Science of the Total Environment 871 (2023) 162034

Contents lists available at ScienceDirect

Science of the Total Environment

journal homepage: www.elsevier.com/locate/scitotenv

Biodegradable mulch films significantly affected rhizosphere microbial

communities and increased peanut yield
Zhirui Zhao a,1, Haimiao Wu a,b,1, Tuo Jin c,d,1, Huiying Liu e, Jianan Men b, Guangxing Cai e, Tomislav Cernava f,
⁎
Guilan Duan b,g, Decai Jin b,g,
a
Hebei Province Key Laboratory of Sustained Utilization and Development of Water Recourse, School of Water Resources and Environment, Hebei GEO University, Shijiazhuang
050031, China
b
Key Laboratory of Environmental Biotechnology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China
c
College of Resources and Environment, Hunan Agricultural University, Changsha 410128, China
d
Rural Energy and Environment Agency, Ministry of Agriculture and Rural Affairs, Beijing 100125, China
e
Liaoning Academy of Agricultural Sciences, Shenyang 110161, China
f
Institute of Environmental Biotechnology, Graz University of Technology, Petersgasse 12, 8010 Graz, Austria
g
College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049, China

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• BDMs promoted the growth and develop-

ment of peanuts.
• BDMs increased bacterial richness and di-
versity of rhizosphere soil.
• BDMs increased microbial interactions in
the rhizosphere.
• Plastic film altered microbial community
composition and enriched specific taxa.
• Impacts of plastic film on bacterial com-
munities in agricultural fields varied de-
pendent on their type.

A R T I C L E I N F O A B S T R A C T

Editor: Zhaozhong Feng Biodegradable mulch films are widely used to replace conventional plastic films in agricultural fields. However, their
ecological effects on different microbial communities that naturally inhabit agricultural fields are scarcely explored.
Keywords: Herein, differences in bacterial communities recovered from biofilms, bulk soil, and rhizosphere soil were compara-
Plastic film
tively assessed for polyethylene film (PE) and biodegradable mulch film (BDM) application in peanut planted fields.
Agricultural fields
The results showed that the plastic film type significantly influenced the bacterial community in different ecological
Bacterial communities
Network analysis
niches of agricultural fields (P < 0.001). Specifically, BDMs significantly increased the diversity and abundance of bac-
Biomarkers teria in the rhizosphere soil. The bacterial communities in each ecological niche were distinguishable from each other;
bacterial communities in the rhizosphere soil showed the most pronounced response among different treatments.
Acidobacteria and Pseudomonas were significantly enriched in the rhizosphere soil when BDMs were used. BDMs
also increased the rhizosphere soil bacterial network complexity and stability. The enrichment of beneficial bacteria
in the rhizosphere soil under BDMs may also have implications for the observed increase in peanut yield. Deepening
analyses indicated that Pseudoxanthomonas and Glutamicibacter are biomarkers in biofilms of PE and BDMs respec-
tively. Our study provides new insights into the consequences of the application of different types of plastic films on
microbial communities in different ecological niches of agricultural fields.

⁎ Corresponding author at: Key Laboratory of Environmental Biotechnology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
E-mail address: dcjin@rcees.ac.cn (D. Jin).
1
These authors contribute equally to the manuscript.

http://dx.doi.org/10.1016/j.scitotenv.2023.162034
Received 17 November 2022; Received in revised form 30 January 2023; Accepted 1 February 2023
Available online 6 February 2023
0048-9697/© 2023 Elsevier B.V. All rights reserved.
Z. Zhao et al.
1. Introduction
Plastic mulch is widely used as an effective agricultural practice to reg-
ulate soil temperature and moisture (Ding et al., 2019), promote crop yields
(Kasirajan and Ngouajio, 2012), and prevent various pests and diseases
(Steinmetz et al., 2016). A recent meta-analysis of 266 published articles re-
lated to plastic films showed that mulching had a significant yield increase
of up to 24 % for most food crops (Gao et al., 2019; Liu et al., 2022). There-
fore, the use of plastic mulch films is an important tool to promote food se-
curity in different regions of the world. However, this also has a less
desirable downside. Plastic film application has steadily increased during
the last years and was followed by the accumulation of large amounts of
plastic residues that can negatively affect crop production and soil proper-
ties. Plastic mulch films are made primarily of polyethylene (PE); and its
stability make it the preferred choice of most plant producers (Kasirajan
and Ngouajio, 2012). The stability of these plastic films greatly exceeds
the growing cycle of crops. However, PE films are often not reused due to
the labor and material required to recycle and clean it. Moreover, due to
practical reasons mulch films often cannot be completely removed from
fields, leading to the accumulation of plastic residues in the soil. It has
been found that when the accumulation of plastic residues in the soil ex-
ceeds 240 kg/ha, crop yield and soil properties such as water use efficiency
are significantly reduced while the soil microbial activity is also negative af-
fected (Qi et al., 2020).
To circumvent the disadvantages of PE and reduce soil contamination
caused by the use of mulch, biodegradable mulch films (BDMs) have
emerged as an alternative for conventional films (Sander, 2019). Currently,
global bioplastic production is growing rapidly and could reach 2.62 mil-
lion tons by 2023 (Chinthapalli et al., 2019). In theory, BDMs can be con-
verted into non-toxic compounds such as carbon dioxide or methane, and
microbial biomass via microbial metabolism (Gross and Kalra, 2002),
greatly reducing the threat to the environment. This is because BDMs are
generally composed of biodegradable polymers such as polylactic acid
(PLA), poly-3-hydroxybutyrate, and poly-(butylene adipate-co-terephthal-
ate) (PBAT) or natural substrates such as cellulose (Brodhagen et al.,
2015). Since blends of PBAT and PLA significantly increases material prop-
erties, the BDMs made from the composite of these two materials are more
durable and more popular in practical applications. It was observed that
certain BDMs do not undergo complete decomposition within the expected
time after the crop growth cycle (Li et al., 2014) and they may thus modify
soil properties. Therefore, the degradation of biodegradable mulch has
attracted the attention of many researchers. Griffin-LaHue et al. (2022)
assessed the degradation of different types of biodegradable mulch and cal-
culated according to their model that it would take 21–58 months to de-
grade 90 % of the mulch. Meanwhile, in another study, a biodegradable
mulch consisting of a PBAT-PLA mixture still had undegraded parts after
36 months of use in the field (Sintim et al., 2020). The short-and long-
term ecological impacts of BDMs on agroecosystems remain largely un-
known (Sintim and Flury, 2017).
In terms of the impact of mulch films on microbial communities, it was
already shown that PE reduces the activity and α-diversity of soil bacteria
and negatively affects the stability and complexity of microbial networks
(Fei et al., 2020; Shi et al., 2020). On the other hand, BDMs can be utilized
as carbon source by specific microorganisms, which can result in composi-
tional changes within exposed bacterial communities and enrichment of
specific taxa (Feng et al., 2022b). In contrast to this observation, Chen
et al. (2020) reported that the accumulation of BDMs residues in soil
does not affect the structural composition or related ecosystem func-
tions and processes. Targeted experiments are therefore needed to deci-
pher the effect of long-term or short-term use of specific BDMs on
microbial communities naturally occurring in agricultural fields and
soil-plant systems.
Previous studies have mostly focused on microbial communities on
plastic films in aquatic environments (Amaral-Zettler et al., 2020). These
studies indicated that the compositions of the bacterial communities differ
between plastic debris in aquatic environments and the surrounding water
2
Science of the Total Environment 871 (2023) 162034
(Zettler et al., 2013). In contrast, only a few studies have targeted microor-
ganisms on plastic films in soil. Plastic films provide a durable substrate for
soil microbes that can be colonized by microorganisms and support the de-
velopment of microbial biofilms (Zhou et al., 2021). Effects of plastic films
on soil microbial communities were primarily assessed in simulated pot or
and shake flask experiments (Hu et al., 2022; Ju et al., 2021). Less studies
were conducted as field experiments, and they mostly targeted surface
bulk soil with less emphasis on the plant rhizosphere (Liu et al., 2021;
Wang et al., 2022). It was found that the addition of plastic residues had
no significant effect on the diversity, structure and biomass of the bulk
soil bacterial communities (Brown et al., 2022). Deepening studies on
biofilms attached to the surface of agricultural mulch films and the micro-
bial communities in mulch-covered soil could provide the means to better
understand the ecological implications of the underlying agricultural
practices.
In this study, a field experiment was designed where peanut planted
fields were covered with PE film and a PBAT-PLA composite BDMs,
which are representative materials for conventional and biodegradable
mulch films currently used in agriculture, respectively. The plastic film ap-
plication covered the entire peanut growing cycle and sampling of mulch
biofilms, bulk soil, and rhizosphere soil were conducted during the harvest
period. The main objectives of this study were as follows: (1) to investigate
the effects of different types of plastic mulch films on peanut yield, bacterial
diversity and community structure in different ecological niches of agricul-
tural fields; (2) to reveal biomarkers to distinguish between biofilms found
on PE and BDMs; and (3) to explore microbial co-occurrence patterns and
key species responses to different types of plastic mulch films. We hypoth-
esized that the bacterial community composition, function, and network
structure of the different compartments would be significantly altered by
plastic films and that the effects of mulching with BDMs would be distin-
guishable from conventional methods.
2. Materials and methods
2.1. Sample collection
The experiment field was located at an agricultural experiment station
in Linghai County, Liaoning Province. The sample site has a temperate
monsoonal continental climate. The annual rainfall in 2021 reached
895.7 mm.
A total of five field plot experiments were established, each of approxi-
mately 60 m2 (Fig. S1): CK (no plastic film cover), area covered with con-
ventional polyethylene film (PE) and three areas covered with three
different thicknesses (QT2: 0.008 mm, QT4: 0.010 mm, and QT3: 0.012
mm) of biodegradable mulch films (BDMs), one of which was the same
thickness as the PE film (Table S1). All sample areas were planted with pea-
nuts (Arachis hypogaea L. variety Tangke 8252) on 8 May 2021 (after the ap-
plication compound fertilizer for peanuts) and mulch and soil samples were
collected during the peanut harvest in September 2021.
Six sampling points were randomly selected in each area to collect pea-
nut roots by using sterile scissors and tweezers and then transferred into 50
mL sterile centrifuge tubes: The subsequent processing steps were carried
out in the laboratory. Briefly, phosphate-buffer (PBS) was added to each
tube and shaken and sonicated, then centrifuged and the resulting pellet
was frozen and stored at −80 °C (Li et al., 2021). Surface soil (depth:
0–20 cm) at least 20–50 cm away from the plant was obtained for the
bulk soil analyses after removal of visible plant and animal residues and
gravel. In this study, “biofilm” refers to the organized microbial aggregates
formed by microorganisms on the surface of plastic film (Zhou et al., 2021).
Biofilms from mulching films were collected according to the method de-
scribed in a previous study (Tarafdar et al., 2021). Briefly, the collected
mulching film was firstly washed three times with sterilized 0.85 % NaCl
solution to remove loosely bound microbes, and then was cut into small
pieces using sterilized scissors for total DNA isolation from the attached bio-
film. In total three sample types were obtained for each treatment: biofilm,
rhizosphere soil, and bulk soil. Part of the bulk soil samples was stored at
Z. Zhao et al.
4 °C for the determination of physical and chemical properties, whiles
samples for DNA extraction were stored at −80 °C.
2.2. Determination of soil physical and chemical parameters and crop yield
Various physical and chemical indicators such as moisture content
(MC), pH, organic Matter (OM), total nitrogen (TN), and total phosphorus
(TP) were evaluated by the national standards of the People's Republic of
China. Specifically, the soil samples were freeze-dried to determine mois-
ture content using a vacuum freeze-dryer (FD-1B-50, Beijing Baku Experi-
mental Instruments Co.); soil pH was determined at a soil-to-water ratio
of 1:2.5 with a pH meter; organic matter was determined via the potassium
dichromate oxidation method; TN and TP were measured using Kjeldahl
method and digestion method, respectively (Kong et al., 2019). Four
months after seeding, plants in each plot were harvested for yield. Har-
vested material was classified as below-ground peanut grain yield and
above-ground straw fresh weight.
2.3. DNA extraction, PCR, and sequencing
Total bulk and rhizosphere soil genomic DNA as well as biofilms DNA
were extracted by using the Fast DNA spin kit for soil (MP Biomedicals
LLC, USA) from 0.5 g dry soil and 2 mm × 2 mm plastic film fragments, re-
spectively. DNA quality and concentration were assessed based on absor-
bance ratios of 260/280 nm (1.8–2.0) and 260/230 nm (>1.7) as
detected by a NanoDrop Spectrophotometer (Nano-100, Aosheng Instru-
ment Co. Ltd., Hangzhou, China). The V4 region of bacterial 16S rRNA
genes was amplified using the primer pair 515F (5′-GTGCCAGCMGCCG
CGGTAA-3′)/806R (5′-GGACTACHVGGGTWTCTAAT-3′). A unique
barcode sequence (12 bp) was used to distinguish among samples. The 50
μL PCR mixture contained 5 μL 10× PCR Buffer, 4 μL dNTPs, 1 μL forward
and reverse primers (10 μM), 0.5 μL Taq DNA polymerase (TaKaRa), 1 μL
template DNA, and 37.5 μL ddH2O. The PCR conditions were as follows:
94 °C for 1 min, 30 cycles of 94 °C for 20 s, 57 °C for 25 s, 72 °C for 45 s,
and a final extension at 72 °C for 10 min. The PCR products were purified
with E.Z.N.A.™ Gel Extraction Kit (Omega Bio-tek, Norcross, GA, USA).
Samples with different barcodes and with equal molar mass were pooled
to create a single MiSeq sequencing library at Guangdong Magigene Bio-
technology Co. Ltd. (Shenzhen, China). Additionally, the copy numbers of
bacterial 16S rRNA gene fragments in all biofilm and soil samples were
quantified via quantitative PCR (qPCR) using primer pairs 515F/806R, re-
spectively on the CFX96 Touch Real-Time PCR Detection System (BioRad,
California, USA), the detailed procedure and condition were selected ac-
cording to Kong et al. (2019).
2.4. Bioinformatic and statistical analyses
Sequencing data were analyzed with a publicly accessible Galaxy pipe-
line (http://mem.rcees.ac.cn:8080) (Feng et al., 2017). In brief, raw se-
quences were split into samples according to their specific barcodes.
Thereafter, sequences with low quality (QC < 20) and invalid length
(<245 bp or >260 bp) were deleted. UPARSE (Edgar, 2013) was used to
Table 1
Soil physical and chemical properties and peanut yield.
Treatment CK PE
pH 7.96 ± 0.04a 7.77 ± 0.08b
MC (%) 22.45 ± 1.25a 22.27 ± 0.31a
OM (‰) 13.98 ± 2.68a 12.97 ± 5.48a
TN (mg/kg) 2046.31 ± 99.09b 2425.89 ± 153.02a
TP (mg/kg) 345.99 ± 36.17a 345.72 ± 82.14a
Humidity (%) 40.78 ± 1.47c 44.12 ± 1.07a, b
Temperature (°C) 22.15 ± 1.36a 22.94 ± 1.68a
Peanut yield (kg/667 m2) 169.68 ± 28.38b 74 ± 13.47c
Strawweight (kg/667 m2) 481.32 ± 67.94b 351.55 ± 59.89c
Different letters indicate that there are differences between the treatment groups of the same physical and chemical factor (P < 0.05).
3
Science of the Total Environment 871 (2023) 162034
remove chimeras and classify the sequences into operational taxonomy
units (OTUs) at a similarity of 97 %.The OTU table consisting of 24 biofilm
samples and 59 soil samples was resampled with 34,846 sequences to guar-
antee the same sequencing depth for further statistical analyses. The se-
quence data were deposited in the National Center for Biotechnology
Information (NCBI) database under the project number PRJNA875578.
Microbial α-diversity was estimated using the Shannon Index and
Chao1 values. The significance of differences for soil physicochemical var-
iable, peanut yield and α-diversity were determined using One-way
ANOVA between multiple groups. Non-metric multidimensional scaling
(NMDS) analysis based on Bray-Curtis distances was used to visualize
changes in community composition among multiple treatments and sample
types. Additionally, the significance of the effects of multiple factors on
the microbial community structure was analyzed by permutational multi-
variate analysis of variance (MRPP). Moreover, the SourceTracker was
used to estimate the proportion of the microbiome transferred between
the mulch film biofilms and soil compartments. This tool is can be specifi-
cally applied to estimate the proportion of a given microbiome that origi-
nates from a set of source-sink environments (Abdelfattah et al., 2019;
Knights et al., 2011). Function annotation of the 16S rRNA gene fragment
dataset was performed with Functional Annotation of Prokaryotic Taxa
(FAPROTAX) (Louca et al., 2016). All the above analyses were carried out
in the aforementioned pipeline (http://mem.rcees.ac.cn:8080).
To explore microbial interactions under different treatments in each
sample type, phylogenetic molecular networks (MENs) were constructed
by using another online pipeline (http://mem.rcees.ac.cn:8081) (Feng
et al., 2022a). Only OTUs that were present in 80 % of the samples in one
group were included in the analysis. The Pearson coefficient was used to
calculate correlation, and the random matrix theory (RMT)-based approach
was applied to delineate the network interactions among different bacteria
groups. The constructed networks were visualized with Gephi 0.9.4.
Network properties including power-law model fitness, average connec-
tivity (avgK), harmonic geodesic distance (HD), average clustering coeffi-
cient (avgCC), and modularity were calculated(He et al., 2021). Keystone
taxa also were assigned based on within-module connectivity (Zi) and
among-module connectivity (Pi) according to a previously described
method (Deng et al., 2012). Nodes (OTUs) were classified into four catego-
ries: (1) Peripheral (Zi ≤ 2.5, Pi ≤ 0.62), indicates nodes interconnected
within modules with a few links; (2) Connectors (Zi ≤ 2.5, Pi > 0.62), indi-
cates nodes linking to several modules; (3) Module hubs (Zi > 2.5, Pi ≤
0.62), indicates nodes highly connected within modules; and (4) Network
hubs (Zi > 2.5, Pi > 0.62), indicates nodes highly connected among the net-
work.
3. Results
3.1. Influence of plastic mulch film on the crop yield and soil physicochemical
properties
We evaluated peanut yield and multiple soil physical and chemical
properties when different types of plastic mulching films were used
(Table 1). Under the two film types (PE and BDMs), pH, TN, humidity
QT2 QT4 QT3
7.76 ± 0.08b 7.84 ± 0.11b 7.75 ± 0.08b
22.82 ± 0.38a 22.33 ± 0.44a 22.89 ± 0.15a
13.02 ± 1.59a 10.8 ± 2.14a 14.64 ± 1.37a
2539.82 ± 101.44a 2557.67 ± 195.65a 2028.37 ± 225.78b
354.83 ± 27.88a 332.25 ± 53.80a 329.88 ± 24.32a
42.15 ± 1.14b, c 42.26 ± 1.67b, c 44.78 ± 1.88a
22.3 ± 0.97a 22.32 ± 1.19a 22.8 ± 2.34a
260.77 ± 22.40a 234.72 ± 14.50a 172.95 ± 17.82b
758.19 ± 96.57a 675.21 ± 20.83a 491.91 ± 86.24b
Z. Zhao et al.
changed significantly (P < 0.05) between the treatment and the control
group. Temperature also increased in the treatment group compared with
the control group. The soil with no plastic film had the highest pH value
7.96 (±0.04). TN content was increased by both PE and BDMs, with the
highest increment (25.0 %) in the QT4 treatment (middle thickness of
BDMs foil used in this study). The temperature and humidity under the
PE treatment were higher than in the other treatment groups. MC, OM,
and TP were not significantly changed by all the treatments.
Peanut yield and strawweight showed significant differences between
treatments after plastic film applications (Table 1). In particular, the control
group had a raw yield of 169.68 (±28.38) kg/667 m2, BDMs of each thick-
ness increased the peanut yield and the growth rate increase was the
highest, reaching 53.7 %. In contrast, PE application resulted in a decrease
in peanut yields. The strawweight showed the same trend as the peanut
yield, and a significant negative correlation with temperature and humidity
was observed (Table S2).
3.2. Changes in bacterial community composition and total bacterial counts
To evaluate the influence of various types of plastic mulch film on the
bacterial biomass in the distinct sample types of the agricultural fields,
qPCR was used to analyze the differences in bacterial gene copies, as
shown in (Fig. S2). According to the qPCR results, we found that the num-
ber of total bacteria was highest in BDMs treatments in ecological niches,
however, it was not significant.
Two α-diversity indices, including the Shannon index and Chao1 index
were calculated (Fig. 1.a–f). We observed that the α-diversity index varied
between different sample types. The plastic film altered bacterial α-
diversity in two compartments of the soil (Shannon and Chao1). All treat-
ment groups (PE and BDMs) resulted in a significant reduction the Chao1
index in the bulk soil while the Shannon index decreased significantly
only in the BQT2 group. The Shannon and Chao1 indices of the rhizosphere
soil showed a significant increase in the BDMs groups and reached the
Fig. 1. Assessment of the diversity of bacterial communities in different samples. a–f: Alpha diversity analysis (Shannon index and Chao1); h–g: Non-Metric Multidimensional
Scaling (NMDS) analysis between treatments based on Bray-Curtis distance matrix (M: biofilm; B: bulk soil; R: rhizosphere; Different letters indicate that there are differences
between the treatment groups (P < 0.05)).
4
Science of the Total Environment 871 (2023) 162034
highest level there, while PE slightly reduced the Chao1 index when com-
pared with the control group.
Microbial communities were compared for their composition across the
three compartments of agricultural fields using a NMDS approach. Bacterial
communities were discriminative according to the environmental niches
that were analyzed (Fig. S3). In detail, bacterial communities from biofilms
were discriminative from those of bulk soil and rhizosphere soil along the
MDS1 and MDS2 axes whereas bulk soil and rhizosphere bacterial commu-
nities were discriminated from one another only along the MDS2 axis. Ad-
ditionally, the distances among samples from the same type were also
significantly different, indicating different levels of stress under the plastic
films. MRPP analysis showed a significant distinction (P = 0.001) between
bacteria colonizing on the two types of plastic films (Fig. 1.h).
Bulk soil and rhizosphere soil were significantly different in the effects
caused by plastic films. Compared with the control group, the distance of
the bulk soil bacterial community in the treatment groups was significantly
reduced, especially in the PE group. However, compared with the PE group,
the distance of the bacterial community distribution in the BDMs group in-
creased along MDS2 (P = 0.001) (Fig. 1.i). The rhizosphere soil compart-
ment had a dissimilarity results (P = 0.001). Compared to the control
group, PE increased the distance of bacterial communities in the rhizo-
sphere soil, but the BDMs resulted in a smaller distance of bacterial commu-
nities in the rhizosphere soil (Fig. 1.g).
The analysis of the correlation between the bacterial community and
environmental factors indicated that the changes within bacterial
community were positively correlated with soil temperature and moisture.
However, the changes of bacterial diversity were not affected by environ-
mental factors (Table S3, S4).
3.3. Differences in bacterial community composition after different treatments
The relative abundances (RA) of bacterial OTUs in the different treat-
ments are visualized in Fig. 2. The top 15 phyla and genera were selected
Z. Zhao et al. Science of the Total Environment 871 (2023) 162034

Fig. 2. Taxonomic composition of bacterial communities and their relative abundance at phylum (a) and genus (b) levels (CK: control group; PE: polyethylene plastic film;
QT2, QT4, QT3: three different thicknesses of BDMs).

to display the taxonomic composition of the bacterial community. Across
all samples, the dominant phyla (RA > 10 %) consisted of Proteobacteria,
Bacteroidetes, Actinobacteria, Acidobacteria, and Cyanobacteria. Never-
theless, the type of plastic film had a highly significant effect on the compo-
sition of bacterial communities in different sample types. In biofilms, the
relative abundance of Proteobacteria, Bacteroidetes, and Actinobacteria
colonized on PE (43.51 %, 18.75 %, and 12.74 %) was higher than that
on BDMs (26.92 %, 9.64 %, and 7.27 %); Cyanobacteria was the most abun-
dant phylum on BDMs (33.72 %). Plastic film application decreased the rel-
ative abundance of Bacteroidetes in bulk soil, while the decrease degree in
the BDMs treatments was much lower than that in the PE group; but the rel-
ative abundance of Acidobacteria and Chloroflexi in the PE group was
significantly increased. However, BDMs applications significantly in-
creased the abundance of Acidobacteria in rhizosphere soil.
Unlike the biofilm and bulk soil samples, the rhizosphere soil showed
higher variations at the genus level between treatments, and was dominated
by Pseudomonas, Flavobacterium, and Nitrososphaera. After plastic film applica-
tion, the relative abundance of Pseudomonas decreased while Nitrososphaera
increased significantly, and both of them were enriched in the rhizosphere
soil of BDMs. However, the relative abundance of a few genera showed differ-
ent trends. For example, the relative abundance of Flavobacterium increased
significantly in the PE group and slightly decreased in the BDMs groups.
Linear discriminant analysis effect size (LEfSe) analysis performed on
biofilm microbiomes revealed that 111 biomarkers were differentially

Fig. 3. SourceTracker and LEfSe analyses. SourceTracker analyses were conducted to determine the origin of (a) PE film and (b) BDMs colonizing bacteria; (c) LEfSe analysis
to identify differentially abundant taxa between two types of the plastic film (P < 0.05; LDA score 3.1).

5
Z. Zhao et al.
abundant between biofilm microbiomes (Fig. 3c). The bacteria with the
highest LDA scores (LDA > 3.1, P < 0.05) under PE and BDMs are visualized
in Fig. S4. Xanthomonadaceae and Pseudoxanthomonas were the main mi-
crobial taxa at the class level that were enriched in biofilms colonized on
PE. On BDMs, the characteristic taxa strains were Armatimonadetes at the
phylum level and Glutamicibacter at the genus level.
3.4. SourceTracker analysis of colonizing microbiota on mulch films
The SourceTracker tool was employed to study the microbiome that was
potentially transferred between the biofilm on different mulch films and
soil compartments. For each treatment group, bulk soil and rhizosphere
soil were set as sources, while the biofilm was set as a sink. The results re-
vealed that low proportions of the microbiome were likely transferred
from soil to the biofilm via the rhizosphere in the BDMs groups (8.81 %).
Moreover, direct microbiome transfer from bulk soil to the biofilm rela-
tively higher (19.60 %) (Fig. 3b). In contrast, a major fraction of the
microbiome transfer identified in the PE group was from rhizosphere soil
to the biofilm, with only a few microbiome members transferred from
bulk soil to the biofilm (14.70 %) (Fig. 3a). The above-mentioned results in-
dicate that biofilms forming on different types of plastic films have different
sources of bacteria that enrich on them during their use, but the majority of
bacteria on plastic films are spontaneously formed from other sources than
soil and plant rhizosphere.
3.5. Impact of plastic films on bacterial functions
To further explore the functional classification in different samples after
using various plastic films, FAPROTAX was carried out and all functional
predictions of the biofilm and soil samples were presented (Fig. 4). The
Fig. 4. Heatmap visualization of variations in the abundance of biodegradation-relevant functions in different compartments of the farmland across treatments (M: biofilm; B:
bulk soil; R: rhizosphere).
6
Science of the Total Environment 871 (2023) 162034
results indicate that both the ecological niche and plastic films make a sig-
nificant influence on the functional distribution and abundance of bacteria.
According to the 16S rRNA gene sequence annotations, a total of 37
functional groups (relative abundance>1.0 %) were obtained and the func-
tions related to carbon, nitrogen, and pathogens were relatively active. By
comparing the bacterial functions in the biofilm, bulk soil, and rhizosphere
soil, we found that the functions associated with carbon and pathogens
were enriched in biofilms; carbon and nitrogen-related functional taxa
were concentrated in the bulk soil. In the rhizosphere soil, the abundance
of a few functional taxa was high, and mainly assigned to carbon-related
bacterial functions (chemoheterotrophy and methylotrophy) and individ-
ual nitrogen-related functions.
Despite the relatively high abundance of carbon-related and nitrogen-
related functional taxa in samples from biofilms of the PE and BDMs groups,
there was still a clear preference observable in terms of the selection of func-
tional groups. For example, cellulolysis, dark reactions, and urea catabolism
were significantly enriched in PE biofilms than in those of BDMs. Meanwhile,
the pathogen-associated functional taxa including animal parasites or symbi-
onts and human pathogens were all also abundant in PE biofilms.
The use of plastic films significantly increased the abundance of carbon
and nitrogen-related functional taxa. For example, microbial functions
such as methane production (methanogenesis and hydrogenotrophic
methanogenesis) were enriched in bulk soils, especially in the PE group.
This may be due to the molecular structure of PE mulch film, which is
dense and lead to the development of a relatively anaerobic soil environ-
ment. Such environments are conducive to the growth of various anaerobic
bacteria. For rhizosphere soil, plastic films increased the abundance of a
few functional groups performing nitrogen-related functions in this com-
partment, such as nitrate reduction, and nitrogen fixation; and this trend
was more obvious in the BDMs group.
Z. Zhao et al.
3.6. Bacterial interspecies relationships and co-occurrence networks
Co-occurrence network analysis was used to reveal the general influ-
ences of the application of various types of plastic films on bacterial inter-
play in biofilms and different soil compartments (Fig. 5). The topological
parameters of the constructed networks were calculated to explore differ-
ences between treatments and presented in (Table S5). The overall topol-
ogy indices showed that all network connectivity distributions fitted well
with the power-law model (R2 of power law from 0.757 to 0.886), indicat-
ing that most nodes had fewer neighbors while fewer nodes had many
neighbors in a network (scale-free). Significant differences in the co-
occurrence patterns of the biofilm and the different soil compartment com-
munities were observed in the two types of plastic film treatments. Com-
pared with soils, biofilms showed a more complex and tighter network
structure, characterized by a higher average degree (avgK) and clustering
coefficient (avgCC), shorter harmonic geodesic distance (HD), and smaller
modularity. It was also observed that the PE biofilm had a higher network
complexity and tightness than those of the BDMs. Furthermore, a more
complex and tighter bacterial network structure was detected in rhizo-
sphere soil after using plastic film, while the opposite trend was found in
the bulk soil networks. The average degree of the rhizosphere soil network
was higher under BDMs than PE, while little difference was observed in
bulk soil networks under BDMs and PE. The number of nodes and links of
rhizosphere soil in the BDMs group were increased 4.7 times higher than
those in the control group while no significant changes were found in
bulk soils. Together, this indicated that microbial interactions in the biofilm
of the PE group and rhizosphere soil of the BDM groups were more clus-
tered and connected than others, as confirmed by the node-level topologi-
cal features of significantly higher degree and closeness centrality of the
two networks. Most of the keystone taxa belonged to connector hubs and
module hubs. In the biofilm samples, BDMs contained 4 keystone taxa (con-
nector hubs: 3, module hubs: 1) while PE did not contain any keystone taxa.
In the bulk soil compartment, 15 of the 37 keystone taxa were found in the
control group, which harbored the most keystone taxa. In contrast, most of
the 21 key species in the rhizosphere soil compartment were derived from
BDMs (Table S6).
4. Discussion
In this study, 16S rRNA gene fragment sequencing was implemented to
assess the impact of structural composition, function, and co-occurrence
networks of bacterial communities in three ecological niches of agricultural
fields following the application of PE and BDMs. Significant differences in
bacterial communities were observed in biofilm, bulk soil, and rhizosphere
soil among different treatments.
Fig. 5. Co-occurrence networks of bacterial communities in different samples. Mulch films: (a) PE, (b) BDMs (QT4). Bulk soil: (c) CK, (d) PE, (e) BDMs (QT4). Rhizosphere
soil: (f) CK, (g) PE, (h) BDMs (QT4). Different colors indicate different modules.
7
Science of the Total Environment 871 (2023) 162034
4.1. Biodegradable mulch films significantly affect rhizosphere soil bacterial
diversity
The Chao1 index provides an estimation of the numbered species (rich-
ness) within a given environment (Roesch et al., 2007), while the Shannon
index estimates the diversity of microbial communities (Bent and Forney,
2008). In the present study, the abundance and diversity of microbial com-
munity were influenced apparently by the ecological niches in agricultural
fields (Fig. 1). The formation of distinct communities is likely due to the
specificity of the plastic surface layer as a space for microbial colonization.
Previous research has demonstrated that the surface layer of plastic films is
a unique “habitat” that is distinct from the environment and natural sub-
strate (De Tender et al., 2015; Li et al., 2022; Mccormick et al., 2016). At
the same time, Miao et al. (2019) observed that the α-diversity of bacterial
communities on polyethylene and polypropylene microplastics was signifi-
cantly lower than that on natural substrates using experimental conditions.
Huang et al. (2019) also confirmed that the diversity of the microbiota on
plastic fragments was significantly lower than those in bulk soils. The
SourceTracker results obtained in the present study also coincide with the
uniqueness of the microbiota colonizing the plastic film (Du et al., 2021).
It was previously suggested that low microbial diversity in the rhizosphere
may be due to selective changes, such as convergence taking place in the
rhizosphere (Yan et al., 2017). Meanwhile, the accumulation of
microplastics in rhizosphere soil also can change the range and spatial dis-
tribution of hydrolases in the hot zone of rhizosphere soil, and limit the sup-
ply of soil available nutrients and the development of plant roots, and thus
reduce microbial diversity (Tong et al., 2022).
Bacterial diversity and community structure in each ecological niche of
an agricultural field respond differently to plastic film types, which may be
attributed to different polymer components that are used for plastic films.
The PE film is synthesized via the polymerization of ethylene, which has
C\\C backbones. Biodegradable petrochemical-based PBAT as well as
bio-based PLA polymers have oxygen atoms in the backbone, and the pres-
ence of atoms other than C and H is very likely to affect changes in the bac-
terial community (Fei et al., 2020; Seeley et al., 2020). The increase in the
total number and diversity of rhizosphere soil bacteria may be due to the
improved moisture and temperature conditions after mulching which is
also known to stimulated root development and increased root secretion
(Bandopadhyay et al., 2018). Compared to BDMs, PE causes mechanical
blocking of crop roots, which can affected root growth and root cell viabil-
ity, thus impeding root development and reducing rhizosphere microbial
diversity (Kalcikova et al., 2017), which was consistent with the results of
Zhu et al. (2022), who pointed out that PE impaired rhizosphere microbial
diversity more than other plastics. At the same time, the decomposition of
BDMs also increased the input of bioavailable carbon (Sun et al., 2022b)
Z. Zhao et al.
and provided favorable conditions for the enrichment of rhizosphere soil
microorganisms.
4.2. Biodegradable mulch films facilitate the accumulation of beneficial microor-
ganisms
In this study, we assessed the bacterial community composition at dif-
ferent levels, including Proteobacteria, Bacteroidetes, and Acidobacteria,
at the phylum level, and mainly Pseudomonas, Flavobacterium, and
Nitrososphaera, at the genus level, which were all predominant in the ana-
lyzed samples. We found significant differences in species composition
under different treatments in different compartments. The relative abun-
dance of phylum Cyanobacteria phylum was high in the biofilm of BDMs.
Previous studies have reported that Cyanobacteria have a role in maintain-
ing soil fertility (Abinandan et al., 2019) and restoring carbon sequestration
in the soil matrix (Munoz-Rojas et al., 2018). Functional prediction showed
that some carbon-related functional taxa were enriched on BDMs. Micro-
bial communities, particularly in bulk soils in direct contact with plastic
mulch, start to function soon after the application of such films (Serrano-
Ruiz et al., 2021). Previous studies have reported that Proteobacteria,
Actinobacteria, and Acidobacteria, accounting for 67 % of the microbial
community, are the main decomposers of organic matter (Liang et al.,
2020) and that their enrichment accelerates the occurrence of damages of
plastic films. This is most likely the reason why the bulk soil in this study
had a high relative abundance of Proteobacteria, Actinobacteria, and
Acidobacteria. The enrichment of Acidobacteria in the soil might result in
their competitive colonization of the rhizosphere, leading to the establish-
ment of beneficial relationships with plants and thus a large increase in pea-
nut yield (Kalam et al., 2020). In the present study, the abundance of
Pseudomonas and Flavobacterium was higher in the rhizosphere soil follow-
ing BDM applications when compared to PE application. Pseudomonas can
use various substrates as nutrients and survive in different conditions that
would be stressful for other bacteria, can metabolize compounds such as an-
tibiotics, polysaccharides, and iron carriers, and has great potential for sup-
pressing plant diseases and degrading plastic-related substances (Santoyo
et al., 2012; Sun et al., 2022a). Flavobacterium is also a common genus
that includes previously identified biocontrol bacteria and it can degrade
complex organic compounds such as starch and cellulose (Bae et al.,
2018; Kim and Yu, 2020). It is consistent with the functional prediction
for the rhizosphere soil where carbon-related functions were found to be
enriched in this study. The enrichment of Pseudomonas and Flavobacterium
is likely connected to a significant increase in the abundance of carbon
degrading functions in the rhizosphere soil. In addition, the fact that Pseu-
domonas and Flavobacterium can substantially enhance crop resistance to
disease and crop yield in general was also likely reflected by the signifi-
cantly higher crop yields under BDMs treatments.
4.3. The potential sources of biofilm formers and identification of biomarkers of
the biofilm
Using SourceTracker, we found that only a small proportion of the mi-
crobiota in the biofilm compartment was transferred from bulk and rhizo-
sphere soils under different types of plastic mulch treatments, and that
the sources of the biofilm microbiota on PE and BDMs differed (Fig. 3a.
b). It has been previously noted that the highest proportions of microorgan-
isms are found in ripe and immature fruits (Olimi et al., 2022), and as pea-
nuts are typically subterranean-fruiting legumes, there is likely a major
microbial transfer process that moves rhizosphere microorganisms to the
biofilm on the surface during fruit development. All plastic films were
used throughout the peanut planting period, but the BDMs were already
damaged and mixed into the bulk soil at harvest, so a high proportion of
the microbiota in the bulk soil was transferred to the BDMs biofilms. In con-
trast, the PE film did not show such damages after use, so the majority of the
microbiota might have been directly transferred from the rhizosphere.
Combining the SourceTracker results with α-diversity indices, NMDS anal-
yses, and composition results, indicates that plastic films may have a strong
8
Science of the Total Environment 871 (2023) 162034
effect in shaping the composition of the bacterial community. Thus, unique
biomarkers were hypothesized to exist on PE and BDMs.
The family Xanthomonadaceae is a widespread family of bacteria be-
longing to the gamma subdivision of the Gram-negative proteobacteria, in-
cluding the two pathogenic genera Xanthomonas and Xylella (Mhedbi-Hajri
et al., 2011). The results from the LEfSe analysis indicated that
Xanthomonadaceae is a biomarker for enriched bacteria on PE biofilms at
the family level. This may be responsible for the observed enrichment of
the PE biofilms with pathogen-associated functional taxa. At the genus
level, the characteristic genus of the PE biofilm compartment was identified
as Pseudoxanthomonas which can degrade polycarbonate plastics and can
grow and multiply using DEHP as a carbon source (Meng et al., 2015;
Yue et al., 2021). The BDM biofilms enriched Glutamicibacter, which has
been reported to be able to degrade aromatic compounds and similar com-
ponents to those that occur in BDMs (Duraisamy et al., 2020).
4.4. Biodegradable mulch films enhance bacterial interactions in the rhizosphere
compartments
There are often complex association networks among microorganisms.
Such networks are of great significance for understanding microbial com-
munity structures and interactions between the prevalent members (de
Vries et al., 2018; Duran et al., 2018). Our network analysis indicated
that compared to the control group, BDMs significantly increased the
total number of links in the rhizosphere compartments microbial network,
and especially positive links. This indicates that mulching plastic films can
promote cooperation between the occurring bacteria (Fig. 5). Importantly,
the BDMs treatment yielded the most positive links in this study, an out-
come that is perhaps related to root exudates under the cover crop, as bac-
teria usually show rapid responses to rhizosphere exudates (Berg and
Smalla, 2009). The presence of more network hubs indicates more concen-
trated interactions within the microbial community. Thus, the interactions
within the microbial community were more common in the rhizosphere
when BDMs were used. Based on these observations, we propose that
BDMs may promote the formation of a complex bacterial community net-
work (certain topological features such as the numbers of nodes and links
were higher) in rhizosphere soil. Complex networks can activate the plant
immune system and accelerate defense mechanisms against pathogens,
thus favoring the growth of the planted crop. The consistency of network
complexity and peanut yield in this study also provides indications for
this. Previous studies have also shown that a complex rhizosphere network
led to a significant increase in forage height, fresh weight, and dry weight
(Yan et al., 2022).
In addition to analyzing functions of microbial communities, the rela-
tionship between soil parameters and crop yield was also assessed. Changes
in soil temperature and humidity were found to be associated with the
yield, indicating that both of them are important factors affecting peanut
yield. Previously, soil temperature and humidity were reported to be
closely related to plant yield. For example, it was shown that reducing
soil temperature can prolong the growth and reproduction time of crops,
delay premature aging of crops, and improve crop yield (Hu et al., 2020).
Soil moisture is another important soil parameter that affects crop yield
(Acharya et al., 2019). In this study, changes in soil temperature and hu-
midity caused significant changes in peanut yield, indicating that suitable
soil temperature and humidity are beneficial to crop production and prop-
agation.
5. Conclusions
In this study, we systematically investigated the differences in bacterial
community diversity and interactions between a polyethylene plastic film
and biodegradable mulch films in different ecological niches of agricultural
fields. Our results show that there are clear differences in the structure and
composition of the bacterial communities between the two types of plastic
film treatments in the three analyzed niches of the peanut field. BDMs sig-
nificantly increased the bacterial diversity in rhizosphere soil and enriched
Z. Zhao et al.
beneficial bacteria such as Acidobacteria and Pseudomonas in rhizosphere
soil. Network changes were more pronounced in rhizosphere soil, where
BDMs resulted in a more complex and stable bacterial network. The occur-
rence of beneficial bacteria, and a more complex and stable network is con-
ducive to increased peanuts yield. In addition, Pseudoxanthomonas and
Glutamicibacter were the main bacteria taxa attached to PE and BDMs, re-
spectively. The overall results contribute to the deeper understanding of
the impact of plastic films on the microbial ecology of agricultural fields
and provide a reference for the safe use of certain plastic films in the future.
CRediT authorship contribution statement
Zhirui Zhao and Haimiao Wu: Conceptualization, Experiments, Date
analysis, Writing-original draft, Writing-review & editing, Visualization.
Tuo Jin: Investigation, Experiments, Writing-review & editing. Huiying
Liu and Jianan Men: Investigation, Experiments, Date analysis, Writing-
review & editing. Guangxin Cai: Investigation, Experiments. Tomislav
Cernava: Formal analysis, Writing-review & editing. Guilan Duan: Inves-
tigation, Writing-review & editing. Decai Jin: Supervision, Conceptualiza-
tion, Writing-original draft, Writing-review & editing, Funding acquisition.
Data availability
Data will be made available on request.
Declaration of competing interest
The authors declare that they have no known competing financial inter-
ests or personal relationships that could have appeared to influence the
work reported in this paper.
Acknowledgements
This study was financially supported by the National Natural Science
Foundation of China (grant No. 41977122), Agricultural Ecology and Envi-
ronment Protection Project (13210118), and the Science and Technology
Innovation Team Project of Hebei GEO University (KJCXTD-2021-09).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2023.162034.
References
Abdelfattah, A., Sanzani, S.M., Wisniewski, M., Berg, G., Cacciola, S.O., Schena, L., 2019. Reveal-
ing cues for fungal interplay in the plant-air interface in vineyards. Front. Plant Sci. 10, 922.
Abinandan, S., Subashchandrabose, S.R., Venkateswarlu, K., Megharaj, M., 2019. Soil
microalgae and cyanobacteria: the biotechnological potential in the maintenance of soil
fertility and health. Crit. Rev. Biotechnol. 39 (8), 981–998.
Acharya, B.S., Dodla, S., Gaston, L.A., Darapuneni, M., Wang, J.J., Sepat, S., Bohara, H., 2019.
Winter cover crops effect on soil moisture and soybean growth and yield under different
tillage systems. Soil Till. Res. 195, 104430.
Amaral-Zettler, L.A., Zettler, E.R., Mincer, T.J., 2020. Ecology of the plastisphere. Nat. Rev.
Microbiol. 18 (3), 139–151.
Bae, S.S., Kim, M.R., Jung, Y., Yang, S.H., Kwon, K.K., Baek, K., 2018. Flavobacterium
sediminis sp. nov., a starch-degrading bacterium isolated from tidal flat sediment. Int.
J. Syst. Evol. Microbiol. 68 (12), 3886–3891.
Bandopadhyay, S., Martin-Closas, L., Pelacho, A.M., DeBruyn, J.M., 2018. Biodegradable plas-
tic mulch films: impacts on soil microbial communities and ecosystem functions. Front.
Microbiol. 9, 819.
Bent, S.J., Forney, L.J., 2008. The tragedy of the uncommon: understanding limitations in the
analysis of microbial diversity. ISME J. 2 (7), 689–695.
Berg, G., Smalla, K., 2009. Plant species and soil type cooperatively shape the structure and func-
tion of microbial communities in the rhizosphere. FEMS Microbiol. Ecol. 68 (1), 1–13.
Brodhagen, M., Peyron, M., Miles, C., Inglis, D.A., 2015. Biodegradable plastic agricultural
mulches and key features of microbial degradation. Appl. Microbiol. Biotechnol. 99 (3),
1039–1056.
Brown, R.W., Chadwick, D.R., Thornton, H., Marshall, M.R., Bei, S., Distaso, M.A., Bargiela,
R., Marsden, K.A., Clode, P.L., Murphy, D.V., Pagella, S., Jones, D.L., 2022. Field applica-
tion of pure polyethylene microplastic has no significant short-term effect on soil biolog-
ical quality and function. Soil Boil. Biochem. 165, 108496.
9
Science of the Total Environment 871 (2023) 162034
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.
Chinthapalli, R., Skoczinski, P., Carus, M., Baltus, W., de Guzman, D., Käb, H., Raschka, A.,
Ravenstijn, J., 2019. Biobased building blocks and polymers—global capacities, produc-
tion and trends 2018–2023. Ind. Biotechnol. 15 (4), 237–241.
De Tender, C.A., Devriese, L.I., Haegeman, A., Maes, S., Ruttink, T., Dawyndt, P., 2015. Bac-
terial community profiling of plastic litter in the Belgian part of the north sea. Environ.
Sci. Technol. 49 (16), 9629–9638.
de Vries, F.T., Griffiths, R.I., Bailey, M., Craig, H., Girlanda, M., Gweon, H.S., Hallin, S.,
Kaisermann, A., Keith, A.M., Kretzschmar, M., Lemanceau, P., Lumini, E., Mason, K.E.,
Oliver, A., Ostle, N., Prosser, J.I., Thion, C., Thomson, B., Bardgett, R.D., 2018. Soil bac-
terial networks are less stable under drought than fungal networks. Nat. Commun. 9 (1),
3033.
Deng, Y., Jiang, Y.-H., Yang, Y., He, Z., Luo, F., Zhou, J., 2012. Molecular ecological network
analyses. BMC Bioinformatics 13 (1), 113.
Ding, F., Li, S., Lü, X.-T., Dijkstra, F.A., Schaeffer, S., An, T., Pei, J., Sun, L., Wang, J., 2019.
Opposite effects of nitrogen fertilization and plastic film mulching on crop N and P stoi-
chiometry in a temperate agroecosystem. J. Plant Ecol. 12 (4), 682–692.
Du, X., Deng, Y., Li, S., Escalas, A., Feng, K., He, Q., Wang, Z., Wu, Y., Wang, D., Peng, X.,
Wang, S., 2021. Steeper spatial scaling patterns of subsoil microbiota are shaped by deter-
ministic assembly process. Mol. Ecol. 30 (4), 1072–1085.
Duraisamy, P., Sekar, J., Arunkumar, A.D., Ramalingam, P.V., 2020. Kinetics of phenol bio-
degradation by heavy metal tolerant rhizobacteria glutamicibacter nicotianae
MSSRFPD35 from distillery effluent contaminated soils. Front. Microbiol. 11, 1573.
Duran, P., Thiergart, T., Garrido-Oter, R., Agler, M., Kemen, E., Schulze-Lefert, P., Hacquard,
S., 2018. Microbial interkingdom interactions in roots promote Arabidopsis survival. Cell
175 (4), 973–983 e914.
Edgar, R.C., 2013. UPARSE: highly accurate OTU sequences from microbial amplicon reads.
Nat. Methods 10 (10), 996–998.
Fei, Y., Huang, S., Zhang, H., Tong, Y., Wen, D., Xia, X., Wang, H., Luo, Y., Barcelo, D., 2020.
Response of soil enzyme activities and bacterial communities to the accumulation of
microplastics in an acid cropped soil. Sci. Total Environ. 707, 135634.
Feng, K., Zhang, Z., Cai, W., Liu, W., Xu, M., Yin, H., Wang, A., He, Z., Deng, Y., 2017. Biodi-
versity and species competition regulate the resilience of microbial biofilm community.
Mol. Ecol. 26 (21), 6170–6182.
Feng, K., Peng, X., Zhang, Z., Gu, S., He, Q., Shen, W., Wang, Z., Wang, D., Hu, Q., Li, Y.,
Wang, S., Deng, Y., 2022. iNAP: An integrated network analysis pipeline for microbiome
studies. iMeta 1 (2).
Feng, X., Wang, Q., Sun, Y., Zhang, S., Wang, F., 2022b. Microplastics change soil properties,
heavy metal availability and bacterial community in a pb-zn-contaminated soil.
J. Hazard. Mater. 424 (Pt A), 127364.
Gao, H., Yan, C., Liu, Q., Ding, W., Chen, B., Li, Z., 2019. Effects of plastic mulching and plastic
residue on agricultural production: a meta-analysis. Sci. Total Environ. 651 (Pt 1),
484–492.
Griffin-LaHue, D., Ghimire, S., Yu, Y., Scheenstra, E.J., Miles, C.A., Flury, M., 2022. In-field
degradation of soil-biodegradable plastic mulch films in a mediterranean climate. Sci.
Total Environ. 806 (Pt 1), 150238.
Gross, R.A., Kalra, B., 2002. Biodegradable polymers for the environment. Science 297
(5582), 803–807.
He, Q., Wang, S., Hou, W., Feng, K., Li, F., Hai, W., Zhang, Y., Sun, Y., Deng, Y., 2021. Tem-
perature and microbial interactions drive the deterministic assembly processes in sedi-
ments of hot springs. Sci. Total Environ. 772, 145465.
Hu, X., Gu, H., Wang, Y., Liu, J., Yu, Z., Li, Y., Jin, J., Liu, X., Dai, Q., Wang, G., 2022. Succession
of soil bacterial communities and network patterns in response to conventional and biode-
gradable microplastics: a microcosmic study in mollisol. J. Hazard. Mater. 436, 129218.
Hu, Y., Ma, P., Duan, C., Wu, S., Feng, H., Zou, Y., 2020. Black plastic film combined with
straw mulching delays senescence and increases summer maize yield in northwest
China. Agr. Water Manag 231.
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. Environ. Pollut. 254
(Pt A), 112983.
Ju, Z., Du, X., Feng, K., Li, S., Gu, S., Jin, D., Deng, Y., 2021. The succession of bacterial com-
munity attached on biodegradable plastic mulches during the degradation in soil. Front.
Microbiol. 12, 785737.
Kalam, S., Basu, A., Ahmad, I., Sayyed, R.Z., El-Enshasy, H.A., Dailin, D.J., Suriani, N.L., 2020.
Recent understanding of soil acidobacteria and their ecological significance: a critical re-
view. Front. Microbiol. 11, 580024.
Kalcikova, G., Zgajnar Gotvajn, A., Kladnik, A., Jemec, A., 2017. Impact of polyethylene
microbeads on the floating freshwater plant duckweed Lemna minor. Environ. Pollut.
230, 1108–1115.
Kasirajan, S., Ngouajio, M., 2012. Polyethylene and biodegradable mulches for agricultural
applications: a review. Agron. Sustain. Dev. 32 (2), 501–529.
Kim, H., Yu, S.M., 2020. Flavobacterium nackdongense sp. nov., a cellulose-degrading bacte-
rium isolated from sediment. Arch. Microbiol. 202 (3), 591–595.
Knights, D., Kuczynski, J., Charlson, E.S., Zaneveld, J., Mozer, M.C., Collman, R.G., Bushman,
F.D., Knight, R., Kelley, S.T., 2011. Bayesian community-wide culture-independent mi-
crobial source tracking. Nat. Methods 8 (9), 761–763.
Kong, X., Jin, D., Tai, X., Yu, H., Duan, G., Yan, X., Pan, J., Song, J., Deng, Y., 2019. Bioreme-
diation of dibutyl phthalate in a simulated agricultural ecosystem by gordonia sp. Strain
QH-11 and the microbial ecological effects in soil. Sci. Total Environ. 667, 691–700.
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 U.S. locations. Agroecol.
Sust. Food. 38 (8), 861–889.
Li, Y., Li, T., Wang, Z., Wang, S., Qin, X., Liao, Y., 2021. Plastic film mulch changes the micro-
bial community in maize root-associated compartments. Plant Soil 470 (1–2), 5–20.
Z. Zhao et al.
Li, Y., Yang, R., Guo, L., Gao, W., Su, P., Xu, Z., Xiao, H., Ma, Z., Liu, X., Gao, P., Li, B., Sun, X.,
Yan, G., Sun, W., 2022. The composition, biotic network, and assembly of plastisphere
protistan taxonomic and functional communities in plastic-mulching croplands.
J. Hazard. Mater. 430, 128390.
Liang, J., Tang, S., Gong, J., Zeng, G., Tang, W., Song, B., Zhang, P., Yang, Z., Luo, Y., 2020.
Responses of enzymatic activity and microbial communities to biochar/compost amend-
ment in sulfamethoxazole polluted wetland soil. J. Hazard. Mater. 385, 121533.
Liu, J., Li, S., Yue, S., Tian, J., Chen, H., Jiang, H., Siddique, K.H.M., Zhan, A., Fang, Q., Yu, Q.,
2021. Soil microbial community and network changes after long-term use of plastic
mulch and nitrogen fertilization on semiarid farmland. Geoderma 396.
Liu, Y., Xiao, M., Shahbaz, M., Hu, Z., Zhu, Z., Lu, S., Yu, Y., Yao, H., Chen, J., Ge, T., 2022.
Microplastics in soil can increase nutrient uptake by wheat. J. Hazard. Mater. 438,
129547.
Louca, S., Parfrey, L.W., Doebeli, M., 2016. Decoupling function and taxonomy in the global
ocean microbiome. Science 353 (6305), 1272–1277.
Mccormick, A.R., Hoellein, T.J., London, M.G., Hittie, J., Scott, J.W., Kelly, J.J., 2016.
Microplastic in surface waters of urban rivers: concentration, sources, and associated bac-
terial assemblages. Ecosphere 7 (11), e01556.
Meng, X., Niu, G., Yang, W., Cao, X., 2015. Di(2-ethylhexyl) phthalate biodegradation and de-
nitrification by a pseudoxanthomonas sp. Strain. Bioresour. Technol. 180, 356–359.
Mhedbi-Hajri, N., Jacques, M.A., Koebnik, R., 2011. Adhesion mechanisms of plant-
pathogenic xanthomonadaceae. Adv. Exp. Med. Biol. 715, 71–89.
Miao, L., Wang, P., Hou, J., Yao, Y., Liu, Z., Liu, S., Li, T., 2019. Distinct community structure
and microbial functions of biofilms colonizing microplastics. Sci. Total Environ. 650 (Pt
2), 2395–2402.
Munoz-Rojas, M., Roman, J.R., Roncero-Ramos, B., Erickson, T.E., Merritt, D.J., Aguila-
Carricondo, P., Canton, Y., 2018. Cyanobacteria inoculation enhances carbon sequestra-
tion in soil substrates used in dryland restoration. Sci. Total Environ. 636, 1149–1154.
Olimi, E., Kusstatscher, P., Wicaksono, W.A., Abdelfattah, A., Cernava, T., Berg, G., 2022. In-
sights into the microbiome assembly during different growth stages and storage of straw-
berry plants. Environ. Microbiome 17 (1), 21.
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.
J. Hazard. Mater. 387, 121711.
Roesch, L.F., Fulthorpe, R.R., Riva, A., Casella, G., Hadwin, A.K., Kent, A.D., Daroub, S.H.,
Camargo, F.A., Farmerie, W.G., Triplett, E.W., 2007. Pyrosequencing enumerates and
contrasts soil microbial diversity. ISME J. 1 (4), 283–290.
Sander, M., 2019. Biodegradation of polymeric mulch films in agricultural soils: concepts,
knowledge gaps, and future research directions. Environ. Sci. Technol. 53 (5),
2304–2315.
Santoyo, G., Orozco-Mosqueda, M.D.C., Govindappa, M., 2012. Mechanisms of biocontrol and
plant growth-promoting activity in soil bacterial species of Bacillus and Pseudomonas: a
review. Biocontrol Sci. Techn. 22 (8), 855–872.
Seeley, M.E., Song, B., Passie, R., Hale, R.C., 2020. Microplastics affect sedimentary microbial
communities and nitrogen cycling. Nat. Commun. 11 (1), 2372.
Serrano-Ruiz, H., Martin-Closas, L., Pelacho, A.M., 2021. Biodegradable plastic mulches: im-
pact on the agricultural biotic environment. Sci. Total Environ. 750, 141228.
10
Science of the Total Environment 871 (2023) 162034
Shi, Y., Zhang, K., Li, Q., Liu, X., He, J.S., Chu, H., 2020. Interannual climate variability and
altered precipitation influence the soil microbial community structure in a Tibetan Pla-
teau grassland. Sci. Total Environ. 714, 136794.
Sintim, H.Y., Flury, M., 2017. Is biodegradable plastic mulch the solution to agriculture's plas-
tic problem? Environ. Sci. Technol. 51 (3), 1068–1069.
Sintim, H.Y., Bary, A.I., Hayes, D.G., Wadsworth, L.C., Anunciado, M.B., English, M.E.,
Bandopadhyay, S., Schaeffer, S.M., DeBruyn, J.M., Miles, C.A., Reganold, J.P., Flury,
M., 2020. In situ degradation of biodegradable plastic mulch films in compost and agri-
cultural soils. Sci. Total Environ. 727, 138668.
Steinmetz, Z., Wollmann, C., Schaefer, M., Buchmann, C., David, J., Troger, J., Munoz, K.,
Fror, O., Schaumann, G.E., 2016. Plastic mulching in agriculture. Trading short-term ag-
ronomic benefits for long-term soil degradation? Sci. Total Environ. 550, 690–705.
Sun, X., Xu, Z., Xie, J., Hesselberg-Thomsen, V., Tan, T., Zheng, D., Strube, M.L., Dragos, A.,
Shen, Q., Zhang, R., Kovacs, A.T., 2022a. Bacillus velezensis stimulates resident rhizo-
sphere Pseudomonas stutzeri for plant health through metabolic interactions. ISME J.
16 (3), 774–787.
Sun, Y., Duan, C., Cao, N., Ding, C., Huang, Y., Wang, J., 2022b. Biodegradable and conven-
tional microplastics exhibit distinct microbiome, functionality, and metabolome changes
in soil. J. Hazard. Mater. 424 (Pt A), 127282.
Tarafdar, A., Lee, J.U., Jeong, J.E., Lee, H., Jung, Y., Oh, H.B., Woo, H.Y., Kwon, J.H., 2021.
Biofilm development of Bacillus siamensis ATKU1 on pristine short chain low-density
polyethylene: a case study on microbe-microplastics interaction. J. Hazard. Mater. 409,
124516.
Tong, Y., Ding, J., Xiao, M., Shahbaz, M., Zhu, Z., Chen, M., Kuzyakov, Y., Deng, Y., Chen, J.,
Ge, T., 2022. Microplastics affect activity and spatial distribution of C, N, and P hydro-
lases in rice rhizosphere. Soil Ecol. Lett. 5 (3).
Wang, P., Song, T., Bu, J., Zhang, Y., Liu, J., Zhao, J., Zhang, T., Xi, J., Xu, J., Li, L., Lin, Y.,
2022. Does bacterial community succession within the polyethylene mulching film
plastisphere drive biodegradation? Sci. Total Environ. 824, 153884.
Yan, H., Gu, S., Li, S., Shen, W., Zhou, X., Yu, H., Ma, K., Zhao, Y., Wang, Y., Zheng, H., Deng,
Y., Lu, G., 2022. Grass-legume mixtures enhance forage production via the bacterial com-
munity. Agric. Ecosyst. Environ. 338.
Yan, Y., Kuramae, E.E., de Hollander, M., Klinkhamer, P.G., van Veen, J.A., 2017. Functional
traits dominate the diversity-related selection of bacterial communities in the rhizo-
sphere. ISME J. 11 (1), 56–66.
Yue, W., Yin, C.F., Sun, L., Zhang, J., Xu, Y., Zhou, N.Y., 2021. Biodegradation of bisphenol-a
polycarbonate plastic by Pseudoxanthomonas sp. Strain NyZ600. J. Hazard. Mater. 416,
125775.
Zettler, E.R., Mincer, T.J., Amaral-Zettler, L.A., 2013. Life in the "plastisphere": microbial com-
munities on plastic marine debris. Environ. Sci. Technol. 47 (13), 7137–7146.
Zhou, J., Gui, H., Banfield, C.C., Wen, Y., Zang, H., Dippold, M.A., Charlton, A., Jones, D.L.,
2021. The microplastisphere: biodegradable microplastics addition alters soil microbial
community structure and function. Soil boilBiochemist 156.
Zhu, J., Liu, S., Wang, H., Wang, D., Zhu, Y., Wang, J., He, Y., Zheng, Q., Zhan, X., 2022.
Microplastic particles alter wheat rhizosphere soil microbial community composition
and function. J. Hazard. Mater. 436, 129176.