Impact of traditional and biodegradable mulching films on plant performance and substrate microbial communities in a small-scale strawberry cultivation

Impact of traditional and biodegradable mulching films on plant performance and substrate microbial communities in a small-scale strawberry cultivation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Impact of traditional and biodegradable mulching films on plant performance and substrate microbial communities in a small-scale strawberry cultivation Agnese Bellabarba, Lorenzo Bini, Costanza Scopetani, Giulia Selvolini, and 9 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9428619/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background and Aim. Traditional plastic mulching provides clear benefits for strawberry cultivation but pose risks of long-term environmental pollution. Besides, a comprehensive investigation on how different mulch materials may impact the overall strawberry cultivation, from crop productivity and quality to soil health, is still lacking. With this aim, this study investigates the influence of conventional and biodegradable mulching films on plant performance, fruit quality, and substrate microbial ecology in a strawberry cultivation. Methods. Four different mulch films were applied and compared to unmulched control: polyethylene (PE), oxo-degradable (OxoPE), polybutylene adipate terephthalate corn starch-blended (PBAT), polypropylene (PP). Assessments included plant morphological and physiological analysis, strawberries yield and quality, substrate physicochemical analysis and substrate bacterial community characterization by high throughput sequencing of the 16S rRNA gene. Results. OxoPE and PP significantly increased fruit yield up to 29% compared to the control, while physiological and growth traits remained unvaried. Fruit quality and nutraceutical properties were consistent across treatments. PBAT promotes nitrogen enrichment, whereas PE is associated with an increase in available micronutrients. Mulching enhanced bacterial community evenness and diversity, and community structural analysis revealed that each material recruits unique microbial niches leading to distinct bacterial community structures driven by humidity and nitrate gradients. Functional predictions suggest that PE mulches boost nitrogen-fixing communities, whereas PBAT promotes nitrification and complex carbon catabolism. Conclusion. PBAT and OxoPE represent effective alternatives to traditional PE, preserving high productivity and fruit quality while promoting specialized substrates bacterial communities, thus enhancing overall sustainability within strawberry production frameworks. Fragaria x ananassa mulching practices microbial diversity yield and fruits quality soilless cultivation Figures Figure 1 Figure 2 Figure 3 Figure 4 1. INTRODUCTION Feeding a rapidly growing global population requires substantial improvements in agricultural productivity. However, the availability of key resources (i.e., freshwater and fertile soil) is rapidly declining due to the impact of climate change (Schewe et al., 2014 ; Smith and Myers, 2018 ). As a result, conventional farming methods alone are no longer sufficient; agriculture must adopt practices that not only improve yields but also safeguard natural resources and minimize environmental impact (Somanathan et al., 2022 ). One of these environmental-practices is mulching, a widely widespread technique in agricultural systems across the globe (Meyer et al., 2021 ; Sintim and Flury, 2017 ). Mulching involves covering the soil surface with a layer of material, ranging from organic residues to synthetic films (Mansoor et al., 2022 ), to create a more favourable microenvironment for soil and crops. Indeed, mulching contributes to an earlier increase in soil temperature (Ham et al., 1993 ), which in turn promotes faster plant growth (El-Beltagi et al., 2022 ; Kasirajan and Ngouajio, 2012 ) and higher yield (Di Mola et al., 2023 ; Hossain et al., 2022 ; Masny & Żurawicz, 2015.) simultaneously decreasing weed emergence (Schonbeck, 1999 ) and then reducing the need for herbicide application. By minimizing water evaporation from the soil surface, the application of mulching films enhances water-use efficiency, resulting in notable economic advantages for farmers (Ingman et al., 2015 ; Jabran et al., 2016 ). Beyond yield enhancement, mulching has been shown to improve product quality (Neri et al., 2012 ; Scarascia-Mugnozza et al., 2011 ) and is widely used in the cultivation of seasonal horticultural crops such as vegetables, asparagus, and strawberries (Neri et al., 2012 ; Scarascia-Mugnozza et al., 2011 ). Many materials with diverse thicknesses can be applied as mulching film, in different arrangements, and periods (Kasirajan and Ngouajio, 2012 ; Liu et al., 2014 ; Zhao et al., 2018 ) based on the crop, soil features, and climatic conditions. Wheat straw and plastic films, primarily black polyethylene (PE), are the most common mulching materials used in strawberry cultivation (Muñoz et al., 2022 ). Indeed, numerous studies have shown that PE films outperform wheat straw, particularly regarding water-use efficiency (Qin et al., 2015 ). Specifically, plastic mulches are widely adopted worldwide due to their economic production, flexibility, durability, and high use capacity (Scopetani et al., 2025 ). Although beneficial in terms of plant yield and quality, these materials have a considerable impact on the soil environment through both indirect microclimate modification and the direct accumulation of plastic residues (Qiang et al., 2023 ; Shi et al., 2022 ). Conventional PE, mulch films, particularly low-density polyethylene (LDPE), exhibit high resistance to chemical degradation, leading to significant long-term legacy effects on the soil microbiome as residues gradually break down into microplastics and nano-plastics (Lee et al., 2025 ; Qiang et al., 2023 ). Mulching creates a physical barrier that reshapes microbial dynamics by increasing soil temperature and moisture while reducing gas exchange, conditions that typically stimulate root development and the release of root exudates (Kader et al., 2017 ; Kapanen et al., 2008 ; Kasirajan and Ngouajio, 2012 ; Li et al., 2025 ; Zhang et al., 2026 ). These hydrothermal changes can alter the composition of bacterial and fungal communities, often promoting the growth of Gram-negative bacteria and selective ecological modules associated with improved crop yields (Li et al., 2025 ). Furthermore, the incorporation of plastic fragments into the soil matrix establishes a unique microbial niche known as the "plastisphere," which recruits distinct microbial groups from the surrounding bulk soil and rhizosphere (Lee et al., 2025 ; Qiang et al., 2023 ; Rillig et al., 2024 ; Shi et al., 2022 ). Exposure to persistent PE residues frequently results in a reduction of bacterial alpha-diversity and shifts in taxonomic composition across various soil types (Dong et al., 2024 ; Li et al., 2023 ). Furthermore, mulching films are usually coupled with additives (i.e., plasticizers, colour pigments, ultraviolet stabilizers), which can be released and contaminate the soil. Among these, the most common are phthalates, addressed also as phthalic acid esters (PAEs), which are lipophilic chemicals added during the plastic production process to increase the malleability of the material (Steinmetz et al., 2016 ; Zhou et al., 2023 ). PE films are not degradable, and the removal procedure is highly expensive and often not totally completed (Morra et al., 2022 ). All these conditions affect the soil safety and the environmental pollution, which need to be addressed for a more sustainable cultivation practice (El-Beltagi et al., 2022 ; Salama and Geyer, 2023 ). In this framework, a possible alternative is based on the application of biodegradable mulches because of their shorter degradation capacity and, therefore, avoiding any removal procedures. Unlike conventional PE films, biodegradable mulch films (BDM), primarily composed of polymers like polybutylene adipate terephthalate (PBAT), polylactic acid (PLA), or corn starch blends, are designed to be tilled directly into the soil, where they serve as a bioavailable carbon source for resident microorganisms (Zhang et al., 2026 ). The impact of these materials on bacterial α-diversity is complex and varies depending on soil type and exposure duration. Some investigations indicate that BDMs significantly increase bacterial richness and diversity, particularly within the rhizosphere, where favourable hydrothermal conditions stimulate root growth and the secretion of exudates (Lee et al., 2025 ; Meng et al., 2025 ; Wang et al., 2025 ). Conversely, other studies observed a reduction in bacterial alpha diversity, which is often attributed to the selective enrichment of specialized polymer degraders that suppress less competitive phyla (Xu et al., 2024 ). The presence of both conventional and biodegradable plastic residues significantly disturbs essential biogeochemical cycling of carbon (C), nitrogen (N), phosphorus (P), and sulfur (S); residues can interfere with nutrient mineralization, and alter the expression of functional genes involved in nutrient transformations (Wu et al., 2024 ; Zhang et al., 2026 ). The strawberry is recognized as one of the most significant and lucrative agricultural crops in Europe (Miller et al., 2019 ). This success is driven by a combination of exquisite sensory qualities, such as its distinct aroma, flavour, and attractive colour, and its role as a functional food (Morales-Quintana and Ramos, 2019 ). Indeed, the strawberry fruit is rich in vitamin C, folic acid, anthocyanins, flavonoids, and phenolic acids. These compounds provide a high antioxidant capacity that is essential for maintaining human wellbeing and preventing the onset of chronic or degenerative diseases when integrated into a daily diet (Hannum, 2004 ; Miller et al., 2019 ). In Europe, strawberry production heavily relies on mulching films, which enable earlier harvests, increase yields, optimize soil temperature and moisture (Biswas et al., 2025 ; Datta et al., 2024 ; Sharma et al., 2024 ; Steinmetz et al., 2016 ), and reduce disease incidence in vegetable crops (Kasirajan and Ngouajio, 2012 ; Menossi et al., 2021 ). However, research is shifting beyond general agronomic performance to investigate how differences among various mulching films may influence plant growth as well as fruit nutraceutical properties (Morra et al., 2022 ). Indeed, it was observed that biodegradable mulching showed similar performance in plant productivity and quality of PE-based materials (Costa et al., 2014 ). Conversely, the application of a new and biodegradable corn starch-based films reduced strawberry fruit quality, even though it reported a similar yield with black PE (Morra et al., 2022 ). However, despite previous research, the effect of mulches on microbial communities in the framework of a strawberry production has not been fully elucidated. Therefore, the aim of the present study was to examine the influence of both conventional and biodegradable mulching films on several levels of a small-scale strawberry cultivation system, spanning from substrate microbial communities, to plant performance, fruit yield and quality. In more detail, the study conducted a comprehensive evaluation of plant physiological and morphological responses, as well as productivity, alongside an assessment of substrate microbial α- and β-diversity under different mulching film treatments. 2. MATERIALS AND METHODS 2.1. Mulching films, plant material and experimental set-up The trial was conducted from April to June 2023 in the experimental area of the Department of Agriculture, Food, Environment and Forestry of the University of Florence located in Sesto Fiorentino, Italy (55 m above sea level; DMS coordinates: 43°81′68’’N, 11°19′99’’E). The climate in Sesto Fiorentino is typically Mediterranean, characterized by hot, dry summers and mild, wet winters. During the trial period, the average temperature was 19.2°C with the minimum value of 9.0°C (April 2023) and the maximum one of 29.1°C (June 2024). The cumulative rainfall and the average humidity during the trial were 150 mm and 59%, respectively. Strawberry plants ( Fragaria x ananassa Duch.) cv. ‘Camarosa’ were purchased from Vivai Fratelli Zanzi (Ferrara, Italy). Plants were transplanted into ceramic pots (size: 45 × 45 × 45 cm; volume: 91 L). Each pot was filled with a commercial growing medium and 4 plants were transplanted. The substrate was composed by a mixture (1:1 v/v) of peat and lapillus (particle size 10–14 mm) with the following physicochemical properties: bulk density = 0.63 g/cm 3 , Porosity = 74.87%; Water Holding Capacity (WHC) = 30.52% (v/v); Total Organic Carbon (TOC) = 4.83%; Total Nitrogen (TN) = 0.08 (%); C/N ratio = 59.23; pH = 6.23; electrical conductivity (EC) = 0.04 dS/m; cation exchange capacity (CEC) = 22.17 meq/100g. Treatments were arranged according to a randomized block design with 5 replicates (pots) per treatment for a total of 20 plants each one. Four different mulching films were used in this study: polyethylene (PE, thickness 0.05 µm), oxo-degradable polyethylene (Oxo-PE, thickness 15 µm), corn starch and polybutylene adipate terephthalate (PBAT, thickness 18 µm), and polypropylene (PP, thickness 450 µm) (Scopetani et al., 2025 ). Mulching films were applied to cover the inner walls of the pots and subsequently used to cover the surface of the substrate, prior to transplanting. As control, five pots were used without mulching application. Each pot was equipped with two drippers, and irrigation was managed automatically using an irrigation controller, supplying freshwater twice daily to maintain soil moisture at approximately 80% of the field capacity. Prior to planting, a slow-release fertilizer (16-8-12-4(Mg); TRIABON, Compo-Expert GmbH, Münster, Germany) was mixed into the substrate at a rate of 135 g per pot. Additionally, calcium nitrate (Haifa Cal Agri, Haifa Chemicals, Haifa, Israel) was applied twice (7 g per pot) throughout the entire duration of the experiment. Subsequently, potassium nitrate (PONI Haifa, Haifa Chemicals, Haifa, Israel) was applied weekly (7 g per pot) until the fruit harvest. 2.2 Substrate sampling and physicochemical characterisation At the end of the experiment (June 2023), substrate samples were collected to assess both physicochemical properties and the characterization of the bacterial communities. For the analysis of physicochemical characteristics, approximately 500–600 g of substrate was taken from each pot (0–20 cm depth), sieved to ≤ 2 mm, and subsequently pooled by treatment, combining the samples from five pots to form three replicates per treatment. The following parameters were analyzed following the methodology previously described (Bini et al., 2024 ): bulk density, porosity, total organic carbon (TOC), total nitrogen, nitrate, ammonium, pH, electrical conductivity (EC), and cation exchange capacity (CEC). Additionally, the carbon-to-nitrogen (C/N) ratio was calculated as the quotient of TOC and total nitrogen. The concentration of elements in each substrate was determined with an ICP-OES (iCAP 7000 Plus ICP- OES; Thermo Fisher Scientific, Waltham, MA, USA) by digesting 500 mg of pre-dried sample with 5 mL of nitric acid in a microwave (Mars Microwave Digestor, CEM, Matthews, NC, USA) at 190°C for 15 min. 2.3 Morphological, physiological and yield analyses Before transplanting (T0), a baseline morphological assessment was performed on 10 plants, recording plant height and leaf number. These parameters were subsequently monitored throughout the experiment, with measurements taken at three time points spaced 21 days each one (from 29th April to 8th June). For each treatment, data were collected from 10 plants (two plants per pot). Gas exchange measurements (n = 7) were conducted on fully expanded leaves of randomly selected plants using a portable infrared gas analyzer (LI-6400 XT, LI-COR Biosciences, Lincoln, NE, USA). Measurements were carried out between 11:00 and 14:00 GMT under a photosynthetic photon flux density of 1600 µmol m⁻² s⁻¹. The chamber conditions were maintained at a CO₂ concentration of 400 µmol mol⁻¹ and a flow rate of 500 µmol s⁻¹. Once leaf gas exchange stabilized, the following parameters were recorded: net photosynthetic rate (Pn), stomatal conductance (gs), and intercellular CO₂ concentration (Ci). The measurements were recorded at the same time reported in the morphological analyses. Fruit harvesting occurred throughout the cropping cycle when strawberries reached commercial ripeness, indicated by full red coloration (from 24th May to 15th June 2023). Fruits were weighed individually to determine total yield per plant over the experimental period. Yield assessments were conducted on five randomly selected plants per treatment. For each plant, the number and individual weight of harvested fruits were recorded, and cumulative yield was calculated by summing all harvests per plant, following the method of Soppelsa and collaborators (Soppelsa et al., 2019 ). In total, three harvests were carried out during the trial (each 7 days). A subset of fruits from each plant was stored at 20°C until the end of the production cycle, yielding at least 20 fruit samples per cultivar and treatment. The remaining fruits were immediately used for pomological analyses. The total yield has been reported in kg/ha by calculating the plant density used in this study (50000 plants/ha) and reported in previous studies (Menzel, 2025 ). 2.4 Quality parameters of strawberries fruits and nutraceutical properties For each plant, fruits were weighted and the size of the fruits (i.e., length and diameter) was also determined. Subsequently, fruits were dried in the oven at 65°C for 48–72 hours and the fruit dry weight was determined. Firmness of each strawberry was measured using a manual fruit penetrometer (TR Turoni, Forlì, Italy). The firmness was measured using a puncture test at the fruit’s equatorial side using a 6 mm diameter cylindrical probe and is expressed as Newton (N). The probe was attached to the stand with a handle to control the penetration speed. The maximum force (N) detected during the puncture test was recorded as the firmness of that particular fruit. For the quality analyses, fruits from the randomized blocks were collected in biological replicates (n = 3). In more detail, total soluble solids (TSS, °Brix) were measured by cutting and squeezing each sample to obtain a few drops of fruit juice and analyzed using an N1 Atago refractometer (Atago Co., Tokyo, Japan). Titratable acids (TA, % of citric acid) were evaluated by titrating with NaOH (0.1 N) a suspension obtained after shredding and blending 15 g of fruit pulp with 150 mL of deionized water until pH 8.1. Regarding nutraceutical properties, 0.3 g (fw) of strawberry fruits were extracted with 10 mL of 70% (v/v) ethanol. The total polyphenols were determined spectrophotometrically using Folin-Ciocalteu protocol and gallic acid as reference standard, as described in previous studies (Bini et al., 2024 ; Doumett et al., 2011 ). Total sugars were determined according to (Georgiadou et al., 2025 ). Organic acids (ascorbic, malic, and citric acids) were detected spectrophotometrically as described by (Tozzi et al., 2020 ). 2.5 Substrate DNA extraction, sequencing, and bioinformatics processing Total genomic DNA was extracted from 0.3 g of substrate samples, stored at − 80°C, using the FastDNA SPIN Kit for Soil (MP Biomedicals, Solon, OH, USA) according to the manufacturer's instructions. DNA purity and quantity were assessed using an ND-1000 Spectrophotometer (NanoDrop Technologies, Labtech, Ringmer, UK) and a Qubit™ 4 Fluorometer and Qubit™ ssDNA Assay Kit (Thermo Fisher Scientific). DNA was then standardized to a concentration of 10 ng/µL. Amplicons preparation and sequencing were carried out at IGATech (Udine, Italy). Libraries were prepared at IGATech (Udine, Italy) using a custom Illumina 16S Metagenomic Sequencing Library Preparation protocol and sequenced by a MiSeq instrument (Illumina, San Diego, CA) using 300-bp paired-end mode. The bacterial V3-V4 hypervariable regions of 16S rDNA were PCR-amplified with primers 341F (5′- CCTACGGGNBGCASCAG − 3′) and 805R (5′- GACTACNVGGGTATCTAATCC − 3′) (Takahashi et al., 2014 ). 2.6 Bioinformatic methodologies and Statistical Analysis To process the Illumina reads of bacterial communities, the DADA2 pipeline v.1.32.0 (Callahan et al., 2016) was used in RStudio software 4.4.1 (R Core Team). 16S rRNA reads, filtering and trimming step was performed with the following settings: maxEE = c(1,1), truncLen = c(270,250), and trimLeft = c(17,21). Sample inference, the merging of paired reads, and the removal of the chimera were performed with default parameters. To align and classify 16S sequences, the SILVA database v.138.2 with 80% confidence (Pruesse et al., 2007 ). Bacterial annotated amplicon sequence variants (ASVs) were processed with the vegan package v.2.6.8 (Oksanen, 2015 ) in RStudio software 4.4.1. ASVs with a relative abundance lower than 0.01% in all the samples were discarded. Rarefied bacterial dataset was generated for alpha-diversity, with subsample sizes of 25000 sequences per sample using the function rrarefy in the vegan package. The rarefaction curve was generated with the function rarefied . For alpha diversity, the richness (Sobs), Pielou’s index (J), Simpson index and the Shannon diversity index were estimated as previously reported (Bellabarba et al., 2024 ) and carried out with the ggplot2 package v.3.5.1 (Wickham, 2016 ). Before each statistical analysis, the Shapiro test was applied to assess the data distribution. According to Shapiro test results, differences in substrates bacterial α-diversity indices among different applied films were detected performing or the non-parametric Kruskal-Wallis tests in stats v.4.1.2 package or the parametric one-way ANOVA test in agricolae package v.1.3.7. For multiple comparisons, Kruskal-Wallis tests were followed by the post hoc Dunn Test in FSA v.0.9.5 (Ogle et al., 2020 ) and rcompanion v.2.4.36 packages (Mangiafico, 2016 ); while the one-way ANOVA was followed by post hoc Tukey’s test with agricolae package v.1.3.7. In both post hoc tests, the Benjamini–Hochberg correction for multiple comparisons was applied. For β-diversity, the overall ASVs abundances (i.e. not rarefied dataset) were analysed for the bacterial community. Non-metric multidimensional scaling NMDS ordination of Bray-Curtis’s distances were carried in the vegan package and plotted with the ggplot2 package. To investigate differences in community structure among substrates covered with different films, permutational multivariate analysis of variance PERMANOVA of Bray-Curtis’s dissimilarity metrics was executed with the adonis2 function in the vegan package. The number of permutations was set to 9999 for all permutational analyses. To test if the average within-groups dispersion was the same in all groups (Anderson, 2006 ), the Analysis of multivariate homogeneity PERMDISP was fulfilled through the functions betadisper and permutest implemented in the vegan package. The number of permutations was set to 9999 for all permutational analyses. On the overall ASVs abundances (i.e. not rarefied dataset), the average relative abundances (%) were calculated at phylum and order level, and bar plots were created with the ggplot2 package (v. 3.3.6). Only genera with a relative abundance of at least 3% in at least one sample were reported for each condition, while the average relative abundances (%) of unknown genera were not reported. On the overall ASVs abundances (i.e. not rarefied dataset), the upset plot was created with packages UpSetR (v. 1.4.0) and gridExtra (v. 2.3). Linear discriminant analysis effect size (LEfSe) (Segata et al., 2011 ) were performed to identify ASVs that were discriminately more abundant in substrates covered with different mulching films, using packages microeco (v.1.15.0) (Liu et al., 2021 ) and file2meco (v.0.9.1) in RStudio software 4.4.3, with default setting (Kruskal-Wallis rank sum test, p -value 2). The LEfSe bar plot was created with the microeco package (Liu et al., 2021 ) setting the LDA score threshold to 3.5 with the microeco package. To identify physicochemical drivers of different bacterial communities, a PERMANOVA analysis based on Bray-Curtis distances was performed, followed by A dbRDA ordination plot using the adonis2 and capscale function in the vegan package v.2.6.8, respectively (Oksanen, 2015 ). Functional inference of substrate bacterial communities was performed with FAPROTAX (Louca et al., 2016 ). Only the metabolic categories with abundance higher than 0 in at least one sample were considered. Categories Human-related were discarded, as well as the chloroplasts group. 3. RESULTS 3.1 Substrate Physicochemical properties With regard to the chemical-physical properties of the substrate the application of different mulching films did not lead to significant variations in TOC, pH, EC, and CEC at the end of the trial (Table 1 ). However, mulching significantly influenced the total nitrogen (N_tot) content. In particular, mulching with PBAT resulted in higher N_tot concentrations compared to the unmulched control. Despite this increase in total nitrogen, no significant differences were observed among the applied films in terms of nitrate and ammonium content. Similarly, the carbon-to-nitrogen (C/N) ratio remained statistically similar across treatments. Overall, mulching films had a substantial impact on the elemental composition of the commercial substrate. Substrates covered with PE films exhibited the highest concentrations of K and Mg, while those coated with PP films showed the lowest ones. Regarding micronutrients and trace metals, the Fe content was significantly higher in substrates covered with PE films. In contrast, the lowest value was recorded in coated substrates with PP. Unmulched substrates, and covered with PBAT and OxoPE, revealed intermediate profiles. Similarly, Co and Ni concentrations were significantly higher in PE compared to the other treatments. In contrast, unmulched control exhibited the greatest levels of Mo and Zn. Table 1 Physicochemical properties of substrates with different mulching films at the end of strawberry cultivation. Within rows, different letters represent statistically significant variation (p < 0.05) per Kruskal−Wallis tests and post hoc multiple Dunn’s test. Parameter Unmulched control OxoPE PBAT PE PP TOC (%) 3.71 ± 0.58 a 4.54 ± 0.53 a 3.51 ± 0.34 a 3.7 ± 0.37 a 2.8 ± 0.08 a N_tot (%) 0.13 ± 0.01 b 0.12 ± 0.03 b 0.23 ± 0.03 a 0.13 ± 0.01 ab 0.17 ± 0.01 ab C/N 27.43 ± 3.67 a 43.93 ± 13.19 a 16.23 ± 4.19 a 28.93 ± 3.38 a 16.53 ± 1.43 a Humidity (%) 10.5 ± 1.51 a 16.03 ± 0.41 a 14.67 ± 1.26 a 16.7 ± 2.32 a 14 ± 1.49 a CEC (meq 100 g − 1 ) 18.43 ± 2.06 a 20.9 ± 1.08 a 20.4 ± 1.62 a 20.47 ± 1.2 a 16.27 ± 0.52 a pH 6.17 ± 0.03 a 6.1 ± 0 a 6.07 ± 0.07 a 6.1 ± 0 a 6.13 ± 0.03 a EC (dS m − 1) 0.26 ± 0.04 a 0.3 ± 0.04 a 0.36 ± 0.03 a 0.26 ± 0.08 a 0.3 ± 0.1 a Nitrate (mg L − 1 ) 18.3 ± 0.75 a 19.77 ± 1.63 a 23.03 ± 3.19 a 12.33 ± 4.35 a 17.87 ± 4.61 a Ammonium (mg L − 1 ) 7.07 ± 2.07 a 3.07 ± 1.12 a 6.43 ± 2.14 a 4.67 ± 3.15 a 8.6 ± 6.44 a Ca (g kg –1 ) 11.23 ± 0.31 a 11.60 ± 0.03 a 12.10 ± 0.04 a 12.43 ± 1.16 a 9.72 ± 0.36 a K (g kg –1 ) 7.05 ± 0.68 b 6.71 ± 0.34 b 7.76 ± 0.17 ab 9.69 ± 0.78 a 5.55 ± 0.14 c Mg (g kg –1 ) 4.13 ± 0.13 b 4.06 ± 0.15 b 4.59 ± 0.13 ab 5.27 ± 0.41 a 3.93 ± 0.04 b Na (g kg –1 ) 1.96 ± 0.06 ab 2.062 ± 0.09 ab 2.28 ± 0.04 ab 2.37 ± 0.20 a 1.85 ± 0.03 b P (g kg –1 ) 1.19 ± 0.08 a 0.82 ± 0.19 a 1.65 ± 0.19 a 1.08 ± 0.15 a 1.61 ± 0.32 a Fe (g kg –1 ) 6.80 ± 0.18 b 6.71 ± 0.20 b 7.04 ± 0.08 b 9.17 ± 0.48 a 4.64 ± 0.25 c B (mg kg –1 ) 18.25 ± 0.53 ab 17.65 ± 1.31 ab 19.32 ± 0.88 ab 22.87 ± 2.06 a 14.35 ± 0.48 b Ba (mg kg –1 ) 204.19 ± 6.87 a 190.71 ± 5.54 a 198.04 ± 3.97 a 203.83 ± 42.32 a 139.9 ± 3.58 a Co (mg kg –1 ) 4.62 ± 0.18 b 4.94 ± 0.15 b 4.97 ± 0.08 b 6.35 ± 0.4 a 3.53 ± 0.08 c Cr (mg kg –1 ) 1.65 ± 0.16 a 1.01 ± 0.1 b 1.67 ± 0.08 a 2.01 ± 0.06 a 1.21 ± 0.12 b Cu (mg kg –1 ) 29.38 ± 0.85 a 23.59 ± 0.77 a 32.36 ± 4.35 a 24.85 ± 2.59 a 25.6 ± 2.37 a Mn (mg kg –1 ) 192.98 ± 8.03 a 181.7 ± 4.8 a 202.48 ± 0.7 a 240.43 ± 15.34 a 212.19 ± 31.35 a Mo (mg kg –1 ) 4.29 ± 1.03 a 1.23 ± 0.22 b 1.37 ± 0.1 b 1.05 ± 0.09 b 3.28 ± 0.44 ab Ni (mg kg –1 ) 1.77 ± 0.03 c 1.34 ± 0.23 c 3.31 ± 0.33 b 4.68 ± 0.37 a 1.38 ± 0.07 c Pb (mg kg –1 ) 9.14 ± 0.26 a 8.27 ± 0.17 a 9.49 ± 0.65 a 10.43 ± 0.51 a 6.21 ± 0.36 b Zn (mg kg –1 ) 10.94 ± 3.66 a 4.57 ± 1.03 ab 6 ± 0.24 ab 5.51 ± 0.18 ab 2.88 ± 0.12 b 3.2 Effect of mulching practice on physiological response and plant growth Mulching practice led to slight differences in physiological traits during the plant cycle (Table S1 ). At the first time point (T1), PE mulch revealed the highest net photosynthetic rate (Pn) value, which was significantly greater than the unmulched control, whilst no variations were noted with the other mulched treatments. At T2 and T3, all treatments displayed similar net Pn values. No differences among treatments were observed for stomatal conductance (gs) in all time points analysed. At T1, the unmulched control revealed the highest value of the intracellular CO 2 concentration (Ci), which was also significantly greater than PE and PBAT. Conversely, an opposite trend was observed at T2, with a significantly lower value of Ci in the unmulched control compared to PP. At T3, no statistical differences in Ci values were observed among treatments. Regarding the plant's growth parameters, at T1, no clear differences in plant height and in the number of leaves were reported among different films applied (Table S2 ). As plant growth evolved (T2 and T3), all treatments showed a similar increment in plant height (p < 0.05). At T3, the PP treatment showed the highest value of number of leaves, which was significantly higher than PE), whereas no variations for this parameter were observed among the unmulched control and all the mulching films. Despite slight differences observed in plant growth parameters, the mulching practice positively affected strawberry yield (Fig. 1 ). The highest yield was observed in OxoPE treatment (4425 ± 243.38 kg ha –1 ), while the lowest yield was in the unmulched control (3145 ± 132.38 kg ha –1 ). Mostly, OxoPE and PP treatments showed a significantly higher productivity than the unmulched control, while no significant variations were observed with PBAT and PE treatments. Concerning the quality parameter of strawberry fruits, there were no significant differences in fruit weight, diameter, height, TSS, or texture among treatments (Table 4). However, the OxoPE treatment showed a significantly higher fruit brightness (L*) and colour intensity (chroma index) compared to the other treatments. Strawberries produced in PBAT treatment showed the highest titratable acidity (1.2%), compared to the lowest value of the unmulched control (0.87%). Strawberries treated with PP films had the lowest texture value, which differed significantly from the PBAT treatment. With respect to the nutraceutical strawberry fruit properties, no significant differences were observed among treatments in polyphenols, total sugars, citric acid, or malic acid content (Table 5). The highest value for ascorbic acid was observed in unmulched control, being significantly higher in fruits than OxoPE which showed the lowest value. The other treatments showed intermediate levels with no significant differences. 3.3 Composition of bacterial communities in substrates covered with different mulching films. Concerning the 16S rRNA sequencing analysis of bacterial communities, a total of 1100496 high-quality sequences were obtained (average of 73396 ± 24204 sequences per sample). An average of about 33% of sequences remained after quality filtering and removal of chimera, enough to accurately describe the biodiversity within the bacterial communities as suggested by the rarefaction curves (Fig. S1 ). The bioinformatic processing of the 16S rRNA sequences yielded 3201 ASVs, assigned to Bacteria kingdom, spanning 28 phyla, 57 classes, 121 orders, 175 families, and 311 genera (File S1). Bacterial communities of different substrates were predominantly composed by Pseudomonadota , with an average relative abundance between 53.3% and 40.6%, followed by Bacteroidota (~ 21.2% - ~12.1%), Acidobacteriota (~ 16.7% - ~6.4%), and, to a lesser extent, by Actinomycetota (~ 9% - ~6.2%), Verrucomicrobiota (~ 7.9% - ~2.8%), Planctomycetota (~ 4.2% - ~1.9%), Patescibacteria (~ 3.4% - ~1.3%), Myxococcota (~ 1.5% - ~1.3%), Cyanobacteriota (~ 1.3% - ~1.2%) (Fig. S2 A). At family level, the bacterial communities of substrates shared a ‘core’ of enriched taxonomic groups with a mean relative abundance higher than 3%, composed of Sphingobacteriaceae (~ 12% - ~7.5%), Caulobacteraceae (~ 11.8% - ~6.7%), Sphingobacteriaceae (~ 12% - ~7.5%), Rhodanobacteraceae (~ 13% - ~3.4%), Acidobacteriaceae (~ 11.8% - ~4.8%), and Micropepsaceae (~ 5.9% - ~4.1%) (Fig. S2 B). The relative abundance of other families shifted across the bacterial communities of substrates mulched with different films: the family of Acidimicrobiaceae was more abundant in OxoPE and PE substrates, while the family of Burkholderiaceae was more abundant in substrates covered with OxoPE, PE and PP (Fig. S2 B). Similarly, the family of Chitinophagaceae was enriched in substrates covered with PBAT or PP and uncovered substrates, while the family of Sphingomonadaceae was more abundant in substrates mulched with PP and unmulched control (Fig. S2 B). Specific bacterial families were uniquely enriched in individual mulch treatments: Lysobacteraceae was more abundant in PBAT-mulched substrates, while Acetobacteraceae and Pedosphaeraceae were specifically associated with PE mulching (Fig. S2 B). Among the most enriched genera, the taxonomic groups shared by all bacterial communities were Asticcacaulis (~ 7.0% - ~4.8%) and Granulicella (~ 7.1% - ~3.5%) (Fig. S2 C). The genera Mucilaginibacter (~ 6.4% - ~3.4%) and Rhodanobacter (~ 11.2% - ~4.0%) were more abundant in all substrates except the PBAT and PE, respectively. Furthermore, Albibacterium (~ 4.1%) and Brevundimonas (~ 4.3%) were uniquely enriched in PBAT-mulched substrates, whereas Sphingomonas (~ 3.9%) were more abundant in PP-mulched substrates. 3.4 Diversity of bacterial communities in substrates covered with different mulching films/ Effect of different mulching films on diversity of substrate bacterial communities Substrate bacterial α-diversity was characterized by high species richness (Sobs) which remained statistically comparable across all substrates, indicating that mulching practices, regardless of film type, did not influence total number of bacterial taxa (Fig. S3 A). However, mulching practices significantly enhanced bacterial community diversity and evenness in respect to the unmulched control, according to Simpson and Shannon indices (Fig. S3 C-D). Specifically, both indices were significantly higher in substrates covered with OxoPE and PE films compared to control. In particular, the higher values of Simpson indices suggest a reduced dominance of some bacterial taxa and, consequently, a higher diversity compared to the control. Moreover, a significant increase of Shannon index was also observed in substrates covered with PP, in line with the slight increasing trend observed for Evenness index of PE and PP communities (Fig. S3 B). Analysis of ꞵ-diversity revealed a clear separation in bacterial community structures between mulched and unmulched substrates. Most notably, bacterial communities associated with plastic-based films (OxoPE, PE, and PP) formed a distinct cluster compared to those found with PBAT (Fig. 2 ). Both the Permutational multivariate analysis of variance (PERMANOVA) and the Analysis of multivariate homogeneity (PERMDISP) confirmed significant differences in community composition, indicating that mulching practices, regardless of film type, distinctly shape the structures of substrate microbial communities (Fig. 2 ). The distribution of bacterial ASVs revealed a distinct partitioning of bacterial taxa across the different substrate treatments (Fig. S4 ). A shared core of only 319 ASVs, representing approximately 10% of the total richness (3,201 ASVs), was identified across all communities. In contrast, the majority of ASVs were unique to specific substrates, likely driving the structural variations observed in the NMDS analysis. Notably, all plastic-mulched substrates harboured a higher proportion of exclusive taxa compared to the unmulched control (232 ASVs; ~7.2%), ranging from ~ 9,6% for OxoPE (306 ASVs) to ~ 7,9% for PP (254 ASVs). To uncover the taxonomic groups significantly associated with the distinct microbial communities of each substrate, a linear discriminant analysis effect size (LEfSe) was performed. The total number of the discriminant ASVs identified was 299 (File S2). Only 19 ASVs, with LDA score higher than 3.5, were considered as highly discriminants (Fig. 3 A-B). Consistent with the relative abundance patterns, the unmulched control was primarily characterized by the genera of Rhodanobacter (ASV_4, ASV_5, and ASV_24), Nocardioides (ASV_41), Chryseobacterium (ASV_59) and Arachidicoccus (ASV_60) (Fig. 3 A-B, Fig. S2 C). In contrast, the OxoPE bacterial community was uniquely distinguished by Asticcacaulis (ASV_9) as the sole highly discriminant ASV. The PE substrate harboured the greatest number of discriminant taxa, including Mucilaginibacter (ASV_37), Candidatus solibacter (ASV_35) and several unclassified genera within the Acidimicrobiaceae (ASV_6), Acetobacteraceae (ASV_14), Acidimicrobiaceae (ASV_31), Pedosphaeraceae (ASV_63) and Xanthobacteraceae (ASV_8) families. Bacterial communities of PP-mulched substrates were primarily distinguished by the genera Cellvibrio (ASV_36) and Sphingomonas (ASV_10), with the latter strongly enriched in these substrates (Fig. S2 C). Finally, the PBAT community was characterized by the genera Brevundimonas (ASV_128), a specific strain of genus Rhodanobacter (ASV_49) and the genus TM7a of Saccharimonadaceae family (ASV_22). 3.5 Relationship between bacterial community structures and Substrate Physicochemical properties To evaluate the influence of soil physicochemical properties on different bacterial community structure, a Permutational Multivariate Analysis of Variance (PERMANOVA) coupled with a distance-based Redundancy Analysis (dbRDA) were performed based on ASVs Bray-Curtis’s dissimilarities. The PERMANOVA model revealed that the soil chemical profile significantly shaped the microbial community (R 2 = 0.77908, p -value = 0.0024). When evaluating the independent contribution of each variable, the substrate humidity and nitrate concentration revealed as the primary significant drivers (Table 1 ). Specifically, humidity explained 15.1% of the total variance (F = 3.43, p = 0.0072), while nitrate levels accounted for 13.9% (F = 3.14, p = 0.0052). The other variables, including Cation Exchange Capacity (CEC) and Electrical Conductivity (EC) did not reach the standard significance threshold, suggesting a secondary role in community differentiation. The dbRDA analysis confirmed a high degree of correlation between environmental constraints and microbial community shifts (R 2 = 0.779). The first two axes of the ordination (CAP1 and CAP2) accounted for 46.8% and 14.3% of the constrained variation, respectively. Along the CAP1 axis, db-RDA plot illustrated a clear separation of bacterial communities among conventional plastic films, PBAT films and unmulched related communities, which was strongly associated with gradients of Humidity and Nitrate. Bacterial communities of substrates mulched with OxoPE, PE and PP were positively correlated with higher humidity compared to the unmulched control, suggesting that this feature was a driver in shaping this community. To understand the possible effect of mulching films on substrate functional activities, putative soil bacterial functional profiles were inferred with the FAPROTAX database from taxonomies. Among the database processes, only 25 were detected. This predictive analysis revealed a distinct separation in terms of both bacterial metabolic pathways and ecological roles across the different substrate treatments, primarily separating PE and PP mulching films from the Unmulched control, PBAT and OxoPE (Figure S5 ). Specifically, traditional polyethylene mulching induced a functional shift compared to other films characterized by the putative enrichment of nitrogen fixation, concurrently with a marked depletion in nitrification and aerobic nitrite oxidation, chemoheterotrophy and aerobic chemoheterotrophy, methanol oxidation and methylotrophy (Figure S5 ). In contrast, the PBAT treatment displayed a unique functional fingerprint defined by the putative increase of nitrification, aerobic nitrite oxidation and nitrate respiration, also in manganese oxidation, and overall sulfur compound respiration, suggesting an increased turnover of nitrogen and sulfur cycles compared to other mulches. In OxoPE films increased the overall fermentation process, while aromatic and hydrocarbon degradation was predictively higher in the PBAT and OxoPE films than in the PE and PP treatments, indicating that biodegradable films may promote microbial guilds capable of complex carbon catabolism. DISCUSSION Previous research on strawberry cultivation has typically focused on the effects of mulching either in terms of soil health and microbial dynamics (Meyer et al., 2021 ; Muñoz et al., 2017a ) or through plant physiological and morphological aspects (Abdelfattah et al., 2025 ; Biswas et al., 2025 ; Dangi et al., 2025 ), often lacking of a more comprehensive and integrated approach. To bridge this gap, our research investigates the impact of conventional versus biodegradable mulching on strawberry cultivation by evaluating both plant performance and microbial ecology within the same experimental framework. Through a detailed evaluation of plant physiological and morphological traits, fruit yield and quality, and substrate microbial communities, we provided a broader understanding of how mulch selection may drive both strawberry plant and substrate health. Soil quality is fundamental to ensuring consistent crop productivity and optimal fruit quality. The impact of mulching films on soil properties remains a subject of debate, as research findings are often controversial. While some studies suggested that both plastic and biodegradable films significantly alter soil physical (e.g., bulk density, moisture, and temperature) and chemical features (e.g., total organic carbon TOC and nitrogen uptake) (Dewi et al., 2024 ; Wu et al., 2022 ) others reported no significant changes in pH, organic matter, or nitrogen content followed the application of different mulch films. Our results aligned with the latter, as most physicochemical properties of the peat-based substrate, including TOC, pH, EC, CEC, and the C/N ratio, were not significantly affected by the tested mulching films. Specifically, in the work of Xu and collaborators (Xu et al., 2025 ), a similar stability in TOC content to our results was observed over a 180-day period in a laboratory experiment, suggesting that more pronounced alterations in substrate properties might be tracked mainly with long-term application of mulching, within different degradation time and environmental conditions (Ding et al., 2023 ). Whereas primary physicochemical characteristics remained stable over the limited cultivation period, the different mulching films significantly influenced specific macro- and micronutrient concentrations. In particular, PBAT film increased total nitrogen content compared to the unmulched control, while PE films lead to higher concentrations of K, Mg, Fe, Co, and Ni. These shifts support previous reports that mulching with plastic films, and the composition of the film itself, could modify nutrient retention and mineral transformations by altering soil hydrothermal conditions (Mo et al., 2020 ; Shan et al., 2022 ; Y. Zhao et al., 2023 ). Several studies suggest that both plastic and biodegradable films can modify soil physical properties (e.g., bulk density, moisture, and temperature) as well as chemical characteristics (e.g., TOC and nitrogen uptake) (Dewi et al., 2024 ; Wu et al., 2022 ). However, other studies have reported no significant changes in pH, organic matter, or nitrogen content (Ni et al., 2016 ; M. Zhang et al., 2022 ). The present research is consistent with the latter findings, as most main physicochemical properties of the peat-based substrate, including TOC, pH, EC, CEC, and C/N ratio, were not significantly affected by the different mulching films tested. Similar results for TOC were reported by Xu et al. (Xu et al., 2025 ), who evaluated these properties over 180 days after the application of plastic or biodegradable mulching films in a laboratory experiment. However, previous studies suggest that the effects of mulching films may depend on degradation time and environmental conditions. In particular, long-term application could lead to more pronounced alterations in substrate properties (Ding et al., 2023 ). Although limited variation was detected in the main physicochemical characteristics in a limited cultivation period (i.e. 3 months), the four mulching films influenced macro- and micronutrient concentrations in the peat-based substrate. In particular, a factor determining these alterations is the different film composition (Bandopadhyay et al., 2018 ; Shan et al., 2022 ). In the present study different mulching films did not significantly influence the physiological or vegetative growth of strawberry plants. Indeed, both photosynthetic rate and stomatal conductance did not show significant variations either compared to the control samples or among the different mulch films. Consistent with our results, previous studies on cucumber and pickling cucumber have shown that plastic mulching does not significantly modify leaf gas exchange compared to bare soil (López-Tolentino et al., 2017 ; Torres-Olivar et al., 2018 ) (Lopez-Tolentino et al., 2017; Torres-Olivar et al., 2018 ). These studies also reported significant increases in yield under mulched conditions, despite the non-significant differences in gas exchange. Similarly, in our study, mulching coverage enhanced fruit yield. Increased fruit production under mulching is a well-established response in strawberry cultivation, as reported in previous studies (Abdelfattah et al., 2025 ; Dangi et al., 2025 ). A key finding of our study was the significant impact of mulch type on productivity, with OxoPE and PP that outperformed the control (+ 29% and + 25%, respectively), then mirroring the 28% yield increase reported by Biswas et al. (Biswas et al., 2025 ) for strawberries grown with biodegradable plastic films. In contrast, PE mulches produced yields comparable to the unmulched control, reflecting an intermediate performance previously reported for both black and white PE (Pinto et al., 2022 ). The impact of PBAT-type mulches on strawberry yield, in our study, differ from some previous works. Indeed, an 49% yield increase in strawberry cv. ‘Albion’ using PBAT-based biodegradable mulch (BDM) was observed compared to bare soil control (Wang et al., 2022 ), and a similar upward trend was noted in two-year open-field cultivation (DeVetter et al., 2017 ). This discrepancy might likely due to the different experimental setups, as our study utilized a peat-based growing substrate, whereas the aforementioned research was conducted in open-field soil conditions. Despite these differences, when directly comparing PBAT to PE, our results remain consistent with other studies showing comparable strawberry yields between the two materials (Costa et al., 2014 ; DeVetter et al., 2017 ; Wang et al., 2022 ). Concerning fruit quality, strawberry fruits showed total soluble solids (TSS) values comparable to those previously reported (DeVetter et al., 2017 ; Supreetha et al., 2025 ; Wang et al., 2022 ). However, mulching can influence strawberry fruit quality by modifying the amount and spectral composition of light reflected into the plant canopy (Kasperbauer et al., 2007 ; Shiukhy et al., 2015 ), which in turn may affect fruits composition and reduce TSS in unmulched plants (DeVetter et al., 2017 ). In our study, although a slight variation in TSS was detected, no significant differences were observed among different films used, implying that mulch application did not affect TSS (Wang et al., 2022 ). In contrast, we observed an increment in titratable acidity (TA) in the PBAT treatment significantly outperforming the control. The influence of different mulches on TA levels has been previously reported across various fruit species (Morra et al., 2021 ; Wang et al., 1998 ). Despite this specific shift, no marked variations in overall fruit quality were recorded among the biodegradable (PBAT, OxoPE) and the PE mulching films, suggesting that replacing PE with biodegradable mulches would not lead to meaningful changes in fruit quality (Costa et al., 2014 ; DeVetter et al., 2017 ). Mulching practices are well-known to influence soil microbial structure and functionality through the regulation of the soil microclimate and physicochemical environment, thereby significantly altering community composition and diversity (Li et al., 2025 ; Sun et al., 2022 ; Zhang et al., 2024 ). In this study, mulching with OxoPE, PE, and PP significantly enhanced α-diversity, as indicated by increased Shannon and Simpson indexes. This variation effectively reduced the dominance of certain taxa, leading to a considerably more even community composition. Overall, these results suggested that mulching with plastic films promotes a more balanced and diverse bacterial assembly without shifting the overall species richness. In previous works, mulching materials showed highly variable outcomes regarding bacterial α-diversity, which seems to be strongly dependent on the type of soil and its interaction with diverse mulch films (Xu et al., 2024 ), the plant species (Gao et al., 2025 ), and micro-habitat taken into account (i.e. bulk soil, rhizosphere, or plastisphere) (Meng et al., 2025 ; Zhang et al., 2026 ). Our findings are strongly supported by studies in which mulching likely alleviates environmental stressors (as salinity or drought) enabling a broader range of taxa to thrive (Wang et al., 2025 ). Conversely, several studies report a decrease in α-diversity, particularly with biodegradable materials (Dong et al., 2024 ; Fang et al., 2025 ; Xu et al., 2024 ; Z. Zhao et al., 2023 ) often attributed to the enrichment of specific polymers-degrading taxa, thereby reducing overall community evenness. However, α-diversity indexes are known to be less sensitive to environmental changes than overall community structure analysis. Indeed, while specific bacterial groups may shift in response to the environmental pressure, these changes are often offset by opposing trends in other guilds, thereby maintaining relatively stable total diversity (Hartman et al., 2018 ; Hartmann and Widmer, 2006 ). In this context, community shifts we detected were more evident at the structural level. Indeed, our results revealed distinct bacterial community structures among mulching materials, implying that substrate bacterial diversity respond differently to the application of different film types as previously observed (Zhao et al., 2023 ). Clear variations in bacterial communities were previously detected in soils both under BDMs (Koitabashi et al., 2012 ; Li et al., 2014 ; Muroi et al., 2016 ), and under non-biodegradable plastic mulches (Farmer et al., 2017 ; Muñoz et al., 2017b , 2015 ). In particular, some recent research consistently highlighted how BDM residues, or generally BDM treatments, elicited more pronounced divergences in community structure than conventional PE or low-density polyethylene (LDPE) (Liu et al., 2022 ; L. Liu et al., 2025 ; Xu et al., 2024 ). A common explanation for this trend lies on different material properties. In more detail, PE is a chemically inert material, with slow biodegradation rate (Khandare et al., 2022 ), performing primarily as a physical barrier (Ammala et al., 2011 ; Koutny et al., 2006 ). Conversely, BDM acts as a bioavailable exogenous carbon source, exerting selective pressure on soil microbial community then reshaping its structure and functionality (Bandopadhyay et al., 2020 ; Zhang et al., 2026 ). We also noticed a slight divergence between the traditional plastic films (OxoPE, PE, and PP) and the PBAT-related communities, that we likely attribute to the presence of corn starch in PBAT films. The corn starch is a key component of several starch-PBAT blends biodegradable films and, as a naturally hydrophilic polysaccharide, is highly sensitive to moisture (Grimaut et al., 2023 ; Küster et al., 2025 ) and undergoes rapid mineralization (Villena et al., 2022 ). Therefore, we hypothesize that, unlike conventional plastics, the gradual degradation of starch provides an additional carbon source that recruits specific starch-utilizing bacterial species. This process may alter the microbial community (Gao et al., 2022 ; Šerá et al., 2020 ) toward copiotrophic and plant-beneficial genera as the PBAT-related Rhodanobacter , which are specialised for nutrient-rich environments (Mahawar et al., 2026 ; Wang et al., 2024 ). In line with this trend, the unique increase of genus Brevundimonas in PBAT-mulched substrates aligns with its ability to metabolize starch, as demonstrated by its use its use in dark fermentation processes for biohydrogen production (Bao et al., 2012 ). Nonetheless, in our work the OxoPE and PE application supported the highest levels of unique bacterial recruitment, whereas the unmulched control exhibited the most restricted set of exclusive ASVs. Beyond chemical composition, the physical presence of plastic mulch may promote specialized microbial niches, with film fragments acting as unique micro-environment that selectively enriches specific microbial guilds (Huang et al., 2019 ; Jacquin et al., 2019 ; Qi et al., 2022 ; Shi et al., 2022 ). Consistent with this observation, our results further highlight the family Sphingomonadaceae , and specifically the genus Sphingomonas , which exhibited a significantly higher abundance in PP-mulched substrates. In particular Sphingomonas , well-known as pollutants degraders (Asaf et al., 2020 ), have been consistently associated to different mulch blends (Xu et al., 2024 ; Liu et al., 2022 ). Moreover, this genus was found enriched in the PP plastisphere of the sorghum rhizosphere (Z.-H. Wang et al., 2026 ) and on PP polypropylene particles used for the biodegradation of crude oil (Vita et al., 2022 ). Consequently, Sphingomonas was recognized as key plastic-degrading bacterium in soil (Xu et al., 2024 ; Liu et al., 2022 ) with proven metabolic potential to break down a wide range of organic pollutants (Gatheru Waigi et al., 2017 ) and synthetic polymers (Chen et al., 2020 ; Y. Zhang et al., 2022 ). According to recent research, different film thickness significantly changed the overall community structure of rhizosphere soil bacteria (G. Wang et al., 2026 ). Moreover, the taxonomic composition of bacterial communities at both phylum and genus varied significantly among different BDM thicknesses (Zhao et al., 2023 ). Specifically, it was observed a significant positive correlation among film thickness and soil water content, soil organic matter, and total nitrogen, but a negative correlation with soil temperature (G. Wang et al., 2026 ). This suggests that thicker films provide a more stable micro-environment in terms of moisture, temperature, and nutrient availability compared to thinner films, which are more susceptible to environmental fluctuations (G. Wang et al., 2026 ). Therefore, film thickness serves as a determining factor for both the physical durability of the mulch and the modulation of the soil microclimate, which in turn shapes the response of the associated bacterial communities. Indeed, our results revealed that the application of plastic films of varying thicknesses produced bacterial communities with unique structures and compositions. This separation followed a gradient corresponding to the thickness of the materials, ranging from the conventional PE as the thinnest material (0.05 µm) to the PP as the thickest one (450 µm). While thinner films (e.g., PE or LDPE < 10 µm) degrade easily to the mechanical action of wind and rain, leading to a release of microplastics and chemical additives (Crossman et al., 2020 ; Kim et al., 2021 ; Qiang et al., 2023 ; Scheurer and Bigalke, 2018 ), thicker mulch films in our study likely acted as more robust physical barriers. These films likely reshaped the microbial communities through long-term microclimatic stabilization. For this reason, such a broad range of thicknesses likely imposed differential selective pressures creating different substrate microclimates. As a further supporting of these speculations/hypothesis, distance-based Redundancy Analysis (dbRDA) identified humidity and nitrate concentrations as the primary significant drivers shaping the substrate microbial communities. Overall, this finding suggests that changes in soil microclimate and nitrogen dynamics induced by mulching act as the main selective pressures shaping the specialized bacterial communities, as reported previously (Gao et al., 2025 ; Wang et al., 2025 ; Zhang et al., 2026 , 2020 ; M. Zhang et al., 2022 ; Y. Zhang et al., 2022 ). In particular, the distinct associations observed between substrate humidity and communities of conventional films (PE, PP, and OxoPE), and between nitrate concentrations and those of PBAT treatments, likely underscoring two contrasting mechanisms: the passive cover effect of conventional films, which primarily modulates soil microclimate and moisture, and the direct chemical and nutrient-input effect of biodegradable films (PBAT) during the degradation process. Distinct functional shifts of bacterial communities based on mulch type were predicted, with the PE communities linked to an enrichment in nitrogen fixation. Conversely, PBAT seems to promote nitrification and sulfur respiration processes. These predictions are consistent with the compositional profiling analysis, which revealed that the family of Burkholderiaceae was more prevalent in substrates covered with conventional plastic mulch (OxoPE, PE, PP). This family, a large group within the Proteobacteria phylum, is recognized for its metabolic versatility and ecological plasticity which encompasses several well-known genera (e.g., Burkholderia and Paraburkholderia ), characterized as Plant Growth-Promoting Rhizobacteria (PGPR) and involved in nitrogen fixation and phosphate solubilization (Eberl and Vandamme, 2016 ). Interestingly, it has been previously observed that LDPE microplastics promote the abundance of Burkholderiaceae potentially to enhance nitrogen fixation (Fei et al., 2020 ). Besides, others genera were previously associated to PE mulch films, as Mucilaginibacter (Tagg et al., 2022 ), with a significant role in the degradation of complex carbon sources (Kumar et al., 2025 ), and Candidatus Solibacter known for its ability to decompose organic matter then promoting soil carbon cycling and nitrogen fixation (Gu et al., 2023 ). The accumulation residues from LDPE and BDM mulches negatively affects soil physical properties and water dynamics (Li YuanQiao et al., 2020 ), while altering carbon and nutrient cycling (Zang et al., 2017 ). These changes frequently lead to a significant nitrogen limitation (Hegan et al., 2015 ) and higher soil C/N ratio (Qi et al., 2020 ). Therefore, it is reasonable to infer that plastic-derived carbon input, together with changes in soil physical structure, can trigger a short-term nitrogen starvation through microbial immobilization, even without observing significant change in C/N ratio. In this regard, the recruitment of nitrogen-fixing bacteria represents a community adaptation to restore the C/N balance. Conversely, in PBAT-mulched substrates the input of bioavailable carbon might promotes the synthesis of organic nitrogen compounds and, consequently, stimulates the nitrification. This process may facilitate the niche stabilization for nitrifying bacteria following the soil physical micro-alterations induced by mulching. For instance, it has been observed that plastic residues from PLA films increase soil porosity, which in turn may enhance soil aeration and accelerate nitrification processes (De Souza Machado et al., 2018 ), ultimately leading to a depletion of soil ammonium levels (Dong et al., 2024 ). The predicted increase in sulfur respiration within PBAT-amended substrates may be due to the overall metabolic remodulation of bacterial communities induced by mulch residues, as sulfur cycling genes are tightly coupled with carbon and nitrogen metabolic pathways (Zheng et al., 2019 ). However, some studies have reported a negative correlation between PBAT mulching and sulfur cycling (X. Liu et al., 2025 ) as well as the negative impact of microplastic on sulfur-related processes in coastal sediments (Wang et al., 2023 ). In our pathway prediction, biodegradable films (PBAT and OxoPE) were also associated with microbial groups capable of complex carbon catabolism, such as aromatic and hydrocarbon compounds degradation. Indeed, TM7a of Saccharimonadaceae family was previously identified as a strong biomarker for the PBAT community since its relative abundance increasing proportionally to the amount of PBAT (Xu et al., 2024 ). Moreover, this taxon is consistently associated to different microplastic materials in both marine (Scales et al., 2021 ) and soil environments (Li et al., 2022 ; Scopetani et al., 2023 ), further highlighting its putative plastic degradation potential. Similarly, Brevundimonas was identified an efficient aromatic degrading bacteria (Uba, 2019 ) and already isolated from oil-contaminated soil (Chaudhary and Kim, 2018 ). Furthermore, the enrichment of Brevundimonas was previously correlated with PBAT degradation during composting, reinforcing its role in the breakdown of biodegradable polymers (Cao et al., 2025 ). CONCLUSION The present study demonstrated that plastic mulch films did not significantly influence strawberry plant physiology or vegetative growth, but they played a critical role in productivity, with OxoPE and PP providing the most substantial yield enhancements. Alongside these agronomic improvements, fruit quality was maintained by the application of films. However, PBAT treatments specifically increased titratable acidity without compromising essential traits as soluble solids. Interestingly, OxoPE treatment improved the aesthetic quality of fruits by increasing lightness and chroma index. Although the main physicochemical properties of growth substrates remained largely unaffected, elemental composition was significantly influenced by mulches. Specifically, PBAT promotes nitrogen enrichment, whereas PE is associated with an increase in available micronutrients. Crucially, plastic-based mulching serves as a key driver of microbial assembly, enhancing the substrate bacterial community evenness and diversity. Structural analysis revealed that each material recruits unique microbial niches, with PP and PBAT that recruited specialized pollutant-degrading taxa. These structural shifts were primarily driven by humidity and nitrate gradients, highlighting a possible divergence between the physical microclimate stabilization of conventional films and the likely chemical input of biodegradable films. Lastly, functional predictions indicated that PE mulches application promoted nitrogen-fixing communities as a possible response to even slight nutrient shifts, whereas biodegradable films enhanced nitrification processes and complex carbon catabolism. By providing a comprehensive evaluation of traditional and biodegradable mulching in a small-scale strawberry cultivation, this study confirms that biodegradable films (OxoPE and PBAT) might be viable substitutes for traditional films, able to simultaneously promote high strawberries yields and the development of a specialized and functional growth substrate bacterial communities. Nevertheless, further investigations are needed to assess the long-term impact of both biodegradable and non-biodegradable plastic material on both strawberry agronomic performance and substrate microbial communities and functions. Declarations COMPETING INTEREST The authors have no relevant financial or non-financial interests to disclose. FUNDING This research was funded by the European Union-Next Generation EU, UNIFI Young Independent Researchers Call, within the MuSC project MuSC (Analysis and sensing of contaminants in agriculture: from Mulches to Soil and Crops). AUTHOR CONTRIBUTIONS All authors contributed to data interpretation, drafted the manuscript, agreed with its final version, and revised the manuscript. Conceptualization: Agnese Bellabarba, Costanza Scopetani, Giulia Selvolini; Methodology: Agnese Bellabarba, William Antonio Petrucci, Ermes Lo Piccolo, Lorenzo Bini; Formal analysis and investigation: Agnese Bellabarba, Lorenzo Bini, Costanza Scopetani, Giulia Selvolini, Ermes Lo Piccolo, Elisabetta Toni; Writing - original draft preparation: Agnese Bellabarba, Lorenzo Bini, Francesca Decorosi; Writing - review and editing: Agnese Bellabarba, Lorenzo Bini, Carlo Viti, Edgardo Giordani, Giulia Selvolini, Costanza Scopetani; Funding acquisition: Agnese Bellabarba, Costanza Scopetani, Giulia Selvolini; Resources: Agnese Bellabarba, Costanza Scopetani, Giulia Selvolini, Carlo Viti, Edgardo Giordani, Giovanna Marrazza, Alessandra Cincinelli, Tania Martinelli; Supervision: Carlo Viti, Edgardo Giordani, Giovanna Marrazza, Alessandra Cincinelli, Tania Martinelli. DATA AVAILABILITY STATEMENT Genomic sequences of this project have been deposited at GenBank under BioProject number PRJNA1450036 References Abdelfattah, A., Awad, M., Sorour, O., 2025. Synergistic effects of magnetic water treatment and mulching on crop and soil moisture-salinity distribution. Sci Rep 15, 15741. https://doi.org/10.1038/s41598-025-98802-6 Ammala, A., Bateman, S., Dean, K., Petinakis, E., Sangwan, P., Wong, S., Yuan, Q., Yu, L., Patrick, C., Leong, K.H., 2011. An overview of degradable and biodegradable polyolefins. Progress in Polymer Science 36, 1015–1049. https://doi.org/10.1016/j.progpolymsci.2010.12.002 Anderson, M.J., 2006. Distance-based tests for homogeneity of multivariate dispersions. Biometrics 62, 245–253. Asaf, S., Numan, M., Khan, A.L., Al-Harrasi, A., 2020. Sphingomonas : from diversity and genomics to functional role in environmental remediation and plant growth. Critical Reviews in Biotechnology 40, 138–152. https://doi.org/10.1080/07388551.2019.1709793 Bandopadhyay, S., Martin-Closas, L., Pelacho, A.M., DeBruyn, J.M., 2018. Biodegradable Plastic Mulch Films: Impacts on Soil Microbial Communities and Ecosystem Functions. Front. Microbiol. 9, 819. https://doi.org/10.3389/fmicb.2018.00819 Bandopadhyay, S., Sintim, H.Y., DeBruyn, J.M., 2020. Effects of biodegradable plastic film mulching on soil microbial communities in two agroecosystems. PeerJ 8, e9015. https://doi.org/10.7717/peerj.9015 Bao, M., Su, H., Tan, T., 2012. Biohydrogen Production by Dark Fermentation of Starch Using Mixed Bacterial Cultures of Bacillus sp and Brevumdimonas sp. Energy Fuels 26, 5872–5878. https://doi.org/10.1021/ef300666m Bellabarba, A., Giagnoni, L., Adessi, A., Marra, E., Laschi, A., Neri, F., Mastrolonardo, G., 2024. Short-term machinery impact on microbial activity and diversity in a compacted forest soil. Applied Soil Ecology 203, 105646. https://doi.org/10.1016/j.apsoil.2024.105646 Bini, L., Renai, L., Fichera, M., Petrucci, W.A., Lenzi, A., Biricolti, S., Giordani, E., Rivoira, L., Bruzzoniti, M.C., Piesik, D., Del Bubba, M., 2024. Assessing the Impact of Sustainable Biochar-Enriched Substrates on Safety and Quality of Tomato ( Solanum lycopersicum L.) as Relevant Model Crop. ACS Agric. Sci. Technol. 4, 681–689. https://doi.org/10.1021/acsagscitech.3c00589 Biswas, B., Timsina, J., Mandal, K.G., Naorem, A., 2025. Effects of different irrigation methods and mulching on yield, growth and water use efficiency of strawberry. New Zealand Journal of Crop and Horticultural Science 53, 1408–1427. https://doi.org/10.1080/01140671.2024.2348138 Cao, L., Wang, L., Qi, Y., Yang, S., Gao, J., Liu, Q., Song, L., Hu, R., Wang, Z., Zhang, H., 2025. Enhanced effect of ferrous sulfate on nitrogen retention and PBAT degradation during co-composting by combing with biochar-loaded FN1 bacterial composites. Journal of Environmental Management 373, 123749. https://doi.org/10.1016/j.jenvman.2024.123749 Chaudhary, D.K., Kim, J., 2018. Brevundimonas mongoliensis sp. nov., A Novel Psychrotolerant Bacterium Isolated from Oil-Contaminated Soil. Curr Microbiol 75, 1530–1536. https://doi.org/10.1007/s00284-018-1555-4 Chen, H., Wang, Y., Sun, X., Peng, Y., Xiao, L., 2020. Mixing effect of polylactic acid microplastic and straw residue on soil property and ecological function. Chemosphere 243, 125271. https://doi.org/10.1016/j.chemosphere.2019.125271 Costa, R., Saraiva, A., Carvalho, L., Duarte, E., 2014. The use of biodegradable mulch films on strawberry crop in Portugal. Scientia Horticulturae 173, 65–70. https://doi.org/10.1016/j.scienta.2014.04.020 Crossman, J., Hurley, R.R., Futter, M., Nizzetto, L., 2020. Transfer and transport of microplastics from biosolids to agricultural soils and the wider environment. Science of The Total Environment 724, 138334. https://doi.org/10.1016/j.scitotenv.2020.138334 Dangi, K.K., Darshan, D., Hitesh, P., Singh Dhawai, D., 2025. Efficacy of Various Mulches for the Modulation of Strawberry (Fragaria × ananassa Duch.) Yield, Quality, and Storage Ability. Applied Fruit Science 67, 233. https://doi.org/10.1007/s10341-025-01452-1 Datta, H.S., Barua, P.C., Kotoky, U., Das, R., Saikia, H., Nath, H.K.D., 2024. Impact of Plant Spacing and Mulch on Growth Parameters of Strawberry (Fragaria x ananassa Duch.). JEAI 46, 926–936. https://doi.org/10.9734/jeai/2024/v46i52448 De Souza Machado, A.A., Lau, C.W., Till, J., Kloas, W., Lehmann, A., Becker, R., Rillig, M.C., 2018. Impacts of Microplastics on the Soil Biophysical Environment. Environ. Sci. Technol. 52, 9656–9665. https://doi.org/10.1021/acs.est.8b02212 DeVetter, L.W., Zhang, H., Ghimire, S., Watkinson, S., Miles, C.A., 2017. Plastic Biodegradable Mulches Reduce Weeds and Promote Crop Growth in Day-neutral Strawberry in Western Washington. horts 52, 1700–1706. https://doi.org/10.21273/HORTSCI12422-17 Dewi, S.K., Han, Z.M., Bhat, S.A., Zhang, F., Wei, Y., Li, F., 2024. Effect of plastic mulch residue on plant growth performance and soil properties. Environmental Pollution 343, 123254. https://doi.org/10.1016/j.envpol.2023.123254 Di Mola, I., Cozzolino, E., Ottaiano, L., Riccardi, R., Spigno, P., Petriccione, M., Fiorentino, N., Fagnano, M., Mori, M., 2023. Biodegradable Mulching Film vs. Traditional Polyethylene: Effects on Yield and Quality of San Marzano Tomato Fruits. Plants 12, 3203. https://doi.org/10.3390/plants12183203 Ding, F., Li, S., Lu, J., Penn, C.J., Wang, Q.-W., Lin, G., Sardans, J., Penuelas, J., Wang, J., Rillig, M.C., 2023. Consequences of 33 Years of Plastic Film Mulching and Nitrogen Fertilization on Maize Growth and Soil Quality. Environ. Sci. Technol. 57, 9174–9183. https://doi.org/10.1021/acs.est.2c08878 Dong, D., Guo, Z., Wu, F., Yang, X., Li, J., 2024. Plastic residues alter soil microbial community compositions and metabolite profiles under realistic conditions. Science of The Total Environment 906, 167352. https://doi.org/10.1016/j.scitotenv.2023.167352 Doumett, S., Fibbi, D., Cincinelli, A., Giordani, E., Nin, S., Del Bubba, M., 2011. Comparison of nutritional and nutraceutical properties in cultivated fruits of Fragaria vesca L. produced in Italy. Food Research International 44, 1209–1216. https://doi.org/10.1016/j.foodres.2010.10.044 Eberl, L., Vandamme, P., 2016. Members of the genus Burkholderia: good and bad guys. F1000Res 5, 1007. https://doi.org/10.12688/f1000research.8221.1 El-Beltagi, H.S., Basit, A., Mohamed, H.I., Ali, I., Ullah, S., Kamel, E.A.R., Shalaby, T.A., Ramadan, K.M.A., Alkhateeb, A.A., Ghazzawy, H.S., 2022. Mulching as a Sustainable Water and Soil Saving Practice in Agriculture: A Review. Agronomy 12, 1881. https://doi.org/10.3390/agronomy12081881 Fang, Y., Lin, C., Zhao, J., Gao, Y., Jia, X., 2025. Dosages of Biodegradable Poly(butylene adipate-co-terephthalate) Microplastics Affect Soil Microbial Community, Function, and Metabolome in Plant–Soil System. Agronomy 15, 990. https://doi.org/10.3390/agronomy15040990 Farmer, J., Zhang, B., Jin, X., Zhang, P., Wang, J., 2017. Long-term effect of plastic film mulching and fertilization on bacterial communities in a brown soil revealed by high through-put sequencing. Archives of Agronomy and Soil Science 63, 230–241. Fei, Y., Huang, S., Zhang, H., Tong, Y., Wen, D., Xia, X., Wang, H., Luo, Y., Barceló, D., 2020. Response of soil enzyme activities and bacterial communities to the accumulation of microplastics in an acid cropped soil. Science of The Total Environment 707, 135634. https://doi.org/10.1016/j.scitotenv.2019.135634 Gao, W., Tu, Z., Yin, X., Ming, S., Cai, K., 2025. Effects of PBAT biodegradable mulch on lettuce (Lactuca sativa L.) physiology and soil microbial community: Based on a long-term degradation trial. Ecotoxicology and Environmental Safety 302, 118734. https://doi.org/10.1016/j.ecoenv.2025.118734 Gao, X., Fu, C., Li, M., Qi, X., Jia, X., 2022. Effects of Biodegradation of Corn-Starch–Sodium-Alginate-Based Liquid Mulch Film on Soil Microbial Functions. IJERPH 19, 8631. https://doi.org/10.3390/ijerph19148631 Gatheru Waigi, M., Sun, K., Gao, Y., 2017. Sphingomonads in Microbe-Assisted Phytoremediation: Tackling Soil Pollution. Trends in Biotechnology 35, 883–899. https://doi.org/10.1016/j.tibtech.2017.06.014 Georgiadou, E.C., García, C.J., Taliadorou, A.M., Gedeon, S., Valanides, N., Varaldo, A., Gohari, G., Balsells-Llauradó, M., Alcázar, R., Hertog, M.L.A.T.M., Tomás-Barberán, F.A., Manganaris, G.A., Fotopoulos, V., 2025. Pre-harvest application of sodium alginate functionalized with melatonin enhances secondary metabolism in strawberry fruit. Current Plant Biology 43, 100515. https://doi.org/10.1016/j.cpb.2025.100515 Grimaut, D.A., Da Silva, J.B.A., Lemos, P.V.F., Correia, P.R.C., Santana, J.S., Pessôa, L.C., Estevez-Areco, S., Famá, L.M., Goyanes, S.N., Marcelino, H.R., De Jesus Assis, D., De Souza, C.O., 2023. Effect of Addition of Cross-Linked Starch on the Properties of Degraded PBAT Poly(butylene adipate-co-terephthalate) Films. Polymers 15, 3106. https://doi.org/10.3390/polym15143106 Gu, J., Guo, F., Lin, L., Zhang, J., Sun, W., Muhammad, R., Liang, H., Duan, D., Deng, X., Lin, Z., Wang, Y., Zhong, Y., Xu, Z., 2023. Microbiological mechanism for “production while remediating” in Cd-contaminated paddy fields: A field experiment. Science of The Total Environment 885, 163896. https://doi.org/10.1016/j.scitotenv.2023.163896 Ham, J.M., Kluitenberg, G.J., Lamont, W.J., 1993. Optical Properties of Plastic Mulches Affect the Field Temperature Regime. jashs 118, 188–193. https://doi.org/10.21273/JASHS.118.2.188 Hannum, S.M., 2004. Potential Impact of Strawberries on Human Health: A Review of the Science. Critical Reviews in Food Science and Nutrition 44, 1–17. https://doi.org/10.1080/10408690490263756 Hartman, K., Van Der Heijden, M.G.A., Wittwer, R.A., Banerjee, S., Walser, J.-C., Schlaeppi, K., 2018. Cropping practices manipulate abundance patterns of root and soil microbiome members paving the way to smart farming. Microbiome 6, 14. https://doi.org/10.1186/s40168-017-0389-9 Hartmann, M., Widmer, F., 2006. Community Structure Analyses Are More Sensitive to Differences in Soil Bacterial Communities than Anonymous Diversity Indices. Appl Environ Microbiol 72, 7804–7812. https://doi.org/10.1128/AEM.01464-06 Hegan, D., Tong, L., Zhiquan, H., Qinming, S., Ru, L., 2015. Determining time limits of continuous film mulching and examining residual effects on cotton yield and soil properties. Journal of Environmental Biology 36, 677. Hossain, M.E., Zhang, Z., Dong, W., Wang, S., Liu, M., Liu, E., Mei, X., 2022. Plastic Film Mulching Improved Maize Yield, Water Use Efficiency, and N Use Efficiency under Dryland Farming System in Northeast China. Plants 11, 1710. https://doi.org/10.3390/plants11131710 Huang, Y., Zhao, Y., Wang, J., Zhang, M., Jia, W., Qin, X., 2019. LDPE microplastic films alter microbial community composition and enzymatic activities in soil. Environmental Pollution 254, 112983. https://doi.org/10.1016/j.envpol.2019.112983 Ingman, M., Santelmann, M.V., Tilt, B., 2015. Agricultural water conservation in china: plastic mulch and traditional irrigation. Ecosyst Health Sustain 1, 1–11. https://doi.org/10.1890/EHS14-0018.1 Jabran, K., Hussain, M., Fahad, S., Farooq, M., Bajwa, A.A., Alharrby, H., Nasim, W., 2016. Economic assessment of different mulches in conventional and water-saving rice production systems. Environ Sci Pollut Res 23, 9156–9163. https://doi.org/10.1007/s11356-016-6162-y Jacquin, J., Cheng, J., Odobel, C., Pandin, C., Conan, P., Pujo-Pay, M., Barbe, V., Meistertzheim, A.-L., Ghiglione, J.-F., 2019. Microbial Ecotoxicology of Marine Plastic Debris: A Review on Colonization and Biodegradation by the “Plastisphere.” Front. Microbiol. 10, 865. https://doi.org/10.3389/fmicb.2019.00865 Kader, M.A., Senge, M., Mojid, M.A., Ito, K., 2017. Recent advances in mulching materials and methods for modifying soil environment. Soil and Tillage Research 168, 155–166. https://doi.org/10.1016/j.still.2017.01.001 Kapanen, A., Schettini, E., Vox, G., Itävaara, M., 2008. Performance and Environmental Impact of Biodegradable Films in Agriculture: A Field Study on Protected Cultivation. J Polym Environ 16, 109–122. https://doi.org/10.1007/s10924-008-0091-x Kasirajan, S., Ngouajio, M., 2012. Polyethylene and biodegradable mulches for agricultural applications: a review. Agron. Sustain. Dev. 32, 501–529. https://doi.org/10.1007/s13593-011-0068-3 Kasperbauer, M.J., Loughrin, J.H., Wang, S.Y., 2007. Light Reflected from Red Mulch to Ripening Strawberries Affects Aroma, Sugar and Organic Acid Concentrations¶. Photochemistry and Photobiology 74, 103–107. https://doi.org/10.1562/0031-8655(2001)0740103LRFRMT2.0.CO2 Khandare, S.D., Agrawal, D., Mehru, N., Chaudhary, D.R., 2022. Marine bacterial based enzymatic degradation of low-density polyethylene (LDPE) plastic. Journal of Environmental Chemical Engineering 10, 107437. https://doi.org/10.1016/j.jece.2022.107437 Kim, S.-K., Kim, J.-S., Lee, H., Lee, H.-J., 2021. Abundance and characteristics of microplastics in soils with different agricultural practices: Importance of sources with internal origin and environmental fate. Journal of Hazardous Materials 403, 123997. https://doi.org/10.1016/j.jhazmat.2020.123997 Koitabashi, M., Noguchi, M.T., Sameshima-Yamashita, Y., Hiradate, S., Suzuki, K., Yoshida, S., Watanabe, T., Shinozaki, Y., Tsushima, S., Kitamoto, H.K., 2012. Degradation of biodegradable plastic mulch films in soil environment by phylloplane fungi isolated from gramineous plants. AMB express 2, 40. Koutny, M., Lemaire, J., Delort, A.-M., 2006. Biodegradation of polyethylene films with prooxidant additives. Chemosphere 64, 1243–1252. https://doi.org/10.1016/j.chemosphere.2005.12.060 Kumar, A., Männistö, M.K., Pätsi, M., Kerkhof, L.J., Häggblom, M.M., 2025. Genome analysis reveals diverse novel psychrotolerant Mucilaginibacter species in Arctic tundra soils. ISME Communications 5, ycaf071. https://doi.org/10.1093/ismeco/ycaf071 Küster, A.N., Paula, C., Azevedo, J., Serra, A.C., Coelho, J.F.J., 2025. Formulations, Processing, and Application of Poly(butylene adipate-co-terephthalate)/Thermoplastic Starch Blends: A Review. Polymers 17, 1457. https://doi.org/10.3390/polym17111457 Lee, D., Lee, E., Lee, Y., Shin, M., Yang, J.S., Kim, M., Sang, M.K., Park, H.J., Jung, H.W., 2025. Agri-plastics in soils drive changes in the rhizosphere bacterial community and plant transcriptome in Arabidopsis. https://doi.org/doi.org/10.1093/jxb/eraf336 Li, C., Cui, Q., Li, Y., Zhang, K., Lu, X., Zhang, Y., 2022. Effect of LDPE and biodegradable PBAT primary microplastics on bacterial community after four months of soil incubation. Journal of Hazardous Materials 429, 128353. https://doi.org/10.1016/j.jhazmat.2022.128353 Li, C., Moore-Kucera, J., Miles, C., Leonas, K., Lee, J., Corbin, A., Inglis, D., 2014. Degradation of potentially biodegradable plastic mulch films at three diverse US locations. Agroecology and sustainable food systems 38, 861–889. Li, H., Li, P., Cao, G., Zhao, M., Zhu, Z., Ma, Y., Wang, W., Cao, S., Xu, Y., Dong, C., 2025. Mulching influences pear yield and quality by changing rhizosphere microbial community structure in the arid region of Northwest China. Front. Plant Sci. 16, 1633540. https://doi.org/10.3389/fpls.2025.1633540 Li, N., Qu, J., Yang, J., 2023. Microplastics distribution and microbial community characteristics of farmland soil under different mulch methods. Journal of Hazardous Materials 445, 130408. https://doi.org/10.1016/j.jhazmat.2022.130408 Li YuanQiao, L.Y., Zhao CaiXia, Z.C., Yan ChangRong, Y.C., Mao LiLi, M.L., Liu Qi, L.Q., Li Zhen, L.Z., He WenQing, H.W., 2020. Effects of agricultural plastic film residues on transportation and distribution of water and nitrate in soil. Liu, C., Cui, Y., Li, X., Yao, M., 2021. microeco : an R package for data mining in microbial community ecology. FEMS Microbiology Ecology 97, fiaa255. https://doi.org/10.1093/femsec/fiaa255 Liu, E.K., He, W.Q., Yan, C.R., 2014. ‘White revolution’ to ‘white pollution’—agricultural plastic film mulch in China. Environ. Res. Lett. 9, 091001. https://doi.org/10.1088/1748-9326/9/9/091001 Liu, L., Li, L., Zou, G., Gu, J., Zuo, Q., Zheng, X., Du, L., Liu, D., 2025. Soil fungi respond more violently to both polyethylene and PBAT biodegradable mulch film residues than bacteria do. Front. Environ. Sci. 13, 1533441. https://doi.org/10.3389/fenvs.2025.1533441 Liu, L., Zou, G., Zuo, Q., Li, C., Gu, J., Kang, L., Ma, M., Liang, K., Liu, D., Du, L., 2022. Soil bacterial community and metabolism showed a more sensitive response to PBAT biodegradable mulch residues than that of LDPE mulch residues. Journal of Hazardous Materials 438, 129507. https://doi.org/10.1016/j.jhazmat.2022.129507 Liu, X., Wen, Z., Zhou, W., Dong, W., Ren, H., Liang, G., Gong, W., 2025. Effect of Multiyear Biodegradable Plastic Mulch on Soil Microbial Community, Assembly, and Functioning. Microorganisms 13, 259. https://doi.org/10.3390/microorganisms13020259 López-Tolentino, G., Ibarra-Jiménez, L., Méndez-Prieto, A., Lozano-del Río, A.J., Lira-Saldivar, R.H., Valenzuela-Soto, J.H., Lozano-Cavazos, C.J., Torres-Olivar, V., 2017. Photosynthesis, growth, and fruit yield of cucumber in response to oxo-degradable plastic mulches. Acta Agriculturae Scandinavica, Section B—Soil & Plant Science 67, 77–84. Louca, S., Parfrey, L.W., Doebeli, M., 2016. Decoupling function and taxonomy in the global ocean microbiome. Science 353, 1272–1277. Mahawar, L., Mishra, A., Tsitouri, A., Albrectsen, B.R., 2026. Straw Mulching Differentially Shapes the Structure and Function of Below‐Ground Bacterial Communities in Potato Depending on eDNA Source and Cultivar. Plant Enviro Interactions 7, e70131. https://doi.org/10.1002/pei3.70131 Mangiafico, S., 2016. rcompanion: Functions to Support Extension Education Program Evaluation. https://doi.org/10.32614/CRAN.package.rcompanion Mansoor, Z., Tchuenbou-Magaia, F., Kowalczuk, M., Adamus, G., Manning, G., Parati, M., Radecka, I., Khan, H., 2022. Polymers Use as Mulch Films in Agriculture—A Review of History, Problems and Current Trends. Polymers 14, 5062. https://doi.org/10.3390/polym14235062 Masny, A., 2015. SEASON EXTENSION POSSIBILITIES IN TWO POLISH JUNE-BEARING STRAWBERRY CULTIVARS. Meng, T., Bu, H., Zhang, X., Chen, X., Wang, W., Zhao, M., Liu, J., Zhang, J., Zhang, D., Lu, Z., Zhao, X., 2025. Degradable film mulching recruited beneficial microbiota and increased rhizosphere bacterial diversity in sunflower. Sci Rep 15, 18522. https://doi.org/10.1038/s41598-025-03213-2 Menossi, M., Cisneros, M., Alvarez, V.A., Casalongué, C., 2021. Current and emerging biodegradable mulch films based on polysaccharide bio-composites. A review. Agron. Sustain. Dev. 41, 53. https://doi.org/10.1007/s13593-021-00685-0 Menzel, C.M., 2025. The relationship between yield and plant density in strawberry: competition does not impose an upper limit to population-level production. The Journal of Horticultural Science and Biotechnology 100, 1–28. https://doi.org/10.1080/14620316.2024.2400127 Meyer, M., Diehl, D., Schaumann, G.E., Muñoz, K., 2021. Multiannual soil mulching in agriculture: analysis of biogeochemical soil processes under plastic and straw mulches in a 3-year field study in strawberry cultivation. J Soils Sediments 21, 3733–3752. https://doi.org/10.1007/s11368-021-03037-3 Miller, K., Feucht, W., Schmid, M., 2019. Bioactive Compounds of Strawberry and Blueberry and Their Potential Health Effects Based on Human Intervention Studies: A Brief Overview. Nutrients 11, 1510. https://doi.org/10.3390/nu11071510 Mo, F., Han, J., Wen, X., Wang, X., Li, P., Vinay, N., Jia, Z., Xiong, Y., Liao, Y., 2020. Quantifying regional effects of plastic mulch on soil nitrogen pools, cycles, and fluxes in rain‐fed agroecosystems of the Loess Plateau. Land Degrad Dev 31, 1675–1687. https://doi.org/10.1002/ldr.3548 Morales-Quintana, L., Ramos, P., 2019. Chilean strawberry (Fragaria chiloensis): An integrative and comprehensive review. Food Research International 119, 769–776. https://doi.org/10.1016/j.foodres.2018.10.059 Morra, L., Bilotto, M., Mignoli, E., Sicignano, M., Magri, A., Cice, D., Cozzolino, R., Malorni, L., Siano, F., Picariello, G., Guerrini, S., Petriccione, M., 2022. New Mater-Bi, Biodegradable Mulching Film for Strawberry (Fragaria × Ananassa Duch.): Effects on Film Duration, Crop Yields, Qualitative, and Nutraceutical Traits of Fruits. Plants 11, 1726. https://doi.org/10.3390/plants11131726 Morra, L., Cozzolino, E., Salluzzo, A., Modestia, F., Bilotto, M., Baiano, S., Del Piano, L., 2021. Plant Growth, Yields and Fruit Quality of Processing Tomato (Solanum lycopersicon L.) as Affected by the Combination of Biodegradable Mulching and Digestate. Agronomy 11, 100. https://doi.org/10.3390/agronomy11010100 Muñoz, K., Buchmann, C., Meyer, M., Schmidt-Heydt, M., Steinmetz, Z., Diehl, D., Thiele-Bruhn, S., Schaumann, G.E., 2017a. Physicochemical and microbial soil quality indicators as affected by the agricultural management system in strawberry cultivation using straw or black polyethylene mulching. Applied Soil Ecology 113, 36–44. https://doi.org/10.1016/j.apsoil.2017.01.014 Muñoz, K., Buchmann, C., Meyer, M., Schmidt-Heydt, M., Steinmetz, Z., Diehl, D., Thiele-Bruhn, S., Schaumann, G.E., 2017b. Physicochemical and microbial soil quality indicators as affected by the agricultural management system in strawberry cultivation using straw or black polyethylene mulching. Applied Soil Ecology 113, 36–44. https://doi.org/10.1016/j.apsoil.2017.01.014 Muñoz, K., Schmidt-Heydt, M., Stoll, D., Diehl, D., Ziegler, J., Geisen, R., Schaumann, G.E., 2015. Effect of plastic mulching on mycotoxin occurrence and mycobiome abundance in soil samples from asparagus crops. Mycotoxin research 31, 191–201. Muñoz, K., Thiele-Bruhn, S., Kenngott, K.G.J., Meyer, M., Diehl, D., Steinmetz, Z., Schaumann, G.E., 2022. Effects of Plastic versus Straw Mulching Systems on Soil Microbial Community Structure and Enzymes in Strawberry Cultivation. Soil Systems 6, 21. https://doi.org/10.3390/soilsystems6010021 Muroi, F., Tachibana, Y., Kobayashi, Y., Sakurai, T., Kasuya, K., 2016. Influences of poly(butylene adipate-co-terephthalate) on soil microbiota and plant growth. Polymer Degradation and Stability 129, 338–346. https://doi.org/10.1016/j.polymdegradstab.2016.05.018 Neri, D., Baruzzi, G., Massetani, F., Faedi, W., 2012. Strawberry production in forced and protected culture in Europe as a response to climate change. Can. J. Plant Sci. 92, 1021–1036. https://doi.org/10.4141/cjps2011-276 Ni, X., Song, W., Zhang, H., Yang, X., Wang, L., 2016. Effects of Mulching on Soil Properties and Growth of Tea Olive (Osmanthus fragrans). PLoS ONE 11, e0158228. https://doi.org/10.1371/journal.pone.0158228 Ogle, D.H., Wheeler, P., Dinno, A., 2020. FSA: fisheries stock analysis. R package version 0.8. 30. Vienna: R Core Team. Oksanen, J., 2015. Vegan: community ecology package. R package version 2, 3. Pinto, J.P., Da Cunha, F.F., Da Silva Adão, A., De Paula, L.B., Ribeiro, M.C., Costa Neto, J.R.R., 2022. Strawberry Production with Different Mulches and Wetted Areas. Horticulturae 8, 930. https://doi.org/10.3390/horticulturae8100930 Pruesse, E., Quast, C., Knittel, K., Fuchs, B.M., Ludwig, W., Peplies, J., Glockner, F.O., 2007. SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Research 35, 7188–7196. https://doi.org/10.1093/nar/gkm864 Qi, Y., Ossowicki, A., Yang, X., Huerta Lwanga, E., Dini-Andreote, F., Geissen, V., Garbeva, P., 2020. Effects of plastic mulch film residues on wheat rhizosphere and soil properties. Journal of Hazardous Materials 387, 121711. https://doi.org/10.1016/j.jhazmat.2019.121711 Qi, Y., Ossowicki, A., Yergeau, É., Vigani, G., Geissen, V., Garbeva, P., 2022. Plastic mulch film residues in agriculture: impact on soil suppressiveness, plant growth, and microbial communities. FEMS Microbiology Ecology 98, fiac017. https://doi.org/10.1093/femsec/fiac017 Qiang, L., Hu, H., Li, G., Xu, J., Cheng, J., Wang, J., Zhang, R., 2023. Plastic mulching, and occurrence, incorporation, degradation, and impacts of polyethylene microplastics in agroecosystems. Ecotoxicology and Environmental Safety 263, 115274. https://doi.org/10.1016/j.ecoenv.2023.115274 Qin, W., Hu, C., Oenema, O., 2015. Soil mulching significantly enhances yields and water and nitrogen use efficiencies of maize and wheat: a meta-analysis. Sci Rep 5, 16210. https://doi.org/10.1038/srep16210 Rillig, M.C., Kim, S.W., Zhu, Y.-G., 2024. The soil plastisphere. Nat Rev Microbiol 22, 64–74. https://doi.org/10.1038/s41579-023-00967-2 Salama, K., Geyer, M., 2023. Plastic Mulch Films in Agriculture: Their Use, Environmental Problems, Recycling and Alternatives. Environments 10, 179. https://doi.org/10.3390/environments10100179 Scales, B.S., Cable, R.N., Duhaime, M.B., Gerdts, G., Fischer, F., Fischer, D., Mothes, S., Hintzki, L., Moldaenke, L., Ruwe, M., Kalinowski, J., Kreikemeyer, B., Pedrotti, M.-L., Gorsky, G., Elineau, A., Labrenz, M., Oberbeckmann, S., 2021. Cross-Hemisphere Study Reveals Geographically Ubiquitous, Plastic-Specific Bacteria Emerging from the Rare and Unexplored Biosphere. mSphere 6, e00851-20. https://doi.org/10.1128/mSphere.00851-20 Scarascia-Mugnozza, G., Sica, C., Russo, G., 2011. PLASTIC MATERIALS IN EUROPEAN AGRICULTURE: ACTUAL USE AND PERSPECTIVES. Scheurer, M., Bigalke, M., 2018. Microplastics in Swiss Floodplain Soils. Environ. Sci. Technol. 52, 3591–3598. https://doi.org/10.1021/acs.est.7b06003 Schewe, J., Heinke, J., Gerten, D., Haddeland, I., Arnell, N.W., Clark, D.B., Dankers, R., Eisner, S., Fekete, B.M., Colón-González, F.J., Gosling, S.N., Kim, H., Liu, X., Masaki, Y., Portmann, F.T., Satoh, Y., Stacke, T., Tang, Q., Wada, Y., Wisser, D., Albrecht, T., Frieler, K., Piontek, F., Warszawski, L., Kabat, P., 2014. Multimodel assessment of water scarcity under climate change. Proc. Natl. Acad. Sci. U.S.A. 111, 3245–3250. https://doi.org/10.1073/pnas.1222460110 Schonbeck, M.W., 1999. Weed Suppression and Labor Costs Associated with Organic, Plastic, and Paper Mulches in Small-Scale Vegetable Production. Journal of Sustainable Agriculture 13, 13–33. https://doi.org/10.1300/J064v13n02_04 Scopetani, C., Bellabarba, A., Selvolini, G., Martellini, T., Viti, C., Cincinelli, A., 2025. Evaluating additive release from conventional and biodegradable mulch films. Science of The Total Environment 975, 179294. https://doi.org/10.1016/j.scitotenv.2025.179294 Scopetani, C., Cincinelli, A., Martellini, T., Rodrigues, A.C.M., 2023. Editorial: Hazardous contaminants associated with plastics: occurrence and environmental effects. Front. Environ. Sci. 11, 1341738. https://doi.org/10.3389/fenvs.2023.1341738 Segata, N., Izard, J., Waldron, L., Gevers, D., Miropolsky, L., Garrett, W.S., Huttenhower, C., 2011. Metagenomic biomarker discovery and explanation. Genome Biol 12, R60. https://doi.org/10.1186/gb-2011-12-6-r60 Šerá, J., Kadlečková, M., Fayyazbakhsh, A., Kučabová, V., Koutný, M., 2020. Occurrence and Analysis of Thermophilic Poly(butylene adipate-co-terephthalate)-Degrading Microorganisms in Temperate Zone Soils. IJMS 21, 7857. https://doi.org/10.3390/ijms21217857 Shan, X., Zhang, W., Dai, Z., Li, J., Mao, W., Yu, F., Ma, J., Wang, S., Zeng, X., 2022. Comparative Analysis of the Effects of Plastic Mulch Films on Soil Nutrient, Yields and Soil Microbiome in Three Vegetable Fields. Agronomy 12, 506. https://doi.org/10.3390/agronomy12020506 Sharma, S., Pant, P., Sangwan, M., Sahrawat, R., 2024. Effect of Organic and Inorganic Mulch on Growth, Yield and Quality of Strawberry CV. Winter Dawn. IJECC 14, 377–382. https://doi.org/10.9734/ijecc/2024/v14i34049 Shi, Z., Xiong, L., Liu, T., Wu, W., 2022. Alteration of bacterial communities and co-occurrence networks as a legacy effect upon exposure to polyethylene residues under field environment. Journal of Hazardous Materials 426, 128126. https://doi.org/10.1016/j.jhazmat.2021.128126 Shiukhy, S., Raeini-Sarjaz, M., Chalavi, V., 2015. Colored plastic mulch microclimates affect strawberry fruit yield and quality. Int J Biometeorol 59, 1061–1066. https://doi.org/10.1007/s00484-014-0919-0 Sintim, H.Y., Flury, M., 2017. Is Biodegradable Plastic Mulch the Solution to Agriculture’s Plastic Problem? Environ. Sci. Technol. 51, 1068–1069. https://doi.org/10.1021/acs.est.6b06042 Smith, M.R., Myers, S.S., 2018. Impact of anthropogenic CO2 emissions on global human nutrition. Nature Clim Change 8, 834–839. https://doi.org/10.1038/s41558-018-0253-3 Somanathan, H., Sathasivam, R., Sivaram, S., Mariappan Kumaresan, S., Muthuraman, M.S., Park, S.U., 2022. An update on polyethylene and biodegradable plastic mulch films and their impact on the environment. Chemosphere 307, 135839. https://doi.org/10.1016/j.chemosphere.2022.135839 Soppelsa, S., Kelderer, M., Casera, C., Bassi, M., Robatscher, P., Matteazzi, A., Andreotti, C., 2019. Foliar Applications of Biostimulants Promote Growth, Yield and Fruit Quality of Strawberry Plants Grown under Nutrient Limitation. Agronomy 9, 483. https://doi.org/10.3390/agronomy9090483 Steinmetz, Z., Wollmann, C., Schaefer, M., Buchmann, C., David, J., Tröger, J., Muñoz, K., Frör, O., Schaumann, G.E., 2016. Plastic mulching in agriculture. Trading short-term agronomic benefits for long-term soil degradation? Science of The Total Environment 550, 690–705. https://doi.org/10.1016/j.scitotenv.2016.01.153 Sun, Y., Duan, C., Cao, N., Ding, C., Huang, Y., Wang, J., 2022. Biodegradable and conventional microplastics exhibit distinct microbiome, functionality, and metabolome changes in soil. Journal of Hazardous Materials 424, 127282. https://doi.org/10.1016/j.jhazmat.2021.127282 Supreetha, B.G., Singh, N.P., Sharma, S., Sharda, R., Sharma, A., 2025. Comparison of fruit colour development and biochemical attributes under different mulching regimes in strawberry. Acta Physiol Plant 47, 7. https://doi.org/10.1007/s11738-024-03751-8 Tagg, A.S., Sperlea, T., Labrenz, M., Harrison, J.P., Ojeda, J.J., Sapp, M., 2022. Year-Long Microbial Succession on Microplastics in Wastewater: Chaotic Dynamics Outweigh Preferential Growth. Microorganisms 10, 1775. https://doi.org/10.3390/microorganisms10091775 Takahashi, S., Tomita, J., Nishioka, K., Hisada, T., Nishijima, M., 2014. Development of a Prokaryotic Universal Primer for Simultaneous Analysis of Bacteria and Archaea Using Next-Generation Sequencing. PLoS ONE 9, e105592. https://doi.org/10.1371/journal.pone.0105592 Torres-Olivar, V., Ibarra-Jiménez, L., Cárdenas-Flores, A., Lira-Saldivar, R.H., Valenzuela-Soto, J.H., Castillo-Campohermoso, M.A., 2018. Changes induced by plastic film mulches on soil temperature and their relevance in growth and fruit yield of pickling cucumber. Acta Agriculturae Scandinavica, Section B—Soil & Plant Science 68, 97–103. Tozzi, F., Del Bubba, M., Petrucci, W.A., Pecchioli, S., Macci, C., Hernández García, F., Martínez Nicolás, J.J., Giordani, E., 2020. Use of a remediated dredged marine sediment as a substrate for food crop cultivation: Sediment characterization and assessment of fruit safety and quality using strawberry (Fragaria x ananassa Duch.) as model species of contamination transfer. Chemosphere 238, 124651. https://doi.org/10.1016/j.chemosphere.2019.124651 Uba, B.O., 2019. Growth Profile and Catabolic Pathways Involved in Degradation of Aromatic Hydrocarbons by Marine Bacteria Isolated from Niger Delta, Nigeria. MRJI 1–18. https://doi.org/10.9734/mrji/2018/v26i530075 Villena, J., Moreno, M., González-Mora, S., López-Perales, J., Morales-Rodríguez, P., Moreno, C., 2022. Degradation Pattern of Five Biodegradable, Potentially Low-Environmental-Impact Mulches under Laboratory Conditions. Agriculture 12, 1910. https://doi.org/10.3390/agriculture12111910 Vita, M.M., Iturbe-Espinoza, P., Bonte, M., Brandt, B.W., Braster, M., Brown, D.M., Van Spanning, R.J.M., 2022. Oil Absorbent Polypropylene Particles Stimulate Biodegradation of Crude Oil by Microbial Consortia. Front. Microbiol. 13, 853285. https://doi.org/10.3389/fmicb.2022.853285 Wang, C., Dong, J., Ji, B., Lv, Y., Song, Y., Li, Q., Sun, C., Zong, R., Zhang, M., 2025. Sub-soil biodegradable film mulching: A sustainable solution for enhancing winter wheat yield and shaping soil microbial communities in saline-alkali soils. Journal of Hazardous Materials 495, 139020. https://doi.org/10.1016/j.jhazmat.2025.139020 Wang, G., Li, Y., Pan, X., Li, A., Wang, J., Yin, L., Zeng, X., Qian, X., 2026. Soil Properties and Microbial Community Assemblages in Response to Plastic Film Mulches with Divergent Degradation Characteristics. Microorganisms 14, 553. https://doi.org/10.3390/microorganisms14030553 Wang, H., Yang, Q., Li, D., Wu, J., Yang, S., Deng, Y., Luo, C., Jia, W., Zhong, Y., Peng, P., 2023. Stable Isotopic and Metagenomic Analyses Reveal Microbial-Mediated Effects of Microplastics on Sulfur Cycling in Coastal Sediments. Environ. Sci. Technol. 57, 1167–1176. https://doi.org/10.1021/acs.est.2c06546 Wang, S.Y., Galletta, G.J., Camp, M.J., Kasperbauer, M.J., 1998. Mulch types affect fruit quality and composition of two strawberry genotypes. HortScience 33, 636–640. Wang, X., Shrestha, S., Tymon, L., Zhang, H., Miles, C., DeVetter, L., 2022. Soil-biodegradable mulch is an alternative to non-biodegradable plastic mulches in a strawberry-lettuce double-cropping system. Front. Sustain. Food Syst. 6, 942645. https://doi.org/10.3389/fsufs.2022.942645 Wang, Z., Wang, S., Bian, T., Wang, T., Fu, H., Sun, Z., 2024. Revealing the Response of Cucumber Soil Microbial Community Composition and Function to Nitrogen Addition in Northern Chinese Greenhouses. Horticulturae 10, 1090. https://doi.org/10.3390/horticulturae10101090 Wang, Z.-H., Gao, S.-S., Yang, L., Meng, Y.-L., Wang, M., Li, B.-L.L., Chen, Z.-J., 2026. Responses of Sorghum Growth and Rhizosphere–Plastisphere Microbiomes to Cadmium and Polypropylene Microplastic Co-Contamination. Agronomy 16, 293. https://doi.org/10.3390/agronomy16030293 Wickham, H., 2016. ggplot2, Use R! Springer International Publishing, Cham. https://doi.org/10.1007/978-3-319-24277-4 Wu, C., Ma, Yajie, Wang, D., Shan, Y., Song, X., Hu, H., Ren, X., Ma, X., Cui, J., Ma, Yan, 2022. Integrated microbiology and metabolomics analysis reveal plastic mulch film residue affects soil microorganisms and their metabolic functions. Journal of Hazardous Materials 423, 127258. https://doi.org/10.1016/j.jhazmat.2021.127258 Wu, C., Song, X., Wang, D., Ma, Yajie, Shan, Y., Ren, X., Hu, H., Cui, J., Ma, Yan, 2024. Combined effects of mulch film-derived microplastics and pesticides on soil microbial communities and element cycling. Journal of Hazardous Materials 466, 133656. https://doi.org/10.1016/j.jhazmat.2024.133656 Xu, Z., Zhang, L., Jiang, G., Ding, X., Guo, Y., Tian, Y., 2025. Degradation of mulch films in different soils and its effects on soil properties and ecotoxicology. Environ Geochem Health 47, 345. https://doi.org/10.1007/s10653-025-02651-1 Xu, Z., Zheng, B., Yang, Yichen, Yang, Yi, Jiang, G., Tian, Y., 2024. Effects of biodegradable (PBAT/PLA) and conventional (LDPE) mulch film residues on bacterial communities and metabolic functions in different agricultural soils. Journal of Hazardous Materials 472, 134425. https://doi.org/10.1016/j.jhazmat.2024.134425 Zang, H., Blagodatskaya, E., Wang, J., Xu, X., Kuzyakov, Y., 2017. Nitrogen fertilization increases rhizodeposit incorporation into microbial biomass and reduces soil organic matter losses. Biology and Fertility of Soils 53, 419–429. Zhang, H., Shu, D., Wang, K., Liu, X., Wang, L., Jiang, R., 2026. Biodegradable film mulching alters soil C, N, P and S cycling via mediating microbial communities in dryland. J Sci Food Agric 106, 632–642. https://doi.org/10.1002/jsfa.70193 Zhang, H., Shu, D., Zhang, J., Liu, X., Wang, K., Jiang, R., 2024. Biodegradable film mulching increases soil microbial network complexity and decreases nitrogen-cycling gene abundance. Science of The Total Environment 933, 172874. https://doi.org/10.1016/j.scitotenv.2024.172874 Zhang, M., Xue, Y., Jin, T., Zhang, K., Li, Z., Sun, C., Mi, Q., Li, Q., 2022. Effect of Long-Term Biodegradable Film Mulch on Soil Physicochemical and Microbial Properties. Toxics 10, 129. https://doi.org/10.3390/toxics10030129 Zhang, S., Wang, Y., Sun, L., Qiu, C., Ding, Y., Gu, H., Wang, L., Wang, Z., Ding, Z., 2020. Organic mulching positively regulates the soil microbial communities and ecosystem functions in tea plantation. BMC Microbiol 20, 103. https://doi.org/10.1186/s12866-020-01794-8 Zhang, Y., Gao, W., Mo, A., Jiang, J., He, D., 2022. Degradation of polylactic acid/polybutylene adipate films in different ratios and the response of bacterial community in soil environments. Environmental Pollution 313, 120167. https://doi.org/10.1016/j.envpol.2022.120167 Zhao, Y., Mao, X., Li, S., Huang, X., Che, J., Ma, C., 2023. A Review of Plastic Film Mulching on Water, Heat, Nitrogen Balance, and Crop Growth in Farmland in China. Agronomy 13, 2515. https://doi.org/10.3390/agronomy13102515 Zhao, Y., Zhai, X., Wang, Z., Li, H., Jiang, R., Lee Hill, R., Si, B., Hao, F., 2018. Simulation of soil water and heat flow in ridge cultivation with plastic film mulching system on the Chinese Loess Plateau. Agricultural Water Management 202, 99–112. https://doi.org/10.1016/j.agwat.2018.02.017 Zhao, Z., Wu, H., Jin, T., Liu, H., Men, J., Cai, G., Cernava, T., Duan, G., Jin, D., 2023. Biodegradable mulch films significantly affected rhizosphere microbial communities and increased peanut yield. Science of The Total Environment 871, 162034. https://doi.org/10.1016/j.scitotenv.2023.162034 Zheng, B.-X., Zhao, Y., Bi, Q.-F., Zhou, G.-W., Wang, H.-J., Hao, X.-L., Ding, K., 2019. How to disentangle microbially functional complexity: an insight from the network analysis of C, N, P and S cycling genes. Science Bulletin 64, 1129–1131. https://doi.org/10.1016/j.scib.2019.06.011 Zhou, J., Jia, R., Brown, R.W., Yang, Y., Zeng, Z., Jones, D.L., Zang, H., 2023. The long-term uncertainty of biodegradable mulch film residues and associated microplastics pollution on plant-soil health. Journal of Hazardous Materials 442, 130055. https://doi.org/10.1016/j.jhazmat.2022.130055 Supplementary Files FigureS1.png FigureS2.png FigureS3.png FigureS4.png FigureS5.png FileS1.xlsx FileS2.xlsx SupplementaryInformations.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9428619","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":624308359,"identity":"b7fd1c88-87de-407d-808e-619dafea2da6","order_by":0,"name":"Agnese Bellabarba","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-5656-5225","institution":"University of Florence: Universita degli Studi di Firenze","correspondingAuthor":true,"prefix":"","firstName":"Agnese","middleName":"","lastName":"Bellabarba","suffix":""},{"id":624308360,"identity":"45f8b221-b114-4ed3-a8cb-2e6cb339adcd","order_by":1,"name":"Lorenzo Bini","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Lorenzo","middleName":"","lastName":"Bini","suffix":""},{"id":624308361,"identity":"68b81b2c-163a-4895-ac83-cefaddd1d674","order_by":2,"name":"Costanza Scopetani","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Costanza","middleName":"","lastName":"Scopetani","suffix":""},{"id":624308362,"identity":"ab527dc7-0eac-49fe-bf37-10d5313e7c5d","order_by":3,"name":"Giulia Selvolini","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Giulia","middleName":"","lastName":"Selvolini","suffix":""},{"id":624308363,"identity":"2542025e-698d-45f6-ae7e-51c9165e953c","order_by":4,"name":"Francesca Decorosi","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Francesca","middleName":"","lastName":"Decorosi","suffix":""},{"id":624308364,"identity":"32960abc-f20c-422a-bf52-3caa53c6052d","order_by":5,"name":"Ermes Lo Piccolo","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Ermes","middleName":"Lo","lastName":"Piccolo","suffix":""},{"id":624308365,"identity":"85d75529-a9fb-4330-8248-bff02ccc4979","order_by":6,"name":"Elisabetta Toni","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Elisabetta","middleName":"","lastName":"Toni","suffix":""},{"id":624308366,"identity":"209d955c-f1d1-49fc-b924-9ec77d324fb3","order_by":7,"name":"William Antonio Petrucci","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"William","middleName":"Antonio","lastName":"Petrucci","suffix":""},{"id":624308367,"identity":"df98c4e8-02b5-40ba-8900-7339f1f67081","order_by":8,"name":"Edgardo Giordani","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Edgardo","middleName":"","lastName":"Giordani","suffix":""},{"id":624308368,"identity":"87d98fc5-94ec-4446-bb99-5c1e3649a922","order_by":9,"name":"Giovanna Marrazza","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Giovanna","middleName":"","lastName":"Marrazza","suffix":""},{"id":624308369,"identity":"99cff0e7-0a03-49eb-bdc2-763009e2c630","order_by":10,"name":"Alessandra Cincinelli","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Alessandra","middleName":"","lastName":"Cincinelli","suffix":""},{"id":624308370,"identity":"5576310b-0175-48a1-9b47-f3c59804f4c8","order_by":11,"name":"Tania Martinelli","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Tania","middleName":"","lastName":"Martinelli","suffix":""},{"id":624308371,"identity":"fb5b0af7-d7fb-4b2b-ab92-46681a79a0e2","order_by":12,"name":"Carlo Viti","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Carlo","middleName":"","lastName":"Viti","suffix":""}],"badges":[],"createdAt":"2026-04-15 15:15:50","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9428619/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9428619/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":107710938,"identity":"7714abb8-a6cf-41a7-a2fb-029cba1e1262","added_by":"auto","created_at":"2026-04-24 09:42:59","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":158789,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eYield of strawberry grown with different mulching films.\u003c/strong\u003e The statistical differences are reported with asterisks (p \u0026lt; 0.05) computed with One-Way ANOVA and Tukey’s post-hoc test.\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/dc79629fd9af91c48318b84c.jpg"},{"id":107710931,"identity":"17eb187b-5152-4bd9-8b41-09fac957a76b","added_by":"auto","created_at":"2026-04-24 09:42:50","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":65818,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eβ-diversity of bacterial communities in substrates covered with different mulching films. \u003c/strong\u003eNon-metric multidimensional scaling plot is based on Bray-Curtis distance on overall ASVs abundances. Stress value is reported for the NMDS, as well as the statistical differences with permutational multivariate analysis of variance (PERMANOVA) and Analysis of multivariate homogeneity (PERMDISP) among different mulching films applied. F-ratio (F) for PERMDISP, the estimation of the variance component (R\u003csup\u003e2\u003c/sup\u003e) for PERMANOVA and the Global R for ANOSIM are reported together with the level of significance (ns, not significant; * P \u0026lt; 0.05; ** P \u0026lt; 0.01, *** P \u0026lt; 0.001).\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/b55889f6c4af09a1073c3641.png"},{"id":107710958,"identity":"ab316365-fb1b-4751-b298-6b23997018dd","added_by":"auto","created_at":"2026-04-24 09:43:12","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":98721,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eHighly discriminant ASVs\u003c/strong\u003e \u003cstrong\u003eof substrate bacterial microbial communities for different mulching films. \u003c/strong\u003eIn panel A,\u003cstrong\u003e \u003c/strong\u003ediscriminant ASVs with LDA score threshold \u0026gt; 3.5 for each different mulching films are reported. In panel B, bars represent the total abundance of discriminant ASVs for each different mulching film, while the whiskers represent the standard deviation.\u003c/p\u003e","description":"","filename":"Figura3.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/d8e04996438becf5b95dfe0d.png"},{"id":107710918,"identity":"59608e0a-4926-4528-b619-539f07e69a16","added_by":"auto","created_at":"2026-04-24 09:42:45","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":68955,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eRelationships between physicochemical features and bacterial community structures. \u003c/strong\u003eA) Distance-based redundancy analysis (RDA) plot with significant physicochemical drivers of substrates bacterial community structures. Vectors indicate physicochemical properties significantly correlated with microbial community structures (PERMANOVA analysis). Dissimilarity of bacterial communities was calculated by Bray-Curtis’s distance. B) Marginal effects of substrates properties on Bray-Curtis’s dissimilarity matrix of ASVs with the Permutational Multivariate Analysis of Variance (PERMANOVA). Degrees of freedom (Df), Sum of squares (SumOfSqs), proportion of variance explained (R2); and the Pseudo-F statistic (F), and p-values (Pr(\u0026gt;F)) were reported (*** p \u0026lt; 0.001; ** p \u0026lt; 0.01; * p \u0026lt; 0.05; p \u0026lt; 0.1).\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/cb3c6cb585139846a1bec831.png"},{"id":109405318,"identity":"62d9bdda-5aa5-47fd-9e99-e3e3f79d417e","added_by":"auto","created_at":"2026-05-17 13:16:43","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":840587,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/b57e8b38-050a-426c-a8ab-3895e6531b87.pdf"},{"id":107710937,"identity":"e502fb46-0657-41d6-b3c9-b9e9a0d32bf9","added_by":"auto","created_at":"2026-04-24 09:42:59","extension":"png","order_by":10,"title":"","display":"","copyAsset":false,"role":"supplement","size":7910,"visible":true,"origin":"","legend":"","description":"","filename":"FigureS1.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/a4f1ba90c2af6ccbb11f0eef.png"},{"id":107710810,"identity":"197708f7-6d0b-47fb-b0a5-feae6b9a7a62","added_by":"auto","created_at":"2026-04-24 09:42:16","extension":"png","order_by":11,"title":"","display":"","copyAsset":false,"role":"supplement","size":191614,"visible":true,"origin":"","legend":"","description":"","filename":"FigureS2.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/85b35e866f01a9c7d88c9516.png"},{"id":107710920,"identity":"26da06fd-b855-4309-8bcb-3bd63d5a673d","added_by":"auto","created_at":"2026-04-24 09:42:45","extension":"png","order_by":12,"title":"","display":"","copyAsset":false,"role":"supplement","size":1502198,"visible":true,"origin":"","legend":"","description":"","filename":"FigureS3.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/c5c446a7b28e9b00256e930c.png"},{"id":107710919,"identity":"ac3bde0c-c9aa-474e-b6af-3048a3f18c7d","added_by":"auto","created_at":"2026-04-24 09:42:45","extension":"png","order_by":13,"title":"","display":"","copyAsset":false,"role":"supplement","size":42528,"visible":true,"origin":"","legend":"","description":"","filename":"FigureS4.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/12f836c0b6dfcb1a87f21ac6.png"},{"id":107710954,"identity":"82d6bcf3-ddf3-4dad-a2d4-9d5fa08a6782","added_by":"auto","created_at":"2026-04-24 09:43:07","extension":"png","order_by":14,"title":"","display":"","copyAsset":false,"role":"supplement","size":47814,"visible":true,"origin":"","legend":"","description":"","filename":"FigureS5.png","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/4bd87b0d65bb6840325aba8d.png"},{"id":107712035,"identity":"2408471c-f3c5-4d60-9fb5-7bec3c991828","added_by":"auto","created_at":"2026-04-24 09:47:15","extension":"xlsx","order_by":15,"title":"","display":"","copyAsset":false,"role":"supplement","size":517753,"visible":true,"origin":"","legend":"","description":"","filename":"FileS1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/b40f1b4af940f0aa443f7eff.xlsx"},{"id":107713202,"identity":"54df2642-e68b-4591-9111-2184a9acfae1","added_by":"auto","created_at":"2026-04-24 09:51:47","extension":"xlsx","order_by":16,"title":"","display":"","copyAsset":false,"role":"supplement","size":37598,"visible":true,"origin":"","legend":"","description":"","filename":"FileS2.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/333036453bfd11c798f176c1.xlsx"},{"id":107712051,"identity":"4d768c4c-d4b6-484e-b184-56094d039606","added_by":"auto","created_at":"2026-04-24 09:47:16","extension":"docx","order_by":17,"title":"","display":"","copyAsset":false,"role":"supplement","size":436523,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryInformations.docx","url":"https://assets-eu.researchsquare.com/files/rs-9428619/v1/36ea7a6a168b6606a407ee6f.docx"}],"financialInterests":"","formattedTitle":"Impact of traditional and biodegradable mulching films on plant performance and substrate microbial communities in a small-scale strawberry cultivation","fulltext":[{"header":"1. INTRODUCTION","content":"\u003cp\u003eFeeding a rapidly growing global population requires substantial improvements in agricultural productivity. However, the availability of key resources (i.e., freshwater and fertile soil) is rapidly declining due to the impact of climate change (Schewe et al., \u003cspan citationid=\"CR102\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Smith and Myers, \u003cspan citationid=\"CR113\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). As a result, conventional farming methods alone are no longer sufficient; agriculture must adopt practices that not only improve yields but also safeguard natural resources and minimize environmental impact (Somanathan et al., \u003cspan citationid=\"CR114\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). One of these environmental-practices is mulching, a widely widespread technique in agricultural systems across the globe (Meyer et al., \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Sintim and Flury, \u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Mulching involves covering the soil surface with a layer of material, ranging from organic residues to synthetic films (Mansoor et al., \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), to create a more favourable microenvironment for soil and crops. Indeed, mulching contributes to an earlier increase in soil temperature (Ham et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), which in turn promotes faster plant growth (El-Beltagi et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Kasirajan and Ngouajio, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) and higher yield (Di Mola et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Hossain et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Masny \u0026amp; Żurawicz, 2015.) simultaneously decreasing weed emergence (Schonbeck, \u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e1999\u003c/span\u003e) and then reducing the need for herbicide application. By minimizing water evaporation from the soil surface, the application of mulching films enhances water-use efficiency, resulting in notable economic advantages for farmers (Ingman et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Jabran et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Beyond yield enhancement, mulching has been shown to improve product quality (Neri et al., \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Scarascia-Mugnozza et al., \u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) and is widely used in the cultivation of seasonal horticultural crops such as vegetables, asparagus, and strawberries (Neri et al., \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Scarascia-Mugnozza et al., \u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMany materials with diverse thicknesses can be applied as mulching film, in different arrangements, and periods (Kasirajan and Ngouajio, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Zhao et al., \u003cspan citationid=\"CR145\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) based on the crop, soil features, and climatic conditions. Wheat straw and plastic films, primarily black polyethylene (PE), are the most common mulching materials used in strawberry cultivation (Mu\u0026ntilde;oz et al., \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Indeed, numerous studies have shown that PE films outperform wheat straw, particularly regarding water-use efficiency (Qin et al., \u003cspan citationid=\"CR96\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Specifically, plastic mulches are widely adopted worldwide due to their economic production, flexibility, durability, and high use capacity (Scopetani et al., \u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAlthough beneficial in terms of plant yield and quality, these materials have a considerable impact on the soil environment through both indirect microclimate modification and the direct accumulation of plastic residues (Qiang et al., \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Shi et al., \u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Conventional PE, mulch films, particularly low-density polyethylene (LDPE), exhibit high resistance to chemical degradation, leading to significant long-term legacy effects on the soil microbiome as residues gradually break down into microplastics and nano-plastics (Lee et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Qiang et al., \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Mulching creates a physical barrier that reshapes microbial dynamics by increasing soil temperature and moisture while reducing gas exchange, conditions that typically stimulate root development and the release of root exudates (Kader et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Kapanen et al., \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Kasirajan and Ngouajio, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Li et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2026\u003c/span\u003e). These hydrothermal changes can alter the composition of bacterial and fungal communities, often promoting the growth of Gram-negative bacteria and selective ecological modules associated with improved crop yields (Li et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Furthermore, the incorporation of plastic fragments into the soil matrix establishes a unique microbial niche known as the \"plastisphere,\" which recruits distinct microbial groups from the surrounding bulk soil and rhizosphere (Lee et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Qiang et al., \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Rillig et al., \u003cspan citationid=\"CR97\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Shi et al., \u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Exposure to persistent PE residues frequently results in a reduction of bacterial alpha-diversity and shifts in taxonomic composition across various soil types (Dong et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Li et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Furthermore, mulching films are usually coupled with additives (i.e., plasticizers, colour pigments, ultraviolet stabilizers), which can be released and contaminate the soil. Among these, the most common are phthalates, addressed also as phthalic acid esters (PAEs), which are lipophilic chemicals added during the plastic production process to increase the malleability of the material (Steinmetz et al., \u003cspan citationid=\"CR116\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Zhou et al., \u003cspan citationid=\"CR148\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). PE films are not degradable, and the removal procedure is highly expensive and often not totally completed (Morra et al., \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). All these conditions affect the soil safety and the environmental pollution, which need to be addressed for a more sustainable cultivation practice (El-Beltagi et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Salama and Geyer, \u003cspan citationid=\"CR98\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn this framework, a possible alternative is based on the application of biodegradable mulches because of their shorter degradation capacity and, therefore, avoiding any removal procedures. Unlike conventional PE films, biodegradable mulch films (BDM), primarily composed of polymers like polybutylene adipate terephthalate (PBAT), polylactic acid (PLA), or corn starch blends, are designed to be tilled directly into the soil, where they serve as a bioavailable carbon source for resident microorganisms (Zhang et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2026\u003c/span\u003e). The impact of these materials on bacterial α-diversity is complex and varies depending on soil type and exposure duration. Some investigations indicate that BDMs significantly increase bacterial richness and diversity, particularly within the rhizosphere, where favourable hydrothermal conditions stimulate root growth and the secretion of exudates (Lee et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Meng et al., \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Conversely, other studies observed a reduction in bacterial alpha diversity, which is often attributed to the selective enrichment of specialized polymer degraders that suppress less competitive phyla (Xu et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The presence of both conventional and biodegradable plastic residues significantly disturbs essential biogeochemical cycling of carbon (C), nitrogen (N), phosphorus (P), and sulfur (S); residues can interfere with nutrient mineralization, and alter the expression of functional genes involved in nutrient transformations (Wu et al., \u003cspan citationid=\"CR135\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2026\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe strawberry is recognized as one of the most significant and lucrative agricultural crops in Europe (Miller et al., \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). This success is driven by a combination of exquisite sensory qualities, such as its distinct aroma, flavour, and attractive colour, and its role as a functional food (Morales-Quintana and Ramos, \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Indeed, the strawberry fruit is rich in vitamin C, folic acid, anthocyanins, flavonoids, and phenolic acids. These compounds provide a high antioxidant capacity that is essential for maintaining human wellbeing and preventing the onset of chronic or degenerative diseases when integrated into a daily diet (Hannum, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Miller et al., \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In Europe, strawberry production heavily relies on mulching films, which enable earlier harvests, increase yields, optimize soil temperature and moisture (Biswas et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Datta et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Sharma et al., \u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Steinmetz et al., \u003cspan citationid=\"CR116\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), and reduce disease incidence in vegetable crops (Kasirajan and Ngouajio, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Menossi et al., \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). However, research is shifting beyond general agronomic performance to investigate how differences among various mulching films may influence plant growth as well as fruit nutraceutical properties (Morra et al., \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Indeed, it was observed that biodegradable mulching showed similar performance in plant productivity and quality of PE-based materials (Costa et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Conversely, the application of a new and biodegradable corn starch-based films reduced strawberry fruit quality, even though it reported a similar yield with black PE (Morra et al., \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, despite previous research, the effect of mulches on microbial communities in the framework of a strawberry production has not been fully elucidated.\u003c/p\u003e \u003cp\u003eTherefore, the aim of the present study was to examine the influence of both conventional and biodegradable mulching films on several levels of a small-scale strawberry cultivation system, spanning from substrate microbial communities, to plant performance, fruit yield and quality. In more detail, the study conducted a comprehensive evaluation of plant physiological and morphological responses, as well as productivity, alongside an assessment of substrate microbial α- and β-diversity under different mulching film treatments.\u003c/p\u003e"},{"header":"2. MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Mulching films, plant material and experimental set-up\u003c/h2\u003e \u003cp\u003eThe trial was conducted from April to June 2023 in the experimental area of the Department of Agriculture, Food, Environment and Forestry of the University of Florence located in Sesto Fiorentino, Italy (55 m above sea level; DMS coordinates: 43\u0026deg;81\u0026prime;68\u0026rsquo;\u0026rsquo;N, 11\u0026deg;19\u0026prime;99\u0026rsquo;\u0026rsquo;E). The climate in Sesto Fiorentino is typically Mediterranean, characterized by hot, dry summers and mild, wet winters. During the trial period, the average temperature was 19.2\u0026deg;C with the minimum value of 9.0\u0026deg;C (April 2023) and the maximum one of 29.1\u0026deg;C (June 2024). The cumulative rainfall and the average humidity during the trial were 150 mm and 59%, respectively. Strawberry plants (\u003cem\u003eFragaria\u003c/em\u003e x \u003cem\u003eananassa\u003c/em\u003e Duch.) cv. \u0026lsquo;Camarosa\u0026rsquo; were purchased from Vivai Fratelli Zanzi (Ferrara, Italy). Plants were transplanted into ceramic pots (size: 45 \u0026times; 45 \u0026times; 45 cm; volume: 91 L). Each pot was filled with a commercial growing medium and 4 plants were transplanted. The substrate was composed by a mixture (1:1 v/v) of peat and lapillus (particle size 10\u0026ndash;14 mm) with the following physicochemical properties: bulk density\u0026thinsp;=\u0026thinsp;0.63 g/cm\u003csup\u003e3\u003c/sup\u003e, Porosity\u0026thinsp;=\u0026thinsp;74.87%; Water Holding Capacity (WHC)\u0026thinsp;=\u0026thinsp;30.52% (v/v); Total Organic Carbon (TOC)\u0026thinsp;=\u0026thinsp;4.83%; Total Nitrogen (TN)\u0026thinsp;=\u0026thinsp;0.08 (%); C/N ratio\u0026thinsp;=\u0026thinsp;59.23; pH\u0026thinsp;=\u0026thinsp;6.23; electrical conductivity (EC)\u0026thinsp;=\u0026thinsp;0.04 dS/m; cation exchange capacity (CEC)\u0026thinsp;=\u0026thinsp;22.17 meq/100g. Treatments were arranged according to a randomized block design with 5 replicates (pots) per treatment for a total of 20 plants each one. Four different mulching films were used in this study: polyethylene (PE, thickness 0.05 \u0026micro;m), oxo-degradable polyethylene (Oxo-PE, thickness 15 \u0026micro;m), corn starch and polybutylene adipate terephthalate (PBAT, thickness 18 \u0026micro;m), and polypropylene (PP, thickness 450 \u0026micro;m) (Scopetani et al., \u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Mulching films were applied to cover the inner walls of the pots and subsequently used to cover the surface of the substrate, prior to transplanting. As control, five pots were used without mulching application. Each pot was equipped with two drippers, and irrigation was managed automatically using an irrigation controller, supplying freshwater twice daily to maintain soil moisture at approximately 80% of the field capacity. Prior to planting, a slow-release fertilizer (16-8-12-4(Mg); TRIABON, Compo-Expert GmbH, M\u0026uuml;nster, Germany) was mixed into the substrate at a rate of 135 g per pot. Additionally, calcium nitrate (Haifa Cal Agri, Haifa Chemicals, Haifa, Israel) was applied twice (7 g per pot) throughout the entire duration of the experiment. Subsequently, potassium nitrate (PONI Haifa, Haifa Chemicals, Haifa, Israel) was applied weekly (7 g per pot) until the fruit harvest.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Substrate sampling and physicochemical characterisation\u003c/h2\u003e \u003cp\u003eAt the end of the experiment (June 2023), substrate samples were collected to assess both physicochemical properties and the characterization of the bacterial communities. For the analysis of physicochemical characteristics, approximately 500\u0026ndash;600 g of substrate was taken from each pot (0\u0026ndash;20 cm depth), sieved to \u0026le;\u0026thinsp;2 mm, and subsequently pooled by treatment, combining the samples from five pots to form three replicates per treatment. The following parameters were analyzed following the methodology previously described (Bini et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2024\u003c/span\u003e): bulk density, porosity, total organic carbon (TOC), total nitrogen, nitrate, ammonium, pH, electrical conductivity (EC), and cation exchange capacity (CEC). Additionally, the carbon-to-nitrogen (C/N) ratio was calculated as the quotient of TOC and total nitrogen. The concentration of elements in each substrate was determined with an ICP-OES (iCAP 7000 Plus ICP- OES; Thermo Fisher Scientific, Waltham, MA, USA) by digesting 500 mg of pre-dried sample with 5 mL of nitric acid in a microwave (Mars Microwave Digestor, CEM, Matthews, NC, USA) at 190\u0026deg;C for 15 min.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Morphological, physiological and yield analyses\u003c/h2\u003e \u003cp\u003eBefore transplanting (T0), a baseline morphological assessment was performed on 10 plants, recording plant height and leaf number. These parameters were subsequently monitored throughout the experiment, with measurements taken at three time points spaced 21 days each one (from 29th April to 8th June). For each treatment, data were collected from 10 plants (two plants per pot). Gas exchange measurements (n\u0026thinsp;=\u0026thinsp;7) were conducted on fully expanded leaves of randomly selected plants using a portable infrared gas analyzer (LI-6400 XT, LI-COR Biosciences, Lincoln, NE, USA). Measurements were carried out between 11:00 and 14:00 GMT under a photosynthetic photon flux density of 1600 \u0026micro;mol m⁻\u0026sup2; s⁻\u0026sup1;. The chamber conditions were maintained at a CO₂ concentration of 400 \u0026micro;mol mol⁻\u0026sup1; and a flow rate of 500 \u0026micro;mol s⁻\u0026sup1;. Once leaf gas exchange stabilized, the following parameters were recorded: net photosynthetic rate (Pn), stomatal conductance (gs), and intercellular CO₂ concentration (Ci). The measurements were recorded at the same time reported in the morphological analyses.\u003c/p\u003e \u003cp\u003eFruit harvesting occurred throughout the cropping cycle when strawberries reached commercial ripeness, indicated by full red coloration (from 24th May to 15th June 2023). Fruits were weighed individually to determine total yield per plant over the experimental period. Yield assessments were conducted on five randomly selected plants per treatment. For each plant, the number and individual weight of harvested fruits were recorded, and cumulative yield was calculated by summing all harvests per plant, following the method of Soppelsa and collaborators (Soppelsa et al., \u003cspan citationid=\"CR115\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In total, three harvests were carried out during the trial (each 7 days). A subset of fruits from each plant was stored at 20\u0026deg;C until the end of the production cycle, yielding at least 20 fruit samples per cultivar and treatment. The remaining fruits were immediately used for pomological analyses. The total yield has been reported in kg/ha by calculating the plant density used in this study (50000 plants/ha) and reported in previous studies (Menzel, \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Quality parameters of strawberries fruits and nutraceutical properties\u003c/h2\u003e \u003cp\u003eFor each plant, fruits were weighted and the size of the fruits (i.e., length and diameter) was also determined. Subsequently, fruits were dried in the oven at 65\u0026deg;C for 48\u0026ndash;72 hours and the fruit dry weight was determined. Firmness of each strawberry was measured using a manual fruit penetrometer (TR Turoni, Forl\u0026igrave;, Italy). The firmness was measured using a puncture test at the fruit\u0026rsquo;s equatorial side using a 6 mm diameter cylindrical probe and is expressed as Newton (N). The probe was attached to the stand with a handle to control the penetration speed. The maximum force (N) detected during the puncture test was recorded as the firmness of that particular fruit. For the quality analyses, fruits from the randomized blocks were collected in biological replicates (n\u0026thinsp;=\u0026thinsp;3). In more detail, total soluble solids (TSS, \u0026deg;Brix) were measured by cutting and squeezing each sample to obtain a few drops of fruit juice and analyzed using an N1 Atago refractometer (Atago Co., Tokyo, Japan). Titratable acids (TA, % of citric acid) were evaluated by titrating with NaOH (0.1 N) a suspension obtained after shredding and blending 15 g of fruit pulp with 150 mL of deionized water until pH 8.1. Regarding nutraceutical properties, 0.3 g (fw) of strawberry fruits were extracted with 10 mL of 70% (v/v) ethanol. The total polyphenols were determined spectrophotometrically using Folin-Ciocalteu protocol and gallic acid as reference standard, as described in previous studies (Bini et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Doumett et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Total sugars were determined according to (Georgiadou et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Organic acids (ascorbic, malic, and citric acids) were detected spectrophotometrically as described by (Tozzi et al., \u003cspan citationid=\"CR122\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Substrate DNA extraction, sequencing, and bioinformatics processing\u003c/h2\u003e \u003cp\u003eTotal genomic DNA was extracted from 0.3 g of substrate samples, stored at \u0026minus;\u0026thinsp;80\u0026deg;C, using the FastDNA SPIN Kit for Soil (MP Biomedicals, Solon, OH, USA) according to the manufacturer's instructions. DNA purity and quantity were assessed using an ND-1000 Spectrophotometer (NanoDrop Technologies, Labtech, Ringmer, UK) and a Qubit\u0026trade; 4 Fluorometer and Qubit\u0026trade; ssDNA Assay Kit (Thermo Fisher Scientific). DNA was then standardized to a concentration of 10 ng/\u0026micro;L. Amplicons preparation and sequencing were carried out at IGATech (Udine, Italy). Libraries were prepared at IGATech (Udine, Italy) using a custom Illumina 16S Metagenomic Sequencing Library Preparation protocol and sequenced by a MiSeq instrument (Illumina, San Diego, CA) using 300-bp paired-end mode. The bacterial V3-V4 hypervariable regions of 16S rDNA were PCR-amplified with primers 341F (5\u0026prime;- CCTACGGGNBGCASCAG\u0026thinsp;\u0026minus;\u0026thinsp;3\u0026prime;) and 805R (5\u0026prime;- GACTACNVGGGTATCTAATCC\u0026thinsp;\u0026minus;\u0026thinsp;3\u0026prime;) (Takahashi et al., \u003cspan citationid=\"CR120\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Bioinformatic methodologies and Statistical Analysis\u003c/h2\u003e \u003cp\u003eTo process the Illumina reads of bacterial communities, the DADA2 pipeline v.1.32.0 (Callahan et al., 2016) was used in RStudio software 4.4.1 (R Core Team). 16S rRNA reads, filtering and trimming step was performed with the following settings: maxEE\u0026thinsp;=\u0026thinsp;c(1,1), truncLen\u0026thinsp;=\u0026thinsp;c(270,250), and trimLeft\u0026thinsp;=\u0026thinsp;c(17,21). Sample inference, the merging of paired reads, and the removal of the chimera were performed with default parameters. To align and classify 16S sequences, the SILVA database v.138.2 with 80% confidence (Pruesse et al., \u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Bacterial annotated amplicon sequence variants (ASVs) were processed with the vegan package v.2.6.8 (Oksanen, \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) in RStudio software 4.4.1. ASVs with a relative abundance lower than 0.01% in all the samples were discarded. Rarefied bacterial dataset was generated for alpha-diversity, with subsample sizes of 25000 sequences per sample using the function \u003cem\u003errarefy\u003c/em\u003e in the vegan package. The rarefaction curve was generated with the function \u003cem\u003erarefied\u003c/em\u003e. For alpha diversity, the richness (Sobs), Pielou\u0026rsquo;s index (J), Simpson index and the Shannon diversity index were estimated as previously reported (Bellabarba et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and carried out with the ggplot2 package v.3.5.1 (Wickham, \u003cspan citationid=\"CR133\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Before each statistical analysis, the Shapiro test was applied to assess the data distribution. According to Shapiro test results, differences in substrates bacterial α-diversity indices among different applied films were detected performing or the non-parametric Kruskal-Wallis tests in stats v.4.1.2 package or the parametric one-way ANOVA test in agricolae package v.1.3.7. For multiple comparisons, Kruskal-Wallis tests were followed by the \u003cem\u003epost hoc\u003c/em\u003e Dunn Test in FSA v.0.9.5 (Ogle et al., \u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) and rcompanion v.2.4.36 packages (Mangiafico, \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2016\u003c/span\u003e); while the one-way ANOVA was followed by \u003cem\u003epost hoc\u003c/em\u003e Tukey\u0026rsquo;s test with agricolae package v.1.3.7. In both post hoc tests, the Benjamini\u0026ndash;Hochberg correction for multiple comparisons was applied. For β-diversity, the overall ASVs abundances (i.e. not rarefied dataset) were analysed for the bacterial community. Non-metric multidimensional scaling NMDS ordination of Bray-Curtis\u0026rsquo;s distances were carried in the vegan package and plotted with the ggplot2 package. To investigate differences in community structure among substrates covered with different films, permutational multivariate analysis of variance PERMANOVA of Bray-Curtis\u0026rsquo;s dissimilarity metrics was executed with the adonis2 function in the vegan package. The number of permutations was set to 9999 for all permutational analyses. To test if the average within-groups dispersion was the same in all groups (Anderson, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), the Analysis of multivariate homogeneity PERMDISP was fulfilled through the functions \u003cem\u003ebetadisper\u003c/em\u003e and \u003cem\u003epermutest\u003c/em\u003e implemented in the vegan package. The number of permutations was set to 9999 for all permutational analyses. On the overall ASVs abundances (i.e. not rarefied dataset), the average relative abundances (%) were calculated at phylum and order level, and bar plots were created with the ggplot2 package (v. 3.3.6). Only genera with a relative abundance of at least 3% in at least one sample were reported for each condition, while the average relative abundances (%) of unknown genera were not reported. On the overall ASVs abundances (i.e. not rarefied dataset), the upset plot was created with packages UpSetR (v. 1.4.0) and gridExtra (v. 2.3). Linear discriminant analysis effect size (LEfSe) (Segata et al., \u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) were performed to identify ASVs that were discriminately more abundant in substrates covered with different mulching films, using packages microeco (v.1.15.0) (Liu et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and file2meco (v.0.9.1) in RStudio software 4.4.3, with default setting (Kruskal-Wallis rank sum test, \u003cem\u003ep\u003c/em\u003e-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05; LDA score\u0026thinsp;\u0026gt;\u0026thinsp;2). The LEfSe bar plot was created with the microeco package (Liu et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) setting the LDA score threshold to 3.5 with the microeco package. To identify physicochemical drivers of different bacterial communities, a PERMANOVA analysis based on Bray-Curtis distances was performed, followed by A dbRDA ordination plot using the adonis2 and capscale function in the vegan package v.2.6.8, respectively (Oksanen, \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Functional inference of substrate bacterial communities was performed with FAPROTAX (Louca et al., \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Only the metabolic categories with abundance higher than 0 in at least one sample were considered. Categories Human-related were discarded, as well as the chloroplasts group.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. RESULTS","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Substrate Physicochemical properties\u003c/h2\u003e \u003cp\u003eWith regard to the chemical-physical properties of the substrate the application of different mulching films did not lead to significant variations in TOC, pH, EC, and CEC at the end of the trial (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). However, mulching significantly influenced the total nitrogen (N_tot) content. In particular, mulching with PBAT resulted in higher N_tot concentrations compared to the unmulched control. Despite this increase in total nitrogen, no significant differences were observed among the applied films in terms of nitrate and ammonium content. Similarly, the carbon-to-nitrogen (C/N) ratio remained statistically similar across treatments. Overall, mulching films had a substantial impact on the elemental composition of the commercial substrate. Substrates covered with PE films exhibited the highest concentrations of K and Mg, while those coated with PP films showed the lowest ones. Regarding micronutrients and trace metals, the Fe content was significantly higher in substrates covered with PE films. In contrast, the lowest value was recorded in coated substrates with PP. Unmulched substrates, and covered with PBAT and OxoPE, revealed intermediate profiles. Similarly, Co and Ni concentrations were significantly higher in PE compared to the other treatments. In contrast, unmulched control exhibited the greatest levels of Mo and Zn.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhysicochemical properties of substrates with different mulching films at the end of strawberry cultivation. Within rows, different letters represent statistically significant variation (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) per Kruskal\u0026minus;Wallis tests and post hoc multiple Dunn\u0026rsquo;s test.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eUnmulched control\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOxoPE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePBAT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTOC (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN_tot (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ab\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC/N\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e27.43\u0026thinsp;\u0026plusmn;\u0026thinsp;3.67 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e43.93\u0026thinsp;\u0026plusmn;\u0026thinsp;13.19 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16.23\u0026thinsp;\u0026plusmn;\u0026thinsp;4.19 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e28.93\u0026thinsp;\u0026plusmn;\u0026thinsp;3.38 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16.53\u0026thinsp;\u0026plusmn;\u0026thinsp;1.43 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHumidity (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.51 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.26 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.32 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14\u0026thinsp;\u0026plusmn;\u0026thinsp;1.49 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCEC (meq 100 g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.43\u0026thinsp;\u0026plusmn;\u0026thinsp;2.06 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.08 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.62 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.47\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEC (dS m\u003csup\u003e\u0026minus;\u0026thinsp;1)\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNitrate (mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.75 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e23.03\u0026thinsp;\u0026plusmn;\u0026thinsp;3.19 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.33\u0026thinsp;\u0026plusmn;\u0026thinsp;4.35 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17.87\u0026thinsp;\u0026plusmn;\u0026thinsp;4.61 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmmonium (mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.07\u0026thinsp;\u0026plusmn;\u0026thinsp;2.07 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.07\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.43\u0026thinsp;\u0026plusmn;\u0026thinsp;2.14 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.15 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.6\u0026thinsp;\u0026plusmn;\u0026thinsp;6.44 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCa (g kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.43\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eK (g kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.68 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.78 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMg (g kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNa (g kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.062\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP (g kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFe (g kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.65\u0026thinsp;\u0026plusmn;\u0026thinsp;1.31 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.87\u0026thinsp;\u0026plusmn;\u0026thinsp;2.06 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBa (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e204.19\u0026thinsp;\u0026plusmn;\u0026thinsp;6.87 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e190.71\u0026thinsp;\u0026plusmn;\u0026thinsp;5.54 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e198.04\u0026thinsp;\u0026plusmn;\u0026thinsp;3.97 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e203.83\u0026thinsp;\u0026plusmn;\u0026thinsp;42.32 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e139.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.58 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCo (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCr (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCu (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.85 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e32.36\u0026thinsp;\u0026plusmn;\u0026thinsp;4.35 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.85\u0026thinsp;\u0026plusmn;\u0026thinsp;2.59 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e25.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.37 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMn (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e192.98\u0026thinsp;\u0026plusmn;\u0026thinsp;8.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e181.7\u0026thinsp;\u0026plusmn;\u0026thinsp;4.8 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e202.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e240.43\u0026thinsp;\u0026plusmn;\u0026thinsp;15.34 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e212.19\u0026thinsp;\u0026plusmn;\u0026thinsp;31.35 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMo (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.29\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44 ab\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNi (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePb (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eZn (mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.94\u0026thinsp;\u0026plusmn;\u0026thinsp;3.66 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.57\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Effect of mulching practice on physiological response and plant growth\u003c/h2\u003e \u003cp\u003eMulching practice led to slight differences in physiological traits during the plant cycle (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). At the first time point (T1), PE mulch revealed the highest net photosynthetic rate (Pn) value, which was significantly greater than the unmulched control, whilst no variations were noted with the other mulched treatments. At T2 and T3, all treatments displayed similar net Pn values. No differences among treatments were observed for stomatal conductance (gs) in all time points analysed. At T1, the unmulched control revealed the highest value of the intracellular CO\u003csub\u003e2\u003c/sub\u003e concentration (Ci), which was also significantly greater than PE and PBAT. Conversely, an opposite trend was observed at T2, with a significantly lower value of Ci in the unmulched control compared to PP. At T3, no statistical differences in Ci values were observed among treatments.\u003c/p\u003e \u003cp\u003eRegarding the plant's growth parameters, at T1, no clear differences in plant height and in the number of leaves were reported among different films applied (Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). As plant growth evolved (T2 and T3), all treatments showed a similar increment in plant height (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). At T3, the PP treatment showed the highest value of number of leaves, which was significantly higher than PE), whereas no variations for this parameter were observed among the unmulched control and all the mulching films.\u003c/p\u003e \u003cp\u003eDespite slight differences observed in plant growth parameters, the mulching practice positively affected strawberry yield (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The highest yield was observed in OxoPE treatment (4425\u0026thinsp;\u0026plusmn;\u0026thinsp;243.38 kg ha\u003csup\u003e\u0026ndash;1\u003c/sup\u003e), while the lowest yield was in the unmulched control (3145\u0026thinsp;\u0026plusmn;\u0026thinsp;132.38 kg ha\u003csup\u003e\u0026ndash;1\u003c/sup\u003e). Mostly, OxoPE and PP treatments showed a significantly higher productivity than the unmulched control, while no significant variations were observed with PBAT and PE treatments.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eConcerning the quality parameter of strawberry fruits, there were no significant differences in fruit weight, diameter, height, TSS, or texture among treatments (Table\u0026nbsp;4). However, the OxoPE treatment showed a significantly higher fruit brightness (L*) and colour intensity (chroma index) compared to the other treatments. Strawberries produced in PBAT treatment showed the highest titratable acidity (1.2%), compared to the lowest value of the unmulched control (0.87%). Strawberries treated with PP films had the lowest texture value, which differed significantly from the PBAT treatment.\u003c/p\u003e \u003cp\u003eWith respect to the nutraceutical strawberry fruit properties, no significant differences were observed among treatments in polyphenols, total sugars, citric acid, or malic acid content (Table\u0026nbsp;5). The highest value for ascorbic acid was observed in unmulched control, being significantly higher in fruits than OxoPE which showed the lowest value. The other treatments showed intermediate levels with no significant differences.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Composition of bacterial communities in substrates covered with different mulching films.\u003c/h2\u003e \u003cp\u003eConcerning the 16S rRNA sequencing analysis of bacterial communities, a total of 1100496 high-quality sequences were obtained (average of 73396\u0026thinsp;\u0026plusmn;\u0026thinsp;24204 sequences per sample). An average of about 33% of sequences remained after quality filtering and removal of chimera, enough to accurately describe the biodiversity within the bacterial communities as suggested by the rarefaction curves (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The bioinformatic processing of the 16S rRNA sequences yielded 3201 ASVs, assigned to Bacteria kingdom, spanning 28 phyla, 57 classes, 121 orders, 175 families, and 311 genera (File S1). Bacterial communities of different substrates were predominantly composed by \u003cem\u003ePseudomonadota\u003c/em\u003e, with an average relative abundance between 53.3% and 40.6%, followed by \u003cem\u003eBacteroidota\u003c/em\u003e (~\u0026thinsp;21.2% - ~12.1%), \u003cem\u003eAcidobacteriota\u003c/em\u003e (~\u0026thinsp;16.7% - ~6.4%), and, to a lesser extent, by \u003cem\u003eActinomycetota\u003c/em\u003e (~\u0026thinsp;9% - ~6.2%), \u003cem\u003eVerrucomicrobiota\u003c/em\u003e (~\u0026thinsp;7.9% - ~2.8%), \u003cem\u003ePlanctomycetota\u003c/em\u003e (~\u0026thinsp;4.2% - ~1.9%), \u003cem\u003ePatescibacteria\u003c/em\u003e (~\u0026thinsp;3.4% - ~1.3%), \u003cem\u003eMyxococcota\u003c/em\u003e (~\u0026thinsp;1.5% - ~1.3%), \u003cem\u003eCyanobacteriota\u003c/em\u003e (~\u0026thinsp;1.3% - ~1.2%) (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eA). At family level, the bacterial communities of substrates shared a \u0026lsquo;core\u0026rsquo; of enriched taxonomic groups with a mean relative abundance higher than 3%, composed of \u003cem\u003eSphingobacteriaceae\u003c/em\u003e (~\u0026thinsp;12% - ~7.5%), \u003cem\u003eCaulobacteraceae\u003c/em\u003e (~\u0026thinsp;11.8% - ~6.7%), \u003cem\u003eSphingobacteriaceae\u003c/em\u003e (~\u0026thinsp;12% - ~7.5%), \u003cem\u003eRhodanobacteraceae\u003c/em\u003e (~\u0026thinsp;13% - ~3.4%), \u003cem\u003eAcidobacteriaceae\u003c/em\u003e (~\u0026thinsp;11.8% - ~4.8%), and \u003cem\u003eMicropepsaceae\u003c/em\u003e (~\u0026thinsp;5.9% - ~4.1%) (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eThe relative abundance of other families shifted across the bacterial communities of substrates mulched with different films: the family of \u003cem\u003eAcidimicrobiaceae\u003c/em\u003e was more abundant in OxoPE and PE substrates, while the family of \u003cem\u003eBurkholderiaceae\u003c/em\u003e was more abundant in substrates covered with OxoPE, PE and PP (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eB). Similarly, the family of \u003cem\u003eChitinophagaceae\u003c/em\u003e was enriched in substrates covered with PBAT or PP and uncovered substrates, while the family of \u003cem\u003eSphingomonadaceae\u003c/em\u003e was more abundant in substrates mulched with PP and unmulched control (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eSpecific bacterial families were uniquely enriched in individual mulch treatments: \u003cem\u003eLysobacteraceae\u003c/em\u003e was more abundant in PBAT-mulched substrates, while \u003cem\u003eAcetobacteraceae\u003c/em\u003e and \u003cem\u003ePedosphaeraceae\u003c/em\u003e were specifically associated with PE mulching (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eAmong the most enriched genera, the taxonomic groups shared by all bacterial communities were \u003cem\u003eAsticcacaulis\u003c/em\u003e (~\u0026thinsp;7.0% - ~4.8%) and \u003cem\u003eGranulicella\u003c/em\u003e (~\u0026thinsp;7.1% - ~3.5%) (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eC). The genera \u003cem\u003eMucilaginibacter\u003c/em\u003e (~\u0026thinsp;6.4% - ~3.4%) and \u003cem\u003eRhodanobacter\u003c/em\u003e (~\u0026thinsp;11.2% - ~4.0%) were more abundant in all substrates except the PBAT and PE, respectively. Furthermore, \u003cem\u003eAlbibacterium\u003c/em\u003e (~\u0026thinsp;4.1%) and \u003cem\u003eBrevundimonas\u003c/em\u003e (~\u0026thinsp;4.3%) were uniquely enriched in PBAT-mulched substrates, whereas \u003cem\u003eSphingomonas\u003c/em\u003e (~\u0026thinsp;3.9%) were more abundant in PP-mulched substrates.\u003c/p\u003e \u003cp\u003e \u003cb\u003e3.4 Diversity of bacterial communities in substrates covered with different mulching films/ Effect of different mulching films on diversity of substrate bacterial communities\u003c/b\u003e \u003c/p\u003e \u003cp\u003eSubstrate bacterial α-diversity was characterized by high species richness (Sobs) which remained statistically comparable across all substrates, indicating that mulching practices, regardless of film type, did not influence total number of bacterial taxa (Fig. \u003cspan refid=\"MOESM3\" class=\"InternalRef\"\u003eS3\u003c/span\u003eA). However, mulching practices significantly enhanced bacterial community diversity and evenness in respect to the unmulched control, according to Simpson and Shannon indices (Fig. \u003cspan refid=\"MOESM3\" class=\"InternalRef\"\u003eS3\u003c/span\u003eC-D). Specifically, both indices were significantly higher in substrates covered with OxoPE and PE films compared to control. In particular, the higher values of Simpson indices suggest a reduced dominance of some bacterial taxa and, consequently, a higher diversity compared to the control. Moreover, a significant increase of Shannon index was also observed in substrates covered with PP, in line with the slight increasing trend observed for Evenness index of PE and PP communities (Fig. \u003cspan refid=\"MOESM3\" class=\"InternalRef\"\u003eS3\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eAnalysis of ꞵ-diversity revealed a clear separation in bacterial community structures between mulched and unmulched substrates. Most notably, bacterial communities associated with plastic-based films (OxoPE, PE, and PP) formed a distinct cluster compared to those found with PBAT (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Both the Permutational multivariate analysis of variance (PERMANOVA) and the Analysis of multivariate homogeneity (PERMDISP) confirmed significant differences in community composition, indicating that mulching practices, regardless of film type, distinctly shape the structures of substrate microbial communities (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe distribution of bacterial ASVs revealed a distinct partitioning of bacterial taxa across the different substrate treatments (Fig. \u003cspan refid=\"MOESM4\" class=\"InternalRef\"\u003eS4\u003c/span\u003e). A shared core of only 319 ASVs, representing approximately 10% of the total richness (3,201 ASVs), was identified across all communities. In contrast, the majority of ASVs were unique to specific substrates, likely driving the structural variations observed in the NMDS analysis. Notably, all plastic-mulched substrates harboured a higher proportion of exclusive taxa compared to the unmulched control (232 ASVs; ~7.2%), ranging from ~\u0026thinsp;9,6% for OxoPE (306 ASVs) to ~\u0026thinsp;7,9% for PP (254 ASVs).\u003c/p\u003e \u003cp\u003eTo uncover the taxonomic groups significantly associated with the distinct microbial communities of each substrate, a linear discriminant analysis effect size (LEfSe) was performed. The total number of the discriminant ASVs identified was 299 (File S2). Only 19 ASVs, with LDA score higher than 3.5, were considered as highly discriminants (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA-B). Consistent with the relative abundance patterns, the unmulched control was primarily characterized by the genera of \u003cem\u003eRhodanobacter\u003c/em\u003e (ASV_4, ASV_5, and ASV_24), \u003cem\u003eNocardioides\u003c/em\u003e (ASV_41), \u003cem\u003eChryseobacterium\u003c/em\u003e (ASV_59) and \u003cem\u003eArachidicoccus\u003c/em\u003e (ASV_60) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA-B, Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eC). In contrast, the OxoPE bacterial community was uniquely distinguished by \u003cem\u003eAsticcacaulis\u003c/em\u003e (ASV_9) as the sole highly discriminant ASV. The PE substrate harboured the greatest number of discriminant taxa, including \u003cem\u003eMucilaginibacter\u003c/em\u003e (ASV_37), \u003cem\u003eCandidatus solibacter\u003c/em\u003e (ASV_35) and several unclassified genera within the \u003cem\u003eAcidimicrobiaceae\u003c/em\u003e (ASV_6), \u003cem\u003eAcetobacteraceae\u003c/em\u003e (ASV_14), \u003cem\u003eAcidimicrobiaceae\u003c/em\u003e (ASV_31), \u003cem\u003ePedosphaeraceae\u003c/em\u003e (ASV_63) and \u003cem\u003eXanthobacteraceae\u003c/em\u003e (ASV_8) families. Bacterial communities of PP-mulched substrates were primarily distinguished by the genera \u003cem\u003eCellvibrio\u003c/em\u003e (ASV_36) and \u003cem\u003eSphingomonas\u003c/em\u003e (ASV_10), with the latter strongly enriched in these substrates (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eC). Finally, the PBAT community was characterized by the genera \u003cem\u003eBrevundimonas\u003c/em\u003e (ASV_128), a specific strain of genus \u003cem\u003eRhodanobacter\u003c/em\u003e (ASV_49) and the genus \u003cem\u003eTM7a\u003c/em\u003e of \u003cem\u003eSaccharimonadaceae\u003c/em\u003e family (ASV_22).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.5 Relationship between bacterial community structures and Substrate Physicochemical properties\u003c/h2\u003e \u003cp\u003eTo evaluate the influence of soil physicochemical properties on different bacterial community structure, a Permutational Multivariate Analysis of Variance (PERMANOVA) coupled with a distance-based Redundancy Analysis (dbRDA) were performed based on ASVs Bray-Curtis\u0026rsquo;s dissimilarities. The PERMANOVA model revealed that the soil chemical profile significantly shaped the microbial community (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.77908, \u003cem\u003ep\u003c/em\u003e-value\u0026thinsp;=\u0026thinsp;0.0024). When evaluating the independent contribution of each variable, the substrate humidity and nitrate concentration revealed as the primary significant drivers (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Specifically, humidity explained 15.1% of the total variance (F\u0026thinsp;=\u0026thinsp;3.43, p\u0026thinsp;=\u0026thinsp;0.0072), while nitrate levels accounted for 13.9% (F\u0026thinsp;=\u0026thinsp;3.14, p\u0026thinsp;=\u0026thinsp;0.0052). The other variables, including Cation Exchange Capacity (CEC) and Electrical Conductivity (EC) did not reach the standard significance threshold, suggesting a secondary role in community differentiation. The dbRDA analysis confirmed a high degree of correlation between environmental constraints and microbial community shifts (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.779). The first two axes of the ordination (CAP1 and CAP2) accounted for 46.8% and 14.3% of the constrained variation, respectively. Along the CAP1 axis, db-RDA plot illustrated a clear separation of bacterial communities among conventional plastic films, PBAT films and unmulched related communities, which was strongly associated with gradients of Humidity and Nitrate. Bacterial communities of substrates mulched with OxoPE, PE and PP were positively correlated with higher humidity compared to the unmulched control, suggesting that this feature was a driver in shaping this community.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo understand the possible effect of mulching films on substrate functional activities, putative soil bacterial functional profiles were inferred with the FAPROTAX database from taxonomies. Among the database processes, only 25 were detected. This predictive analysis revealed a distinct separation in terms of both bacterial metabolic pathways and ecological roles across the different substrate treatments, primarily separating PE and PP mulching films from the Unmulched control, PBAT and OxoPE (Figure \u003cspan refid=\"MOESM5\" class=\"InternalRef\"\u003eS5\u003c/span\u003e). Specifically, traditional polyethylene mulching induced a functional shift compared to other films characterized by the putative enrichment of nitrogen fixation, concurrently with a marked depletion in nitrification and aerobic nitrite oxidation, chemoheterotrophy and aerobic chemoheterotrophy, methanol oxidation and methylotrophy (Figure \u003cspan refid=\"MOESM5\" class=\"InternalRef\"\u003eS5\u003c/span\u003e). In contrast, the PBAT treatment displayed a unique functional fingerprint defined by the putative increase of nitrification, aerobic nitrite oxidation and nitrate respiration, also in manganese oxidation, and overall sulfur compound respiration, suggesting an increased turnover of nitrogen and sulfur cycles compared to other mulches. In OxoPE films increased the overall fermentation process, while aromatic and hydrocarbon degradation was predictively higher in the PBAT and OxoPE films than in the PE and PP treatments, indicating that biodegradable films may promote microbial guilds capable of complex carbon catabolism.\u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003ePrevious research on strawberry cultivation has typically focused on the effects of mulching either in terms of soil health and microbial dynamics (Meyer et al., \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Mu\u0026ntilde;oz et al., \u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e2017a\u003c/span\u003e) or through plant physiological and morphological aspects (Abdelfattah et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Biswas et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Dangi et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), often lacking of a more comprehensive and integrated approach. To bridge this gap, our research investigates the impact of conventional \u003cem\u003eversus\u003c/em\u003e biodegradable mulching on strawberry cultivation by evaluating both plant performance and microbial ecology within the same experimental framework. Through a detailed evaluation of plant physiological and morphological traits, fruit yield and quality, and substrate microbial communities, we provided a broader understanding of how mulch selection may drive both strawberry plant and substrate health.\u003c/p\u003e \u003cp\u003eSoil quality is fundamental to ensuring consistent crop productivity and optimal fruit quality. The impact of mulching films on soil properties remains a subject of debate, as research findings are often controversial. While some studies suggested that both plastic and biodegradable films significantly alter soil physical (e.g., bulk density, moisture, and temperature) and chemical features (e.g., total organic carbon TOC and nitrogen uptake) (Dewi et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Wu et al., \u003cspan citationid=\"CR134\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) others reported no significant changes in pH, organic matter, or nitrogen content followed the application of different mulch films. Our results aligned with the latter, as most physicochemical properties of the peat-based substrate, including TOC, pH, EC, CEC, and the C/N ratio, were not significantly affected by the tested mulching films. Specifically, in the work of Xu and collaborators (Xu et al., \u003cspan citationid=\"CR136\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), a similar stability in TOC content to our results was observed over a 180-day period in a laboratory experiment, suggesting that more pronounced alterations in substrate properties might be tracked mainly with long-term application of mulching, within different degradation time and environmental conditions (Ding et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Whereas primary physicochemical characteristics remained stable over the limited cultivation period, the different mulching films significantly influenced specific macro- and micronutrient concentrations. In particular, PBAT film increased total nitrogen content compared to the unmulched control, while PE films lead to higher concentrations of K, Mg, Fe, Co, and Ni. These shifts support previous reports that mulching with plastic films, and the composition of the film itself, could modify nutrient retention and mineral transformations by altering soil hydrothermal conditions (Mo et al., \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Shan et al., \u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Y. Zhao et al., \u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Several studies suggest that both plastic and biodegradable films can modify soil physical properties (e.g., bulk density, moisture, and temperature) as well as chemical characteristics (e.g., TOC and nitrogen uptake) (Dewi et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Wu et al., \u003cspan citationid=\"CR134\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, other studies have reported no significant changes in pH, organic matter, or nitrogen content (Ni et al., \u003cspan citationid=\"CR88\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; M. Zhang et al., \u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The present research is consistent with the latter findings, as most main physicochemical properties of the peat-based substrate, including TOC, pH, EC, CEC, and C/N ratio, were not significantly affected by the different mulching films tested. Similar results for TOC were reported by Xu et al. (Xu et al., \u003cspan citationid=\"CR136\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), who evaluated these properties over 180 days after the application of plastic or biodegradable mulching films in a laboratory experiment. However, previous studies suggest that the effects of mulching films may depend on degradation time and environmental conditions. In particular, long-term application could lead to more pronounced alterations in substrate properties (Ding et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Although limited variation was detected in the main physicochemical characteristics in a limited cultivation period (i.e. 3 months), the four mulching films influenced macro- and micronutrient concentrations in the peat-based substrate. In particular, a factor determining these alterations is the different film composition (Bandopadhyay et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Shan et al., \u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the present study different mulching films did not significantly influence the physiological or vegetative growth of strawberry plants. Indeed, both photosynthetic rate and stomatal conductance did not show significant variations either compared to the control samples or among the different mulch films. Consistent with our results, previous studies on cucumber and pickling cucumber have shown that plastic mulching does not significantly modify leaf gas exchange compared to bare soil (L\u0026oacute;pez-Tolentino et al., \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Torres-Olivar et al., \u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) (Lopez-Tolentino et al., 2017; Torres-Olivar et al., \u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). These studies also reported significant increases in yield under mulched conditions, despite the non-significant differences in gas exchange. Similarly, in our study, mulching coverage enhanced fruit yield. Increased fruit production under mulching is a well-established response in strawberry cultivation, as reported in previous studies (Abdelfattah et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Dangi et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eA key finding of our study was the significant impact of mulch type on productivity, with OxoPE and PP that outperformed the control (+\u0026thinsp;29% and +\u0026thinsp;25%, respectively), then mirroring the 28% yield increase reported by Biswas et al. (Biswas et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) for strawberries grown with biodegradable plastic films. In contrast, PE mulches produced yields comparable to the unmulched control, reflecting an intermediate performance previously reported for both black and white PE (Pinto et al., \u003cspan citationid=\"CR91\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe impact of PBAT-type mulches on strawberry yield, in our study, differ from some previous works. Indeed, an 49% yield increase in strawberry cv. \u0026lsquo;Albion\u0026rsquo; using PBAT-based biodegradable mulch (BDM) was observed compared to bare soil control (Wang et al., \u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), and a similar upward trend was noted in two-year open-field cultivation (DeVetter et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). This discrepancy might likely due to the different experimental setups, as our study utilized a peat-based growing substrate, whereas the aforementioned research was conducted in open-field soil conditions. Despite these differences, when directly comparing PBAT to PE, our results remain consistent with other studies showing comparable strawberry yields between the two materials (Costa et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; DeVetter et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eConcerning fruit quality, strawberry fruits showed total soluble solids (TSS) values comparable to those previously reported (DeVetter et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Supreetha et al., \u003cspan citationid=\"CR118\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, mulching can influence strawberry fruit quality by modifying the amount and spectral composition of light reflected into the plant canopy (Kasperbauer et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Shiukhy et al., \u003cspan citationid=\"CR111\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), which in turn may affect fruits composition and reduce TSS in unmulched plants (DeVetter et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). In our study, although a slight variation in TSS was detected, no significant differences were observed among different films used, implying that mulch application did not affect TSS (Wang et al., \u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In contrast, we observed an increment in titratable acidity (TA) in the PBAT treatment significantly outperforming the control. The influence of different mulches on TA levels has been previously reported across various fruit species (Morra et al., \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR129\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). Despite this specific shift, no marked variations in overall fruit quality were recorded among the biodegradable (PBAT, OxoPE) and the PE mulching films, suggesting that replacing PE with biodegradable mulches would not lead to meaningful changes in fruit quality (Costa et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; DeVetter et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMulching practices are well-known to influence soil microbial structure and functionality through the regulation of the soil microclimate and physicochemical environment, thereby significantly altering community composition and diversity (Li et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Sun et al., \u003cspan citationid=\"CR117\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR140\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). In this study, mulching with OxoPE, PE, and PP significantly enhanced α-diversity, as indicated by increased Shannon and Simpson indexes. This variation effectively reduced the dominance of certain taxa, leading to a considerably more even community composition. Overall, these results suggested that mulching with plastic films promotes a more balanced and diverse bacterial assembly without shifting the overall species richness. In previous works, mulching materials showed highly variable outcomes regarding bacterial α-diversity, which seems to be strongly dependent on the type of soil and its interaction with diverse mulch films (Xu et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), the plant species (Gao et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), and micro-habitat taken into account (i.e. bulk soil, rhizosphere, or plastisphere) (Meng et al., \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2026\u003c/span\u003e). Our findings are strongly supported by studies in which mulching likely alleviates environmental stressors (as salinity or drought) enabling a broader range of taxa to thrive (Wang et al., \u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Conversely, several studies report a decrease in α-diversity, particularly with biodegradable materials (Dong et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Fang et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Xu et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Z. Zhao et al., \u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) often attributed to the enrichment of specific polymers-degrading taxa, thereby reducing overall community evenness. However, α-diversity indexes are known to be less sensitive to environmental changes than overall community structure analysis. Indeed, while specific bacterial groups may shift in response to the environmental pressure, these changes are often offset by opposing trends in other guilds, thereby maintaining relatively stable total diversity (Hartman et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Hartmann and Widmer, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2006\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn this context, community shifts we detected were more evident at the structural level. Indeed, our results revealed distinct bacterial community structures among mulching materials, implying that substrate bacterial diversity respond differently to the application of different film types as previously observed (Zhao et al., \u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Clear variations in bacterial communities were previously detected in soils both under BDMs (Koitabashi et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Li et al., \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Muroi et al., \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), and under non-biodegradable plastic mulches (Farmer et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Mu\u0026ntilde;oz et al., \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2017b\u003c/span\u003e, \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In particular, some recent research consistently highlighted how BDM residues, or generally BDM treatments, elicited more pronounced divergences in community structure than conventional PE or low-density polyethylene (LDPE) (Liu et al., \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; L. Liu et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Xu et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). A common explanation for this trend lies on different material properties. In more detail, PE is a chemically inert material, with slow biodegradation rate (Khandare et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), performing primarily as a physical barrier (Ammala et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Koutny et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Conversely, BDM acts as a bioavailable exogenous carbon source, exerting selective pressure on soil microbial community then reshaping its structure and functionality (Bandopadhyay et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2026\u003c/span\u003e). We also noticed a slight divergence between the traditional plastic films (OxoPE, PE, and PP) and the PBAT-related communities, that we likely attribute to the presence of corn starch in PBAT films. The corn starch is a key component of several starch-PBAT blends biodegradable films and, as a naturally hydrophilic polysaccharide, is highly sensitive to moisture (Grimaut et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; K\u0026uuml;ster et al., \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) and undergoes rapid mineralization (Villena et al., \u003cspan citationid=\"CR124\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Therefore, we hypothesize that, unlike conventional plastics, the gradual degradation of starch provides an additional carbon source that recruits specific starch-utilizing bacterial species. This process may alter the microbial community (Gao et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Šer\u0026aacute; et al., \u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) toward copiotrophic and plant-beneficial genera as the PBAT-related \u003cem\u003eRhodanobacter\u003c/em\u003e, which are specialised for nutrient-rich environments (Mahawar et al., \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2026\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR131\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). In line with this trend, the unique increase of genus \u003cem\u003eBrevundimonas\u003c/em\u003e in PBAT-mulched substrates aligns with its ability to metabolize starch, as demonstrated by its use its use in dark fermentation processes for biohydrogen production (Bao et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNonetheless, in our work the OxoPE and PE application supported the highest levels of unique bacterial recruitment, whereas the unmulched control exhibited the most restricted set of exclusive ASVs. Beyond chemical composition, the physical presence of plastic mulch may promote specialized microbial niches, with film fragments acting as unique micro-environment that selectively enriches specific microbial guilds (Huang et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Jacquin et al., \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Qi et al., \u003cspan citationid=\"CR94\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Shi et al., \u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Consistent with this observation, our results further highlight the family \u003cem\u003eSphingomonadaceae\u003c/em\u003e, and specifically the genus \u003cem\u003eSphingomonas\u003c/em\u003e, which exhibited a significantly higher abundance in PP-mulched substrates. In particular \u003cem\u003eSphingomonas\u003c/em\u003e, well-known as pollutants degraders (Asaf et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), have been consistently associated to different mulch blends (Xu et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Moreover, this genus was found enriched in the PP plastisphere of the sorghum rhizosphere (Z.-H. Wang et al., \u003cspan citationid=\"CR127\" class=\"CitationRef\"\u003e2026\u003c/span\u003e) and on PP polypropylene particles used for the biodegradation of crude oil (Vita et al., \u003cspan citationid=\"CR125\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Consequently, \u003cem\u003eSphingomonas\u003c/em\u003e was recognized as key plastic-degrading bacterium in soil (Xu et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) with proven metabolic potential to break down a wide range of organic pollutants (Gatheru Waigi et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and synthetic polymers (Chen et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Y. Zhang et al., \u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAccording to recent research, different film thickness significantly changed the overall community structure of rhizosphere soil bacteria (G. Wang et al., \u003cspan citationid=\"CR127\" class=\"CitationRef\"\u003e2026\u003c/span\u003e). Moreover, the taxonomic composition of bacterial communities at both phylum and genus varied significantly among different BDM thicknesses (Zhao et al., \u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Specifically, it was observed a significant positive correlation among film thickness and soil water content, soil organic matter, and total nitrogen, but a negative correlation with soil temperature (G. Wang et al., \u003cspan citationid=\"CR127\" class=\"CitationRef\"\u003e2026\u003c/span\u003e). This suggests that thicker films provide a more stable micro-environment in terms of moisture, temperature, and nutrient availability compared to thinner films, which are more susceptible to environmental fluctuations (G. Wang et al., \u003cspan citationid=\"CR127\" class=\"CitationRef\"\u003e2026\u003c/span\u003e). Therefore, film thickness serves as a determining factor for both the physical durability of the mulch and the modulation of the soil microclimate, which in turn shapes the response of the associated bacterial communities. Indeed, our results revealed that the application of plastic films of varying thicknesses produced bacterial communities with unique structures and compositions. This separation followed a gradient corresponding to the thickness of the materials, ranging from the conventional PE as the thinnest material (0.05 \u0026micro;m) to the PP as the thickest one (450 \u0026micro;m). While thinner films (e.g., PE or LDPE\u0026thinsp;\u0026lt;\u0026thinsp;10 \u0026micro;m) degrade easily to the mechanical action of wind and rain, leading to a release of microplastics and chemical additives (Crossman et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Kim et al., \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Qiang et al., \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Scheurer and Bigalke, \u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), thicker mulch films in our study likely acted as more robust physical barriers. These films likely reshaped the microbial communities through long-term microclimatic stabilization. For this reason, such a broad range of thicknesses likely imposed differential selective pressures creating different substrate microclimates. As a further supporting of these speculations/hypothesis, distance-based Redundancy Analysis (dbRDA) identified humidity and nitrate concentrations as the primary significant drivers shaping the substrate microbial communities. Overall, this finding suggests that changes in soil microclimate and nitrogen dynamics induced by mulching act as the main selective pressures shaping the specialized bacterial communities, as reported previously (Gao et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2026\u003c/span\u003e, \u003cspan citationid=\"CR142\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; M. Zhang et al., \u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Y. Zhang et al., \u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In particular, the distinct associations observed between substrate humidity and communities of conventional films (PE, PP, and OxoPE), and between nitrate concentrations and those of PBAT treatments, likely underscoring two contrasting mechanisms: the passive cover effect of conventional films, which primarily modulates soil microclimate and moisture, and the direct chemical and nutrient-input effect of biodegradable films (PBAT) during the degradation process.\u003c/p\u003e \u003cp\u003eDistinct functional shifts of bacterial communities based on mulch type were predicted, with the PE communities linked to an enrichment in nitrogen fixation. Conversely, PBAT seems to promote nitrification and sulfur respiration processes. These predictions are consistent with the compositional profiling analysis, which revealed that the family of \u003cem\u003eBurkholderiaceae\u003c/em\u003e was more prevalent in substrates covered with conventional plastic mulch (OxoPE, PE, PP). This family, a large group within the Proteobacteria phylum, is recognized for its metabolic versatility and ecological plasticity which encompasses several well-known genera (e.g., \u003cem\u003eBurkholderia\u003c/em\u003e and \u003cem\u003eParaburkholderia\u003c/em\u003e), characterized as Plant Growth-Promoting Rhizobacteria (PGPR) and involved in nitrogen fixation and phosphate solubilization (Eberl and Vandamme, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Interestingly, it has been previously observed that LDPE microplastics promote the abundance of \u003cem\u003eBurkholderiaceae\u003c/em\u003e potentially to enhance nitrogen fixation (Fei et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Besides, others genera were previously associated to PE mulch films, as \u003cem\u003eMucilaginibacter\u003c/em\u003e (Tagg et al., \u003cspan citationid=\"CR119\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), with a significant role in the degradation of complex carbon sources (Kumar et al., \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), and \u003cem\u003eCandidatus Solibacter\u003c/em\u003e known for its ability to decompose organic matter then promoting soil carbon cycling and nitrogen fixation (Gu et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The accumulation residues from LDPE and BDM mulches negatively affects soil physical properties and water dynamics (Li YuanQiao et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), while altering carbon and nutrient cycling (Zang et al., \u003cspan citationid=\"CR138\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). These changes frequently lead to a significant nitrogen limitation (Hegan et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) and higher soil C/N ratio (Qi et al., \u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Therefore, it is reasonable to infer that plastic-derived carbon input, together with changes in soil physical structure, can trigger a short-term nitrogen starvation through microbial immobilization, even without observing significant change in C/N ratio. In this regard, the recruitment of nitrogen-fixing bacteria represents a community adaptation to restore the C/N balance.\u003c/p\u003e \u003cp\u003eConversely, in PBAT-mulched substrates the input of bioavailable carbon might promotes the synthesis of organic nitrogen compounds and, consequently, stimulates the nitrification. This process may facilitate the niche stabilization for nitrifying bacteria following the soil physical micro-alterations induced by mulching. For instance, it has been observed that plastic residues from PLA films increase soil porosity, which in turn may enhance soil aeration and accelerate nitrification processes (De Souza Machado et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), ultimately leading to a depletion of soil ammonium levels (Dong et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The predicted increase in sulfur respiration within PBAT-amended substrates may be due to the overall metabolic remodulation of bacterial communities induced by mulch residues, as sulfur cycling genes are tightly coupled with carbon and nitrogen metabolic pathways (Zheng et al., \u003cspan citationid=\"CR147\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). However, some studies have reported a negative correlation between PBAT mulching and sulfur cycling (X. Liu et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) as well as the negative impact of microplastic on sulfur-related processes in coastal sediments (Wang et al., \u003cspan citationid=\"CR128\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In our pathway prediction, biodegradable films (PBAT and OxoPE) were also associated with microbial groups capable of complex carbon catabolism, such as aromatic and hydrocarbon compounds degradation. Indeed, \u003cem\u003eTM7a\u003c/em\u003e of \u003cem\u003eSaccharimonadaceae\u003c/em\u003e family was previously identified as a strong biomarker for the PBAT community since its relative abundance increasing proportionally to the amount of PBAT (Xu et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Moreover, this taxon is consistently associated to different microplastic materials in both marine (Scales et al., \u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and soil environments (Li et al., \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Scopetani et al., \u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), further highlighting its putative plastic degradation potential. Similarly, \u003cem\u003eBrevundimonas\u003c/em\u003e was identified an efficient aromatic degrading bacteria (Uba, \u003cspan citationid=\"CR123\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) and already isolated from oil-contaminated soil (Chaudhary and Kim, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Furthermore, the enrichment of \u003cem\u003eBrevundimonas\u003c/em\u003e was previously correlated with PBAT degradation during composting, reinforcing its role in the breakdown of biodegradable polymers (Cao et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThe present study demonstrated that plastic mulch films did not significantly influence strawberry plant physiology or vegetative growth, but they played a critical role in productivity, with OxoPE and PP providing the most substantial yield enhancements. Alongside these agronomic improvements, fruit quality was maintained by the application of films. However, PBAT treatments specifically increased titratable acidity without compromising essential traits as soluble solids. Interestingly, OxoPE treatment improved the aesthetic quality of fruits by increasing lightness and chroma index. Although the main physicochemical properties of growth substrates remained largely unaffected, elemental composition was significantly influenced by mulches. Specifically, PBAT promotes nitrogen enrichment, whereas PE is associated with an increase in available micronutrients. Crucially, plastic-based mulching serves as a key driver of microbial assembly, enhancing the substrate bacterial community evenness and diversity. Structural analysis revealed that each material recruits unique microbial niches, with PP and PBAT that recruited specialized pollutant-degrading taxa. These structural shifts were primarily driven by humidity and nitrate gradients, highlighting a possible divergence between the physical microclimate stabilization of conventional films and the likely chemical input of biodegradable films. Lastly, functional predictions indicated that PE mulches application promoted nitrogen-fixing communities as a possible response to even slight nutrient shifts, whereas biodegradable films enhanced nitrification processes and complex carbon catabolism. By providing a comprehensive evaluation of traditional and biodegradable mulching in a small-scale strawberry cultivation, this study confirms that biodegradable films (OxoPE and PBAT) might be viable substitutes for traditional films, able to simultaneously promote high strawberries yields and the development of a specialized and functional growth substrate bacterial communities. Nevertheless, further investigations are needed to assess the long-term impact of both biodegradable and non-biodegradable plastic material on both strawberry agronomic performance and substrate microbial communities and functions.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eCOMPETING INTEREST\u003c/h2\u003e \u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e \u003ch2\u003eFUNDING\u003c/h2\u003e \u003cp\u003eThis research was funded by the European Union-Next Generation EU, UNIFI Young Independent Researchers Call, within the MuSC project MuSC (Analysis and sensing of contaminants in agriculture: from Mulches to Soil and Crops).\u003c/p\u003e\u003ch2\u003eAUTHOR CONTRIBUTIONS\u003c/h2\u003e \u003cp\u003eAll authors contributed to data interpretation, drafted the manuscript, agreed with its final version, and revised the manuscript. Conceptualization: Agnese Bellabarba, Costanza Scopetani, Giulia Selvolini; Methodology: Agnese Bellabarba, William Antonio Petrucci, Ermes Lo Piccolo, Lorenzo Bini; Formal analysis and investigation: Agnese Bellabarba, Lorenzo Bini, Costanza Scopetani, Giulia Selvolini, Ermes Lo Piccolo, Elisabetta Toni; Writing - original draft preparation: Agnese Bellabarba, Lorenzo Bini, Francesca Decorosi; Writing - review and editing: Agnese Bellabarba, Lorenzo Bini, Carlo Viti, Edgardo Giordani, Giulia Selvolini, Costanza Scopetani; Funding acquisition: Agnese Bellabarba, Costanza Scopetani, Giulia Selvolini; Resources: Agnese Bellabarba, Costanza Scopetani, Giulia Selvolini, Carlo Viti, Edgardo Giordani, Giovanna Marrazza, Alessandra Cincinelli, Tania Martinelli; Supervision: Carlo Viti, Edgardo Giordani, Giovanna Marrazza, Alessandra Cincinelli, Tania Martinelli.\u003c/p\u003e\n\u003ch3\u003eDATA AVAILABILITY STATEMENT\u003c/h3\u003e\n\u003cp\u003eGenomic sequences of this project have been deposited at GenBank under BioProject number PRJNA1450036\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbdelfattah, A., Awad, M., Sorour, O., 2025. Synergistic effects of magnetic water treatment and mulching on crop and soil moisture-salinity distribution. Sci Rep 15, 15741. https://doi.org/10.1038/s41598-025-98802-6\u003c/li\u003e\n\u003cli\u003eAmmala, A., Bateman, S., Dean, K., Petinakis, E., Sangwan, P., Wong, S., Yuan, Q., Yu, L., Patrick, C., Leong, K.H., 2011. An overview of degradable and biodegradable polyolefins. Progress in Polymer Science 36, 1015\u0026ndash;1049. https://doi.org/10.1016/j.progpolymsci.2010.12.002\u003c/li\u003e\n\u003cli\u003eAnderson, M.J., 2006. Distance-based tests for homogeneity of multivariate dispersions. Biometrics 62, 245\u0026ndash;253.\u003c/li\u003e\n\u003cli\u003eAsaf, S., Numan, M., Khan, A.L., Al-Harrasi, A., 2020. \u003cem\u003eSphingomonas\u003c/em\u003e : from diversity and genomics to functional role in environmental remediation and plant growth. Critical Reviews in Biotechnology 40, 138\u0026ndash;152. https://doi.org/10.1080/07388551.2019.1709793\u003c/li\u003e\n\u003cli\u003eBandopadhyay, S., Martin-Closas, L., Pelacho, A.M., DeBruyn, J.M., 2018. Biodegradable Plastic Mulch Films: Impacts on Soil Microbial Communities and Ecosystem Functions. Front. Microbiol. 9, 819. https://doi.org/10.3389/fmicb.2018.00819\u003c/li\u003e\n\u003cli\u003eBandopadhyay, S., Sintim, H.Y., DeBruyn, J.M., 2020. Effects of biodegradable plastic film mulching on soil microbial communities in two agroecosystems. PeerJ 8, e9015. https://doi.org/10.7717/peerj.9015\u003c/li\u003e\n\u003cli\u003eBao, M., Su, H., Tan, T., 2012. Biohydrogen Production by Dark Fermentation of Starch Using Mixed Bacterial Cultures of Bacillus sp and Brevumdimonas sp. Energy Fuels 26, 5872\u0026ndash;5878. https://doi.org/10.1021/ef300666m\u003c/li\u003e\n\u003cli\u003eBellabarba, A., Giagnoni, L., Adessi, A., Marra, E., Laschi, A., Neri, F., Mastrolonardo, G., 2024. Short-term machinery impact on microbial activity and diversity in a compacted forest soil. Applied Soil Ecology 203, 105646. https://doi.org/10.1016/j.apsoil.2024.105646\u003c/li\u003e\n\u003cli\u003eBini, L., Renai, L., Fichera, M., Petrucci, W.A., Lenzi, A., Biricolti, S., Giordani, E., Rivoira, L., Bruzzoniti, M.C., Piesik, D., Del Bubba, M., 2024. Assessing the Impact of Sustainable Biochar-Enriched Substrates on Safety and Quality of Tomato (\u003cem\u003eSolanum lycopersicum\u003c/em\u003e L.) as Relevant Model Crop. ACS Agric. Sci. Technol. 4, 681\u0026ndash;689. https://doi.org/10.1021/acsagscitech.3c00589\u003c/li\u003e\n\u003cli\u003eBiswas, B., Timsina, J., Mandal, K.G., Naorem, A., 2025. Effects of different irrigation methods and mulching on yield, growth and water use efficiency of strawberry. New Zealand Journal of Crop and Horticultural Science 53, 1408\u0026ndash;1427. https://doi.org/10.1080/01140671.2024.2348138\u003c/li\u003e\n\u003cli\u003eCao, L., Wang, L., Qi, Y., Yang, S., Gao, J., Liu, Q., Song, L., Hu, R., Wang, Z., Zhang, H., 2025. Enhanced effect of ferrous sulfate on nitrogen retention and PBAT degradation during co-composting by combing with biochar-loaded FN1 bacterial composites. Journal of Environmental Management 373, 123749. https://doi.org/10.1016/j.jenvman.2024.123749\u003c/li\u003e\n\u003cli\u003eChaudhary, D.K., Kim, J., 2018. Brevundimonas mongoliensis sp. nov., A Novel Psychrotolerant Bacterium Isolated from Oil-Contaminated Soil. Curr Microbiol 75, 1530\u0026ndash;1536. https://doi.org/10.1007/s00284-018-1555-4\u003c/li\u003e\n\u003cli\u003eChen, H., Wang, Y., Sun, X., Peng, Y., Xiao, L., 2020. Mixing effect of polylactic acid microplastic and straw residue on soil property and ecological function. Chemosphere 243, 125271. https://doi.org/10.1016/j.chemosphere.2019.125271\u003c/li\u003e\n\u003cli\u003eCosta, R., Saraiva, A., Carvalho, L., Duarte, E., 2014. The use of biodegradable mulch films on strawberry crop in Portugal. Scientia Horticulturae 173, 65\u0026ndash;70. https://doi.org/10.1016/j.scienta.2014.04.020\u003c/li\u003e\n\u003cli\u003eCrossman, J., Hurley, R.R., Futter, M., Nizzetto, L., 2020. Transfer and transport of microplastics from biosolids to agricultural soils and the wider environment. Science of The Total Environment 724, 138334. https://doi.org/10.1016/j.scitotenv.2020.138334\u003c/li\u003e\n\u003cli\u003eDangi, K.K., Darshan, D., Hitesh, P., Singh Dhawai, D., 2025. Efficacy of Various Mulches for the Modulation of Strawberry (Fragaria \u0026times; ananassa Duch.) Yield, Quality, and Storage Ability. Applied Fruit Science 67, 233. https://doi.org/10.1007/s10341-025-01452-1\u003c/li\u003e\n\u003cli\u003eDatta, H.S., Barua, P.C., Kotoky, U., Das, R., Saikia, H., Nath, H.K.D., 2024. Impact of Plant Spacing and Mulch on Growth Parameters of Strawberry (Fragaria x ananassa Duch.). JEAI 46, 926\u0026ndash;936. https://doi.org/10.9734/jeai/2024/v46i52448\u003c/li\u003e\n\u003cli\u003eDe Souza Machado, A.A., Lau, C.W., Till, J., Kloas, W., Lehmann, A., Becker, R., Rillig, M.C., 2018. Impacts of Microplastics on the Soil Biophysical Environment. Environ. Sci. Technol. 52, 9656\u0026ndash;9665. https://doi.org/10.1021/acs.est.8b02212\u003c/li\u003e\n\u003cli\u003eDeVetter, L.W., Zhang, H., Ghimire, S., Watkinson, S., Miles, C.A., 2017. Plastic Biodegradable Mulches Reduce Weeds and Promote Crop Growth in Day-neutral Strawberry in Western Washington. horts 52, 1700\u0026ndash;1706. https://doi.org/10.21273/HORTSCI12422-17\u003c/li\u003e\n\u003cli\u003eDewi, S.K., Han, Z.M., Bhat, S.A., Zhang, F., Wei, Y., Li, F., 2024. Effect of plastic mulch residue on plant growth performance and soil properties. Environmental Pollution 343, 123254. https://doi.org/10.1016/j.envpol.2023.123254\u003c/li\u003e\n\u003cli\u003eDi Mola, I., Cozzolino, E., Ottaiano, L., Riccardi, R., Spigno, P., Petriccione, M., Fiorentino, N., Fagnano, M., Mori, M., 2023. Biodegradable Mulching Film vs. Traditional Polyethylene: Effects on Yield and Quality of San Marzano Tomato Fruits. Plants 12, 3203. https://doi.org/10.3390/plants12183203\u003c/li\u003e\n\u003cli\u003eDing, F., Li, S., Lu, J., Penn, C.J., Wang, Q.-W., Lin, G., Sardans, J., Penuelas, J., Wang, J., Rillig, M.C., 2023. Consequences of 33 Years of Plastic Film Mulching and Nitrogen Fertilization on Maize Growth and Soil Quality. Environ. Sci. Technol. 57, 9174\u0026ndash;9183. https://doi.org/10.1021/acs.est.2c08878\u003c/li\u003e\n\u003cli\u003eDong, D., Guo, Z., Wu, F., Yang, X., Li, J., 2024. Plastic residues alter soil microbial community compositions and metabolite profiles under realistic conditions. Science of The Total Environment 906, 167352. https://doi.org/10.1016/j.scitotenv.2023.167352\u003c/li\u003e\n\u003cli\u003eDoumett, S., Fibbi, D., Cincinelli, A., Giordani, E., Nin, S., Del Bubba, M., 2011. Comparison of nutritional and nutraceutical properties in cultivated fruits of Fragaria vesca L. produced in Italy. Food Research International 44, 1209\u0026ndash;1216. https://doi.org/10.1016/j.foodres.2010.10.044\u003c/li\u003e\n\u003cli\u003eEberl, L., Vandamme, P., 2016. Members of the genus Burkholderia: good and bad guys. F1000Res 5, 1007. https://doi.org/10.12688/f1000research.8221.1\u003c/li\u003e\n\u003cli\u003eEl-Beltagi, H.S., Basit, A., Mohamed, H.I., Ali, I., Ullah, S., Kamel, E.A.R., Shalaby, T.A., Ramadan, K.M.A., Alkhateeb, A.A., Ghazzawy, H.S., 2022. Mulching as a Sustainable Water and Soil Saving Practice in Agriculture: A Review. Agronomy 12, 1881. https://doi.org/10.3390/agronomy12081881\u003c/li\u003e\n\u003cli\u003eFang, Y., Lin, C., Zhao, J., Gao, Y., Jia, X., 2025. Dosages of Biodegradable Poly(butylene adipate-co-terephthalate) Microplastics Affect Soil Microbial Community, Function, and Metabolome in Plant\u0026ndash;Soil System. Agronomy 15, 990. https://doi.org/10.3390/agronomy15040990\u003c/li\u003e\n\u003cli\u003eFarmer, J., Zhang, B., Jin, X., Zhang, P., Wang, J., 2017. Long-term effect of plastic film mulching and fertilization on bacterial communities in a brown soil revealed by high through-put sequencing. Archives of Agronomy and Soil Science 63, 230\u0026ndash;241.\u003c/li\u003e\n\u003cli\u003eFei, Y., Huang, S., Zhang, H., Tong, Y., Wen, D., Xia, X., Wang, H., Luo, Y., Barcel\u0026oacute;, D., 2020. Response of soil enzyme activities and bacterial communities to the accumulation of microplastics in an acid cropped soil. Science of The Total Environment 707, 135634. https://doi.org/10.1016/j.scitotenv.2019.135634\u003c/li\u003e\n\u003cli\u003eGao, W., Tu, Z., Yin, X., Ming, S., Cai, K., 2025. Effects of PBAT biodegradable mulch on lettuce (Lactuca sativa L.) physiology and soil microbial community: Based on a long-term degradation trial. Ecotoxicology and Environmental Safety 302, 118734. https://doi.org/10.1016/j.ecoenv.2025.118734\u003c/li\u003e\n\u003cli\u003eGao, X., Fu, C., Li, M., Qi, X., Jia, X., 2022. Effects of Biodegradation of Corn-Starch\u0026ndash;Sodium-Alginate-Based Liquid Mulch Film on Soil Microbial Functions. IJERPH 19, 8631. https://doi.org/10.3390/ijerph19148631\u003c/li\u003e\n\u003cli\u003eGatheru Waigi, M., Sun, K., Gao, Y., 2017. Sphingomonads in Microbe-Assisted Phytoremediation: Tackling Soil Pollution. Trends in Biotechnology 35, 883\u0026ndash;899. https://doi.org/10.1016/j.tibtech.2017.06.014\u003c/li\u003e\n\u003cli\u003eGeorgiadou, E.C., Garc\u0026iacute;a, C.J., Taliadorou, A.M., Gedeon, S., Valanides, N., Varaldo, A., Gohari, G., Balsells-Llaurad\u0026oacute;, M., Alc\u0026aacute;zar, R., Hertog, M.L.A.T.M., Tom\u0026aacute;s-Barber\u0026aacute;n, F.A., Manganaris, G.A., Fotopoulos, V., 2025. Pre-harvest application of sodium alginate functionalized with melatonin enhances secondary metabolism in strawberry fruit. Current Plant Biology 43, 100515. https://doi.org/10.1016/j.cpb.2025.100515\u003c/li\u003e\n\u003cli\u003eGrimaut, D.A., Da Silva, J.B.A., Lemos, P.V.F., Correia, P.R.C., Santana, J.S., Pess\u0026ocirc;a, L.C., Estevez-Areco, S., Fam\u0026aacute;, L.M., Goyanes, S.N., Marcelino, H.R., De Jesus Assis, D., De Souza, C.O., 2023. Effect of Addition of Cross-Linked Starch on the Properties of Degraded PBAT Poly(butylene adipate-co-terephthalate) Films. Polymers 15, 3106. https://doi.org/10.3390/polym15143106\u003c/li\u003e\n\u003cli\u003eGu, J., Guo, F., Lin, L., Zhang, J., Sun, W., Muhammad, R., Liang, H., Duan, D., Deng, X., Lin, Z., Wang, Y., Zhong, Y., Xu, Z., 2023. Microbiological mechanism for \u0026ldquo;production while remediating\u0026rdquo; in Cd-contaminated paddy fields: A field experiment. Science of The Total Environment 885, 163896. https://doi.org/10.1016/j.scitotenv.2023.163896\u003c/li\u003e\n\u003cli\u003eHam, J.M., Kluitenberg, G.J., Lamont, W.J., 1993. Optical Properties of Plastic Mulches Affect the Field Temperature Regime. jashs 118, 188\u0026ndash;193. https://doi.org/10.21273/JASHS.118.2.188\u003c/li\u003e\n\u003cli\u003eHannum, S.M., 2004. Potential Impact of Strawberries on Human Health: A Review of the Science. Critical Reviews in Food Science and Nutrition 44, 1\u0026ndash;17. https://doi.org/10.1080/10408690490263756\u003c/li\u003e\n\u003cli\u003eHartman, K., Van Der Heijden, M.G.A., Wittwer, R.A., Banerjee, S., Walser, J.-C., Schlaeppi, K., 2018. Cropping practices manipulate abundance patterns of root and soil microbiome members paving the way to smart farming. Microbiome 6, 14. https://doi.org/10.1186/s40168-017-0389-9\u003c/li\u003e\n\u003cli\u003eHartmann, M., Widmer, F., 2006. Community Structure Analyses Are More Sensitive to Differences in Soil Bacterial Communities than Anonymous Diversity Indices. Appl Environ Microbiol 72, 7804\u0026ndash;7812. https://doi.org/10.1128/AEM.01464-06\u003c/li\u003e\n\u003cli\u003eHegan, D., Tong, L., Zhiquan, H., Qinming, S., Ru, L., 2015. Determining time limits of continuous film mulching and examining residual effects on cotton yield and soil properties. Journal of Environmental Biology 36, 677.\u003c/li\u003e\n\u003cli\u003eHossain, M.E., Zhang, Z., Dong, W., Wang, S., Liu, M., Liu, E., Mei, X., 2022. Plastic Film Mulching Improved Maize Yield, Water Use Efficiency, and N Use Efficiency under Dryland Farming System in Northeast China. Plants 11, 1710. https://doi.org/10.3390/plants11131710\u003c/li\u003e\n\u003cli\u003eHuang, Y., Zhao, Y., Wang, J., Zhang, M., Jia, W., Qin, X., 2019. LDPE microplastic films alter microbial community composition and enzymatic activities in soil. Environmental Pollution 254, 112983. https://doi.org/10.1016/j.envpol.2019.112983\u003c/li\u003e\n\u003cli\u003eIngman, M., Santelmann, M.V., Tilt, B., 2015. Agricultural water conservation in china: plastic mulch and traditional irrigation. Ecosyst Health Sustain 1, 1\u0026ndash;11. https://doi.org/10.1890/EHS14-0018.1\u003c/li\u003e\n\u003cli\u003eJabran, K., Hussain, M., Fahad, S., Farooq, M., Bajwa, A.A., Alharrby, H., Nasim, W., 2016. Economic assessment of different mulches in conventional and water-saving rice production systems. Environ Sci Pollut Res 23, 9156\u0026ndash;9163. https://doi.org/10.1007/s11356-016-6162-y\u003c/li\u003e\n\u003cli\u003eJacquin, J., Cheng, J., Odobel, C., Pandin, C., Conan, P., Pujo-Pay, M., Barbe, V., Meistertzheim, A.-L., Ghiglione, J.-F., 2019. Microbial Ecotoxicology of Marine Plastic Debris: A Review on Colonization and Biodegradation by the \u0026ldquo;Plastisphere.\u0026rdquo; Front. Microbiol. 10, 865. https://doi.org/10.3389/fmicb.2019.00865\u003c/li\u003e\n\u003cli\u003eKader, M.A., Senge, M., Mojid, M.A., Ito, K., 2017. Recent advances in mulching materials and methods for modifying soil environment. Soil and Tillage Research 168, 155\u0026ndash;166. https://doi.org/10.1016/j.still.2017.01.001\u003c/li\u003e\n\u003cli\u003eKapanen, A., Schettini, E., Vox, G., It\u0026auml;vaara, M., 2008. Performance and Environmental Impact of Biodegradable Films in Agriculture: A Field Study on Protected Cultivation. J Polym Environ 16, 109\u0026ndash;122. https://doi.org/10.1007/s10924-008-0091-x\u003c/li\u003e\n\u003cli\u003eKasirajan, S., Ngouajio, M., 2012. Polyethylene and biodegradable mulches for agricultural applications: a review. Agron. Sustain. Dev. 32, 501\u0026ndash;529. https://doi.org/10.1007/s13593-011-0068-3\u003c/li\u003e\n\u003cli\u003eKasperbauer, M.J., Loughrin, J.H., Wang, S.Y., 2007. Light Reflected from Red Mulch to Ripening Strawberries Affects Aroma, Sugar and Organic Acid Concentrations\u0026para;. Photochemistry and Photobiology 74, 103\u0026ndash;107. https://doi.org/10.1562/0031-8655(2001)0740103LRFRMT2.0.CO2\u003c/li\u003e\n\u003cli\u003eKhandare, S.D., Agrawal, D., Mehru, N., Chaudhary, D.R., 2022. Marine bacterial based enzymatic degradation of low-density polyethylene (LDPE) plastic. Journal of Environmental Chemical Engineering 10, 107437. https://doi.org/10.1016/j.jece.2022.107437\u003c/li\u003e\n\u003cli\u003eKim, S.-K., Kim, J.-S., Lee, H., Lee, H.-J., 2021. Abundance and characteristics of microplastics in soils with different agricultural practices: Importance of sources with internal origin and environmental fate. Journal of Hazardous Materials 403, 123997. https://doi.org/10.1016/j.jhazmat.2020.123997\u003c/li\u003e\n\u003cli\u003eKoitabashi, M., Noguchi, M.T., Sameshima-Yamashita, Y., Hiradate, S., Suzuki, K., Yoshida, S., Watanabe, T., Shinozaki, Y., Tsushima, S., Kitamoto, H.K., 2012. Degradation of biodegradable plastic mulch films in soil environment by phylloplane fungi isolated from gramineous plants. AMB express 2, 40.\u003c/li\u003e\n\u003cli\u003eKoutny, M., Lemaire, J., Delort, A.-M., 2006. Biodegradation of polyethylene films with prooxidant additives. Chemosphere 64, 1243\u0026ndash;1252. https://doi.org/10.1016/j.chemosphere.2005.12.060\u003c/li\u003e\n\u003cli\u003eKumar, A., M\u0026auml;nnist\u0026ouml;, M.K., P\u0026auml;tsi, M., Kerkhof, L.J., H\u0026auml;ggblom, M.M., 2025. Genome analysis reveals diverse novel psychrotolerant \u003cem\u003eMucilaginibacter\u003c/em\u003e species in Arctic tundra soils. ISME Communications 5, ycaf071. https://doi.org/10.1093/ismeco/ycaf071\u003c/li\u003e\n\u003cli\u003eK\u0026uuml;ster, A.N., Paula, C., Azevedo, J., Serra, A.C., Coelho, J.F.J., 2025. Formulations, Processing, and Application of Poly(butylene adipate-co-terephthalate)/Thermoplastic Starch Blends: A Review. Polymers 17, 1457. https://doi.org/10.3390/polym17111457\u003c/li\u003e\n\u003cli\u003eLee, D., Lee, E., Lee, Y., Shin, M., Yang, J.S., Kim, M., Sang, M.K., Park, H.J., Jung, H.W., 2025. Agri-plastics in soils drive changes in the rhizosphere bacterial community and plant transcriptome in Arabidopsis. https://doi.org/doi.org/10.1093/jxb/eraf336\u003c/li\u003e\n\u003cli\u003eLi, C., Cui, Q., Li, Y., Zhang, K., Lu, X., Zhang, Y., 2022. Effect of LDPE and biodegradable PBAT primary microplastics on bacterial community after four months of soil incubation. Journal of Hazardous Materials 429, 128353. https://doi.org/10.1016/j.jhazmat.2022.128353\u003c/li\u003e\n\u003cli\u003eLi, C., Moore-Kucera, J., Miles, C., Leonas, K., Lee, J., Corbin, A., Inglis, D., 2014. Degradation of potentially biodegradable plastic mulch films at three diverse US locations. Agroecology and sustainable food systems 38, 861\u0026ndash;889.\u003c/li\u003e\n\u003cli\u003eLi, H., Li, P., Cao, G., Zhao, M., Zhu, Z., Ma, Y., Wang, W., Cao, S., Xu, Y., Dong, C., 2025. Mulching influences pear yield and quality by changing rhizosphere microbial community structure in the arid region of Northwest China. Front. Plant Sci. 16, 1633540. https://doi.org/10.3389/fpls.2025.1633540\u003c/li\u003e\n\u003cli\u003eLi, N., Qu, J., Yang, J., 2023. Microplastics distribution and microbial community characteristics of farmland soil under different mulch methods. Journal of Hazardous Materials 445, 130408. https://doi.org/10.1016/j.jhazmat.2022.130408\u003c/li\u003e\n\u003cli\u003eLi YuanQiao, L.Y., Zhao CaiXia, Z.C., Yan ChangRong, Y.C., Mao LiLi, M.L., Liu Qi, L.Q., Li Zhen, L.Z., He WenQing, H.W., 2020. Effects of agricultural plastic film residues on transportation and distribution of water and nitrate in soil.\u003c/li\u003e\n\u003cli\u003eLiu, C., Cui, Y., Li, X., Yao, M., 2021. \u003cem\u003emicroeco\u003c/em\u003e : an R package for data mining in microbial community ecology. FEMS Microbiology Ecology 97, fiaa255. https://doi.org/10.1093/femsec/fiaa255\u003c/li\u003e\n\u003cli\u003eLiu, E.K., He, W.Q., Yan, C.R., 2014. \u0026lsquo;White revolution\u0026rsquo; to \u0026lsquo;white pollution\u0026rsquo;\u0026mdash;agricultural plastic film mulch in China. Environ. Res. Lett. 9, 091001. https://doi.org/10.1088/1748-9326/9/9/091001\u003c/li\u003e\n\u003cli\u003eLiu, L., Li, L., Zou, G., Gu, J., Zuo, Q., Zheng, X., Du, L., Liu, D., 2025. Soil fungi respond more violently to both polyethylene and PBAT biodegradable mulch film residues than bacteria do. Front. Environ. Sci. 13, 1533441. https://doi.org/10.3389/fenvs.2025.1533441\u003c/li\u003e\n\u003cli\u003eLiu, L., Zou, G., Zuo, Q., Li, C., Gu, J., Kang, L., Ma, M., Liang, K., Liu, D., Du, L., 2022. Soil bacterial community and metabolism showed a more sensitive response to PBAT biodegradable mulch residues than that of LDPE mulch residues. Journal of Hazardous Materials 438, 129507. https://doi.org/10.1016/j.jhazmat.2022.129507\u003c/li\u003e\n\u003cli\u003eLiu, X., Wen, Z., Zhou, W., Dong, W., Ren, H., Liang, G., Gong, W., 2025. Effect of Multiyear Biodegradable Plastic Mulch on Soil Microbial Community, Assembly, and Functioning. Microorganisms 13, 259. https://doi.org/10.3390/microorganisms13020259\u003c/li\u003e\n\u003cli\u003eL\u0026oacute;pez-Tolentino, G., Ibarra-Jim\u0026eacute;nez, L., M\u0026eacute;ndez-Prieto, A., Lozano-del R\u0026iacute;o, A.J., Lira-Saldivar, R.H., Valenzuela-Soto, J.H., Lozano-Cavazos, C.J., Torres-Olivar, V., 2017. Photosynthesis, growth, and fruit yield of cucumber in response to oxo-degradable plastic mulches. Acta Agriculturae Scandinavica, Section B\u0026mdash;Soil \u0026amp; Plant Science 67, 77\u0026ndash;84.\u003c/li\u003e\n\u003cli\u003eLouca, S., Parfrey, L.W., Doebeli, M., 2016. Decoupling function and taxonomy in the global ocean microbiome. Science 353, 1272\u0026ndash;1277.\u003c/li\u003e\n\u003cli\u003eMahawar, L., Mishra, A., Tsitouri, A., Albrectsen, B.R., 2026. Straw Mulching Differentially Shapes the Structure and Function of Below‐Ground Bacterial Communities in Potato Depending on eDNA Source and Cultivar. Plant Enviro Interactions 7, e70131. https://doi.org/10.1002/pei3.70131\u003c/li\u003e\n\u003cli\u003eMangiafico, S., 2016. rcompanion: Functions to Support Extension Education Program Evaluation. https://doi.org/10.32614/CRAN.package.rcompanion\u003c/li\u003e\n\u003cli\u003eMansoor, Z., Tchuenbou-Magaia, F., Kowalczuk, M., Adamus, G., Manning, G., Parati, M., Radecka, I., Khan, H., 2022. Polymers Use as Mulch Films in Agriculture\u0026mdash;A Review of History, Problems and Current Trends. Polymers 14, 5062. https://doi.org/10.3390/polym14235062\u003c/li\u003e\n\u003cli\u003eMasny, A., 2015. SEASON EXTENSION POSSIBILITIES IN TWO POLISH JUNE-BEARING STRAWBERRY CULTIVARS.\u003c/li\u003e\n\u003cli\u003eMeng, T., Bu, H., Zhang, X., Chen, X., Wang, W., Zhao, M., Liu, J., Zhang, J., Zhang, D., Lu, Z., Zhao, X., 2025. Degradable film mulching recruited beneficial microbiota and increased rhizosphere bacterial diversity in sunflower. Sci Rep 15, 18522. https://doi.org/10.1038/s41598-025-03213-2\u003c/li\u003e\n\u003cli\u003eMenossi, M., Cisneros, M., Alvarez, V.A., Casalongu\u0026eacute;, C., 2021. Current and emerging biodegradable mulch films based on polysaccharide bio-composites. A review. Agron. Sustain. Dev. 41, 53. https://doi.org/10.1007/s13593-021-00685-0\u003c/li\u003e\n\u003cli\u003eMenzel, C.M., 2025. The relationship between yield and plant density in strawberry: competition does not impose an upper limit to population-level production. The Journal of Horticultural Science and Biotechnology 100, 1\u0026ndash;28. https://doi.org/10.1080/14620316.2024.2400127\u003c/li\u003e\n\u003cli\u003eMeyer, M., Diehl, D., Schaumann, G.E., Mu\u0026ntilde;oz, K., 2021. Multiannual soil mulching in agriculture: analysis of biogeochemical soil processes under plastic and straw mulches in a 3-year field study in strawberry cultivation. J Soils Sediments 21, 3733\u0026ndash;3752. https://doi.org/10.1007/s11368-021-03037-3\u003c/li\u003e\n\u003cli\u003eMiller, K., Feucht, W., Schmid, M., 2019. Bioactive Compounds of Strawberry and Blueberry and Their Potential Health Effects Based on Human Intervention Studies: A Brief Overview. Nutrients 11, 1510. https://doi.org/10.3390/nu11071510\u003c/li\u003e\n\u003cli\u003eMo, F., Han, J., Wen, X., Wang, X., Li, P., Vinay, N., Jia, Z., Xiong, Y., Liao, Y., 2020. Quantifying regional effects of plastic mulch on soil nitrogen pools, cycles, and fluxes in rain‐fed agroecosystems of the Loess Plateau. Land Degrad Dev 31, 1675\u0026ndash;1687. https://doi.org/10.1002/ldr.3548\u003c/li\u003e\n\u003cli\u003eMorales-Quintana, L., Ramos, P., 2019. Chilean strawberry (Fragaria chiloensis): An integrative and comprehensive review. Food Research International 119, 769\u0026ndash;776. https://doi.org/10.1016/j.foodres.2018.10.059\u003c/li\u003e\n\u003cli\u003eMorra, L., Bilotto, M., Mignoli, E., Sicignano, M., Magri, A., Cice, D., Cozzolino, R., Malorni, L., Siano, F., Picariello, G., Guerrini, S., Petriccione, M., 2022. New Mater-Bi, Biodegradable Mulching Film for Strawberry (Fragaria \u0026times; Ananassa Duch.): Effects on Film Duration, Crop Yields, Qualitative, and Nutraceutical Traits of Fruits. Plants 11, 1726. https://doi.org/10.3390/plants11131726\u003c/li\u003e\n\u003cli\u003eMorra, L., Cozzolino, E., Salluzzo, A., Modestia, F., Bilotto, M., Baiano, S., Del Piano, L., 2021. Plant Growth, Yields and Fruit Quality of Processing Tomato (Solanum lycopersicon L.) as Affected by the Combination of Biodegradable Mulching and Digestate. Agronomy 11, 100. https://doi.org/10.3390/agronomy11010100\u003c/li\u003e\n\u003cli\u003eMu\u0026ntilde;oz, K., Buchmann, C., Meyer, M., Schmidt-Heydt, M., Steinmetz, Z., Diehl, D., Thiele-Bruhn, S., Schaumann, G.E., 2017a. Physicochemical and microbial soil quality indicators as affected by the agricultural management system in strawberry cultivation using straw or black polyethylene mulching. Applied Soil Ecology 113, 36\u0026ndash;44. https://doi.org/10.1016/j.apsoil.2017.01.014\u003c/li\u003e\n\u003cli\u003eMu\u0026ntilde;oz, K., Buchmann, C., Meyer, M., Schmidt-Heydt, M., Steinmetz, Z., Diehl, D., Thiele-Bruhn, S., Schaumann, G.E., 2017b. Physicochemical and microbial soil quality indicators as affected by the agricultural management system in strawberry cultivation using straw or black polyethylene mulching. Applied Soil Ecology 113, 36\u0026ndash;44. https://doi.org/10.1016/j.apsoil.2017.01.014\u003c/li\u003e\n\u003cli\u003eMu\u0026ntilde;oz, K., Schmidt-Heydt, M., Stoll, D., Diehl, D., Ziegler, J., Geisen, R., Schaumann, G.E., 2015. Effect of plastic mulching on mycotoxin occurrence and mycobiome abundance in soil samples from asparagus crops. Mycotoxin research 31, 191\u0026ndash;201.\u003c/li\u003e\n\u003cli\u003eMu\u0026ntilde;oz, K., Thiele-Bruhn, S., Kenngott, K.G.J., Meyer, M., Diehl, D., Steinmetz, Z., Schaumann, G.E., 2022. Effects of Plastic versus Straw Mulching Systems on Soil Microbial Community Structure and Enzymes in Strawberry Cultivation. Soil Systems 6, 21. https://doi.org/10.3390/soilsystems6010021\u003c/li\u003e\n\u003cli\u003eMuroi, F., Tachibana, Y., Kobayashi, Y., Sakurai, T., Kasuya, K., 2016. Influences of poly(butylene adipate-co-terephthalate) on soil microbiota and plant growth. Polymer Degradation and Stability 129, 338\u0026ndash;346. https://doi.org/10.1016/j.polymdegradstab.2016.05.018\u003c/li\u003e\n\u003cli\u003eNeri, D., Baruzzi, G., Massetani, F., Faedi, W., 2012. Strawberry production in forced and protected culture in Europe as a response to climate change. Can. J. Plant Sci. 92, 1021\u0026ndash;1036. https://doi.org/10.4141/cjps2011-276\u003c/li\u003e\n\u003cli\u003eNi, X., Song, W., Zhang, H., Yang, X., Wang, L., 2016. Effects of Mulching on Soil Properties and Growth of Tea Olive (Osmanthus fragrans). PLoS ONE 11, e0158228. https://doi.org/10.1371/journal.pone.0158228\u003c/li\u003e\n\u003cli\u003eOgle, D.H., Wheeler, P., Dinno, A., 2020. FSA: fisheries stock analysis. R package version 0.8. 30. Vienna: R Core Team.\u003c/li\u003e\n\u003cli\u003eOksanen, J., 2015. Vegan: community ecology package. R package version 2, 3.\u003c/li\u003e\n\u003cli\u003ePinto, J.P., Da Cunha, F.F., Da Silva Ad\u0026atilde;o, A., De Paula, L.B., Ribeiro, M.C., Costa Neto, J.R.R., 2022. Strawberry Production with Different Mulches and Wetted Areas. Horticulturae 8, 930. https://doi.org/10.3390/horticulturae8100930\u003c/li\u003e\n\u003cli\u003ePruesse, E., Quast, C., Knittel, K., Fuchs, B.M., Ludwig, W., Peplies, J., Glockner, F.O., 2007. SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Research 35, 7188\u0026ndash;7196. https://doi.org/10.1093/nar/gkm864\u003c/li\u003e\n\u003cli\u003eQi, Y., Ossowicki, A., Yang, X., Huerta Lwanga, E., Dini-Andreote, F., Geissen, V., Garbeva, P., 2020. Effects of plastic mulch film residues on wheat rhizosphere and soil properties. Journal of Hazardous Materials 387, 121711. https://doi.org/10.1016/j.jhazmat.2019.121711\u003c/li\u003e\n\u003cli\u003eQi, Y., Ossowicki, A., Yergeau, \u0026Eacute;., Vigani, G., Geissen, V., Garbeva, P., 2022. Plastic mulch film residues in agriculture: impact on soil suppressiveness, plant growth, and microbial communities. FEMS Microbiology Ecology 98, fiac017. https://doi.org/10.1093/femsec/fiac017\u003c/li\u003e\n\u003cli\u003eQiang, L., Hu, H., Li, G., Xu, J., Cheng, J., Wang, J., Zhang, R., 2023. Plastic mulching, and occurrence, incorporation, degradation, and impacts of polyethylene microplastics in agroecosystems. Ecotoxicology and Environmental Safety 263, 115274. https://doi.org/10.1016/j.ecoenv.2023.115274\u003c/li\u003e\n\u003cli\u003eQin, W., Hu, C., Oenema, O., 2015. Soil mulching significantly enhances yields and water and nitrogen use efficiencies of maize and wheat: a meta-analysis. Sci Rep 5, 16210. https://doi.org/10.1038/srep16210\u003c/li\u003e\n\u003cli\u003eRillig, M.C., Kim, S.W., Zhu, Y.-G., 2024. The soil plastisphere. Nat Rev Microbiol 22, 64\u0026ndash;74. https://doi.org/10.1038/s41579-023-00967-2\u003c/li\u003e\n\u003cli\u003eSalama, K., Geyer, M., 2023. Plastic Mulch Films in Agriculture: Their Use, Environmental Problems, Recycling and Alternatives. Environments 10, 179. https://doi.org/10.3390/environments10100179\u003c/li\u003e\n\u003cli\u003eScales, B.S., Cable, R.N., Duhaime, M.B., Gerdts, G., Fischer, F., Fischer, D., Mothes, S., Hintzki, L., Moldaenke, L., Ruwe, M., Kalinowski, J., Kreikemeyer, B., Pedrotti, M.-L., Gorsky, G., Elineau, A., Labrenz, M., Oberbeckmann, S., 2021. Cross-Hemisphere Study Reveals Geographically Ubiquitous, Plastic-Specific Bacteria Emerging from the Rare and Unexplored Biosphere. mSphere 6, e00851-20. https://doi.org/10.1128/mSphere.00851-20\u003c/li\u003e\n\u003cli\u003eScarascia-Mugnozza, G., Sica, C., Russo, G., 2011. PLASTIC MATERIALS IN EUROPEAN AGRICULTURE: ACTUAL USE AND PERSPECTIVES.\u003c/li\u003e\n\u003cli\u003eScheurer, M., Bigalke, M., 2018. Microplastics in Swiss Floodplain Soils. Environ. Sci. Technol. 52, 3591\u0026ndash;3598. https://doi.org/10.1021/acs.est.7b06003\u003c/li\u003e\n\u003cli\u003eSchewe, J., Heinke, J., Gerten, D., Haddeland, I., Arnell, N.W., Clark, D.B., Dankers, R., Eisner, S., Fekete, B.M., Col\u0026oacute;n-Gonz\u0026aacute;lez, F.J., Gosling, S.N., Kim, H., Liu, X., Masaki, Y., Portmann, F.T., Satoh, Y., Stacke, T., Tang, Q., Wada, Y., Wisser, D., Albrecht, T., Frieler, K., Piontek, F., Warszawski, L., Kabat, P., 2014. Multimodel assessment of water scarcity under climate change. Proc. Natl. Acad. Sci. U.S.A. 111, 3245\u0026ndash;3250. https://doi.org/10.1073/pnas.1222460110\u003c/li\u003e\n\u003cli\u003eSchonbeck, M.W., 1999. Weed Suppression and Labor Costs Associated with Organic, Plastic, and Paper Mulches in Small-Scale Vegetable Production. Journal of Sustainable Agriculture 13, 13\u0026ndash;33. https://doi.org/10.1300/J064v13n02_04\u003c/li\u003e\n\u003cli\u003eScopetani, C., Bellabarba, A., Selvolini, G., Martellini, T., Viti, C., Cincinelli, A., 2025. Evaluating additive release from conventional and biodegradable mulch films. Science of The Total Environment 975, 179294. https://doi.org/10.1016/j.scitotenv.2025.179294\u003c/li\u003e\n\u003cli\u003eScopetani, C., Cincinelli, A., Martellini, T., Rodrigues, A.C.M., 2023. Editorial: Hazardous contaminants associated with plastics: occurrence and environmental effects. Front. Environ. Sci. 11, 1341738. https://doi.org/10.3389/fenvs.2023.1341738\u003c/li\u003e\n\u003cli\u003eSegata, N., Izard, J., Waldron, L., Gevers, D., Miropolsky, L., Garrett, W.S., Huttenhower, C., 2011. Metagenomic biomarker discovery and explanation. Genome Biol 12, R60. https://doi.org/10.1186/gb-2011-12-6-r60\u003c/li\u003e\n\u003cli\u003e\u0026Scaron;er\u0026aacute;, J., Kadlečkov\u0026aacute;, M., Fayyazbakhsh, A., Kučabov\u0026aacute;, V., Koutn\u0026yacute;, M., 2020. Occurrence and Analysis of Thermophilic Poly(butylene adipate-co-terephthalate)-Degrading Microorganisms in Temperate Zone Soils. IJMS 21, 7857. https://doi.org/10.3390/ijms21217857\u003c/li\u003e\n\u003cli\u003eShan, X., Zhang, W., Dai, Z., Li, J., Mao, W., Yu, F., Ma, J., Wang, S., Zeng, X., 2022. Comparative Analysis of the Effects of Plastic Mulch Films on Soil Nutrient, Yields and Soil Microbiome in Three Vegetable Fields. Agronomy 12, 506. https://doi.org/10.3390/agronomy12020506\u003c/li\u003e\n\u003cli\u003eSharma, S., Pant, P., Sangwan, M., Sahrawat, R., 2024. Effect of Organic and Inorganic Mulch on Growth, Yield and Quality of Strawberry CV. Winter Dawn. IJECC 14, 377\u0026ndash;382. https://doi.org/10.9734/ijecc/2024/v14i34049\u003c/li\u003e\n\u003cli\u003eShi, Z., Xiong, L., Liu, T., Wu, W., 2022. Alteration of bacterial communities and co-occurrence networks as a legacy effect upon exposure to polyethylene residues under field environment. Journal of Hazardous Materials 426, 128126. https://doi.org/10.1016/j.jhazmat.2021.128126\u003c/li\u003e\n\u003cli\u003eShiukhy, S., Raeini-Sarjaz, M., Chalavi, V., 2015. Colored plastic mulch microclimates affect strawberry fruit yield and quality. Int J Biometeorol 59, 1061\u0026ndash;1066. https://doi.org/10.1007/s00484-014-0919-0\u003c/li\u003e\n\u003cli\u003eSintim, H.Y., Flury, M., 2017. Is Biodegradable Plastic Mulch the Solution to Agriculture\u0026rsquo;s Plastic Problem? Environ. Sci. Technol. 51, 1068\u0026ndash;1069. https://doi.org/10.1021/acs.est.6b06042\u003c/li\u003e\n\u003cli\u003eSmith, M.R., Myers, S.S., 2018. Impact of anthropogenic CO2 emissions on global human nutrition. Nature Clim Change 8, 834\u0026ndash;839. https://doi.org/10.1038/s41558-018-0253-3\u003c/li\u003e\n\u003cli\u003eSomanathan, H., Sathasivam, R., Sivaram, S., Mariappan Kumaresan, S., Muthuraman, M.S., Park, S.U., 2022. An update on polyethylene and biodegradable plastic mulch films and their impact on the environment. Chemosphere 307, 135839. https://doi.org/10.1016/j.chemosphere.2022.135839\u003c/li\u003e\n\u003cli\u003eSoppelsa, S., Kelderer, M., Casera, C., Bassi, M., Robatscher, P., Matteazzi, A., Andreotti, C., 2019. Foliar Applications of Biostimulants Promote Growth, Yield and Fruit Quality of Strawberry Plants Grown under Nutrient Limitation. Agronomy 9, 483. https://doi.org/10.3390/agronomy9090483\u003c/li\u003e\n\u003cli\u003eSteinmetz, Z., Wollmann, C., Schaefer, M., Buchmann, C., David, J., Tr\u0026ouml;ger, J., Mu\u0026ntilde;oz, K., Fr\u0026ouml;r, O., Schaumann, G.E., 2016. Plastic mulching in agriculture. Trading short-term agronomic benefits for long-term soil degradation? Science of The Total Environment 550, 690\u0026ndash;705. https://doi.org/10.1016/j.scitotenv.2016.01.153\u003c/li\u003e\n\u003cli\u003eSun, Y., Duan, C., Cao, N., Ding, C., Huang, Y., Wang, J., 2022. Biodegradable and conventional microplastics exhibit distinct microbiome, functionality, and metabolome changes in soil. Journal of Hazardous Materials 424, 127282. https://doi.org/10.1016/j.jhazmat.2021.127282\u003c/li\u003e\n\u003cli\u003eSupreetha, B.G., Singh, N.P., Sharma, S., Sharda, R., Sharma, A., 2025. Comparison of fruit colour development and biochemical attributes under different mulching regimes in strawberry. Acta Physiol Plant 47, 7. https://doi.org/10.1007/s11738-024-03751-8\u003c/li\u003e\n\u003cli\u003eTagg, A.S., Sperlea, T., Labrenz, M., Harrison, J.P., Ojeda, J.J., Sapp, M., 2022. Year-Long Microbial Succession on Microplastics in Wastewater: Chaotic Dynamics Outweigh Preferential Growth. Microorganisms 10, 1775. https://doi.org/10.3390/microorganisms10091775\u003c/li\u003e\n\u003cli\u003eTakahashi, S., Tomita, J., Nishioka, K., Hisada, T., Nishijima, M., 2014. Development of a Prokaryotic Universal Primer for Simultaneous Analysis of Bacteria and Archaea Using Next-Generation Sequencing. PLoS ONE 9, e105592. https://doi.org/10.1371/journal.pone.0105592\u003c/li\u003e\n\u003cli\u003eTorres-Olivar, V., Ibarra-Jim\u0026eacute;nez, L., C\u0026aacute;rdenas-Flores, A., Lira-Saldivar, R.H., Valenzuela-Soto, J.H., Castillo-Campohermoso, M.A., 2018. Changes induced by plastic film mulches on soil temperature and their relevance in growth and fruit yield of pickling cucumber. Acta Agriculturae Scandinavica, Section B\u0026mdash;Soil \u0026amp; Plant Science 68, 97\u0026ndash;103.\u003c/li\u003e\n\u003cli\u003eTozzi, F., Del Bubba, M., Petrucci, W.A., Pecchioli, S., Macci, C., Hern\u0026aacute;ndez Garc\u0026iacute;a, F., Mart\u0026iacute;nez Nicol\u0026aacute;s, J.J., Giordani, E., 2020. Use of a remediated dredged marine sediment as a substrate for food crop cultivation: Sediment characterization and assessment of fruit safety and quality using strawberry (Fragaria x ananassa Duch.) as model species of contamination transfer. Chemosphere 238, 124651. https://doi.org/10.1016/j.chemosphere.2019.124651\u003c/li\u003e\n\u003cli\u003eUba, B.O., 2019. Growth Profile and Catabolic Pathways Involved in Degradation of Aromatic Hydrocarbons by Marine Bacteria Isolated from Niger Delta, Nigeria. MRJI 1\u0026ndash;18. https://doi.org/10.9734/mrji/2018/v26i530075\u003c/li\u003e\n\u003cli\u003eVillena, J., Moreno, M., Gonz\u0026aacute;lez-Mora, S., L\u0026oacute;pez-Perales, J., Morales-Rodr\u0026iacute;guez, P., Moreno, C., 2022. Degradation Pattern of Five Biodegradable, Potentially Low-Environmental-Impact Mulches under Laboratory Conditions. Agriculture 12, 1910. https://doi.org/10.3390/agriculture12111910\u003c/li\u003e\n\u003cli\u003eVita, M.M., Iturbe-Espinoza, P., Bonte, M., Brandt, B.W., Braster, M., Brown, D.M., Van Spanning, R.J.M., 2022. Oil Absorbent Polypropylene Particles Stimulate Biodegradation of Crude Oil by Microbial Consortia. Front. Microbiol. 13, 853285. https://doi.org/10.3389/fmicb.2022.853285\u003c/li\u003e\n\u003cli\u003eWang, C., Dong, J., Ji, B., Lv, Y., Song, Y., Li, Q., Sun, C., Zong, R., Zhang, M., 2025. Sub-soil biodegradable film mulching: A sustainable solution for enhancing winter wheat yield and shaping soil microbial communities in saline-alkali soils. Journal of Hazardous Materials 495, 139020. https://doi.org/10.1016/j.jhazmat.2025.139020\u003c/li\u003e\n\u003cli\u003eWang, G., Li, Y., Pan, X., Li, A., Wang, J., Yin, L., Zeng, X., Qian, X., 2026. Soil Properties and Microbial Community Assemblages in Response to Plastic Film Mulches with Divergent Degradation Characteristics. Microorganisms 14, 553. https://doi.org/10.3390/microorganisms14030553\u003c/li\u003e\n\u003cli\u003eWang, H., Yang, Q., Li, D., Wu, J., Yang, S., Deng, Y., Luo, C., Jia, W., Zhong, Y., Peng, P., 2023. Stable Isotopic and Metagenomic Analyses Reveal Microbial-Mediated Effects of Microplastics on Sulfur Cycling in Coastal Sediments. Environ. Sci. Technol. 57, 1167\u0026ndash;1176. https://doi.org/10.1021/acs.est.2c06546\u003c/li\u003e\n\u003cli\u003eWang, S.Y., Galletta, G.J., Camp, M.J., Kasperbauer, M.J., 1998. Mulch types affect fruit quality and composition of two strawberry genotypes. HortScience 33, 636\u0026ndash;640.\u003c/li\u003e\n\u003cli\u003eWang, X., Shrestha, S., Tymon, L., Zhang, H., Miles, C., DeVetter, L., 2022. Soil-biodegradable mulch is an alternative to non-biodegradable plastic mulches in a strawberry-lettuce double-cropping system. Front. Sustain. Food Syst. 6, 942645. https://doi.org/10.3389/fsufs.2022.942645\u003c/li\u003e\n\u003cli\u003eWang, Z., Wang, S., Bian, T., Wang, T., Fu, H., Sun, Z., 2024. Revealing the Response of Cucumber Soil Microbial Community Composition and Function to Nitrogen Addition in Northern Chinese Greenhouses. Horticulturae 10, 1090. https://doi.org/10.3390/horticulturae10101090\u003c/li\u003e\n\u003cli\u003eWang, Z.-H., Gao, S.-S., Yang, L., Meng, Y.-L., Wang, M., Li, B.-L.L., Chen, Z.-J., 2026. Responses of Sorghum Growth and Rhizosphere\u0026ndash;Plastisphere Microbiomes to Cadmium and Polypropylene Microplastic Co-Contamination. Agronomy 16, 293. https://doi.org/10.3390/agronomy16030293\u003c/li\u003e\n\u003cli\u003eWickham, H., 2016. ggplot2, Use R! Springer International Publishing, Cham. https://doi.org/10.1007/978-3-319-24277-4\u003c/li\u003e\n\u003cli\u003eWu, C., Ma, Yajie, Wang, D., Shan, Y., Song, X., Hu, H., Ren, X., Ma, X., Cui, J., Ma, Yan, 2022. Integrated microbiology and metabolomics analysis reveal plastic mulch film residue affects soil microorganisms and their metabolic functions. Journal of Hazardous Materials 423, 127258. https://doi.org/10.1016/j.jhazmat.2021.127258\u003c/li\u003e\n\u003cli\u003eWu, C., Song, X., Wang, D., Ma, Yajie, Shan, Y., Ren, X., Hu, H., Cui, J., Ma, Yan, 2024. Combined effects of mulch film-derived microplastics and pesticides on soil microbial communities and element cycling. Journal of Hazardous Materials 466, 133656. https://doi.org/10.1016/j.jhazmat.2024.133656\u003c/li\u003e\n\u003cli\u003eXu, Z., Zhang, L., Jiang, G., Ding, X., Guo, Y., Tian, Y., 2025. Degradation of mulch films in different soils and its effects on soil properties and ecotoxicology. Environ Geochem Health 47, 345. https://doi.org/10.1007/s10653-025-02651-1\u003c/li\u003e\n\u003cli\u003eXu, Z., Zheng, B., Yang, Yichen, Yang, Yi, Jiang, G., Tian, Y., 2024. Effects of biodegradable (PBAT/PLA) and conventional (LDPE) mulch film residues on bacterial communities and metabolic functions in different agricultural soils. Journal of Hazardous Materials 472, 134425. https://doi.org/10.1016/j.jhazmat.2024.134425\u003c/li\u003e\n\u003cli\u003eZang, H., Blagodatskaya, E., Wang, J., Xu, X., Kuzyakov, Y., 2017. Nitrogen fertilization increases rhizodeposit incorporation into microbial biomass and reduces soil organic matter losses. Biology and Fertility of Soils 53, 419\u0026ndash;429.\u003c/li\u003e\n\u003cli\u003eZhang, H., Shu, D., Wang, K., Liu, X., Wang, L., Jiang, R., 2026. Biodegradable film mulching alters soil C, N, P and S cycling via mediating microbial communities in dryland. J Sci Food Agric 106, 632\u0026ndash;642. https://doi.org/10.1002/jsfa.70193\u003c/li\u003e\n\u003cli\u003eZhang, H., Shu, D., Zhang, J., Liu, X., Wang, K., Jiang, R., 2024. Biodegradable film mulching increases soil microbial network complexity and decreases nitrogen-cycling gene abundance. Science of The Total Environment 933, 172874. https://doi.org/10.1016/j.scitotenv.2024.172874\u003c/li\u003e\n\u003cli\u003eZhang, M., Xue, Y., Jin, T., Zhang, K., Li, Z., Sun, C., Mi, Q., Li, Q., 2022. Effect of Long-Term Biodegradable Film Mulch on Soil Physicochemical and Microbial Properties. Toxics 10, 129. https://doi.org/10.3390/toxics10030129\u003c/li\u003e\n\u003cli\u003eZhang, S., Wang, Y., Sun, L., Qiu, C., Ding, Y., Gu, H., Wang, L., Wang, Z., Ding, Z., 2020. Organic mulching positively regulates the soil microbial communities and ecosystem functions in tea plantation. BMC Microbiol 20, 103. https://doi.org/10.1186/s12866-020-01794-8\u003c/li\u003e\n\u003cli\u003eZhang, Y., Gao, W., Mo, A., Jiang, J., He, D., 2022. Degradation of polylactic acid/polybutylene adipate films in different ratios and the response of bacterial community in soil environments. Environmental Pollution 313, 120167. https://doi.org/10.1016/j.envpol.2022.120167\u003c/li\u003e\n\u003cli\u003eZhao, Y., Mao, X., Li, S., Huang, X., Che, J., Ma, C., 2023. A Review of Plastic Film Mulching on Water, Heat, Nitrogen Balance, and Crop Growth in Farmland in China. Agronomy 13, 2515. https://doi.org/10.3390/agronomy13102515\u003c/li\u003e\n\u003cli\u003eZhao, Y., Zhai, X., Wang, Z., Li, H., Jiang, R., Lee Hill, R., Si, B., Hao, F., 2018. Simulation of soil water and heat flow in ridge cultivation with plastic film mulching system on the Chinese Loess Plateau. Agricultural Water Management 202, 99\u0026ndash;112. https://doi.org/10.1016/j.agwat.2018.02.017\u003c/li\u003e\n\u003cli\u003eZhao, Z., Wu, H., Jin, T., Liu, H., Men, J., Cai, G., Cernava, T., Duan, G., Jin, D., 2023. Biodegradable mulch films significantly affected rhizosphere microbial communities and increased peanut yield. Science of The Total Environment 871, 162034. https://doi.org/10.1016/j.scitotenv.2023.162034\u003c/li\u003e\n\u003cli\u003eZheng, B.-X., Zhao, Y., Bi, Q.-F., Zhou, G.-W., Wang, H.-J., Hao, X.-L., Ding, K., 2019. How to disentangle microbially functional complexity: an insight from the network analysis of C, N, P and S cycling genes. Science Bulletin 64, 1129\u0026ndash;1131. https://doi.org/10.1016/j.scib.2019.06.011\u003c/li\u003e\n\u003cli\u003eZhou, J., Jia, R., Brown, R.W., Yang, Y., Zeng, Z., Jones, D.L., Zang, H., 2023. The long-term uncertainty of biodegradable mulch film residues and associated microplastics pollution on plant-soil health. Journal of Hazardous Materials 442, 130055. https://doi.org/10.1016/j.jhazmat.2022.130055\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Fragaria x ananassa, mulching practices, microbial diversity, yield and fruits quality, soilless cultivation","lastPublishedDoi":"10.21203/rs.3.rs-9428619/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9428619/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground and Aim.\u003c/h2\u003e \u003cp\u003eTraditional plastic mulching provides clear benefits for strawberry cultivation but pose risks of long-term environmental pollution. Besides, a comprehensive investigation on how different mulch materials may impact the overall strawberry cultivation, from crop productivity and quality to soil health, is still lacking. With this aim, this study investigates the influence of conventional and biodegradable mulching films on plant performance, fruit quality, and substrate microbial ecology in a strawberry cultivation.\u003c/p\u003e\u003ch2\u003eMethods.\u003c/h2\u003e \u003cp\u003eFour different mulch films were applied and compared to unmulched control: polyethylene (PE), oxo-degradable (OxoPE), polybutylene adipate terephthalate corn starch-blended (PBAT), polypropylene (PP). Assessments included plant morphological and physiological analysis, strawberries yield and quality, substrate physicochemical analysis and substrate bacterial community characterization by high throughput sequencing of the 16S rRNA gene.\u003c/p\u003e\u003ch2\u003eResults.\u003c/h2\u003e \u003cp\u003eOxoPE and PP significantly increased fruit yield up to 29% compared to the control, while physiological and growth traits remained unvaried. Fruit quality and nutraceutical properties were consistent across treatments. PBAT promotes nitrogen enrichment, whereas PE is associated with an increase in available micronutrients. Mulching enhanced bacterial community evenness and diversity, and community structural analysis revealed that each material recruits unique microbial niches leading to distinct bacterial community structures driven by humidity and nitrate gradients. Functional predictions suggest that PE mulches boost nitrogen-fixing communities, whereas PBAT promotes nitrification and complex carbon catabolism.\u003c/p\u003e\u003ch2\u003eConclusion.\u003c/h2\u003e \u003cp\u003ePBAT and OxoPE represent effective alternatives to traditional PE, preserving high productivity and fruit quality while promoting specialized substrates bacterial communities, thus enhancing overall sustainability within strawberry production frameworks.\u003c/p\u003e","manuscriptTitle":"Impact of traditional and biodegradable mulching films on plant performance and substrate microbial communities in a small-scale strawberry cultivation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-24 09:24:02","doi":"10.21203/rs.3.rs-9428619/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0ab5b125-2f49-4aca-bb84-91388c81408b","owner":[],"postedDate":"April 24th, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Reject after review","date":"2026-05-17T04:46:21+00:00","index":"","fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-05-17T08:46:43+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-24 09:24:02","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9428619","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9428619","identity":"rs-9428619","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}