Highly efficient screening and optimal combination of functional isolates for bioremediation of hydrocarbon-polluted soil.

The key to enhancing the efficacy of bioremediation of hydrocarbon-contaminated soil is the precise and highly efficient screening of functional isolates. Low screening effectiveness, narrow screening range and an unstable structure of the constructed microflora during bioremediation are the shortcomings of the traditional shaking culture (TSC) method. To improve the secondary screening of isolates and microflora implemented for alkane degradation, this work evaluated the characterization relationship between bacterial function and enzyme activity and devised an enzyme activity assay (EAA) method. The results indicated a substantial positive correlation (r = 0.97) between 24 candidate isolates and their whole enzymes, proving that whole enzyme activity properly reflects the metabolic functions of microorganisms. The functional analysis of the isolates demonstrated that the EAA method in conjunction with microbial abundance and metabolite determination could broaden the screening range of functional isolates, including aliphatic acid-metabolizing isolates (isolates H4 and H7) and aliphatic acid-sensitive isolates (isolate H2) with n-hexadecane degradation ability. The EAA method also guided the construction of functional microflora and optimized the mode of application using combinations of alkane-degrading bacteria and aliphatic acid-degrading bacteria successively (e.g., F1+H7+H7). The combinations maintained a high abundance of functional isolates and stable α diversity and community composition throughout the experiment, which contributed to more advanced alkane degradation and mineralization ability (p < 0.01). Assuming a workload of 100 tests, the screening efficiency of the EAA method is more than 16 times that of the TSC method, and the greater the quantity of isolates, the higher the screening efficiency, enabling high-throughput screening. In conclusion, the EAA method has a broad-spectrum, accurate and highly efficient screening ability for functional isolates and microflora, which can provide intensive technical support for the development of bioremediation materials and the application of bioremediation technology.

Catabolic profiles dynamics during the bioremediation process of chlorimuron-ethyl contaminated soil by Methanolivorans CHL1T.

Excessive application of the long-term herbicide chlorimuron-ethyl has resulted in series of environmental problems. Bioaugmentation usually a useful method in contaminated-environment remediation. In this study, the strain Methanolivorans CHL1T with highly chlorimuron-ethyl degrading efficiency was employed to assess its remediation effects on chlorimuron-ethyl-contaminated soil. The chlorimuron-ethyl residues in the soils and the survival condition of strain CHL1T were detected. Meanwhile, the shifts of soil microbial catabolic profile were investigated by MicroResp™ analysis for the first time. The results indicated that strain CHL1T significantly shorten the half-life (6-17 days) of chlorimuron-ethyl and removed 95-100% of chlorimuron-ethyl by the end of the experiment. Meanwhile, the strain CHL1 could inhabit in soil steadily (4.2-4.7 × 107 per g dry soil) for a long time. The inoculation with strain CHL1 significantly shorten and relieved the disturbance effects of chlorimuron-ethyl on soil CLPPs. After inoculation with strain CHL1 60 days, the basal respiration rates and Shannon-Wiener indices of groups S10+ and S30+ had recovered to the control level. Even in the high chlorimuron-ethyl-treated groups (S100), the basal respiration rates and Shannon-Wiener indices were significantly higher in S100+ than that in S100-. These results show the outstanding remediation effects of strain CHL1 and provide new insights into the assessment of the remediation process of chlorimuron-ethyl contaminated soils.

Temporal dynamic of parasite-mediated linkages between the forest canopy and soil processes and the microbial community.

Parasitic plants are important drivers of community and ecosystem properties. In this study, we identify different mechanisms by which mistletoe (Viscum album subsp. austriacum) can affect soil chemical and biological properties at different temporal stages of parasitism. We quantified the effect of parasitism on host growth and the number of frugivorous mutualists visiting the host canopy. Then we collected, identified, and weighed the organic matter input underneath tree canopies and analyzed its nutrient content. Simultaneously, we analyzed soil samples under tree canopies and examined the chemical properties, microbial abundance, and functional evenness of heterotrophic microbial communities. Mistletoe increased the amount, quality, and diversity of organic matter input beneath the host canopy, directly through its nutrient-rich litter and indirectly through a reduction in host litterfall and an increase in bird-derived debris. All these effects gave rise to enriched hotspots able to support larger and more functionally even soil microbial communities beneath parasitized hosts, the effects of which were accentuated after host death. We conclude that mistletoe, together with the biotic interactions it mediates, plays a key role in intensifying soil resource availability, regulating the functional evenness, abundance, and spatial distribution of soil microbial communities.

Nitrogen supply modulates the effect of changes in drying-rewetting frequency on soil C and N cycling and greenhouse gas exchange.

Climate change and atmospheric nitrogen (N) deposition are two of the most important global change drivers. However, the interactions of these drivers have not been well studied. We aimed to assess how the combined effect of soil N additions and more frequent soil drying-rewetting events affects carbon (C) and N cycling, soil:atmosphere greenhouse gas (GHG) exchange, and functional microbial diversity. We manipulated the frequency of soil drying-rewetting events in soils from ambient and N-treated plots in a temperate forest and calculated the Orwin & Wardle Resistance index to compare the response of the different treatments. Increases in drying-rewetting cycles led to reductions in soil NO3- levels, potential net nitrification rate, and soil : atmosphere GHG exchange, and increases in NH4+ and total soil inorganic N levels. N-treated soils were more resistant to changes in the frequency of drying-rewetting cycles, and this resistance was stronger for C- than for N-related variables. Both the long-term N addition and the drying-rewetting treatment altered the functionality of the soil microbial population and its functional diversity. Our results suggest that increasing the frequency of drying-rewetting cycles can affect the ability of soil to cycle C and N and soil : atmosphere GHG exchange and that the response to this increase is modulated by soil N enrichment.

Application of MicroResp™ for quick and easy detection of plastic degradation by marine bacterial isolates.

Microplastic debris in the marine environment is a global problem. Biodegradable polymers are being developed as alternatives to petroleum-based plastics, and quick and easy methods for screening for bacterial strains that can degrade such polymers are needed. As a screening method, the clear zone method has been widely used but has technical difficulties such as plate preparation and interpretation of results. In this study, we adapted the MicroResp™ system to easily detect biodegradation activity of marine bacteria in a 3-day assay. Among the 6 bacterial strains tested, 3, 2 and 1 strain degraded poly (butylene succinate-co-adipate) (PBSA), poly (ε-caprolactone) (PCL) and poly (3-hydroxybutyrate-co-3-hydroxyhexanoate), respectively. Only one strain that showed degradation activity of PBSA and PCL in the MicroResp™ system was also positive in the clear zone assay on the respective emulsion plates. Our results show that the adapted MicroResp™ system can screen for bacterial strains that degrade plastic.

Denaturing gradient gel electrophoresis and multi-SIR profiles of soil microbial communities from a karst doline at Aggtelek National Park, Hungary.

Soils play an important role in the ecosystem of karstic landscapes both as a buffer zone and as a source of acidity to belowground water. Although the microbiota of karstic soils is known to have a great effect on karstification processes, the activity and composition of these communities are largely unknown. This study gives a comparative analysis of soil microbial profiles from different parts of a doline located at Aggtelek, Hungary. The aim was to reveal the relationships between the vegetation type and genetic fingerprints and substrate utilisation (multi-SIR) profiles of the soil microbiota. Soil samples were collected in early and late springs along a transect in a doline covered with different types of vegetation. Genetic fingerprints of bacterial communities were examined by denaturing gradient gel electrophoresis (DGGE) based on the 16S rRNA gene, along with multi-SIR profiles of the microbial communities measured by the MicroResp method using 15 different carbon sources. Genetic fingerprinting indicated that vegetation cover had a strong effect on the composition of soil bacterial communities. Procrustean analysis showed only a weak connection between DGGE and multi-SIR profiles, probably due to the high functional redundancy of the communities. Seasonality had a significant effect on substrate usage, which can be an important factor to consider in future studies.

Erratum: Microplastic Shape, Polymer Type, and Concentration Affect Soil Properties and Plant Biomass.

[This corrects the article DOI: 10.3389/fpls.2021.616645.].

Microplastic Shape, Polymer Type, and Concentration Affect Soil Properties and Plant Biomass.

Microplastics may enter the soil in a wide range of shapes and polymers. However, little is known about the effects that microplastics of different shapes, polymers, and concentration may have on soil properties and plant performance. To address this, we selected 12 microplastics representing different shapes (fibers, films, foams, and fragments) and polymers, and mixed them each with soil at a concentration of 0.1, 0.2, 0.3, and 0.4%. A phytometer (Daucus carota) grew in each pot during 4 weeks. Shoot, root mass, soil aggregation, and microbial activity were measured. All shapes increased plant biomass. Shoot mass increased by ∼27% with fibers, ∼60% with films, ∼45% with foams, and by ∼54% with fragments, as fibers hold water in the soil for longer, films decrease soil bulk density, and foams and fragments can increase soil aeration and macroporosity, which overall promote plant performance. By contrast, all shapes decreased soil aggregation by ∼25% as microplastics may introduce fracture points into aggregates and due to potential negative effects on soil biota. The latter may also explain the decrease in microbial activity with, for example, polyethylene films. Our findings show that shape, polymer type, and concentration are key properties when studying microplastic effects on terrestrial systems.

Enzyme activities and microbial functional diversity in metal(loid) contaminated soils near to a copper smelter.

The monitoring of soil metal(loid) contamination is of global significance due to deleterious effects that metal(loid)s have on living organisms. Soil biological properties such as enzyme activities (EAs) are good indicators of metal(loid) contamination due to their high sensitivity, fast response, and low-cost. Here, the effect of metal(loid) contamination on physicochemical properties and microbial functionality in soils sampled from within 10 km of a Cu smelter is investigated. Soil composite samples were randomly taken within 2, 4, 6, 8 and10 km zones from a mining industry Cu smelter. The EAs of dehydrogenase (DHA), arylsulfatase (ARY), ß-glucosidase, urease, and arginine ammonification (AA) were studied as indicators of metal(loid) contamination, which included the ecological dose (ED50) with respect to Cu and As contents. The community level physiological profile (CLPP), functional diversity, and catabolic evenness were evaluated based on the C-substrate utilisation. All EAs decreased in zones with high degrees of metal(loid) contamination, which also had low TOC and clay contents, reflecting long term processes of soil degradation. Positive and strong relationships between EAs and TOC were found. DHA and ARY activities decreased by approximately 85-90% in highly metal(loid) contaminated soils. DHA and AA showed significant ED50 values associated with available Cu (112.8 and 121.6 mg CuDTPA kg-1, respectively) and total As contents (30.8 and 31.8 mg As kg-1, respectively). The CLPP showed different metabolic profiles along the metal(loid) contamination gradients. Long-term stress conditions in soils close to industrial areas resulted in the decreasing of general biological activity, catabolic capacity, and functional diversity.

Distance-dependent varieties of microbial community structure and metabolic functions in the rhizosphere of Sedum alfredii Hance during phytoextraction of a cadmium-contaminated soil.

The recovery of microbial community and activities is crucial to the remediation of contaminated soils. Distance-dependent variations of microbial community composition and metabolic characteristics in the rhizospheric soil of hyperaccumulator during phytoextraction are poorly understood. A 12-month phytoextraction experiment with Sedum alfredii in a Cd-contaminated soil was conducted. A pre-stratified rhizobox was used for separating sub-layer rhizospheric (0-2, 2-4, 4-6, 6-8, 8-10 mm from the root mat)/bulk soils. Soil microbial structure and function were analyzed by phospholipid fatty acid (PLFA) and MicroResp™ methods. The concentrations of total and specified PLFA biomarkers and the utilization rates for the 14 substrates (organic carbon) in the 0-2-mm sub-layer rhizospheric soil were significantly increased, as well as decreased with the increase in the distance from the root mat. Microbial structure measured by the ratios of different groups of PLFAs such as fungal/bacterial, monounsaturated/saturated, ratios of Gram-positive to Gram-negative (GP/GN) bacterial, and cyclopropyl/monoenoic precursors and 19:0 cyclo/18:1ω7c were significantly changed in the 0-2-mm soil. The PLFA contents and substrate utilization rates were negatively correlated with pH and total, acid-soluble, and reducible fractions of Cd, while positively correlated with labile carbon. The dynamics of microbial community were likely due to root exudates and Cd uptake by S. alfredii. This study revealed the stimulations and gradient changes of rhizosphere microbial community through phytoextraction, as reduced Cd concentration, pH, and increased labile carbons are due to the microbial community responses.

Do temperate tree species diversity and identity influence soil microbial community function and composition?

Studies of biodiversity-ecosystem function in treed ecosystems have generally focused on aboveground functions. This study investigates intertrophic links between tree diversity and soil microbial community function and composition. We examined how microbial communities in surface mineral soil responded to experimental gradients of tree species richness (SR), functional diversity (FD), community-weighted mean trait value (CWM), and tree identity. The site was a 4-year-old common garden experiment near Montreal, Canada, consisting of deciduous and evergreen tree species mixtures. Microbial community composition, community-level physiological profiles, and respiration were evaluated using phospholipid fatty acid (PLFA) analysis and the MicroResp™ system, respectively. The relationship between tree species richness and glucose-induced respiration (GIR), basal respiration (BR), metabolic quotient (qCO 2) followed a positive but saturating shape. Microbial communities associated with species mixtures were more active (basal respiration [BR]), with higher biomass (glucose-induced respiration [GIR]), and used a greater number of carbon sources than monocultures. Communities associated with deciduous tree species used a greater number of carbon sources than those associated with evergreen species, suggesting a greater soil carbon storage capacity. There were no differences in microbial composition (PLFA) between monocultures and SR mixtures. The FD and the CWM of several functional traits affected both BR and GIR. In general, the CWM of traits had stronger effects than did FD, suggesting that certain traits of dominant species have more effect on ecosystem processes than does FD. Both the functions of GIR and BR were positively related to aboveground tree community productivity. Both tree diversity (SR) and identity (species and functional identity-leaf habit) affected soil microbial community respiration, biomass, and composition. For the first time, we identified functional traits related to life-history strategy, as well as root traits that influence another trophic level, soil microbial community function, via effects on BR and GIR.

Effect of rice straw application on microbial community and activity in paddy soil under different water status.

Rice straw application and flooding are common practices in rice production, both of which can induce changes in the microbial community. This study used soil microcosms to investigate the impact of water status (saturated and nonsaturated) and straw application (10 g kg(-1) soil) on soil microbial composition (phospholipid fatty acid analysis) and activity (MicroResp(™) method). Straw application significantly increased total PLFA amount and individual PLFA components independent of soil moisture level. The amount of soil fungal PLFA was less than Gram-negative, Gram-positive, and actinomycete PLFA, except the drained treatment with rice straw application, which had higher fungal PLFA than actinomycete PLFA at the initial incubation stage. Straw amendment and waterlogging had different effects on microbial community structure and substrate-induced pattern. PLFA profiles were primarily influenced by straw application, whereas soil water status had the greater influence on microbial respiration. Of the variation in PLFA and respiration data, straw accounted for 30.1 and 16.7 %, while soil water status explained 7.5 and 29.1 %, respectively. Our results suggest that (1) the size of microbial communities in paddy soil is more limited by carbon substrate availability rather than by the anaerobic conditions due to waterlogging and (2) that soil water status is more important as a control of fungal growth and microbial community activity.

Soil microbial respiration and PICT responses to an industrial and historic lead pollution: a field study.

We performed a field investigation to study the long-term impacts of Pb soil contamination on soil microbial communities and their catabolic structure in the context of an industrial site consisting of a plot of land surrounding a secondary lead smelter. Microbial biomass, catabolic profiles, and ecotoxicological responses (PICT) were monitored on soils sampled at selected locations along 110-m transects established on the site. We confirmed the high toxicity of Pb on respirations and microbial and fungal biomasses by measuring positive correlations with distance from the wall factory and negative correlation with total Pb concentrations. Pb contamination also induced changes in microbial and fungal catabolic structure (from carbohydrates to amino acids through carboxylic malic acid). Moreover, PICT measurement allowed to establish causal linkages between lead and its effect on biological communities taking into account the contamination history of the ecosystem at community level. The positive correlation between qCO2 (based on respiration and substrate use) and PICT suggested that the Pb stress-induced acquisition of tolerance came at a greater energy cost for microbial communities in order to cope with the toxicity of the metal. In this industrial context of long-term polymetallic contamination dominated by Pb in a field experiment, we confirmed impacts of this metal on soil functioning through microbial communities, as previously observed for earthworm communities.

Metabolic activity and genetic diversity of microbial communities inhabiting the rhizosphere of halophyton plants.

A preliminary study was conducted to compare the community level physiological profile (CLPP) and genetic diversity of rhizosphere microbial communities of four plant species growing nearby Kiskunság soda ponds, namely Böddi-szék, Kelemen-szék and Zab-szék. CLPP was assessed by MicroResp method using 15 different substrates while Denaturing Gradient Gel Electrophoresis (DGGE) was used to analyse genetic diversity of bacterial communities. The soil physical and chemical properties were quite different at the three sampling sites. Multivariate statistics (PCA and UPGMA) revealed that Zab-szék samples could be separated according to their genetic profile from the two others which might be attributed to the geographical location and perhaps the differences in soil physical properties. Böddi-szék samples could be separated from the two others considering the metabolic activity which could be explained by their high salt and low humus contents. The number of bands in DGGE gels was related to the metabolic activity, and positively correlated with soil humus content, but negatively with soil salt content. The main finding was that geographical location, soil physical and chemical properties and the type of vegetation were all important factors influencing the metabolic activity and genetic diversity of rhizosphere microbial communities.

Soil microbial communities respond differently to three chemically defined polyphenols.

High molecular weight polyphenols (e.g. tannins) that enter the soil may affect microbial populations, by serving as substrates for microbial respiration or by selecting for certain microbes. In this study we examined how three phenolic compounds that represent some environmentally widespread tannins or their constituent functional groups were respired by soil microorganisms and how the compounds affected the abundance and diversity of soil bacteria and archaea, including ammonia oxidizers. An acidic, silt loam soil from a pine forest was incubated for two weeks with the monomeric phenol methyl gallate, the small polyphenol epigallocatechin gallate, or the large polyphenol oenothein B. Respiration of the polyphenols during the incubation was measured using the Microresp™ system. After incubation, metabolic diversity was determined by community level physiological profiling (CLPP), and genetic diversity was determined using denaturing gradient gel electrophoresis (DGGE) analysis on DNA extracted from the soil samples. Total microbial populations and ammonia-oxidizing populations were measured using real time quantitative polymerase chain reaction (qPCR). Methyl gallate was respired more efficiently than the higher molecular weight tannins but not as efficiently as glucose. Methyl gallate and epigallocatechin gallate selected for genetically or physiologically unique populations compared to glucose. None of the polyphenols supported microbial growth, and none of the polyphenols affected ammonia-oxidizing bacterial populations or ammonia-oxidizing archaea. Additional studies using both a wider range of polyphenols and a wider range of soils and environments are needed to elucidate the role of polyphenols in determining soil microbiological diversity.