Plastic waste in the environment is a significant threat due to its resistance to biological processes. Here we report the ability of fungal strains found on floating plastic debris to degrade ...plastics. In particular, we wanted to know which fungi grow on plastic debris floating in the shoreline, whether these fungi have the ability to degrade plastics, whether the plastic-degrading fungi can degrade other complex C-polymers such as lignin, and whether lignin-degraders vice versa can also break down plastics. Overall, more than a hundred fungal strains were isolated from plastic debris of the shoreline of Lake Zurich, Switzerland, and grouped morphologically. Representative strains of these groups were then selected and genetically identified, altogether twelve different fungal species and one species of Oomycota. The list of fungi included commonly occurring saprotrophic fungi but also some plant pathogens. These fungal strains were then used to test the ability to degrade polyethylene and polyurethane. The tests showed that none of the strains were able to degrade polyethylene. However, four strains were able to degrade polyurethane, the three litter-saprotrophic fungi Cladosporium cladosporioides, Xepiculopsis graminea, and Penicillium griseofulvum and the plant pathogen Leptosphaeria sp. A series of additional fungi with an origin other than from plastic debris were tested as well. Here, only the two litter-saprotrophic fungi Agaricus bisporus and Marasmius oreades showed the capability to degrade polyurethane. In contrast, wood-saprotrophic fungi and ectomycorrhizal fungi were unable to degrade polyurethane. Overall, it seems that in majority only a few litter-saprotrophic fungi, which possess a wide variety of enzymes, have the ability to degrade polyurethane. None of the fungi tested was able to degrade polyethylene.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Summary
Due to climate warming, alpine ecosystems are changing rapidly. Ongoing upward migrations of plants and thus an increase of easily decomposable substrates will strongly affect the soil ...microbiome. To understand how belowground communities will respond to such changes, we set up an incubation experiment with permafrost and active soil layers from northern (NW) and southern (SE) slopes of a mountain ridge on Muot da Barba Peider in the Swiss Alps and incubated them with or without artificial root exudates (AREs) at two temperatures, 4°C or 15°C. The addition of AREs resulted in elevated respiration across all soil types. Bacterial and fungal alpha diversity decreased significantly, coinciding with strong shifts in microbial community structure in ARE‐treated soils. These shifts in bacterial community structure were driven by an increased abundance of fast‐growing copiotrophic taxa. Fungal communities were predominantly affected by AREs in SE active layer soils and shifted towards fast‐growing opportunistic yeast. In contrast, in the colder NW facing active layer and permafrost soils fungal communities were more influenced by temperature changes. These findings demonstrate the sensitivity of soil microbial communities in high alpine ecosystems to climate change and how shifts in these communities may lead to functional changes impacting biogeochemical processes.
Plastic is exceedingly abundant in soils, but little is known about its ecological consequences for soil microbiome functioning. Here we report the impacts of polyethylene and biodegradable Ecovio ...and BI-OPL plastic films buried in alpine soils for 5 months on the genetic potential of the soil microbiome using shotgun metagenomics. The microbiome was more affected by Ecovio and BI-OPL than by polyethylene. Fungi, α- and β-Proteobacteria dominated on the biodegradable films. Ecovio and BI-OPL showed signs of degradation after the incubation, whereas polyethylene did not. Genes involved in cellular processes and signaling (intracellular trafficking, secretion, vesicular transport), as well as metabolism (carbohydrate, lipid and secondary metabolism), were enriched in the plastisphere. Several α/β-hydrolase gene families (cutinase_like, polyesterase-lipase-cutinase, carboxylesterase), which encode enzymes essential to plastic degradation, and carbohydrate-active genes involved in lignin and murein degradation increased on Ecovio and BI-OPL films. Enriched nitrogen fixation and organic N degradation and synthesis genes and decreased nitrification genes on Ecovio altered the biogeochemical cycling, leading to higher ammonium concentrations and depletion of nitrite and nitrate in the soil. Our results indicate that plastics affect the alpine soil microbiome and its functions and suggest that the plastisphere has an untapped microbial potential for plastic biodegradation.
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•We studied the microbial genetic potential of the plastisphere in alpine soil.•Biodegradable plastics led to strong changes in the gene structure.•α/β-hydrolase genes were enriched in the plastisphere of biodegradable plastics.•Biodegradable plastics affected C- and N-cycling in the soil.•The alpine soil plastisphere is an untapped source of plastic-degrading enzymes.
Plastic pollution poses a threat to terrestrial ecosystems, even impacting soils from remote alpine and arctic areas. Biodegradable plastics are a promising solution to prevent long-term accumulation ...of plastic litter. However, little is known about the decomposition of biodegradable plastics in soils from alpine and polar ecosystems or the microorganisms involved in the process. Plastics in aquatic environments have previously been shown to form a microbial community on the surface of the plastic distinct from that in the surrounding water, constituting the so-called ‘plastisphere’. Comparable studies in terrestrial environments are scarce. Here, we aimed to characterize the plastisphere microbiome of three types of plastics differing in their biodegradability in soil using DNA metabarcoding. Polylactic acid (PLA), polybutylene adipate terephthalate (PBAT) and polyethylene (PE) were buried in two different soils, from the Swiss Alps and from Northern Greenland, at 15°C for eight weeks. While physico-chemical characteristics of the polymers only showed minor (PLA, PBAT) or no (PE) changes after incubation, a considerably lower α-diversity was observed on the plastic surfaces and prominent shifts occurred in the bacterial and fungal community structures between the plastisphere and the adjacent bulk soil not affected by the plastic. Effects on the plastisphere microbiome decreased with greater biodegradability of the plastics, from PLA to PE. Copiotrophic taxa within the phyla Proteobacteria and Actinobacteria benefitted the most from plastic input. Especially taxa with a known potential to degrade xenobiotics, including Burkholderiales, Caulobacterales, Pseudomonas, Rhodococcus and Streptomyces, thrived in the plastisphere of the Alpine and Arctic soils. In addition, Saccharimonadales (superphylum Patescibacteria) was identified as a key taxon associated with PLA. The association of Saccharibacteria with plastic has not been reported before, and pursuing this finding further may shed light on the lifestyle of this obscure candidate phylum. Plastic addition affected fungal taxa to a lesser extent since only few fungal genera such as Phlebia and Alternaria were increased on the plastisphere. Our findings suggest that the soil microbiome can be strongly influenced by plastic pollution in terrestrial cryoenvironments. Further research is required to fully understand microbial colonization on plastic surfaces and the biodegradation of plastic in soils.
Plastic materials, including microplastics, accumulate in all types of ecosystems, even in remote and cold environments such as the European Alps. This pollution poses a risk for the environment and ...humans and needs to be addressed. Using shotgun DNA metagenomics of soils collected in the eastern Swiss Alps at about 3,000 m a.s.l., we identified genes and their proteins that potentially can degrade plastics. We screened the metagenomes of the plastisphere and the bulk soil with a differential abundance analysis, conducted similarity-based screening with specific databases dedicated to putative plastic-degrading genes, and selected those genes with a high probability of signal peptides for extracellular export and a high confidence for functional domains. This procedure resulted in a final list of nine candidate genes. The lengths of the predicted proteins were between 425 and 845 amino acids, and the predicted genera producing these proteins belonged mainly to Caballeronia and Bradyrhizobium. We applied functional validation, using heterologous expression followed by enzymatic assays of the supernatant. Five of the nine proteins tested showed significantly increased activities when we used an esterase assay, and one of these five proteins from candidate genes, a hydrolase-type esterase, clearly had the highest activity, by more than double. We performed the fluorescence assays for plastic degradation of the plastic types BI-OPL and ecovio® only with proteins from the five candidate genes that were positively active in the esterase assay, but like the negative controls, these did not show any significantly increased activity. In contrast, the activity of the positive control, which contained a PLA-degrading gene insert known from the literature, was more than 20 times higher than that of the negative controls. These findings suggest that in silico screening followed by functional validation is suitable for finding new plastic-degrading enzymes. Although we only found one new esterase enzyme, our approach has the potential to be applied to any type of soil and to plastics in various ecosystems to search rapidly and efficiently for new plastic-degrading enzymes.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Increasing plastic production and the release of some plastic in to the environment highlight the need for circular plastic economy. Microorganisms have a great potential to enable a more sustainable ...plastic economy by biodegradation and enzymatic recycling of polymers. Temperature is a crucial parameter affecting biodegradation rates, but so far microbial plastic degradation has mostly been studied at temperatures above 20°C. Here, we isolated 34 cold-adapted microbial strains from the plastisphere using plastics buried in alpine and Arctic soils during laboratory incubations as well as plastics collected directly from Arctic terrestrial environments. We tested their ability to degrade, at 15°C, conventional polyethylene (PE) and the biodegradable plastics polyester-polyurethane (PUR; Impranil
); ecovio
and BI-OPL, two commercial plastic films made of polybutylene adipate-co-terephthalate (PBAT) and polylactic acid (PLA); pure PBAT; and pure PLA. Agar clearing tests indicated that 19 strains had the ability to degrade the dispersed PUR. Weight-loss analysis showed degradation of the polyester plastic films ecovio
and BI-OPL by 12 and 5 strains, respectively, whereas no strain was able to break down PE. NMR analysis revealed significant mass reduction of the PBAT and PLA components in the biodegradable plastic films by 8 and 7 strains, respectively. Co-hydrolysis experiments with a polymer-embedded fluorogenic probe revealed the potential of many strains to depolymerize PBAT.
and
strains were able to degrade all the tested biodegradable plastic materials, making these strains especially promising for future applications. Further, the composition of the culturing medium strongly affected the microbial plastic degradation, with different strains having different optimal conditions. In our study we discovered many novel microbial taxa with the ability to break down biodegradable plastic films, dispersed PUR, and PBAT, providing a strong foundation to underline the role of biodegradable polymers in a circular plastic economy.
Global warming is affecting all cold environments, including the European Alps and Arctic regions. Here, permafrost may be considered a unique ecosystem harboring a distinct microbiome. The frequent ...freeze-thaw cycles occurring in permafrost-affected soils, and mainly in the seasonally active top layers, modify microbial communities and consequently ecosystem processes. Although taxonomic responses of the microbiomes in permafrost-affected soils have been widely documented, studies about how the microbial genetic potential, especially pathways involved in C and N cycling, changes between active-layer soils and permafrost soils are rare. Here, we used shotgun metagenomics to analyze the microbial and functional diversity and the metabolic potential of permafrost-affected soil collected from an alpine site (Val Lavirun, Engadin area, Switzerland) and a High Arctic site (Station Nord, Villum Research Station, Greenland). The main goal was to discover the key genes abundant in the active-layer and permafrost soils, with the purpose to highlight the potential role of the functional genes found.
We observed differences between the alpine and High Arctic sites in alpha- and beta-diversity, and in EggNOG, CAZy, and NCyc datasets. In the High Arctic site, the metagenome in permafrost soil had an overrepresentation (relative to that in active-layer soil) of genes involved in lipid transport by fatty acid desaturate and ABC transporters, i.e. genes that are useful in preventing microorganisms from freezing by increasing membrane fluidity, and genes involved in cell defense mechanisms. The majority of CAZy and NCyc genes were overrepresented in permafrost soils relative to active-layer soils in both localities, with genes involved in the degradation of carbon substrates and in the degradation of N compounds indicating high microbial activity in permafrost in response to climate warming.
Our study on the functional characteristics of permafrost microbiomes underlines the remarkably high functional gene diversity of the High Arctic and temperate mountain permafrost, including a broad range of C- and N-cycling genes, and multiple survival and energetic metabolisms. Their metabolic versatility in using organic materials from ancient soils undergoing microbial degradation determine organic matter decomposition and greenhouse gas emissions upon permafrost thawing. Attention to their functional genes is therefore essential to predict potential soil-climate feedbacks to the future warmer climate.
Indolyloxazole alkaloids occur in diverse micro‐ and macroorganisms and exhibit a wide range of pharmacological activities. Despite their ubiquitous occurrence and simple structures, the biosynthetic ...pathway remained unknown. Here, we used transposon mutagenesis in the labradorin producer Pseudomonas entomophila to identify a cryptic biosynthetic locus encoding an N‐acyltransferase and a non‐heme diiron desaturase‐like enzyme. Heterologous expression in E. coli demonstrates that both enzymes are sufficient to produce indolyloxazoles. Probing their function in stable‐isotope feeding experiments, we provide evidence for an unusual desaturase mechanism that generates the oxazole by decarboxylative cyclization.
Indolyloxazole alkaloids are widespread natural products with diverse bioactivities but previously unknown biosynthesis. Transposon mutagenesis, heterologous pathway expression, and labeling experiments provide evidence for a Pseudomonas pathway involving oxazole formation by cyclization as a new reaction type catalyzed by a desaturase‐like enzyme.
Indolyloxazole alkaloids occur in diverse micro‐ and macroorganisms and exhibit a wide range of pharmacological activities. Despite their ubiquitous occurrence and simple structures, the biosynthetic ...pathway remained unknown. Here, we used transposon mutagenesis in the labradorin producer Pseudomonas entomophila to identify a cryptic biosynthetic locus encoding an N‐acyltransferase and a non‐heme diiron desaturase‐like enzyme. Heterologous expression in E. coli demonstrates that both enzymes are sufficient to produce indolyloxazoles. Probing their function in stable‐isotope feeding experiments, we provide evidence for an unusual desaturase mechanism that generates the oxazole by decarboxylative cyclization.
Indolyloxazole alkaloids are widespread natural products with diverse bioactivities but previously unknown biosynthesis. Transposon mutagenesis, heterologous pathway expression, and labeling experiments provide evidence for a Pseudomonas pathway involving oxazole formation by cyclization as a new reaction type catalyzed by a desaturase‐like enzyme.
Initial enteropathogen growth in the microbiota-colonized gut is poorly understood. Salmonella Typhimurium is metabolically adaptable and can harvest energy by anaerobic respiration using ...microbiota-derived hydrogen (H
) as an electron donor and fumarate as an electron acceptor. As fumarate is scarce in the gut, the source of this electron acceptor is unclear. Here, transposon sequencing analysis along the colonization trajectory of S. Typhimurium implicates the C4-dicarboxylate antiporter DcuABC in early murine gut colonization. In competitive colonization assays, DcuABC and enzymes that convert the C4-dicarboxylates aspartate and malate into fumarate (AspA, FumABC), are required for fumarate/H
-dependent initial growth. Thus, S. Typhimurium obtains fumarate by DcuABC-mediated import and conversion of L-malate and L-aspartate. Fumarate reduction yields succinate, which is exported by DcuABC in exchange for L-aspartate and L-malate. This cycle allows S. Typhimurium to harvest energy by H
/fumarate respiration in the microbiota-colonized gut. This strategy may also be relevant for commensal E. coli diminishing the S. Typhimurium infection.
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