Biochar modulating soil biological health: A review.

Biochar can be used for multifunctional applications including the improvement of soil health and carbon storage, remediation of contaminated soil and water resources, mitigation of greenhouse gas emissions and odorous compounds, and feed supplementation to improve animal health. A healthy soil preserves microbial biodiversity that is effective in supressing plant pathogens and pests, recycling nutrients for plant growth, promoting positive symbiotic associations with plant roots, improving soil structure to supply water and nutrients, and ultimately enhancing soil productivity and plant growth. As a soil amendment, biochar assures soil biological health through different processes. First, biochar supports habitats for microorganisms due to its porous nature and by promoting the formation of stable soil micro-aggregates. Biochar also serves as a carbon and nutrient source. Biochar alters soil physical and chemical properties, creating optimum soil conditions for microbial diversity. Biochar can also immobilize soil pollutants and reduce their bioavailability that would otherwise inhibit microbial growth. However, depending on the pyrolysis settings and feedstock resources, biochar can be comprised of contaminants including polycyclic aromatic hydrocarbons and potentially toxic elements that can inhibit microbial activity, thereby impacting soil health.

Combining frass and fatty acid co-products derived from Black soldier fly larvae farming shows potential as a slow release fertiliser.

Use of black soldier fly larvae (BSFL) to process large volumes of organic waste is an emerging industry to produce protein. A co-product of this industry, the larval faeces (frass), has potential to be used as an organic fertiliser in a circular economy. However, BSFL frass has a high ammonium (N-NH4+) content which could result in nitrogen (N) loss following its application to land. One solution is to process the frass by combining it with solid fatty acids (FA) that have previously been used to manufacture slow-release inorganic fertilisers. We investigated the slow-releasing effect of N after combining BSFL frass with three FAs - lauric, myristic and stearic acid. Soil was amended with the three forms of FA processed (FA-P) frass, unprocessed frass or a control and incubated for 28 days. The impact of treatments on soil properties and soil bacterial communities were characterised during the incubation. Lower N-NH4+ concentrations occurred in soil treated with FA-P frass compared to unprocessed frass, and N-NH4+ release was slowest for lauric acid processed frass. Initially, all frass treatments caused a large shift in the soil bacterial community towards a dominance of fast-growing r-strategists that were correlated with increased organic carbon levels. FA-P frass appeared to enhance the immobilisation of N-NH4+ (from frass) by diverting it into microbial biomass. Unprocessed and stearic acid processed frass became enriched by slow-growing K-strategist bacteria at the latter stages of the incubation. Consequently, when frass was combined with FAs, FA chain length played an important role in regulating the composition of r-/K- strategists in soil and N and carbon cycling. Modifying frass with FAs could be developed into a slow release fertiliser leading to reduced soil N loss, improved fertiliser use efficiency, increased profitability and lower production costs.

Impacts of Rock Mineral and Traditional Phosphate Fertilizers on Mycorrhizal Communities in Pasture Plants.

Intensive fertilizer use can constrain contributions from soil biological processes in pastures, including those associated with arbuscular mycorrhizal (AM) fungi. We evaluated the effect of fertilizers of different P solubility on the colonization of the roots of two common pasture plants by a community of AM fungi in a pasture soil. The treatments were a rock mineral fertilizer, a chemical fertilizer and a microbial inoculant. Subterranean clover and annual ryegrass were grown in pots for 10 weeks. Both fertilizers reduced the proportion and length of roots colonized by naturally occurring AM fungi. However, by 10 weeks, there was a much greater length of mycorrhizal root for annual ryegrass than for subterranean clover. The relative abundance of mycorrhizal fungi in the families Glomeraceae and Acaulosporaceae in roots was not affected by the form of fertilizer, but diversity indices of AM fungi in roots were altered. The chemical fertilizer had a greater negative effect on AM fungal diversity indices in the annual ryegrass roots compared with the subterranean clover roots. The reduction in OTU richness of AM fungi with fertilizer application corresponded with reduced soil pH. Differential effects of P fertilizers on naturally occurring AM fungi in this agricultural soil have the potential to influence the efficacy of P fertilizer use and dominance of plant species in grasslands.

Establishing a quality management framework for commercial inoculants containing arbuscular mycorrhizal fungi.

Microbial inoculants containing arbuscular mycorrhizal (AM) fungi are potential tools in increasing the sustainability of our food production systems. Given the demand for sustainable agriculture, the production of such inoculants has potential economic value and has resulted in a variety of commercial inoculants currently being advertised. However, their use is limited by inconsistent product efficacy and lack of consumer confidence. Here, we propose a framework that can be used to assess the quality and reliability of AM inoculants. First, we set out a range of basic quality criteria which are required to achieve reliable inoculants. This is followed by a standardized bioassay which can be used to test inoculum viability and efficacy under controlled conditions. Implementation of these measurements would contribute to the adoption of AM inoculants by producers with the potential to increase sustainability in food production systems.

Co-application of a biosolids product and biochar to two coarse-textured pasture soils influenced microbial N cycling genes and potential for N leaching.

Co-application of biochar and biosolids to soil has potential to mitigate N leaching due to physical and chemical properties of biochar. Changes in N cycling pathways in soil induced by co-application of biological amendments could further mitigate N loss, but this is largely unexplored. The aim of this study was to determine whether co-application of a biochar and a modified biosolids product to three pasture soils differing in texture could alter the relative abundance of N cycling genes in soil sown with subterranean clover. The biosolids product contained lime and clay and increased subterranean clover shoot biomass in parallel with increases in soil pH and soil nitrate. Its co-application with biochar similarly increased plant growth and soil pH with a marked reduction in nitrate in two coarse textured soils but not in a clayey soil. While application of the biosolids product altered in silico predicted N cycling functional genes, there was no additional change when applied to soil in combination with biochar. This supports the conclusion that co-application of the biochar and biosolids product used here has potential to mitigate loss of N in coarse textured soils due to N adsoption by the biochar and independently of microbial N pathways.

Plant-Dependent Soil Bacterial Responses Following Amendment With a Multispecies Microbial Biostimulant Compared to Rock Mineral and Chemical Fertilizers.

Biostimulants are gaining momentum as potential soil amendments to increase plant health and productivity. Plant growth responses to some biostimulants and poorly soluble fertilizers could increase soil microbial diversity and provide greater plant access to less soluble nutrients. We assessed an agricultural soil amended with a multispecies microbial biostimulant in comparison with two fertilizers that differed in elemental solubilities to identify effects on soil bacterial communities associated with two annual pasture species (subterranean clover and Wimmera ryegrass). The treatments applied were: a multispecies microbial biostimulant, a poorly soluble rock mineral fertilizer at a rate of 5.6 kg P ha-1, a chemical fertilizer at a rate of 5.6 kg P ha-1, and a negative control with no fertilizer or microbial biostimulant. The two annual pasture species were grown separately for 10 weeks in a glasshouse with soil maintained at 70% of field capacity. Soil bacteria were studied using 16S rRNA with 27F and 519R bacterial primers on the Mi-seq platform. The microbial biostimulant had no effect on growth of either of the pasture species. However, it did influence soil biodiversity in a way that was dependent on the plant species. While application of the fertilizers increased plant growth, they were both associated with the lowest diversity of the soil bacterial community based on Fisher and Inverse Simpson indices. Additionally, these responses were plant-dependent; soil bacterial richness was highly correlated with soil pH for subterranean clover but not for Wimmera ryegrass. Soil bacterial richness was lowest following application of each fertilizer when subterranean clover was grown. In contrast, for Wimmera ryegrass, soil bacterial richness was lowest for the control and rock mineral fertilizer. Beta diversity at the bacterial OTU level of resolution by permanova demonstrated a significant impact of soil amendments, plant species and an interaction between plant type and soil amendments. This experiment highlights the complexity of how soil amendments, including microbial biostimulants, may influence soil bacterial communities associated with different plant species, and shows that caution is required when linking soil biodiversity to plant growth. In this case, the microbial biostimulant influenced soil biodiversity without influencing plant growth.

Dairy soil bacterial responses to nitrogen application in simulated Italian ryegrass and white clover pasture.

Through clearing and use of fertilizer and legumes, areas of southwestern Australia's unique coastal sand plains can support relatively low-cost dairies. However, the ancient, highly weathered nature of the soils in this region makes the dairies susceptible to a range of threats, including nutrient leaching and erosion. Despite this, Western Australian dairy cows typically produce up to 5,500 L of milk per head annually supported by inorganic nitrogen (N) fertilizer (commonly 50:50 urea and ammonium sulfate) at rates up to <320 kg of N/ha per year. Where hotspots exist (up to 2,000 kg of N/ha per year), total N exceeds pasture requirements. We investigated plant and soil bacteria responses to N fertilizer rates consistent with Australian legislated production practices on dairy farms for pure and mixed swards of white clover (Trifolium repens) and Italian ryegrass (Lolium multiflorum) in a long-term pasture experiment in controlled glasshouse conditions. Although the soil bacterial community structure at phylum level was similar for white clover and Italian ryegrass, relative abundances of specific subgroups of bacteria differed among plant species according to the N fertilizer regimen. Marked increases in relative abundance of some bacterial phyla and subphyla indicated potential inhibition of N cycling, especially for N hotspots in soil. Ammonium concentration in soil was less correlated with dominance of some N-cycling bacterial phyla than was nitrate concentration. Changes in bacterial community structure related to altered nutrient cycling highlight the potential for considering this area of research in policy assessment frameworks related to nutrient loads in dairy soils, especially for N.

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.

Response of Wheat to a Multiple Species Microbial Inoculant Compared to Fertilizer Application.

Microbial inoculants, including those formed from multiple species, may have dual functions as biostimulants and/or biocontrol agents, and claimed agricultural benefits are instrumental for regulatory categorisation. Biostimulants include commercial products containing substances or microorganisms that stimulate plant growth. Biostimulant microbes can be involved in a range of processes that affect N and P transformations in soil and thus influence nutrient availability, and N and P fertilizers can influence soil microbial diversity and function. A glasshouse experiment was conducted to investigate the effect of a multiple species microbial inoculant relative to a rock-based mineral fertilizer and a chemical fertilizer on wheat growth and yield, and on microbial diversity in the rhizosphere. The microbial inoculant was compared to the mineral fertilizer (equivalent to 5.6 kg N ha-1 and 5.6 kg P ha-1), and to the chemical fertilizer applied at three rates equivalent to: (i) 7.3 kg N ha-1 and 8.4 kg P ha-1 as recommended for on-farm use, (ii) 5.6 kg N ha-1 and 6.5 kg P ha-1 which matched the N in the mineral fertilizer, and (iii) 4.9 kg N ha-1 and 5.6 kg P ha-1 which matched P content in the mineral fertilizer. Despite an early reduction in plant growth, the microbial inoculant treatment increased shoot growth at maturity compared to the control. Similarly, grain yield was higher after application of the microbial inoculant when compared to control, and it was similar to that of plants receiving the fertilizer treatments. Using 16S rRNA sequencing, the microbial inoculant and fertilizer treatments were shown to influence the diversity of rhizosphere bacteria. The microbial inoculant increased the relative abundance of the phylum Actinobacteria. At tillering, the proportion of roots colonized by arbuscular mycorrhizal (AM) fungi increased with the microbial inoculant and mineral fertilizer treatments, but decreased with the chemical fertilizer treatments. At maturity, there were no treatment effects on the proportion of wheat roots colonized by AM fungi. Overall, the multiple species microbial inoculant had beneficial effects in terms of wheat yield relative to the commercial mineral and chemical fertilizers applied at the level recommended for on-farm use in south-western Australia.

Amending Poultry Broiler Litter to Prevent the Development of Stable Fly, Stomoxys calcitrans (Diptera: Muscidae) and Other Nuisance Flies.

Spent poultry litter use as a fertilizer in horticulture supports stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae) development. Stable fly continues to have an economic impact on livestock production and rural lifestyle in south-western Australia. The use of raw poultry manure is banned in 12 Shires surrounding Perth. The loss of market options for West Australian broiler growers has caused economic hardship. Hence, this study examined a range of chemical and biological amendments to spent poultry broiler litter in preventing stable fly and nuisance fly development. These included alkalizers (i.e., lime sand, quicklime, soda ash, and shell grit), acidifiers (aluminum sulfate, sodium bisulfate), gypsum, zeolite, spongolite, calcium cyanamide, and two fungal agents. The treated litters were placed under irrigation in horticulture with amendments added prior to them being exposed in the field as replicate 1-liter pads. In total, 19,559 stable flies developed from the spent litters exposed over five field experiments (88.7% of all flies recovered). House flies (Musca domestica L. (Diptera: Muscidae); 2,067 or 9.4%), false stable flies (Muscina stabulans Fallén (Diptera: Muscidae); 414 or 1.9%), and two sarcophagids (flesh fly) also developed from the litter. Borax completely prevented any fly development from the litter. Calcium cyanamide (1-2.5% v/v) and sodium bisulfate (10%) reduced stable fly numbers by as much as 99-100% when added to litter. Alkalizers, zeolite, spongolite, and entomopathogenic fungi had no significant impact on stable fly development. The addition of either calcium cyanamide or sodium bisulfate to raw litter can boost the fertilizer value of the litter while preventing stable fly development.

Arbuscular mycorrhizal fungus responses to disturbance are context-dependent.

Anthropogenic disturbance is one of the most important forces shaping soil ecosystems. While organisms that live in the soil, such as arbuscular mycorrhizal (AM) fungi, are sensitive to disturbance, their response is not always predictable. Given the range of disturbance types and differences among AM fungi in their growth strategies, the unpredictability of the responses of AM fungi to disturbance is not surprising. We investigated the role of disturbance type (i.e., soil disruption, agriculture, host perturbation, and chemical disturbance) and fungus identity on disturbance response in the AM symbiosis. Using meta-analysis, we found evidence for differential disturbance response among AM fungal species, as well as evidence that particular fungal species are especially susceptible to certain disturbance types, perhaps because of their life history strategies.

Microbial phylogenetic and functional responses within acidified wastewater communities exhibiting enhanced phosphate uptake.

Acid stimulated accumulation of insoluble phosphorus within microbial cells is highly beneficial to wastewater treatment but remains largely unexplored. Using single cell analyses and next generation sequencing, the response of active polyphosphate accumulating microbial communities under conditions of enhanced phosphorus uptake under both acidic and aerobic conditions was characterised. Phosphorus accumulation activities were highest under acidic conditions (pH 5.5>8.5), where a significant positive effect on bioaccumulation was observed at pH 5.5 when compared to pH 8.5. In contrast to the Betaproteobacteria and Actinobacteria dominated enhanced biological phosphorus removal process, the functionally active polyP accumulators at pH 5.5 belonged to the Gammaproteobacteria, with key accumulators identified as members of the families Aeromonadaceae and Enterobacteriaceae. This study demonstrated a significant enrichment of key polyphosphate kinase and exopolyphosphatase genes within the community metagenome after acidification, concomitant with an increase in P accumulation kinetics.

Interactions between biochar and mycorrhizal fungi in a water-stressed agricultural soil.

Biochar may alleviate plant water stress in association with arbuscular mycorrhizal (AM) fungi but research has not been conclusive. Therefore, a glasshouse experiment was conducted to understand how interactions between AM fungi and plants respond to biochar application under water-stressed conditions. A twin chamber pot system was used to determine whether a woody biochar increased root colonisation by a natural AM fungal population in a pasture soil ('field' chamber) and whether this was associated with increased growth of extraradical AM fungal hyphae detected by plants growing in an adjacent ('bait') chamber containing irradiated soil. The two chambers were separated by a mesh that excluded roots. Subterranean clover was grown with and without water stress and harvested after 35, 49 and 63 days from each chamber. When biochar was applied to the field chamber under water-stressed conditions, shoot mass increased in parallel with mycorrhizal colonisation, extraradical hyphal length and shoot phosphorus concentration. AM fungal colonisation of roots in the bait chamber indicated an increase in extraradical mycorrhizal hyphae in the field chamber. Biochar had little effect on AM fungi or plant growth under well-watered conditions. The biochar-induced increase in mycorrhizal colonisation was associated with increased growth of extraradical AM fungal hyphae in the pasture soil under water-stressed conditions.

Tolerance and induction of tolerance to Ni of arbuscular mycorrhizal fungi from New Caledonian ultramafic soils.

The influence of Ni on arbuscular mycorrhizal fungi (AMF) has not been studied yet. We tested the tolerance to Ni of five AMF isolates from New Caledonian ultramafic soils. Spore germination indicated that these isolates were clearly more tolerant to Ni than three other isolates from non-ultramafic soils. They were able to germinate at 30 microg g(-1) Ni, whereas spores of the non-ultramafic isolates were totally inhibited at 15 microg g(-1) Ni. Among the ultramafic isolates, two were obtained from roots of Ni-hyperaccumulating plants. Their tolerance to Ni was clearly higher than all the other isolates. The proportion of germinated spores of the different isolates in contact with ultramafic soils showed the same tendencies as those observed with Ni solutions. Tolerance to Ni increased when spores were produced from mycorrhiza on plants grown on sand containing 20 microg g(-1) Ni, in comparison with those produced on sand without Ni. These results indicate that the tolerance to Ni of AMF spores can be induced by the presence of this metal in the substrate.

The promise and the potential consequences of the global transport of mycorrhizal fungal inoculum.

Advances in ecology during the past decade have led to a much more detailed understanding of the potential negative consequences of species' introductions. Moreover, recent studies of mycorrhizal symbionts have led to an increased knowledge of the potential utility of fungal inoculations in agricultural, horticultural and ecological management. The intentional movement of mycorrhizal fungal species is growing, but the concomitant potential for negative ecological consequences of invasions by mycorrhizal fungi is poorly understood. We assess the degree to which introductions of mycorrhizal fungi may lead to unintended negative, and potentially costly, consequences. Our purpose is to make recommendations regarding appropriate management guidelines and highlight top priority research needs. Given the difficulty in discerning invasive species problems associated with mycorrhizal inoculations, we recommend the following. First, careful assessment documenting the need for inoculation, and the likelihood of success, should be conducted prior to inoculation because inoculations are not universally beneficial. Second, invasive species problems are costly and often impossible to control by the time they are recognized. We recommend using local inoculum sources whenever possible. Third, non-sterile cultures of inoculum can result in the movement of saprobes and pathogens as well as mutualists. We recommend using material that has been produced through sterile culture when local inoculum is not available. Finally, life-history characteristics of inoculated fungi may provide general guidelines relative to the likelihood of establishment and spread. We recommend that, when using non-local fungi, managers choose fungal taxa that carry life-history traits that may minimize the likelihood of deleterious invasive species problems. Additional research is needed on the potential of mycorrhizal fungi to spread to non-target areas and cause ecological damage.

Invasion of spores of the arbuscular mycorrhizal fungus Gigaspora decipiens by Burkholderia spp.

Burkholderia species are bacterial soil inhabitants that are capable of interacting with a variety of eukaryotes, in some cases occupying intracellular habitats. Pathogenic and nonpathogenic Burkholderia spp., including B. vietnamiensis, B. cepacia, and B. pseudomallei, were grown on germinating spores of the arbuscular mycorrhizal fungus Gigaspora decipiens. Spore lysis assays revealed that all Burkholderia spp. tested were able to colonize the interior of G. decipiens spores. Amplification of specific DNA sequences and transmission electron microscopy confirmed the intracellular presence of B. vietnamiensis. Twelve percent of all spores were invaded by B. vietnamiensis, with an average of 1.5 x 10(6) CFU recovered from individual infected spores. Of those spores inoculated with B. pseudomallei, 7% were invaded, with an average of 5.5 x 10(5) CFU recovered from individual infected spores. Scanning electron and fluorescence microscopy provided insights into the morphology of surfaces of spores and hyphae of G. decipiens and the attachment of bacteria. Burkholderia spp. colonized both hyphae and spores, attaching to surfaces in either an end-on or side-on fashion. Adherence of Burkholderia spp. to eukaryotic surfaces also involved the formation of numerous fibrillar structures.