Humans are dependent upon soil which supplies food, fuel, chemicals, medicine, sequesters pollutants, purifies and conveys water, and supports the built environment. In short, we need soil, but it has little or no need of us. Agriculture, mining, urbanization and other human activities result in temporary land-use and once complete, used and degraded land should be rehabilitated and restored to minimize loss of soil carbon. It is generally accepted that the most effective strategy is phyto-remediation. Typically, phytoremediation involves re-invigoration of soil fertility, physicochemical properties, and its microbiome to facilitate establishment of appropriate climax cover vegetation. A myco-phytoremediation technology called Fungcoal was developed in South Africa to achieve these outcomes for land disturbed by coal mining. Here we outline the contemporary and expanded rationale that underpins Fungcoal, which relies on in situ bio-conversion of carbonaceous waste coal or discard, in order to explore the probable origin of humic substances (HS) and soil organic matter (SOM). To achieve this, microbial processing of low-grade coal and discard, including bio-liquefaction and bio-conversion, is examined in some detail. The significance, origin, structure, and mode of action of coal-derived humics are recounted to emphasize the dynamic equilibrium, that is, humification and the derivation of soil organic matter (SOM). The contribution of plant exudate, extracellular vesicles (EV), extra polymeric substances (EPS), and other small molecules as components of the dynamic equilibrium that sustains SOM is highlighted. Arbuscular mycorrhizal fungi (AMF), saprophytic ectomycorrhizal fungi (EMF), and plant growth promoting rhizobacteria (PGPR) are considered essential microbial biocatalysts that provide mutualistic support to sustain plant growth following soil reclamation and restoration. Finally, we posit that de novo synthesis of SOM is by specialized microbial consortia (or ‘humifiers’) which use molecular components from the root metabolome; and, that combinations of functional biocatalyst act to re-establish and maintain the soil dynamic. It is concluded that a bio-scaffold is necessary for functional phytoremediation including maintenance of the SOM dynamic and overall biogeochemistry of organic carbon in the global ecosystem

During wastewater treatment by integrated algal pond systems (IAPS), microalgal-bacterial flocs (MaB-flocs) form naturally but periodically disaggregate, resulting in poor settling, low biomass recovery, and reduced effluent quality. This study investigates biotic/abiotic-induced changes in microbial community structure in high-rate algal oxidation ponds (HRAOP) of an IAPS on MaB-floc formation and stability during sewage treatment. Results show that dominance by Pseudopediastrum, Desmodesmus and Micractinium species in spring and summer and the chytrids, Paraphysoderma sp. in spring and Sanchytrium sp. in summer, occurred coincident with enhanced MaB-floc formation and biomass recovery (≥90%). In winter, poor floc formation and low biomass recovery were associated with dominance by Desmodesmus, Chlorella, and the Chlorella-like genus Micractinium. A principal components analysis (PCA) confirmed that combinations of colonial microalgae and associated parasitic chytrids underpin MaB-floc formation and stability in spring and summer and that unicells dominated in winter. Dominance by Thiothrix sp. coincided with floc disaggregation. Thus, changes in season, composition and abundance of colonial microalgae and associated parasitic fungi appeared to impact MaB-floc formation, whereas species composition of the bacterial population and emergence of Thiothrix coincided with floc instability and disaggregation.

Inasmuch as coal remains the linchpin for the generation of electricity and liquid petroleum products in South Africa, hydrocarbon waste and coal discard will continue to pose a threat to the environment. Therefore, the onus is on the associated industries to develop and implement efficient and sustainable strategies to mitigate the negative impacts of energy generating activities on the environment. Most conventional efforts in this regard, although successful for soil repair and the initiation of vegetation, have been deemed unsustainable. In an effort to find a sustainable remediation strategy a novel technology termed “FungCoal” was conceptualized and patented as a strategy for the rehabilitation of open cast coal mines, carbonaceous-rich spoils and coal wastes. This biotechnology, which exploits plant-fungal mutualism to achieve effective biodegradation of coal on discard dumps and the breakdown of the carbonaceous component in spoils, promotes revegetation to facilitate rehabilitation of mining-disturbed land. However, one limiting factor of the FungCoal bioprocess is that it requires oxidized weathered coal, a highly complex and variable resource for use as a co-substrate, for growth and proliferation of the coal degrading microorganisms. To fully exploit the potential of plant-fungal mutualism and its interaction for use in the remediation of coal contaminated soils, this study investigated the proposed relationship between plant roots, root exudate and the coal degrading fungus “Aspergillus sp.” (previously Neosartorya fischeri) strain 84 in more detail, in an effort to gain further insight into the mechanisms underpinning plant-fungal mutualism as a strategy for re-vegetation of coal discard dumps and the rehabilitation of hydrocarbon-contaminated soil using the FungCoal approach. A pot-on-beaker (PoB) method was developed for the easy cultivation and collection of extracellular polymeric substance (EPS)-containing exudates from Zea mays L. (maize) and Abelmuschus esculentus (okra). Characterisation of the EPS material from these exudates was carried out using a combination of physicochemical and biochemical methods. The results from analysis of phenolics and indoles showed that exudates contain some form of indoles and phenolic compounds, although in little proportions, which may fulfil a signalling function, responsible for attracting soil microorganisms into the rhizosphere. Spectroscopic analysis of the exudates using FT-IR revealed vibrations corresponding to functional groups of alkanes, alkenes, alkynes, and carboxylic acids. These compounds likely provide an easily accessible source of carbon to soil microorganisms and are also a better alternative to the poly-aromatics which are an inherent component locked-up in the supposed recalcitrant coal material. The results from biochemical analyses also revealed the presence of carbohydrate, proteins, lipids, and low amounts of α-amino-nitrogen in the EPS of maize and okra. These components of EPS are all essential for the stimulation of enzymatic activities in soil microorganisms and, which may in turn aid biodegradation. The action of the root EPS from maize was further tested on three coal-degrading fungal isolates identified as Aspergillus strain ECCN 84, Aspergillus strain ECCN 225 and Penicillium strain ECCN 243 for manganese peroxidase (MnP) and laccase (LAC) activities. The results revealed that the Aspergillus species, strains ECCN 84 and ECCN 225, showed with or without EPS, observable black halos surrounding each of the colonies after 7d incubation indicative of positive MnP activity, while no activity was observed for the Penicillium sp. strain ECCN 243. Analysis for LAC revealed little or no activity in any of the coal degrading fungi following addition of pulverized coal to the growth medium. Interestingly, the addition of EPS-containing exudate to the coal-containing medium resulted in increased LAC activity for all fungal isolates. This finding affirmed the positive contribution of EPS to extracellular LAC activity, purported as an important enzyme in the coal biodegradation process. Finally, the impact of plant-derived exudate on the colonisation and biodegradation of coal was investigated in situ using rhizoboxes, to simulate a coal environment, and was carried out for 16 weeks. Microscopic examination of coal samples after termination of the experiment showed fungal proliferation and attachment to coal particles. All of the rhizoboxes that contained plants had higher medium pH and EC, and the concentration of phenolics, indoles and humic acids was greater than that of control treatments. These observations indicated better rhizosphere colonisation, substrate biodegradation and humification. Therefore, root exudate appears to play a significant role in coordination of soil microorganisms within the rhizosphere and likely serves both as a scaffold for rhizospheric interactions by providing microorganisms with accessible carbon and as a likely ‘trigger’ for induction of coal-degrading enzymes such as fungal LAC for mobilisation of recalcitrant carbon. This study has shown that EPS exuded from roots of Zea mays together with coal degrading fungus Aspergillus strain ECCN 84 can alkalinise the coal substrate and facilitate introduction of oxygen, possibly as a result of increased laccase activity, and increase availability of nutrients (as indicated by higher EC) in a coal-polluted rhizosphere, to provide plants and their associated mycorrhizae and presumably other beneficial microorganisms a more mesic environment for sustained phytoremediation with enhanced rehabilitation potential. In conclusion, this study confirms the positive role of root exudate in mediating a mutualistic rehabilitation strategy involving plants and fungi such as the FungCoal bioprocess.