Blending bauxite residues with multiple byproducts improves capping materials for tailings storage facilities.

Amelioration and management of large volumes of tailings resulting from alumina refining is a major challenge owing to the high alkalinity and salinity of residues. Blended byproduct caps are a potential new and more cost-effective approach to tailings management, where tailings are blended with other local byproducts in order to reduce pH, salinity and toxic elements. Here, alkaline bauxite residue was blended with four byproducts (waste acid, sewage water, fly ash and eucalypt mulch) to create a range of potential capping materials. We leached and weathered materials in the glasshouse with deionized water over nine weeks to investigate if byproducts on their own or in combination improved cap conditions. Combining all four byproducts (10 wt % waste acid, 5 wt % sewage water, 20 wt % fly ash and 10 wt % eucalypt mulch) achieved lower pH (9.60) compared to any byproduct applied individually, or un-remediated bauxite residue (pH 10.7). Leaching decreased EC by dissolving and exporting salts and minerals from the bauxite residue. Fly ash addition increased organic carbon (likely from non-combusted organic material) and nitrogen, while eucalypt mulch increased inorganic phosphorus. Addition of byproducts also decreased the concentration of potentially toxic elements (e.g., Al, Na, Mo and V) and enhanced pH neutralisation. Initial pH with single byproduct treatments was 10.4-10.5, which decreased to between 9.9-10.0. Further lowering of pH and salinity as well as increased nutrient concentrations may be possible through higher addition rates of byproducts, incorporation of other materials such as gypsum, and increasing leaching/weathering time of tailings in situ.

Leaching of phthalate acid esters from plastic mulch films and their degradation in response to UV irradiation and contrasting soil conditions.

Phthalate acid esters (PAEs) are commonly used plastic additives, not chemically bound to the plastic that migrate into surrounding environments, posing a threat to environmental and human health. Dibutyl phthalate (DBP) and di(2-ethylhexyl) phthalate (DEHP) are two common PAEs found in agricultural soils, where degradation is attributed to microbial decomposition. Yet the impact of the plastic matrix on PAE degradation rates is poorly understood. Using 14C-labelled DBP and DEHP we show that migration from the plastic matrix into soil represents a key rate limiting step in their bioavailability and subsequent degradation. Incorporating PAEs into plastic film decreased their degradation in soil, DBP (DEHP) from 79% to 21% (9% to <1%), over four months when compared to direct application of PAEs. Mimicking surface soil conditions, we demonstrated that exposure to ultraviolet radiation accelerated PAE mineralisation twofold. Turnover of PAE was promoted by the addition of biosolids, while the presence of plants and other organic residues failed to promote degradation. We conclude that PAEs persist in soil for longer than previously thought due to physical trapping within the plastic matrix, suggesting PAEs released from plastics over very long time periods lead to increasing levels of contamination.

Revealing the transfer pathways of cyanobacterial-fixed N into the boreal forest through the feather-moss microbiome.

Introduction: Biological N2 fixation in feather-mosses is one of the largest inputs of new nitrogen (N) to boreal forest ecosystems; however, revealing the fate of newly fixed N within the bryosphere (i.e. bryophytes and their associated organisms) remains uncertain. Methods: Herein, we combined 15N tracers, high resolution secondary ion mass-spectrometry (NanoSIMS) and a molecular survey of bacterial, fungal and diazotrophic communities, to determine the origin and transfer pathways of newly fixed N2 within feather-moss (Pleurozium schreberi) and its associated microbiome. Results: NanoSIMS images reveal that newly fixed N2, derived from cyanobacteria, is incorporated into moss tissues and associated bacteria, fungi and micro-algae. Discussion: These images demonstrate that previous assumptions that newly fixed N2 is sequestered into moss tissue and only released by decomposition are not correct. We provide the first empirical evidence of new pathways for N2 fixed in feather-mosses to enter the boreal forest ecosystem (i.e. through its microbiome) and discuss the implications for wider ecosystem function.

Developing Bioprospecting Strategies for Bioplastics Through the Large-Scale Mining of Microbial Genomes.

The accumulation of petroleum-based plastic waste has become a major issue for the environment. A sustainable and biodegradable solution can be found in Polyhydroxyalkanoates (PHAs), a microbially produced biopolymer. An analysis of the global phylogenetic and ecological distribution of potential PHA producing bacteria and archaea was carried out by mining a global genome repository for PHA synthase (PhaC), a key enzyme involved in PHA biosynthesis. Bacteria from the phylum Actinobacteria were found to contain the PhaC Class II genotype which produces medium-chain length PHAs, a physiology until now only found within a few Pseudomonas species. Further, several PhaC genotypes were discovered within Thaumarchaeota, an archaeal phylum with poly-extremophiles and the ability to efficiently use CO2 as a carbon source, a significant ecological group which have thus far been little studied for PHA production. Bacterial and archaeal PhaC genotypes were also observed in high salinity and alkalinity conditions, as well as high-temperature geothermal ecosystems. These genome mining efforts uncovered previously unknown candidate taxa for biopolymer production, as well as microbes from environmental niches with properties that could potentially improve PHA production. This in silico study provides valuable insights into unique PHA producing candidates, supporting future bioprospecting efforts toward better targeted and relevant taxa to further enhance the diversity of exploitable PHA production systems.

Angiosperm symbioses with non-mycorrhizal fungal partners enhance N acquisition from ancient organic matter in a warming maritime Antarctic.

In contrast to the situation in plants inhabiting most of the world's ecosystems, mycorrhizal fungi are usually absent from roots of the only two native vascular plant species of maritime Antarctica, Deschampsia antarctica and Colobanthus quitensis. Instead, a range of ascomycete fungi, termed dark septate endophytes (DSEs), frequently colonise the roots of these plant species. We demonstrate that colonisation of Antarctic vascular plants by DSEs facilitates not only the acquisition of organic nitrogen as early protein breakdown products, but also as non-proteinaceous d-amino acids and their short peptides, accumulated in slowly-decomposing organic matter, such as moss peat. Our findings suggest that, in a warming maritime Antarctic, this symbiosis has a key role in accelerating the replacement of formerly dominant moss communities by vascular plants, and in increasing the rate at which ancient carbon stores laid down as moss peat over centuries or millennia are returned to the atmosphere as CO2 .

Regulatory T cells differ from conventional CD4+ T cells in their recirculatory behavior and lymph node transit times.

Regulatory T cells (Tregs) continuously suppress autoreactive immune responses within tissues to prevent autoimmunity, yet the recirculatory behavior of Tregs between and within tissues enabling the maintenance of peripheral tolerance remains incompletely defined. Here, we quantified homing efficiency to and the dwell time of Tregs within secondary lymphoid organs (SLOs) and used intravital two-photon microscopy to measure Treg surveillance behavior of dendritic cells. Tregs homed substantially less efficiently to SLOs compared with conventional CD4+ T cells (Tconvs), despite similar expression of homing receptors. Tregs remained on average 2-3 times longer within the LN than Tconvs before exiting, and retained Tregs differed from recirculating Tregs in phenotype, motility and interaction duration with dendritic cells. Taken together, these data revealed fundamental differences in Treg versus conventional T cell in vivo recirculation and migration behaviors, identified a Treg population with prolonged LN dwell time, and provided quantitative insight into their spatiotemporal behavior within LNs.

Biochar phosphorus concentration dictates mycorrhizal colonisation, plant growth and soil phosphorus cycling.

We aimed to determine the relationship between biochar properties and colonisation of roots by arbuscular mycorrhizal (AM) fungi in agricultural soil. We used a range of biochars that differed in pH, water holding capacity, C, N and P concentrations to investigate interactions between biochar and AM fungi. A glasshouse experiment was conducted with subterranean clover and wheat, amended separately with 34 sources of biochar (applied at 1% w/w), to investigate potential responses in a phosphorus (P) deficient agricultural soil. Plant growth responses to biochar ranged from positive to negative and were dependent on biochar P concentration, available soil P and AM root colonisation. The higher the nutrient P concentration in biochar, the lower was AM colonisation. Growth responses of wheat and clover to the application of various biochars were mostly positive, and their growth was correlated, but biochar contributions to soil fertility varied with biochar properties. When nutrient concentrations are higher in biochars, especially for P and N, plants can gain access to nutrients via the plant roots and mycorrhizal hyphae. Thus biochar amendments can increase both plant nutrient uptake and crop production in nutrient deficient soil.

Linking bacterial community composition to soil salinity along environmental gradients.

Salinization is recognized as a threat to soil fertility worldwide. A challenge in understanding the effects of salinity on soil microbial communities is the fact that it can be difficult to disentangle the effects of salinity from those of other variables that may co-vary with salinity. Here we use a trait-based approach to identify direct effects of salinity on soil bacterial communities across two salinity gradients. Through dose-response relationships between salinity and bacterial growth, we quantified distributions of the trait salt tolerance within the communities. Community salt tolerance was closely correlated with soil salinity, indicating a strong filtering effect of salinity on the bacterial communities. Accompanying the increases in salt tolerance were consistent shifts in bacterial community composition. We identified specific bacterial taxa that increased in relative abundances with community salt tolerance, which could be used as bioindicators for high community salt tolerance. A strong filtering effect was also observed for pH across the gradients, with pH tolerance of bacterial communities correlated to soil pH. We propose phenotypic trait distributions aggregated at the community level as a useful approach to study the role of environmental factors as filters of microbial community composition.

Copolymers enhance selective bacterial community colonization for potential root zone applications.

Managing the impact of anthropogenic and climate induced stress on plant growth remains a challenge. Here we show that polymeric hydrogels, which maintain their hydrous state, can be designed to exploit functional interactions with soil microorganisms. This microbial enhancement may mitigate biotic and abiotic stresses limiting productivity. The presence of mannan chains within synthetic polyacrylic acid (PAA) enhanced the dynamics and selectivity of bacterial ingress in model microbial systems and soil microcosms. Pseudomonas fluorescens exhibiting high mannan binding adhesins showed higher ingress and localised microcolonies throughout the polymeric network. In contrast, ingress of Bacillus subtilis, lacking adhesins, was unaltered by mannan showing motility comparable to bulk liquids. Incubation within microcosms of an agricultural soil yielded hydrogel populations significantly increased from the corresponding soil. Bacterial diversity was markedly higher in mannan containing hydrogels compared to both control polymer and soil, indicating enhanced selectivity towards microbial families that contain plant beneficial species. Here we propose functional polymers applied to the potential root zone which can positively influence rhizobacteria colonization and potentially plant growth as a new approach to stress tolerance.

Response of microbial biomass and CO2-C loss to wetting patterns are temperature dependent in a semi-arid soil.

One of the greatest contemporary challenges in terrestrial ecology is to determine the impact of climate change on the world's ecosystems. Here we investigated how wetting patterns (frequency and intensity) and nutrient additions altered microbial biomass and CO2-C loss from a semi-arid soil. South-western Australia is predicted to experience declining annual rainfall but increased frequency of summer rainfall events when soil is fallow. Agricultural soils (0-10 cm at 10 °C or 25 °C) received the same total amount of water (15 mL over 30 days) applied at different frequency; with either nil or added nitrogen and phosphorus. Smaller more frequent wetting applications resulted in less CO2-C loss (P < 0.001); with cumulative CO2-C loss 35% lower than a single wetting event. This coincided with increased microbial biomass C at 25 °C but a decline at 10 °C. Increasing nutrient availability decreased CO2-C loss only under a single larger wetting event. While bacterial and fungal abundance remained unchanged, archaeal abundance and laccase-like copper monooxidase gene abundance increased with more frequent wetting at 25 °C. Our findings suggest smaller more frequent summer rainfall may decrease CO2 emissions compared to infrequent larger events; and enhance microbial C use efficiency where sufficient background soil organic matter and nutrients are available.

Microbial respiration, but not biomass, responded linearly to increasing light fraction organic matter input: Consequences for carbon sequestration.

Rebuilding 'lost' soil carbon (C) is a priority in mitigating climate change and underpinning key soil functions that support ecosystem services. Microorganisms determine if fresh C input is converted into stable soil organic matter (SOM) or lost as CO2. Here we quantified if microbial biomass and respiration responded positively to addition of light fraction organic matter (LFOM, representing recent inputs of plant residue) in an infertile semi-arid agricultural soil. Field trial soil with different historical plant residue inputs [soil C content: control (tilled) = 9.6 t C ha-1 versus tilled + plant residue treatment (tilled + OM) = 18.0 t C ha-1] were incubated in the laboratory with a gradient of LFOM equivalent to 0 to 3.8 t C ha-1 (0 to 500% LFOM). Microbial biomass C significantly declined under increased rates of LFOM addition while microbial respiration increased linearly, leading to a decrease in the microbial C use efficiency. We hypothesise this was due to insufficient nutrients to form new microbial biomass as LFOM input increased the ratio of C to nitrogen, phosphorus and sulphur of soil. Increased CO2 efflux but constrained microbial growth in response to LFOM input demonstrated the difficulty for C storage in this environment.

Spatially governed climate factors dominate management in determining the quantity and distribution of soil organic carbon in dryland agricultural systems.

Few studies describe the primary drivers influencing soil organic carbon (SOC) stocks and the distribution of carbon (C) fractions in agricultural systems from semi-arid regions; yet these soils comprise one fifth of the global land area. Here we identified the primary drivers for changes in total SOC and associated particulate (POC), humus (HOC) and resistant (ROC) organic C fractions for 1347 sample points in the semi-arid agricultural region of Western Australia. Total SOC stock (0-0.3 m) varied from 4 to 209 t C ha(-1) with 79% of variation explained by measured variables. The proportion of C in POC, HOC and ROC fractions averaged 28%, 45% and 27% respectively. Climate (43%) and land management practices (32%) had the largest relative influence on variation in total SOC. Carbon accumulation was constrained where average daily temperature was above 17.2 °C and annual rainfall below 450 mm, representing approximately 42% of the 197,300 km(2) agricultural region. As such large proportions of this region are not suited to C sequestration strategies. For the remainder of the region a strong influence of management practices on SOC indicate opportunities for C sequestration strategies associated with incorporation of longer pasture phases and adequate fertilisation.

Ancient landscapes and the relationship with microbial nitrification.

Ammonia oxidizing archaea (AOA) and bacteria (AOB) drive nitrification and their population dynamics impact directly on the global nitrogen cycle. AOA predominate in the majority of soils but an increasing number of studies have found that nitrification is largely attributed to AOB. The reasons for this remain poorly understood. Here, amoA gene abundance was used to study the distribution of AOA and AOB in agricultural soils on different parent materials and in contrasting geologic landscapes across Australia (n = 135 sites). AOA and AOB abundances separated according to the geologic age of the parent rock with AOB higher in the more weathered, semi-arid soils of Western Australia. AOA dominated the younger, higher pH soils of Eastern Australia, independent of any effect of land management and fertilization. This differentiation reflects the age of the underlying parent material and has implications for our understanding of global patterns of nitrification and soil microbial diversity. Western Australian soils are derived from weathered archaean laterite and are acidic and copper deficient. Copper is a co-factor in the oxidation of ammonia by AOA but not AOB. Thus, copper deficiency could explain the unexpectedly low populations of AOA in Western Australian soils.

Carbon and Nitrogen Mineralization in Relation to Soil Particle-Size Fractions after 32 Years of Chemical and Manure Application in a Continuous Maize Cropping System.

Long-term manure application is recognized as an efficient management practice to enhance soil organic carbon (SOC) accumulation and nitrogen (N) mineralization capacity. A field study was established in 1979 to understand the impact of long-term manure and/or chemical fertilizer application on soil fertility in a continuous maize cropping system. Soil samples were collected from field plots in 2012 from 9 fertilization treatments (M0CK, M0N, M0NPK, M30CK, M30N, M30NPK, M60CK, M60N, and M60NPK) where M0, M30, and M60 refer to manure applied at rates of 0, 30, and 60 t ha(-1) yr(-1), respectively; CK indicates no fertilizer; N and NPK refer to chemical fertilizer in the forms of either N or N plus phosphorus (P) and potassium (K). Soils were separated into three particle-size fractions (2000-250, 250-53, and <53 µm) by dry- and wet-sieving. A laboratory incubation study of these separated particle-size fractions was used to evaluate the effect of long-term manure, in combination with/without chemical fertilization application, on the accumulation and mineralization of SOC and total N in each fraction. Results showed that long-term manure application significantly increased SOC and total N content and enhanced C and N mineralization in the three particle-size fractions. The content of SOC and total N followed the order 2000-250 µm > 250-53 µm > 53 µm fraction, whereas the amount of C and N mineralization followed the reverse order. In the <53 µm fraction, the M60NPK treatment significantly increased the amount of C and N mineralized (7.0 and 10.1 times, respectively) compared to the M0CK treatment. Long-term manure application, especially when combined with chemical fertilizers, resulted in increased soil microbial biomass C and N, and a decreased microbial metabolic quotient. Consequently, long-term manure fertilization was beneficial to both soil C and N turnover and microbial activity, and had significant effect on the microbial metabolic quotient.

Ammonia-oxidising bacteria not archaea dominate nitrification activity in semi-arid agricultural soil.

Ammonia-oxidising archaea (AOA) and bacteria (AOB) are responsible for the rate limiting step in nitrification; a key nitrogen (N) loss pathway in agricultural systems. Dominance of AOA relative to AOB in the amoA gene pool has been reported in many ecosystems, although their relative contributions to nitrification activity are less clear. Here we examined the distribution of AOA and AOB with depth in semi-arid agricultural soils in which soil organic matter content or pH had been altered, and related their distribution to gross nitrification rates. Soil depth had a significant effect on gene abundances, irrespective of management history. Contrary to reports of AOA dominance in soils elsewhere, AOA gene copy numbers were four-fold lower than AOB in the surface (0-10 cm). AOA gene abundance increased with depth while AOB decreased, and sub-soil abundances were approximately equal (10-90 cm). The depth profile of total archaea did not mirror that of AOA, indicating the likely presence of archaea without nitrification capacity in the surface. Gross nitrification rates declined significantly with depth and were positively correlated to AOB but negatively correlated to AOA gene abundances. We conclude that AOB are most likely responsible for regulating nitrification in these semi-arid soils.

Relative contribution of maize and external manure amendment to soil carbon sequestration in a long-term intensive maize cropping system.

We aimed to quantify the relative contributions of plant residue and organic manure to soil carbon sequestration. Using a 27-year-long inorganic fertilizer and manure amendment experiment in a maize (Zea mays L.) double-cropping system, we quantified changes in harvestable maize biomass and soil organic carbon stocks (0-20 cm depth) between 1986-2012. By employing natural (13)C tracing techniques, we derived the proportional contributions of below-ground crop biomass return (maize-derived carbon) and external manure amendment (manure-derived carbon) to the total soil organic carbon stock. The average retention of maize-derived carbon plus manure-derived carbon during the early period of the trial (up to 11 years) was relatively high (10%) compared to the later period (22 to 27 years, 5.1-6.3%). About 11% of maize-derived carbon was converted to soil organic carbon, which was double the retention of manure-derived carbon (4.4-5.1%). This result emphasized that organic amendments were necessary to a win-win strategy for both SOC sequestration and maize production.

Exploring the transfer of recent plant photosynthates to soil microbes: mycorrhizal pathway vs direct root exudation.

Plants rapidly release photoassimilated carbon (C) to the soil via direct root exudation and associated mycorrhizal fungi, with both pathways promoting plant nutrient availability. This study aimed to explore these pathways from the root's vascular bundle to soil microbial communities. Using nanoscale secondary ion mass spectrometry (NanoSIMS) imaging and (13) C-phospho- and neutral lipid fatty acids, we traced in-situ flows of recently photoassimilated C of (13) CO2 -exposed wheat (Triticum aestivum) through arbuscular mycorrhiza (AM) into root- and hyphae-associated soil microbial communities. Intraradical hyphae of AM fungi were significantly (13) C-enriched compared to other root-cortex areas after 8 h of labelling. Immature fine root areas close to the root tip, where AM features were absent, showed signs of passive C loss and co-location of photoassimilates with nitrogen taken up from the soil solution. A significant and exclusively fresh proportion of (13) C-photosynthates was delivered through the AM pathway and was utilised by different microbial groups compared to C directly released by roots. Our results indicate that a major release of recent photosynthates into soil leave plant roots via AM intraradical hyphae already upstream of passive root exudations. AM fungi may act as a rapid hub for translocating fresh plant C to soil microbes.

Competition between plant and bacterial cells at the microscale regulates the dynamics of nitrogen acquisition in wheat (Triticum aestivum).

The ability of plants to compete effectively for nitrogen (N) resources is critical to plant survival. However, controversy surrounds the importance of organic and inorganic sources of N in plant nutrition because of our poor ability to visualize and understand processes happening at the root-microbial-soil interface. Using high-resolution nano-scale secondary ion mass spectrometry stable isotope imaging (NanoSIMS-SII), we quantified the fate of ¹5N over both space and time within the rhizosphere. We pulse-labelled the soil surrounding wheat (Triticum aestivum) roots with either ¹5NH4⁺ or ¹5N-glutamate and traced the movement of ¹5N over 24 h. Imaging revealed that glutamate was rapidly depleted from the rhizosphere and that most ¹5N was captured by rhizobacteria, leading to very high ¹5N microbial enrichment. After microbial capture, approximately half of the ¹5N-glutamate was rapidly mineralized, leading to the excretion of NH4⁺, which became available for plant capture. Roots proved to be poor competitors for ¹5N-glutamate and took up N mainly as ¹5NH4⁺. Spatial mapping of ¹5N revealed differential patterns of ¹5N uptake within bacteria and the rapid uptake and redistribution of ¹5N within roots. In conclusion, we demonstrate the rapid cycling and transformation of N at the soil-root interface and that wheat capture of organic N is low in comparison to inorganic N under the conditions tested.

Microbial utilisation of biochar-derived carbon.

Whilst largely considered an inert material, biochar has been documented to contain a small yet significant fraction of microbially available labile organic carbon (C). Biochar addition to soil has also been reported to alter soil microbial community structure, and to both stimulate and retard the decomposition of native soil organic matter (SOM). We conducted a short-term incubation experiment using two (13)C-labelled biochars produced from wheat or eucalypt shoots, which were incorporated in an aridic arenosol to examine the fate of the labile fraction of biochar-C through the microbial community. This was achieved using compound specific isotopic analysis (CSIA) of phospholipid fatty acids (PLFAs). A proportion of the biologically-available fraction of both biochars was rapidly (within three days) utilised by gram positive bacteria. There was a sharp peak in CO2 evolution shortly after biochar addition, resulting from rapid turnover of labile C components in biochars and through positive priming of native SOM. Our results demonstrate that this CO2 evolution was at least partially microbially mediated, and that biochar application to soil can cause significant and rapid changes in the soil microbial community; likely due to addition of labile C and increases in soil pH.

Afforestation alters community structure of soil fungi.

Relatively little is known about the effect of afforestation on soil fungal communities. This study demonstrated that afforestation altered fungal community structure and that changes were correlated to pools of soil C. Pasture at three locations on the same soil type was afforested with Eucalyptus globulus or Pinus pinaster. The structure of fungal communities under the three land uses was measured after 13y using automated ribosomal intergenic spacer analysis (ARISA). Afforestation significantly altered the structure of fungal communities. The effect of location on the structure of fungal communities was limited to pasture soils; although these contained the same plant species, the relative composition of each species varied between locations. Differences in the structure of fungal communities between pasture, E. globulus and P. pinaster were significantly correlated with changes in the amount of total organic C and microbial biomass-C in soil. Afforestation of patches of agricultural land may contribute to conserving soil fungi in agricultural landscapes by supporting fungal communities with different composition to agricultural soils.