct extraction of DNA from soil, terminal restriction fragment length polymorphism (T-RFLP) analysis of rDNA internal transcribed spacer (ITS) regions was conducted. Ordination analysis of the T-RFLP data revealed significant separation of the fungal communities according to forest type along the first canonical axis, with the native rainforest samples separating from the three Araucariaceae plantations along the second axis. Overall, the most abundant ITS sequences in clone assemblages from the four forest types were Ascomycota, followed by Basidiomycota, Zygomycota and Chitridomycota, however their relative importance varied in individual forest types. The results indicate that conversion of tropical rainforest to monoculture plantations of native trees can significantly alter soil fungal diversity.     

铜污染土壤线虫多样性的PCR-DGGE分析

A wheat pot experiment was conducted under greenhouse conditions to assess the effect of copper contamination on soil nematode diversity by polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） method and morphological analysis. The soil was treated with CuSO4.5H2O at the following concentrations： 0, 50, 100, 200, 400, and 800 mg kg^-1 dry soil, and the soil samples were collected at wheat jointing and ripening stages. Nematode diversity index （H′） from morphological analysis showed no difference between the control and the treated samples in either of the sampling dates. At the wheat ripening stage, nematode diversity obtained by the PCR-DGGE method decreased noticeably in the Cu800 treatment in comparison with the control. With optimization of the method of nematode DNA extraction, PCR-DGGE could give more information on nematode genera, and the intensity of the bands could reflect the abundance of nematode genera in the assemblage. The PCR-DGGE method proved promising in distinguishing nematode diversity in heavy metal coritaminated soil.     

Organic nitrogen cycling microbial communities are abundant in a dry Australian agricultural soil

Some microbial nitrogen (N) cycling processes continue under low soil moisture levels in drought-adapted ecosystems. These processes are of particular importance in winter cropping systems, where N availability during autumn sowing informs fertilizer practices and impacts crop productivity. We evaluated the organic and inorganic N-cycling communities in a key cropping soil (Vertosol), using a controlled-environment incubation study that was designed to model the autumn break in south Australian grain growing regions. Soils from wheat, lucerne, and green manure (disced-in vetch) rotations of the Sustainable Cropping Rotations in Mediterranean Environments trial (Victoria, Australia) were collected during the summer when soil moisture was low. Microbial community structure and functional capacity were measured both before and after wetting (21, 49, and 77 days post-wetting) using terminal restriction fragment length polymorphism measures of bacterial and fungal communities, and quantitative PCR of nitrogen cycling genes. Quantified genes included those associated with organic matter decomposition (laccase, cellobiohydrolase), mineralization of N from organic matter (peptidases) and nitrification (bacterial and archaeal ammonia monooxygenase and nitrite oxidoreductase). In general, the N cycling functional capacity decreased with soil wetting, and there was an apparent shift from organic-N cycling dominance to autotrophic mineral-N cycling dominance. Soil nitrate levels were best predicted by laccase and neutral peptidase genes under drought conditions, but by neutral peptidase and bacterial ammonia monooxygenase genes under moist conditions. Rotation history affected both the structural and functional resilience of the soil microbial communities to changing soil moisture. Discing in green manure (vetch) residues promoted a resilient microbial community, with a high organic-N cycling capacity in dry soils. Although this was a small-scale microcosm study, our results suggest that management strategies could be developed to control microbial organic-N processing during the summer fallow period and thus improve crop-available N levels at sowing.     

Urbanization alters the functional composition,but not taxonomic diversity,of the soil nematode community

We evaluated the response of riparian forest soil nematode community structure to the physico-chemical environment associated with urban land use. Soils were sampled seasonally between December 2000 and October 2002 along an urban—rural transect in Asheville, North Carolina. We characterized the taxonomic (to genus) and functional composition (trophic groups) of the nematode community of forest soils, as well as several nematode ecological indicators (maturity index, channel index, weighted faunal index). The diversity of nematode genera was not affected by urban land use. However, there tended to be functional differences in the nematode communities along the land use gradient. The urban soils tended to have lower abundances of predatory and omnivorous nematodes. Differences in channel index scores indicated that there was less fungal dominance in the soil food webs of the urban soils. Our results indicate that the functional composition of the soil food web is an important component of soil biodiversity that can be affected by land use practices. This study was conducted in a relatively small city; hence the influence of pollutants on the soil environment was not as great as in larger cities. Correspondingly, the impact on the soil nematode community was not very severe. The utilization of the nematode community assemblage as an indicator of soil conditions should be further explored in urban places of differing magnitudes of environmental effects.     

Microbial community development and unseen diversity recovery in inoculated sterile soil

Soil is considered as one of the most biodiverse environments on Earth; yet, the taxonomy, occurrence, and role of its different microbial populations are largely unknown. Here, two sterilized soils (from England and Italy) were inoculated with a subsample of their initial microbial communities and/or those from the other soil to study their microbial community evolution. This approach compared two driving factors (original community and soil physico-chemical characteristics) for microbial community definition. After 2 months of incubation and based on metagenomic datasets, the two inoculated communities (from an English grassland and an Italian forest) possessed similar functional and taxonomical structures when inoculated in the same sterile soil. For example, the newly colonized Italian soil was dominated by Actinobacteria related organisms (>66 % of the detected community) with a functional distribution independent of the inoculated soil origin. In addition, some of the organisms that dominated the different inoculated communities after 2 months were similar for a given sterile soil whether they came from the English grassland or the Italian forest, and they had not been detected in the original microbial community from either soil. Thus, similar microorganisms with low representation from the two distinct communities emerged in each sterilized soil, thus increasing the microbial diversity recovered from the microbial community of the donor soil. So far, these observations support the idea that different temperate soil microbial communities have different evenness due to environmental physico-chemical variations, yet have similar community composition (richness), and thus develop similarly when colonizing the same habitat.     

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.     

Influence of repeated prescribed burning on incorporation of C from cellulose by forest soil fungi as determined by RNA stable isotope probing

Repeated prescribed burning is frequently used as a forest management tool and can influence soil microbial diversity and activity. Soil fungi play key roles in carbon and nutrient cycling processes and soil fungal community structure has been shown to alter with increasing burning frequency. Such changes are accompanied by changes to soil carbon and nitrogen pools, however, we know little regarding how repeated prescribed burning alters functional diversity in soil fungal communities. We amended soil with ¹³C-cellulose and used RNA stable isotope probing to investigate the effect of biennial repeated prescribed burning over a 34-year period on cellulolytic soil fungi. Results indicated that repeated burning altered fungal community structure. Moreover, fungal community structure and diversity in ¹²C and ¹³C fractions from the unburned soil were not significantly different from each other, while those from the biennial burned soils differed from each other. The data indicate that fewer active fungi in the biennially burned soil incorporated ¹³C from the labelled cellulose and that repeated prescribed burning had a significant impact on the diversity of an important functional group of soil fungi (cellulolytic fungi) that are key drivers of forest soil decomposition and carbon cycling processes.     

Non-parametric multivariate comparisons of soil fungal composition: Sensitivity to thresholds and indications of structural redundancy in T-RFLP data

Complex soil microbial data produced by molecular profiling techniques, such as terminal restriction fragment length polymorphism (T-RFLP), are often analysed using multivariate statistical methods. Despite this, there has been little evaluation of the sensitivity of multivariate methods to routine data manipulations such as the application of noise thresholds. We examine effects of three percentage area thresholds (0.1%, 0.5% and 1.0%; i.e. ranging from low to commonly applied) on non-metric multidimensional scaling (NMDS) ordinations of soil fungal T-RFLP profiles. We then interpret threshold effects by introducing the concept of structural redundancy in T-RFLP data.NMDS ordinations compared 20 soil samples (encompassing seven sites, two forest types, and three locations) for T-RFLP presence/absence data, which were produced using primers specific to the rDNA internal transcribed spacer (ITS) region, and three separate restriction enzymes (HinfI, AluI and TaqI). Increasing thresholds from 0.1% to 1.0% led to decreases in average number of detected reproducible fragments per site of 57% (HinfI), 75% (AluI), and 69% (TaqI). However, despite the removal of many fragments unique to site/forest type/location, the application of increasing thresholds did not significantly change NMDS patterns for any enzyme in both the complete data and in a more weakly structured sub-group (involving one forest type and two locations). Thus, our data indicate that application of area thresholds up to 1.0% would have minimal impact on NMDS-based comparisons of soil fungal composition.Robustness of NMDS patterns to considerable fragment loss with increasing threshold suggested a degree of ‘structural redundancy’ in our T-RFLP data – that is, multiple fragments per soil sample were interchangeable in the way they contributed to between-sample NMDS patterns. Indeed, for each restriction enzyme, we found 3–4 peels of data (i.e. mutually exclusive sub-sets) that were capable of reproducing overall patterns in presence/absence data at the 0.1% threshold. Despite potential in the T-RFLP method for production of more than one fragment per fungal species/phylotype, we present arguments to support at least some translation of structural redundancy in T-RFLP data to structural redundancy in fungal community comparisons (i.e. the between-soil similarity patterns can be explained by, or are imprinted in, multiple groups of fungal phylotypes). Our analyses highlight structural redundancy as an efficient method for identifying key phylotypes responsible for differences in community composition among soils, and as a promising starting point for improved understanding of many aspects of spatial soil ecology.     

Structural and functional diversity of soil microbes is affected by elevated [CO<Subscript>2</Subscript>] and N addition in a poplar plantation

Background, Aims, and Scope  The genetic structure and the functionality of soil microbes are both important when studying the role of soil in the C cycle in elevated CO₂ scenarios. The aim of this work was to investigate the genetic composition of the fungal community by means of PCR-DGGE and the functional diversity of soil micro-organisms in general with MicroResp-based community level physiological profiling (CLPP) in a poplar plantation (POPFACE) grown under elevated [CO₂] with and without nitrogen fertilization. Materials and Methods  The POPFACE experimental plantation and FACE facility are located in central Italy, Tuscania (VT). Clones of Populus alba, Populus nigra and Populus x euramericana were grown, from 1999 to 2004, in six 314 m² plots treated either with atmospheric (control) or enriched (550 μmol mol⁻¹) CO₂ with FACE (Free Air CO₂ Enrichment) technology in each growing season. Each plot is divided into six triangular sectors, with two sectors per poplar genotype: three species × two nitrogen levels. After removal of the litter layer one soil core per genotype (10 cm wide, 20 cm depth) was taken inside each of the three sectors in each plot, for a total of 36 soil cores (3 replicates × 2 [CO₂] × 2 fertilization × 3 species) in October 2004 and in July 2005. DNA was extracted with a bead beating procedure. 18S rDNA gene fragments were amplified with PCR using fungal primers (FR1 GC and FF390). Analysis of CLPP was performed using the MicroResp method. Carbon substrates were selected depending on their ecological relevance to soil and their solubility in water. In particular rhizospheric C sources (carboxylic acids and carbohydrates) were chosen considering the importance of root inputs for microbial metabolism. Results  The fertilization treatment differentiated the fungal community composition regardless of elevated [CO₂] or the poplar species; moreover the number of fungal species was lower in fertilized soil. The effect of elevated [CO₂] on the fungal community composition was evident only as interaction with the fertilization treatment as, in N-sufficient soils, the elevated [CO₂] selected a different microbial community. For CLPP, the differ ent poplar species were the main factors of variation. The FACE treatment, on average, resulted in lower C utilization rates in un-fertilized soils and higher in fertilized soils. Discussion  Fungal biomass and fungal composition depend on different factors: from previous studies we know that the greater quantity and the higher C/N ratio of organic inputs under elevated [CO₂] influenced positively the fungal biomass both in fertilized and in un-fertilized soil, whereas nitrogen availability resulted to be the main determinant of fungal community composition in this work. Whole active microbial community was directly influenced by the soil nutrient availability and the poplar species. Under elevated CO₂ the competition for N with plants strongly affected the microbial communities, which were not able to benefit from added rhizospheric substrates. Under Nsufficient conditions, the increase of microbial activity due to [CO₂] enrichment was related to a more active microbial community, favoured by the current availability of C and N. Conclusions  Different factors influenced the microbial community at different levels: poplar species and root exudates affected the functional properties of the microbial community, while the fungal specific composition (as seen with DGGE) remained unaffected. On the other hand, factors such as N and C availability had a strong impact on the community functionality and composition. Fungal community structure reflected the availability of N in soils and the effect of elevated [CO₂] on community structure and function was evident only in N-sufficient soils. The simultaneous availability of C and N was therefore the main driving force for microbial structure and function in this plantation. Recommendations and Perspectives  Using the soil instead of soil extracts for CLPP determination provides a direct measurement of substrate catabolism by microbial communities and reflects activity rather than growth because more immediate responses to substrates are measured. Further applications of this approach could include selective inhibition of different microbial functional groups to investigate specific CLPPs. To combine the structural analysis and the catabolic responses of specific microbial communities (i.e. fungi or bacteria) could provide new outlooks on the role of microbes on SOM decomposition. ESS-Submission Editor: Dr. Kirk Semple (k.semple@lancaster.ac.uk)     

Spatial distribution of fungal communities in a coastal grassland soil

In grasslands, saprotrophic fungi, including basidiomycetes, are major decomposers of dead organic matter, although spatial distributions of their mycelial assemblages are little described. The aim of this study was to characterise the scale and distribution of saprotrophic fungal communities in a coastal grassland soil using terminal restriction fragment length polymorphism (T-RFLP).Soil fungi were sampled at Point Reyes, California, USA, by taking forty-five 26 mm diam. cores in a spatially defined manner. Within each sampled core, complete core sections at 1-2 cm and 14-15 cm depths were removed and sub-sampled for DNA extraction and amplification using the primer pairs ITS1F-FAM/ITS4 (general fungi) or ITS1F-FAM/ITS4B (basidiomycete-specific).Nonmetric Multidimensional Scaling showed that general fungal communities could be clearly separated by depth, although basidiomycete communities could not. There were no strong patterns of community similarity or dissimilarity for general or basidiomycete fungal communities at horizontal geographical distances from 25 cm to 96 m in the upper horizon. These results show considerable vertical, but little horizontal, variability in fungal community structure in a semi-natural grassland at the spatial scales measured here.     

Bottom–up or top–down control in forest soil microcosms? Effects of soil fauna on fungal biomass and C/N mineralisation

A major question in soil ecology is whether soil food webs are regulated by resources or by predators, i.e. bottom–up (donor) or top–down controlled. We tested the hypothesis that meso- and macrofaunal soil predators can regulate fungivore populations and, thereby cause a top–down cascade effect on fungal biomass and decomposition/mineralisation processes in boreal forest soils. The study was performed as a microcosm experiment with two contrasting soils (humus layers), one poor and one rich in N, and with different combinations of fungivore and predator soil fauna added to “defaunated” soil. In comparison with control microcosms lacking mesofauna (but with nematodes and protozoans), the presence of a diverse Collembola and Oribatida fungivore community significantly reduced the FDA-active fungal biomass or tended to reduce the ergosterol fraction of the fungal biomass in the N-poor humus, but no clear effect could be detected in the N-rich humus. Fungivores as well as fungivores plus predators (a predator community consisting of gamasids, spiders and beetles or a subset thereof) reduced C mineralisation and increased net N mineralisation in both soils. The presence of predators (particularly gamasid mites) reduced collembolan numbers and alleviated the negative effect of fungivores on fungal biomass in the N-poor soil. In the N-rich soil, the presence of predators increased fungal biomass (ergosterol) in relation to the “defaunated” soil. Therefore, a top–down trophic cascade could be detected in the N-poor humus but not in the N-rich humus. Our results suggest that the degree of top–down control in soil fauna communities depends on resource quality and soil fertility.