Filter centrifugation as a sampling method for miniaturization of extracellular fungal enzyme activity measurements in solid media

Heinonsalo, Jussi; Kabiersch, Grit; Niemi, R. Maarit; Simpanen, Suvi; Ilvesniemi, Hannu; Hofrichter, Martin; Hatakka, Annele; Steffen, Kari

doi:10.1016/j.funeco.2011.07.008

Cited by 19 publications

(12 citation statements)

References 16 publications

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“…For the ectomycorrhizal fungi in our study, peroxidase activity may either have been generated by intracellular or extracellular processes. Peroxidase activity has been observed in the culture filtrates of C. geophilum in the presence of soil humus (Heinonsalo et al, 2012), as well as in the culture filtrates of many other ectomycorrhizal fungi (Gramss et al, 1999), including Piloderma species (Heinonsalo et al, 2012), which had the second highest peroxidase activity of ectomycorrhizal fungi in our study. Whether generated intracellularly or extracellularly, the production of peroxidase activity by certain ectomycorrhizal fungi in soils is substantial (Table 6), and could be due in part to turnover of ectomycorrhizal hyphae and release of intracellular products into the soil matrix.…”

Section: Factorsupporting

confidence: 60%

Functional guild classification predicts the enzymatic role of fungi in litter and soil biogeochemistry

Talbot

Martin

Kohler

et al. 2015

Soil Biology and Biochemistry

130

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a b s t r a c tLinking community composition to ecosystem function is a challenge in complex microbial communities. We tested the hypothesis that key biological features of fungi -evolutionary history, functional guild, and abundance of functional genes e can predict the biogeochemical activity of fungal species during decay. We measured the activity of 10 different enzymes produced by 48 model fungal species on leaf litter in laboratory microcosms. Taxa included closely related species with different ecologies (i.e. species in different "functional guilds") and species with publicly available genomes. Decomposition capabilities differed less among phylogenetic lineages of fungi than among different functional guilds. Activity of carbohydrases and acid phosphatase was significantly higher in litter colonized by saprotrophs compared to ectomycorrhizal species. By contrast, oxidoreductase activities per unit fungal biomass were statistically similar across functional guilds, with white rot fungi having highest polyphenol oxidase activity and ectomycorrhizal fungi having highest peroxidase activity. On the ecosystem level, polyphenol oxidase activity in soil correlated with the abundance of white rot fungi, while soil peroxidase activity correlated with the abundance of ectomycorrhizal fungi in soil. Copy numbers of genes coding for different enzymes explained the activity of some carbohydrases and polyphenol oxidase produced by fungi in culture, but were not significantly better predictors of activity than specific functional guild. Collectively, our data suggest that quantifying the specific functional guilds of fungi in soil, potentially through environmental sequencing approaches, allows us to predict activity of enzymes that drive soil biogeochemical cycles.

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Section: Factorsupporting

confidence: 60%

Functional guild classification predicts the enzymatic role of fungi in litter and soil biogeochemistry

Talbot

Martin

Kohler

et al. 2015

Soil Biology and Biochemistry

130

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“…To determine the oxidative enzyme response of T. versicolor to the control and PyOM treatments, a 2,2′-azino-bis-(3-ethylbenzo thiazoline-6-sulfonate) (ABTS) assay was performed on enzymes extracted via micro-centrifugation of growth media, modified from the methods described in Heinonsalo et al (2012) and Berry et al (2014). To recover enzymes from the MEA layer, 4 mm square plugs of media plus fungi were extracted from each tube of which replicates within the same treatment were combined and homogenized.…”

Section: Methodsmentioning

confidence: 99%

Weathering of pyrogenic organic matter induces fungal oxidative enzyme response in single culture inoculation experiments

Gibson

Berry

Wang

et al. 2016

Organic Geochemistry

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The addition of pyrogenic organic matter (PyOM), the aromatic carbon-rich product of the incomplete combustion of plant biomass or fossil fuels, to soil can influence the rate of microbial metabolism of native soil carbon. The interaction of soil heterotrophs with PyOM may be governed by the surficial chemical and physical properties of PyOM that evolve with environmental exposure. We present results of a 36-day laboratory incubation investigating the interaction of a common white-rot fungus, Trametes versicolor, with three forms of 13C-enriched (2.08 atom% 13C) PyOM derived from Pinus ponderosa (450 °C): one freshly produced, and two artificially weathered (254 nm, UV light-water treatment and water-leaching alone). Analysis (FTIR, XPS) of the UV-weathered PyOM showed increased aliphatic C–H content and oxidation of aromatic carbon relative to both the original and water-leached PyOM. The addition of both weathered forms of PyOM stimulated (positively primed) fungal respiration of the growth media, while the unaltered PyOM mildly inhibited (negatively primed) respiration. Artificial weathering resulted in higher oxidative (laccase and peroxidase) enzyme activity than unaltered PyOM, possibly the result of a diminished capacity to bind reactive substrates and extracellular enzymes after weathering. However, and contrary to expectations, simple water-leached weathering resulted in a relatively higher enzyme activity and respiration than that of UV-weathering. The 13C content of respired CO2 indicated negligible fungal oxidation of PyOM for all treatments, demonstrating the overall low microbial reactivity of this high temperature PyOM. The increased enzymatic and positive priming response of T. versicolor to weathered PyOM highlights the importance of weathering-induced chemistry in controlling PyOM–microbe–soil carbon interactions.

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“…Valášková and Baldrian (2006) noticed that a significant amount of P. ostreatus laccase, but not MnP, was bound to the solid fraction during liquid cultivation. In addition, Heinonsalo et al (2012) noticed different binding between laccase and MnP to various agar media. Table 9.…”

Section: Lme Activities In Fungal Bark Inoculum and In Pcdd/f-soil (Amentioning

confidence: 99%

“…Samples for enzyme activity measurements were taken from the bottles on a daily basis. MnP activity was measured with the filter centrifugation method (Heinonsalo et al, 2012). There were three parallel bottles filled with contaminated soil and two parallel bottles filled with control soil.…”

Section: Degradation Of Pcdd/fs By Fungal Enzymesmentioning

confidence: 99%

Fungal enzyme production and biodegradation of polychlorinated dibenzo-p-dioxins and dibenzofurans in contaminated sawmill soil

et al. 2014

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Soils that are contaminated with the most recalcitrant organic contaminants, such as high molecular weight polyaromatic hydrocarbons (HMW PAH) and polychlorinated dibenzo-pdioxins and dibenzofurans (PCDD/F), cannot be degraded efficiently by conventional composting. The only available treatment method for these soils, which destroys the contaminants, is combustion at high temperature. This thesis examines three alternative fungal methods to treat these soils: 1) treatment with fungal enzymes, 2) treatment with fungal inoculum, and 3) fungal treatment used as a pre-treatment to improve the energy efficacy in combustion. Manganese peroxidase (MnP), which belongs to lignin-modifying enzymes (LME), was produced and used to treat PCDD/F-contaminated soil in the laboratory scale. Nevertheless, no degradation with a MnP preparation was observed, although a substantial amount of MnP activity was found in the soil still after 10 days of incubation. Both PAH-and PCDD/Fcontaminated soils were treated with fungal inoculum in the laboratory scale. HMW PAHs were degraded significantly more by the fungi than by the indigenous microbes alone in the laboratory experiments, where the PAH concentration of soil was 3500 mg kg -1 (sum of 16 PAH). Treatment with Phanerochaete velutina (inoculum) resulted to degradation of 96 % of 4-ring PAHs and 39 % of 5-and 6-ring PAHs in three months. With PCDD/F-contaminated soil, no degradation was observed in the control, but the degradation of PCDD/Fs with fungal treatments was significant (P. velutina: 62 %, Stropharia rugosoannulata: 64 % of WHO-TEQ value). Fungal treatment of PAH-contaminated soil was also applied in the field scale (2 t). However, both P. velutina (inoculum) and control treatment resulted in equal degradation in soil with lower PAH concentration (1400 mg kg -1 , sum of 16 PAH): 94 % of the 16 PAHs were degraded in three months. Fungal treatment was even applied as a pre-treatment for contaminated soil with high organic matter content, and which will be later combusted. In the pilot-scale (300 kg), 13 % degradation of the original organic matter content was obtained in 6 months. To conclude, fungal treatment is reasonable to apply for soils with organic contaminants that cannot be bioremediated by composting. With soils contaminated by chlorinated dioxins, this is always the case, but also PAH-contaminated soils with high total concentration or high proportion of HMW-PAHs. In addition, with fungal treatment the amount of organic matter in the soil can be reduced and the efficacy of the combustion process is improved. Tiivistelmä Kaikkein vaikeimmin hajoavilla orgaanisilla yhdisteillä kuten monirenkaisilla polyaromaattisilla hiilivedyillä (PAH) sekä polyklooratuilla dibenzo-p-dioksiineilla ja -furaaneilla (PCDD/F) pilaantuneita maita ei pystytä tehokkaasti kunnostamaan perinteisellä kompostoinnilla. Ainoa käytössä oleva puhdistusmenetelmä, joka hajottaa pilaavat yhdisteet, on maan poltto korkeassa lämpötilassa. Tämä väitöskirja esittelee kolme vaihtoehtoista valkolah...

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Filter centrifugation as a sampling method for miniaturization of extracellular fungal enzyme activity measurements in solid media

Cited by 19 publications

References 16 publications

Functional guild classification predicts the enzymatic role of fungi in litter and soil biogeochemistry

Functional guild classification predicts the enzymatic role of fungi in litter and soil biogeochemistry

Weathering of pyrogenic organic matter induces fungal oxidative enzyme response in single culture inoculation experiments

Fungal enzyme production and biodegradation of polychlorinated dibenzo-p-dioxins and dibenzofurans in contaminated sawmill soil

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