Advances in Pleurotus ostreatus and Agaricus bisporus plastic mycoremediation: Focus on the isoenzymes of the lignin degradation complex.

This study investigates P. ostreatus and A. bisporus biodegradation capacity of low density polyethylene (LDPE) oxidised to simulate environmental weathering. Fourier transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) were used to analyse the degradation of LDPE treated with fungal cultures. Molecular implications of LDPE degradation by P. ostreatus and A. bisporus were evaluated by Reverse transcription followed by quantitative PCR (qRT-PCR) of lac, mnp and lip genes expression. After 90 days of incubation, FT-IR analysis showed, for both fungal treatments, an increasing in the intensity of peaks related to the asymmetric C-C-O stretching (1160 to 1000 cm-1) and the -OH stretching (3700 to 3200 cm-1) due to the formation of alcohols and carboxylic acid, indicating depolymerisation of LDPE. This was confirmed by the SEM analysis, where a widespread alteration of the surface morphology was observed for treated LDPE fragments. Results revealed that the exposure of P. ostreatus to oxidised LDPE treatment led to a significant increase in the expression of the lac6, lac7, lac9, lac10 and mnp2 genes, while A. bisporus showed an over-expression in lac2 and lac12 genes. The obtained results offer new perspectives for a biotechnological use of P. ostreatus and A. bisporus for plastic bioremediation.

Rhus coriaria L. in tradition and innovation like natural dye.

Nowadays, secondary raw materials (SRM) obtained from plant matrices are of great interest for circular economy, suitable for sustainable measures to reduce environmental impact. This work focused on the extraction, characterization and quantification of compounds obtained from leaves and fruits of the Sicilian sumac, Rhus coriaria L. and their application as natural dyes on textile fibres. Extractions were performed with Extractor Naviglio®, maceration and ultrasound assisted methods and food-grade solvents (aqueous and hydroalcoholic) to evaluate the yields for dye compounds. The presence of colouring molecules was evaluated by UV-Vis spectrophotometer, and the extracts selected for colouring were quantified and characterized by LC-MS. The results showed that Extractor Naviglio® achieved the best extraction yield, and the ethanol-water mixture extracts had a higher amount of total phenolic compounds (TPC) and a higher content of total colouring compounds (TCC). These extracts were selected for subsequent applications as dyes for linen, cotton and wool. The chemical profile of selected extracts was rich in compounds such as gallotannin and anthocyanin class. Fibre dyeing was verified by recording CIELAB colouring coordinates. The results suggest that the dyes obtained from R. coriaria can be of great interest for artisanal and industrial processes, in accordance with environmental sustainability.

Microplastics affect soil-plant system: Implications for rhizosphere biology and fitness of sage (Salvia officinalis L.).

A mesocosm experiment was set-up to investigate the effects of low-density polyethylene (LDPE) fragments deriving from plastic film on soil ecology, rhizosphere and plant (Salvia officinalis L.) fitness. The internal transcribed spacer (ITS) and 16S metagenomic analysis was adopted to evaluate taxonomic and functional shifts of both soil and rhizosphere under the influence of microplastics (MPs). Photosynthetic parameters and enzymes involved in oxidative stress were assessed to unveil the plant physiological state. MP fragments were analysed by scanning electron microscope (SEM) and metagenomics to investigate the plastisphere. Microbial biomarkers of MPs pollution were identified in soil and rhizosphere, reinforcing the concept of molecular biomonitoring. Overall, Bacillus, Nocardioides and Streptomyces genera are bacterial biomarkers of MPs pollution in soil whereas Aspergillus, Fusarium and Trichoderma genera, and Nectriaceae family are fungal biomarkers of MPs polluted soil. The data show that the presence of MPs promotes the abundance of taxa involved in the soil N cycle, but simultaneously reduces the endophytic interaction capability and enhances pathogen related functions at the rhizosphere level. A significant decrease in chlorophyll levels and increase of oxidative stress enzymes was observed in plants grown in MPs-polluted soil. The SEM observations of MPs fragments revealed a complex colonisation, where bacteria (Bacillus in MPSo and Microvirga in MPRz) and fungi (Aspergillus in MPSo and Trichoderma in MPRz) represent the main colonisers. The results demonstrate that the presence of MPs causes changes in the soil and rhizosphere microbial community and functions leading to negative effects on plant fitness.

Effects of microplastics on microbial community dynamics in sediments from the Volturno River ecosystem, Italy.

In this study, the sources, abundance, and ecological implications of microplastic (MP) pollution in Volturno, one of the main rivers in southern Italy, were explored by investigating the MP concentration levels in sediments collected along the watercourse. The samples were sieved through 5- and 2-mm sieves and treated with selective organic solvents. The polymer classes polystyrene (PS), polyethylene (PE), polypropylene (PP), polyethylene terephthalate (PET), polycarbonate (PC), nylon 6 (PA6), and nylon 6,6 (PA66) were quantified using pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS) and high-performance liquid chromatography (HPLC). Furthermore, a 16S rRNA metagenomic analysis was performed using next-generation sequencing in Ion Torrent™ to explore the bacterial taxonomy and ecological dynamics of sediment samples. The MPs were detected in all samples collected from the study area. PP and PET were the most abundant and frequently detected polymer types in the analysed samples. The total MP concentration ranged from 1.05 to 14.55 ppm (parts per million), identifying two distinct data populations: high- and low-MP-contaminated sediments. According to the Polymer Hazard Index (PHI), MP pollution was categorised as hazard levels III and IV (corresponding to the danger category). Metagenomic data revealed that the presence of MPs significantly affected the abundance of bacterial taxa; Flavobacteraceae and Nocardiaceae, which are known to degrade polymeric substances, were present in high-MP-contaminated sediments. This study provides new insights into the ecological relevance of MP pollution and suggests that microorganisms may serve as biomarkers of MP pollution.

The rootstock shape microbial diversity and functionality in the rhizosphere of Vitis vinifera L. cultivar Falanghina.

The rhizosphere effect occurring at the root-soil interface has increasingly been shown to play a key role in plant fitness and soil functionality, influencing plants resilience. Here, for the first time, we investigated whether the rootstock genotype on which Vitis vinifera L. cultivar Falanghina is grafted can influence the rhizosphere microbiome. Specifically, we evaluated to which extent the 5BB and 1103P rootstocks are able to shape microbial diversity of rhizosphere environment. Moreover, we explored the potential function of microbial community and its shift under plant genotype influence. We investigated seven vineyards subjected to the same pedo-climatic conditions, similar age, training system and management and collected twelve rhizosphere soil samples for metagenomic analyses and composite soil samples for physical-chemical properties. In this study, we used 16S rRNA gene-based metagenomic analysis to investigate the rhizosphere bacterial diversity and composition. Liner discriminant analysis effect size (LEFSe) was conducted for metagenomic biomarker discovery. The functional composition of sampled communities was determined using PICRUSt, which is based on marker gene sequencing profiles. Soil analyses involved the determination of texture, pH, Cation Exchange Capacity (CSC), Organic Carbon (OC), electrical conductivity (EC), calcium (Ca), magnesium (Mg), potassium (K) content, Phosphorous (P), nitrogen (N). The latter revealed that soil features were quite homogenous. The metagenomic data showed that the bacterial alpha-diversity (Observed OTUs) significantly increased in 1103P rhizosphere microbiota. Irrespective of cultivar, Pseudomonadota was the dominant phylum, followed by Actinomycetota > Bacteroidota > Thermoproteota. However, Actinomycetota was the major marker phyla differentiating the rhizosphere microbial communities associated with the different rootstock types. At the genus level, several taxa belonging to Actinomycetota and Alphaproteobacteria classes were enriched in 1103P genotype rhizosphere. Investigating the potential functional profile, we found that most key enzyme-encoding genes involved in N cycling were significantly more abundant in 5BB rootstock rhizosphere soil. However, we found that 1103P rhizosphere was enriched in genes involved in C cycle and Plant Growth Promotion (PGP) functionality. Our results suggest that the different rootstocks not only recruit specific bacterial communities, but also specific functional traits within the same environment.

Quercus ilex Phyllosphere Microbiome Environmental-Driven Structure and Composition Shifts in a Mediterranean Contex.

The intra- and interdomain phyllosphere microbiome features of Quercus ilex L. in a Mediterranean context is reported. We hypothesized that the main driver of the phyllosphere microbiome might be the season and that atmospheric pollutants might have a co-effect. Hence, we investigated the composition of epiphytic bacteria and fungi of leaves sampled in urban and natural areas (in Southern Italy) in summer and winter, using microscopy and metagenomic analysis. To assess possible co-effects on the composition of the phyllosphere microbiome, concentrations of particulate matter and polycyclic aromatic hydrocarbons (PAHs) were determined from sampled leaves. We found that environmental factors had a significative influence on the phyllosphere biodiversity, altering the taxa relative abundances. Ascomycota and Firmicutes were higher in summer and in urban areas, whereas a significant increase in Proteobacteria was observed in the winter season, with higher abundance in natural areas. Network analysis suggested that OTUs belonging to Acidobacteria, Cytophagia, unkn. Firmicutes(p), Actinobacteria are keystone of the Q. ilex phyllosphere microbiome. In addition, 83 genes coding for 5 enzymes involved in PAH degradation pathways were identified. Given that the phyllosphere microbiome can be considered an extension of the ecosystem services offered by trees, our results can be exploited in the framework of Next-Generation Biomonitoring.

Influence of sediment texture on HDPE microplastics recovery by density separation.

Microplastics (MPs) are an emerging environmental pollutant, threatening marine and terrestrial ecosystems. Because of their properties and their widely varying size (5mm-0.1 µm), it is still difficult to define a valid and efficient method for extracting MPs from solid matrices. Among the several methods proposed, density separation is the most practical and cost-effective one. Progress is still ongoing towards a deeper understanding of the advantages and limitations related to the application of density separation for MPs extraction, the recovery yields and the factors that may influence it. In this context, we introduce the following work, which provides an early-stage insight into how the sediment texture may influence the efficiency of this extraction method, and how parameters, such as sedimentation time and extraction cycles, can be modified to always achieve the best recovery. Our focus has been directed on evaluating the extraction efficiency of HDPE MPs by density separation using NaCl, from three types of sediment: sandy (SS), sandy loam (SLS) and sandy-clay loam (SCLS). We investigated the impact of sedimentation time (1, 6, 12, 24 h) and extraction cycles (3 cycles for each sedimentation time) on MPs recovery. Finally, we determined the minimum amount of MPs (MPs g/g sediment) below which it is not possible to quantify MPs with the method used. The results have shown that the recovery efficiency of MPs from sediment is structure dependent. The highest recoveries are reached after a settling time of 1 and 6 h. Furthermore, for samples with minimum clay content (SS), only one extraction cycle is needed, whereas two extraction cycles are required for SLS and SCLS. The outcomes about the detection limit (LOD) of the method, showed the existence of an interaction MPs-clay/sediment, which allowed us to understand how far this extraction method is suitable in field, thus defining the minimum grade of MPs pollution (MPs g/g sediment) below which this method is no longer capable to extract MPs from contaminated samples.

Molecular effects on spermatozoa of Mytilus galloprovincialis exposed to hyposaline conditions.

Salinity represents a critical environmental and an ecological factor in the reproduction of marine species. As global climate changes and anthropogenic factors affect salinity, in this study, we have analyzed the responses of Mytilus galloprovincialis spermatozoa to hyposaline stress. We exposed mussels, in laboratory tanks, for 24 hr at 18°C to control (35.9 psu) and three hyposaline (17.1, 22.6, and 26.2 psu) conditions, and evaluated the expression of sperm hsp70 and protamine-like proteins genes. Further we analyzed the electrophoretic pattern, the DNA binding and the release from sperm nuclei of protamine-like proteins. For all experimental approaches used, the results obtained at 17.1 psu condition were very similar to those obtained in the control condition, while alterations were always recorded at 22.6 and 26.2 psu conditions. Particularly, at 22.6 and 26.2 psu, was observed: 42.5- and 17.1-fold increase in hsp70 expression, respectively, and hypoexpression of PL-II/PLIV protamine-like proteins genes. Further, electrophoretic mobility shift assays and salt-induced release of nuclear proteins from sperm nuclei, revealed alterations in the PL proteins/DNA binding, in these two hyposaline conditions. The similarity between the results obtained in control and in the more severe hyposaline condition (17.1 psu) could indicate a phenomenon of fertility preservation strategy due to gamete plasticity.