Plant diversity predicts beta but not alpha diversity of soil microbes across grasslands worldwide

Aboveground–belowground interactions exert critical controls on the composition and function of terrestrial ecosystems, yet the fundamental relationships between plant diversity and soil microbial diversity remain elusive. Theory predicts predominantly positive associations but tests within single sites have shown variable relationships, and associations between plant and microbial diversity across broad spatial scales remain largely unexplored. We compared the diversity of plant, bacterial, archaeal and fungal communities in one hundred and forty-five 1 m2 plots across 25 temperate grassland sites from four continents. Across sites, the plant alpha diversity patterns were poorly related to those observed for any soil microbial group. However, plant beta diversity (compositional dissimilarity between sites) was significantly correlated with the beta diversity of bacterial and fungal communities, even after controlling for environmental factors. Thus, across a global range of temperate grasslands, plant diversity can predict patterns in the composition of soil microbial communities, but not patterns in alpha diversity.

Bioavailability of atrazine to soil microbes in the presence of the earthworm Lumbricus terrestrius (L.)

Experiments were conducted to investigate the interactions between an earthworm species (Lumbricus terrestrius) and soil microflora with respect to the bioavailability and mineralisation of ¹⁴C ring-labelled atrazine. Presence of earthworms had no affect on atrazine in soil solution (assayed by soil centrifugation). This soil solution pool was highly time dependent, decreasing considerably as the experiment proceeded. KCl-extractable label was, however, affected by the presence of earthworms, with this pool initially increasing in the presence of the worms. This pool was also highly time-dependent although, the pattern of this dependence did not follow that for label in soil solution. Mineralisation of the atrazine closely followed the KCl exchangeable pool and not that of the soil solution pool. However, label sorbed to the surface of the worms was closely correlated to the soil solution pool. Mineralisation in the presence of earthworms was double that of the controls. By the end of the experiment 6% of added radioactivity was present in the earthworm biomass.

Soil microbes and community coalescence

Community coalescence is a recently introduced term describing the interaction of entire communities and their environments. We here explicitly place the concept of community coalescence in a soil microbial context, exploring intrinsic and extrinsic drivers of such coalescence events. Examples of intrinsic events include the action of earthworms and the dynamics of soil aggregates, while extrinsic events are exemplified by tillage, flooding, litter-fall, outplanting, and the addition of materials containing microbial communities. Aspects of global change may alter the frequency or severity of coalescence events. We highlight functional consequences of community coalescence in soil, and suggest ways to experimentally tackle this phenomenon. Soil ecology as a whole stands to benefit from conceptualizing soil biodiversity in terms of dynamic coalescent microbial assemblages.

The role of soil microbes in the global carbon cycle: tracking the below-ground microbial processing of plant-derived carbon for manipulating carbon dynamics in agricultural systems.

It is well known that atmospheric concentrations of carbon dioxide (CO2) (and other greenhouse gases) have increased markedly as a result of human activity since the industrial revolution. It is perhaps less appreciated that natural and managed soils are an important source and sink for atmospheric CO2 and that, primarily as a result of the activities of soil microorganisms, there is a soil-derived respiratory flux of CO2 to the atmosphere that overshadows by tenfold the annual CO2 flux from fossil fuel emissions. Therefore small changes in the soil carbon cycle could have large impacts on atmospheric CO2 concentrations. Here we discuss the role of soil microbes in the global carbon cycle and review the main methods that have been used to identify the microorganisms responsible for the processing of plant photosynthetic carbon inputs to soil. We discuss whether application of these techniques can provide the information required to underpin the management of agro-ecosystems for carbon sequestration and increased agricultural sustainability. We conclude that, although crucial in enabling the identification of plant-derived carbon-utilising microbes, current technologies lack the high-throughput ability to quantitatively apportion carbon use by phylogentic groups and its use efficiency and destination within the microbial metabolome. It is this information that is required to inform rational manipulation of the plant–soil system to favour organisms or physiologies most important for promoting soil carbon storage in agricultural soil.

The impact of land-use practices on soil microbes

The extensive use of land resources for food production, fibre for construction, wood pulp for paper, removal for extractive industries, sealing for urban and industrial development and as a receiver (either deliberate or accidental) of polluting substances has wrought huge changes in the chemistry, structure and biology of soils, away from their natural state.

Do arbuscular mycorrhizas or heterotrophic soil microbes contribute toward plant acquisition of a pulse of mineral phosphate?

Aims: We investigated the role of arbuscular mycorrhizal fungi (AMF) and heterotrophic soil microbes in the uptake of phosphorus (P) by Trifolium subterraneum from a pulse. Methods: Plants were grown in sterilised pasture field soil with a realistic level of available P. There were five treatments, two of which involved AMF: 1) unsterilised field soil containing a community of AMF and heterotrophic organisms; 2) Scutellospora calospora inoculum (AMF); 3) microbes added as filtrate from the field soil; 4) microbes added as filtrate from the S. calospora inoculum; 5) no additions, i.e. sterilised field soil. After 11 weeks, plants were harvested: 1 day before (day 0), 1 day after (day 2) and 7 days after (day 8) the pulse of P (10 mg kg−1). Results: There was no difference among treatments in shoot and root dry weight, which increased from day 0 to day 8. At day 0, shoots and roots of plants in the colonised treatments had higher P and lower Mn concentrations. After the pulse, the rate of increase in P concentration in the shoots was slower for the colonised plants, and the root Mn concentration declined by up to 50 % by day 2. Conclusions: Plants colonised by AMF had a lower rate of increase in shoot P concentration after a pulse, perhaps because intraradical hyphae accumulated P and thus reduced its transport to the shoots.

Autoclaving kills soil microbes yet soil enzymes remain active

Biocidal treatment of soil is used to remove or inhibit soil microbial activity, and thus provides insight into the relationship between soil biology and soil processes. Chemical (soil pH, phosphodiesterase, protease) and biological (substrate induced respiration) characteristics of three contrasting soils from tropical savanna ecosystems in north Queensland, Australia were measured in field fresh samples and following autoclaving (121 °C/103 kPa for 30 min on two consecutive days). Autoclaving treatment killed the active soil microbial biomass and significantly decreased protease activity (∼90%) in all three soils. Phosphodiesterase activity in kaolinitic soils also significantly decreased by 78% and 92%. However, autoclave treatment of smectitic soil only decreased phosphodiesterase activity by 4% only. This study demonstrates phosphodiesterase can remain stable in extreme conditions. This might be a characteristic vital to the cycling of phosphorus in shrink–swell clays in Australian tropical savanna ecosystems.

Effect of nitrification inhibitors (DMPP and 3MP+ TZ) on soil nitrous oxide emissions from a sub-tropical vegetable production system in Queensland, Australia

The use of nitrification inhibitors, in combination with ammonium based fertilisers, has been promoted recently as an effective method to reduce nitrous oxide (N2O) emissions from fertilised agricultural fields, whilst increasing yield and nitrogen use efficiency. Vegetable cropping systems are often characterised by high inputs of nitrogen fertiliser and consequently elevated emissions of nitrous oxide (N2O) can be expected. However, to date only limited data is available on the use of nitrification inhibitors in sub-tropical vegetable systems. A field experiment investigated the effect of the nitrification inhibitors (DMPP & 3MP+TZ) on N2O emissions and yield from a typical vegetable production system in sub-tropical Australia. Soil N2O fluxes were monitored continuously over an entire year with a fully automated system. Measurements were taken from three subplots for each treatment within a randomized complete blocks design. There was a significant inhibition effect of DMPP and 3MP+TZ on N2O emissions and soil mineral N content directly following the application of the fertiliser over the vegetable cropping phase. However this mitigation was offset by elevated N2O emissions from the inhibitor treatments over the post-harvest fallow period. Cumulative annual N2O emissions amounted to 1.22 kg-N/ha, 1.16 kg-N/ha, 1.50 kg-N/ha and 0.86 kg-N/ha in the conventional fertiliser (CONV), the DMPP treatment, the 3MP+TZ treatment and the zero fertiliser (0N) respectively. Corresponding fertiliser induced emission factors (EFs) were low with only 0.09 - 0.20% of the total applied fertiliser lost as N2O. There was no significant effect of the nitrification inhibitors on yield compared to the CONV treatment for the three vegetable crops (green beans, broccoli, lettuce) grown over the experimental period. This study highlights that N2O emissions from such vegetable cropping system are primarily controlled by post-harvest emissions following the incorporation of vegetable crop residues into the soil. It also shows that the use of nitrification inhibitors can lead to elevated N2O emissions by storing N in the soil profile that is available to soil microbes during the decomposition of the vegetable residues over the post-harvest phase. Hence the use of nitrification inhibitors in vegetable systems has to be treated carefully and fertiliser rates need to be adjusted to avoid excess soil nitrogen during the postharvest phase.

Comparative phytotoxicity of nitrophenolic soil pollutants and their microbial metabolites to the growth of cucumber

2,4-Dinitrophenol and paranitrophenol are two major soil pollutants which are known to be metabolized by different soil microbes. Relative phytotoxicities of these parent compounds and their metabolic transformation products to the growth of cucumber seedlings were assessed. It was evident that such microbial transformations widely occurring in the soil are effective detoxification reactions and are beneficial for the plants.

Assessing the relationship between patch type and soil mites: A molecular approach

An urgent need exists for indicators of soil health and patch functionality in extensive rangelands that can be measured efficiently and at low cost. Soil mites are candidate indicators, but their identification and handling is so specialised and time-consuming that their inclusion in routine monitoring is unlikely. The aim of this study was to measure the relationship between patch type and mite assemblages using a conventional approach. An additional aim was to determine if a molecular approach traditionally used for soil microbes could be adapted for soil mites to overcome some of the bottlenecks associated with soil fauna diversity assessment. Soil mite species abundance and diversity were measured using conventional ecological methods in soil from patches with perennial grass and litter cover (PGL), and compared to soil from bare patches with annual grasses and/or litter cover (BAL). Soil mite assemblages were also assessed using a molecular method called terminal-restriction fragment length polymorphism (T-RFLP) analysis. The conventional data showed a relationship between patch type and mite assemblage. The Prostigmata and Oribatida were well represented in the PGL sites, particularly the Aphelacaridae (Oribatida). For T-RFLP analysis, the mite community was represented by a series of DNA fragment lengths that reflected mite sequence diversity. The T-RFLP data showed a distinct difference in the mite assemblage between the patch types. Where possible, T-RFLP peaks were matched to mite families using a reference 18S rDNA database, and the Aphelacaridae prevalent in the conventional samples at PGL sites were identified, as were prostigmatids and oribatids. We identified limits to the T-RFLP approach and this included an inability to distinguish some species whose DNA sequences were similar. Despite these limitations, the data still showed a clear difference between sites, and the molecular taxonomic inferences also compared well with the conventional ecological data. The results from this study indicated that the T-RFLP approach was effective in measuring mite assemblages in this system. The power of this technique lies in the fact that species diversity and abundance data can be obtained quickly because of the time taken to process hundreds of samples, from soil DNA extraction to data output on the gene analyser, can be as little as 4 days.

Plant secondary compounds and soil microbial processes in carbon and nitrogen cycling in relation to tree species

The aim of this study was to explore soil microbial activities related to C and N cycling and the occurrence and concentrations of two important groups of plant secondary compounds, terpenes and phenolic compounds, under silver birch (Betula pendula Roth), Norway spruce (Picea abies (L.) Karst) and Scots pine (Pinus sylvestris L.) as well as to study the effects of volatile monoterpenes and tannins on soil microbial activities. The study site, located in Kivalo, northern Finland, included ca. 70-year-old adjacent stands dominated by silver birch, Norway spruce and Scots pine. Originally the soil was very probably similar in all three stands. All forest floor layers (litter (L), fermentation layer (F) and humified layer (H)) under birch and spruce showed higher rates of CO2 production, greater net mineralisation of nitrogen and higher amounts of carbon and nitrogen in microbial biomass than did the forest floor layers under pine. Concentrations of mono-, sesqui-, di- and triterpenes were higher under both conifers than under birch, while the concentration of total water-soluble phenolic compounds as well as the concentration of condensed tannins tended to be higher or at least as high under spruce as under birch or pine. In general, differences between tree species in soil microbial activities and in concentrations of secondary compounds were smaller in the H layer than in the upper layers. The rate of CO2 production and the amount of carbon in the microbial biomass correlated highly positively with the concentration of total water-soluble phenolic compounds and positively with the concentration of condensed tannins. Exposure of soil to volatile monoterpenes and tannins extracted and fractionated from spruce and pine needles affected carbon and nitrogen transformations in soil, but the effects were dependent on the compound and its molecular structure. Monoterpenes decreased net mineralisation of nitrogen and probably had a toxic effect on part of the microbial population in soil, while another part of the microbes seemed to be able to use monoterpenes as a carbon source. With tannins, low-molecular-weight compounds (also compounds other than tannins) increased soil CO2 production and nitrogen immobilisation by soil microbes while the higher-molecular-weight condensed tannins had inhibitory effects. In conclusion, plant secondary compounds may have a great potential in regulation of C and N transformations in forest soils, but the real magnitude of their significance in soil processes is impossible to estimate.

Responses of soil microbial communities to clonal variation of Norway spruce

In boreal forests, microorganisms have a pivotal role in nutrient and water supply of trees as well as in litter decomposition and nutrient cycling. This reinforces the link between above-ground and below-ground communities in the context of sustainable productivity of forest ecosystems. In northern boreal forests, the diversity of microbes associated with the trees is high compared to the number of distinct tree species. In this thesis, the aim was to study whether conspecific tree individuals harbour different soil microbes and whether the growth of the trees and the community structure of the associated microbes are connected. The study was performed in a clonal field trial of Norway spruce, which was established in a randomized block design in a clear-cut area. Since out-planting in 1994, the spruce clones showed two-fold growth differences. The fast-growing spruce clones were associated with a more diverse community of ectomycorrhizal fungi than the slow-growing spruce clones. These growth performance groups also differed with respect to other aspects of the associated soil microorganisms: the species composition of ectomycorrhizal fungi, in the amount of extraradical fungal mycelium, in the structure of bacterial community associated with the mycelium, and in the structure of microbial community in the organic layer. The communities of fungi colonizing needle litter of the spruce clones in the field did not differ and the loss of litter mass after two-years decomposition was equal. In vitro, needles of the slow-growing spruce clones were colonized by a more diverse community of endophytic fungi that were shown to be significant needle decomposers. This study showed a relationship between the growth of Norway spruce clones and the community structure of the associated soil microbes. Spatial heterogeneity in soil microbial community was connected with intraspecific variation of trees. The latter may therefore influence soil biodiversity in monospecific forests.

Two ex situ fungal technologies to treat contaminated soil

Wood-degrading fungi are able to degrade a large range of recalcitrant pollutants which resemble the lignin biopolymer. This ability is attributed to the production of lignin-modifying enzymes, which are extracellular and non-specific. Despite the potential of fungi in bioremediation, there is still an understanding gap in terms of the technology. In this thesis, the feasibility of two ex situ fungal bioremediation methods to treat contaminated soil was evaluated. Treatment of polycyclic aromatic hydrocarbons (PAHs)-contaminated marsh soil was studied in a stirred slurry-phase reactor. Due to the salt content in marsh soil, fungi were screened for their halotolerance, and the white-rot fungi Lentinus tigrinus, Irpex lacteus and Bjerkandera adusta were selected for further studies. These fungi degraded 40 - 60% of a PAH mixture (phenanthrene, fluoranthene, pyrene and chrysene) in a slurry-phase reactor (100 ml) during 30 days of incubation. Thereafter, B. adusta was selected to scale-up and optimize the process in a 5 L reactor. Maximum degradation of dibenzothiophene (93%), fluoranthene (82%), pyrene (81%) and chrysene (83%) was achieved with the free mycelium inoculum of the highest initial biomass (2.2 g/l). In autoclaved soil, MnP was the most important enzyme involved in PAH degradation. In non-sterile soil, endogenous soil microbes together with B. adusta also degraded the PAHs extensively, suggesting a synergic action between soil microbes and the fungus. A fungal solid-phase cultivation method to pretreat contaminated sawmill soil with high organic matter content was developed to enhance the effectiveness of the subsequent soil combustion. In a preliminary screening of 146 fungal strains, 28 out of 52 fungi, which extensively colonized non-sterile contaminated soil, were litter-decomposing fungi. The 18 strains further selected were characterized by their production of lignin-modifying and hydrolytic enzymes, of which MnP and endo-1,4-β-glucanase were the main enzymes during cultivation on Scots pine (Pinus sylvestris) bark. Of the six fungi selected for further tests, Gymnopilus luteofolius, Phanerochaete velutina, and Stropharia rugosoannulata were the most active soil organic matter degraders. The results showed that a six-month pretreatment of sawmill soil would result in a 3.5 - 9.5% loss of organic matter, depending on the fungus applied. The pretreatment process was scaled-up for a 0.56 m3 reactor, in which perforated plastic tubes filled with S. rugosoannulata growing on pine bark were introduced into the soil. The fungal pretreatment resulted in a soil mass loss of 30.5 kg, which represents 10% of the original soil mass (308 kg). Despite the fact that Scots pine bark contains several antimicrobial compounds, it was a suitable substrate for fungal growth and promoter of the production of oxidative enzymes, as well as an excellent and cheap natural carrier of fungal mycelium. This thesis successfully developed two novel fungal ex situ bioremediation technologies and introduce new insights for their further full-scale application. Ex situ slurry-phase fungal reactors might be applied in cases when the soil has a high water content or when the contaminant bioavailability is low; for example, in wastewater treatment plants to remove pharmaceutical residues. Fungal solid-phase bioremediation is a promising remediation technology to ex situ or in situ treat contaminated soil.