Science of the Total Environment 881 (2023) 163398

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Science of the Total Environment

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

Black carbon derived from pyrolysis of maize straw and polystyrene

microplastics affects soil biodiversity
Changlu Hu a, Fadan Lei a,1, Xue Zhang a, Jiabao Shi a, Jing Li b, Xin Zuo a, Shuxing Li a,
⁎ ⁎
Zhuozhi Ouyang a,c, , Xuetao Guo a,c,
a
College of Natural Resources and Environment, Northwest A&F University, Yangling, Shaanxi 712100, China
b
School of Ecology and Environment, Beijing Technology and Business University, Beijing 100048, China
c
Key Laboratory of Plant Nutrition and the Agro-environment in Northwest China, Ministry of Agriculture and Rural Affairs, Yangling, Shaanxi 712100, 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

• The different ratios of PS and maize straw

were used for preparing BC by pyrolysis.
• The yield, value of O/C and content of N
element of BC decreased with more PS.
• The main component of filtrate from BC
was humic-like and fulvic-like acid.
• BC affected the abundance, composition
and metabolic pathways of microorgan-
isms.

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

Editor: Damia Barcelo Understanding the environmental correlation of microbial community under external stimulation is significant for eco-
logical restoration. However, few studies focused on the response of soil biodiversity induced by black carbon (BC) de-
Keywords: rived from pyrolysis of straw and microplastics (MPs) due to their widespread existence in natural environment. In this
Black carbon study, polystyrene MPs (PS) and maize straw with different mass ratios were used as raw materials to prepare BC by
Pyrolysis
pyrolysis. The surface morphology, chemical composition and sequential variations of different functional groups of
Microplastics
Biochar
BC were systematically analyzed. The leachate from BC was identified by three-dimensional excitation emission ma-
Biodiversity trice (3D-EEM). The corresponding results showed that yield, value of O/C and N element content of BC decreased
with more PS. The changed C content and oxygen-containing functional groups occurred. The order of functional
groups of BC formed by co-pyrolysis was: C=C > C-O > C-H > Si-O-Si. The main component of leaching from BC
was humic-like and fulvic-like acid. Simultaneously, the input of exogenous BC into soil affected abundance, compo-
sition and metabolic pathways of microorganisms. The study helps to understand environmental implication of BC
which was pyrolyzed from maize straw and MPs, providing an idea for improving biogeochemical cycle process in soil.

1. Introduction
⁎ Corresponding authors at: College of Natural Resources and Environment, Northwest A&F
University, Yangling, Shaanxi 712100, China.
E-mail addresses: [email protected] (Z. Ouyang), [email protected]
Microplastics (MPs) are defined as particles with diameter less than
(X. Guo). 5 mm (Thompson et al., 2004). As a type of emerging contaminants, the
1
Contributed equally to this work. MPs pollution has become a global environmental issue (Kane et al.,

http://dx.doi.org/10.1016/j.scitotenv.2023.163398
Received 10 February 2023; Received in revised form 18 March 2023; Accepted 5 April 2023
Available online 13 April 2023
0048-9697/© 2023 Elsevier B.V. All rights reserved.
C. Hu et al. Science of the Total Environment 881 (2023) 163398

2020; Nizzetto et al., 2016b; Rochman, 2018). Because of its high concen- 2. Materials and methods
tration, long-range transport and small particle size, MPs easily cause
harm to the ecological environment and health (Ding et al., 2022; Ge 2.1. Chemicals and reagents
et al., 2022; Ouyang et al., 2022a; Ouyang et al., 2022b). A large number
of MPs have been found in the water, soil, atmosphere and other ecosystem The experimental materials used in this experiment included polysty-
component. Surprisingly, MPs can also enter animals (Li et al., 2018; Lv rene MPs (PS) and maize straw. The PS powder was purchased from
et al., 2019; Wang et al., 2019), plants (Li et al., 2020b; Mao et al., 2018) Thailand Petrochemical Industry Co., Ltd. and its particle size was
and in human embryos (Ragusa et al., 2021). Besides, the terrestrial envi- 100.35 ± 6.78 μm. Before the experiment, MPs were cleaned with absolute
ronment is most vulnerable to MPs pollution as the main sink of MPs ethyl alcohol and deionized water. All other reagents used were standard
(Huang et al., 2021). And the corresponding pollution is worse in terrestrial reagents (GR) and needed no further purification.
environment than in marine environment (Horton et al., 2017; Mao et al.,
2021). MPs easily enter the terrestrial environment through a variety of 2.2. Preparation of BC
sources (Nizzetto et al., 2016a). As a result, the soil certainly suffers from
serious MPs pollution due to the sewage, fertilizers, and plastics used in ag- BC was pyrolyzed by mixing different mass ratio of maize straw and PS
riculture (Li et al., 2021). Although the persistence and accumulation of under an oxygen limited condition. The mass ratio of straw powder to PS
MPs in the soil environment occurs, the effect of their transformation be- was 100:0, 80:20 and 20:80, which was called BC, 4:1 BC, and 1:4 BC, re-
havior and fate on microbial community is scarcely documented. As the spectively. The heating rate of was set as 5 °C/min. And the holding time
most important components of soil microenvironment, microbes lead to was 180 min and terminal temperature was 400 °C.
the transformation of soil nutrients, decomposition of organic matter, and
take part in a series of biochemical reactions in soil (Pu et al., 2022), 2.3. Microcosm incubation in soil
which is considered as an indispensable indicator of soil environmental
quality (Ho and Chambers, 2019). The soil used in the soil microbial experiment was taken from the unpol-
Black carbon (BC) is generated from the incomplete combustion and py- luted topsoil of 0–20 cm in the south campus of Northwest A&F University
rolysis of fossil fuels and biomass to refractory char, soot, and graphite, in Yangling, Shaanxi Province, China. Impurities such as stones, the broken
which has highly aromatic structure of carbon particles (Kelesidis et al., branches and fallen leaves were removed after retrieval. Fully mixing and
2022). Despite the ubiquitous persistence of BC in the particulate organic air-dry in a ventilated place was necessary. The soil was ground and sieved
matter and dissolved organic carbon pools (Malits et al., 2015), its effect with 10 mesh (2 mm) (Ameloot et al., 2014). The environmental condition
on soil environment and its role in the terrestrial carbon cycle remain of colonization and physicochemical property of soil had important effects
largely enigmatic. It has been verified that the BC acts as vector due to its on microbial activity (Qin et al., 2021; Wang et al., 2020). In order to sim-
porous structure and abundant functional groups (Cattaneo et al., 2010). ulate the natural environment of soil, the ultrapure water was adding into
Besides, it can stimulate the aggregation of organic particles and bacterial the soil to keep the certain content of moisture (Manirakiza et al., 2019).
production (Mari et al., 2014). BC is supposed to be significant in the regu- The soil was pre-cultured for 7 d to ensure the microbial activity in soil.
lation of element transformation and redox reactions, which greatly influ- BC, 4:1 BC and 1:4 BC were added into the pre-cultured soil environment
ence the various biogeochemical processes (e.g., carbon sequestration, at a 1.8 % ratio of the total soil, and a blank control was set up. Samples
nutrient availability, microbial activity, migration and transformation of were collected at the incubation time of 30 d and 50 d. Before the testing,
pollutants) (Beesley et al., 2011; Lehmann, 2007). It is estimated that all the samples were placed in a refrigerator at −80 °C. The microbial sam-
80 % of BC comes from burning vegetation. The global annual production ples were named as following: K1–0 d soil blank, K2–30 d soil blank, K3–50
of BC continues to increase every year with the global fuel use and some ag- d soil blank, C1–30 d BC, C2–30 d 4:1 BC, C3–30 d 1:4 BC, C4–50 d BC,
ricultural activities increasing. Especially, the formation of BC is related to C5–50 d 4:1 BC and C6–50 d 1:4 BC.
the raw materials, pyrolysis temperature and duration under natural condi-
tions (Dai et al., 2020; Lian and Xing, 2017). BC finally entered in soil and 2.4. Characterization methods of BC
sediments, frequently makes up 1–20 % of the total organic carbon
(Lehmann et al., 2008; Lian and Xing, 2017). The massive emissions of Fourier transform infrared spectrometer (FTIR, Vetex70, Bruker) was
BC lead to its ubiquity in the environment. And the formation of BC cer- applied to determine the functional groups of BC. According to the FTIR
tainly will be affected by other coexistence components (Forbes et al., spectra of different BC, the two-dimensional correlation Fourier transform
2006). The elemental composition of MPs is mainly dominated by C and infrared spectroscopy (2D-FTIR-COS) were conducted (Noda et al., 2000;
H elements. As a waste of polymer materials, MPs have low water content Ouyang et al., 2020; Zhou et al., 2017b). The crystal structure of BC was de-
and high calorific value, which are good additives for pyrolysis reaction tected by X-ray diffraction (XRD, Ultima IV, Rigaku). Chemical composition
(Zhang et al., 2022). Therefore, the effect of MPs on the carbon components changes of BC were determined by X-ray photoelectron spectroscopy (XPS,
in BC during pyrolysis cannot be ignored. Simultaneously, the physico- Escalab Xi+, Thermo Fischer).
chemical properties of obtained BC from different sources and different py-
rolysis preparation conditions differ greatly, which further altered its 2.5. Filtrate analysis from BC
environmental utilization (Li et al., 2020a; Wang et al., 2018). As BC parti-
cles are a relevant type of organic particle, knowledge on the potential im- The BC was dissolved in deionized water and oscillated, and then the fil-
pact of different sources of BC on production and loss of microorganisms trate was analyzed by fluorescence spectrometer (RF-6000, Shimadzu) for
and the consequences for biogeochemical cycles in the soil is necessary. three-dimensional excitation emission matrix spectrum. The detailed infor-
In this study, the BC formed by co-pyrolysis of MPs and straw was pre- mation of index is illustrated in the Text S1.
pared. And the effect of different BC on microbial community was studied
systematically. Owing to being frequently detected in natural environment, 2.6. Microbial community analysis
polystyrene (PS) was selected as a representative. The aims of this research
were to (1) analyze the physicochemical properties of BC formed by The testing samples were prepared for DNA extraction, 16S rDNA spe-
co-pyrolysis with different proportions of PS and straw; (2) access the cific primer PCR amplification and sequencing at Biomarker Technologies
component leaching from the different BC; (3) investigate the effect of dif- Corporation (Beijing, China). Based on Illumina Novaseq sequencing plat-
ferent BC on the abundance, diversity and metabolic pathways of microbial form, a small fragment library was constructed and sequenced by using
community in soil. The study provides a theoretical basis for improving soil the method of Paired-End. By clustering high-quality sequences and divid-
ecological environment and the application of BC. ing the operation taxon (OTUs), the species classification was obtained.

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And the species abundance, Alpha diversity and differences in microbial 3.2. Chemical composition of different BC
metabolic pathways of each community were analyzed based on the results
of feature analysis. The Kyoto Encyclopedia of Genes and Genomes (KEGG) One-dimensional infrared spectroscopy was used to analyze the BC
indicated an intuitive distribution of the functional metabolic pathways of doped under different conditions, and the differences of chemical bonds
soil microorganisms in the 30th day (the most active and representative mi- of BC prepared under different conditions were observed. From Fig. S1,
croorganisms) after the addition of different BC. the -OH bond at 3400 cm−1 of BC vibrated in the BC prepared by calcina-
tion (Mandal et al., 2017). The peak at 2923 cm−1 was found to be caused
3. Results and discussion by asymmetric stretching vibration of -CHX bond and the signals of
1600 cm−1 was the vibration of C_C bond (Tang et al., 2021). The peak
3.1. Morphology characterization of different BC at 1439 cm−1 was caused by the vibration of C\\H bond. The fluctuation
at 1090 cm−1 was caused by the extension of C\\O bond, and the oxidation
The yields of BC, 4:1 BC and 1:4 BC were 44.53 %, 36.17 % and 9.15 %, of organic matter resulted in a large number of C\\O bond stretching at the
among which BC had the highest yield (Fig. 1a). The coexistence of PS ob- final pyrolysis temperature of 400 °C. Signals of stretching of Si-O-Si bond
viously reduced the yield of BC. The yield of BC was also lower with more appeared nearly at 473 cm−1 (Tang et al., 2021).
dopant of PS, which was determined by the pyrolysis characteristics of PS. The FTIR spectrum provides little information about the pyrolysis pro-
PS is a common MPs with 140–180 °C melting temperature and a decompo- cess, especially the changing sequence of functional groups (Ding et al.,
sition temperature above 300 °C (Onwudili et al., 2009). Due to the high 2020). We applied 2D-FTIR-COS analysis based on FTIR spectrum in
temperature, the yield of BC decreased with the increase of PS content. In order to further explain the changing order of a large number of functional
order to further investigate the changed surface morphology of BC medi- groups and the mechanism of BC formation under different conditions.
ated by PS. Fig. 1b shows the XRD analysis results of BC formed by co- From the FTIR spectrum, the vibration amplitude of BC was larger in the
pyrolysis with different proportions of PS. The phase structure of BC with range of 400–2000 cm−1. Fig. 1 also shows the 2D-FTIR-COS spectra of
three proportions had little change, and the change of crystal structure BC. Four automatic peak diagonals were found at 473, 1090, 1439 and
was basically the same. Three kinds of BC had obvious broad peaks at 1600 cm−1. The peak intensity of C_C bond at 1600 cm−1 and C\\H
22°, which were related to graphitic carbon structure of BC. The sharp bond at 1090 cm−1 were the largest (Nan et al., 2021). As for the synchro-
peaks at the 2θ around 16° and 22° related to the crystal structure of cellu- nous, the positive signals were shown in all cross peaks (Table S1). It indi-
lose were disappeared at high temperatures above 300 °C (Keiluweit et al., cated that most of the total cross peaks were synchronized, which meant
2010; Kim et al., 2012). The peak at 26° was the diffraction peak of SiO2, that almost all the functional groups were synchronously changed. Also,
which is the main mineral composition in straw (Jindo et al., 2014). The the automatic peak at 1439 cm−1 had higher intensity than the automatic
peak at 28° represented CaCO3 in BC, which are the main components of peak at 473 cm−1 as the increasing of PS, which showed that the functional
ash in BC (Liu et al., 2014). The diffraction peak intensity of the three BC groups C\\H bond changed before the Si-O-Si bond during the formation of
has no obvious change. Therefore, the pyrolytic process in the presence of BC. According to the rules of 2D-FTIR-COS, the asynchronous correlation
PS had little effect on the crystal structure of BC. spectrum showed the continuous sequence of reactions in the process of

Fig. 1. (a) The yield and (b) XRD image of BC; (c) synchronous and (d) asynchronous 2D-FTIR-COS maps of BC at different conditions. Note: red color denotes positive
correlation, and blue color means negative correlation; darker color demon-strates a higher intensity and thus a stronger positive or negative correlation. The peaks
located at the diagonal line are called auto-peaks while cross peaks are located off-diagonal positions.

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generating BC by pyrolysis. Asynchronous mapping was antisymmetric almost disappeared. Therefore, the group of C\\O and C_O increased ac-
with respect to diagonal, so there is no automatic peak. Individual differ- cordingly. This result was also consistent with the above FTIR results. Anal-
ences in cross peak markers were observed during the three treatments. ysis above further demonstrated that the greatly changed C content and
As shown in Table S1, as the mass fraction of PS in the material increased, oxygen-containing functional groups of BC occurred in the presence of PS.
a large number of the cross-over peaks showed positive markers, except for
negative cross-over peaks at ψ (1439, 1090). The results showed that the 3.3. Identification of filtrate from different BC
functional groups at 1600, 1090, 1439 and 473 cm−1 changed firstly.
The changing order of functional groups was: C_C bond at 1600 cm−1 > The three-dimensional fluorescence spectrum of leaching from BC is il-
C-O bond at 1090 cm−1 > C-H bond at 1439 cm−1 > Si-O-Si bond at lustrated in Fig. 3. Two peaks C1 (Ex/Em: 300 nm/405 nm) and C2 (Ex/Em:
473 cm−1. 225 nm/415 nm) observably presented in the three-dimensional excitation
The changed contents of C and O in three BC were analyzed by XPS. emission matrice (3D-EEM) were identified as humic-like and fulvic-like,
Table S2 shows the contents of C, O and N in BC. Compared with the orig- respectively (Lin and Guo, 2020; Osburn et al., 2016; Osburn et al.,
inal BC, the content of C gradually increased with more PS, while the con- 2011). The related study demonstrated the DOM from BC was mostly com-
tent of N, O contents and the O/C ratio gradually decreased. The XPS posed of humic-like components (Wei et al., 2020). The other study also
spectra of three kinds of BC are fitted by C1s and O1s, and the fitting results verified that BC-derived water-soluble organic carbon mainly included
are shown in Fig. 2. It could be seen that the main functional groups of BC humic-like, fulvic-like substances. These characteristics were same as typi-
C1s were C\\C (284.8 eV), C-O-C (286.00 eV) and O-C=O (289.00 eV), cal terrestrial humic-like fluorophores (Zhou et al., 2017a). The released
among which the signal of C\\C was the strongest, followed by the C-O-C material from BC was generally composed of both components in the orig-
and O-C=O. The stronger the C\\C peak and the weaker C-O-C and O- inal feedstock and products from pyrolysis, including humic- or protein-like
C=O peak occurred in the presence of more PS. And the area of O-C=O substances. The increased charring PS led to weaken the fluorescence inten-
in 1:4 BC was very small. The main functional groups of O1s were C_O sity of the two peaks. BC formed solely from the pyrolysis of straw has the
(533.51 eV), -COO (531.33 eV) and C\\O (532.39 eV). The -COO peak in- highest fluorescence intensity (Yu et al., 2023). The fluorescence index of
tensity of BC and 4:1 BC were almost the same, while the peak in 1:4 BC was 3D-EEM could provide more effective information about the substances

Fig. 2. XPS spectra of (a, b, c) C1s and (d, e, f) O1s of three BC; (g) The content of surface element of BC; and proportion of surface functional groups revealed by (h) C1 s and
(i) O1s.

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Fig. 3. The (a, b, c) fluorescent components of 3D-EEM and (d) index of filtrate from different BC.

released by BC. The humification index (HIX) was used to determine the de- the changes of microbial community. The Venn diagram based on the
gree of humification and the biogenic index (BIX) indicated the contribu- whole OTUs identified under different treatments showed that most of
tion of dissolved substances from biogenesis (Birdwell and Engel, 2010). them had a common phenomenon in all treatments (Fig. S3). Compared
From the Fig. 3, the HIX index of all the three BC was less than 1.5, with the number of OTUs in the presence of BC, the blank samples in-
which indicated that the humification degree of was very low (Gao et al., creased with the time. For the soil samples in presence of different BC, the
2019; Huguet et al., 2009), indicating relatively low stability with simple number of OTUs of samples (30 d) was higher than that of the blank at
structure (Wilson and Xenopoulos, 2009). The leachate of BC had the the same period, which was due to the porous structure of BC providing aer-
highest value of HIX index. The BIX of BC and 4:1 BC ranged from 0.8 to ation, humidity and better habitat for aerobic microorganisms (Muhammad
1, indicating that the released substances were similar to terrigenous and et al., 2014). The input of exogenous BC into the soil would disturb the
endogenous inputs. The BIX of 1:4 BC was greater than 1, showing mainly input-output balance of material and energy in the soil-microbial system
the source of endogeny (Jaffé et al., 2004; Shimizu et al., 2018). Simulta- and directly affect the community abundance and metabolic activity of mi-
neously, the freshness index (β: α) also increased when more PS were in- croorganisms. Further, while the number of OTUs decreased as the incuba-
volved in pyrolysis. tion time increasing. The presence of BC in the early stage provided a
habitat for microbial communities and abundant carbon source, which
3.4. The effects of BC on soil microbial communities was conducive to growth and reproduction of microorganisms (Ameloot
et al., 2014). Simultaneously, some carbon sources were taken up and de-
3.4.1. Changed abundance and diversity of microbial communities graded by microorganisms. The growth and reproduction of microbial com-
According to the high-throughput sequencing, the changed abundance munities would be affected under the condition of less extraneous carbon
and diversity of microbial communities caused by BC have observed source. Therefore, BC could be used as one of the sources for the growth
high-quality sequencing readings of 3 % sequence dissimilarity in of soil microorganisms (Wang et al., 2020), which affect the biogeochemi-
different samples (Nielsen et al., 2014). The sparse curve of features in cal cycle process in the soil ecosystem by changing the microbial commu-
the total sample tended to be flat (Fig. S2), which showed that the sequence nity structure.
was sufficient to explain the diversity of species communities in this exper- The alpha diversity also reflected the species richness and diversity of
iment (Huang et al., 2021). From the Table S3, the coverage index exceeded microorganisms (Huang et al., 2021; Li et al., 2022). The alpha diversity
99.86 %, reflecting that the results could represent real situation of micro- values of different treatments were shown in Table S3. The indexes of
bial communities. The different treatments in the presence of BC caused Chao1 and Ace indicated the species abundance, which increased with

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the cultivation time for blank samples. The index of the soil samples with Bacteroidetes, Gemmatimonadetes, Acidobacteria and Proteobacteria. The
three different BC was greater than that of the blank control K2 in the 30 Proteobacteria and Acidobacteria was dominant bacteria in a relative abun-
d. The Chao1 and Ace index of C6 were higher than that of K3 in the dance appeared in the nine soil samples, accounting for more than 70 %
same period (50 d), while the indexes of C4 and C5 was lower than that of the total abundance. Proteobacteria commonly existed in soil (Chen
of K3. The different properties of BC resulted in the changed soil microbial et al., 2015), which was the higher in the blank (K1, K2, and K3) samples.
community. The toxic elements or adsorbed nutrients leaching from BC Moreover, the abundance of the Nitrospirae, Latescibacteria and Chloroflexi
around in the soil environment, which could further inhibit or increase spe- in the soil sample added with the BC was generally higher than the blank,
cific microbial community (Muhammad et al., 2014). Moreover, the Shan- which indicated that these soil conditions were suitable for the growth of
non and Simpson index can be used to identify the sample species diversity the Nitrospirae, Latescibacteria and Chloroflexi (Chen et al., 2015). BC easily
as well as the impact on species abundance and evenness. With the same experienced the aging process in soil, and the changed physicochemical
abundance of species, the greater the evenness of each species was, the properties further affected the bacterial community in soil samples
greater diversity of the community would be. The higher of the Shannon (Ameloot et al., 2014). The abundance of Actinobacteria, Bacteroidetes and
index and Simpson index represented the higher the species diversity Proteobacteria decreased with the cultivation time, while the abundance
(Huang et al., 2021). Especially, the Shannon index and Simpson index of of Latescibacteria, Chloroflexi, and Gemmatimonadetes increased. The above
C4 and C5 soil samples had little difference from the K3 index of blank in results showed that the different BC changed the bacterial composition
the 50 d. The above results indicated that the exposure of 1:4 BC which and abundance, and the cultivation time was an important factor.
was pyrolyzed from more PS led to the highest microbial community diver-
sity and richness compared with other BC. The presence of BC improved the 3.4.3. Differences in metabolic pathways
abundance and diversity of soil microbial community in early stage, while Based on the analysis of the composition and difference of Kyoto Ency-
the effects of different BC treatments differed with the increase of culture clopedia of Genes and Genomes (KEGG) metabolic pathways, the differ-
time. The result also provided a new idea for better improving the soil eco- ences in the metabolic pathways of the functional genes of the microbial
logical environment. community between different groups of samples were observed (Huang
et al., 2022). As shown in Fig. 5a, the functional metabolic pathways at
3.4.2. Changed composition of microbial communities the two levels were mainly concentrated on global and overview maps, car-
In addition to the changed diversity and abundance of microbial com- bohydrate metabolism, amino acid metabolism, energy metabolism, me-
munity caused by BC, the community composition also was affected. tabolism of cofactors and vitamins, membrane transport, nucleotide
Fig. 4 shows the abundance of the composition and abundance at phylum metabolism, translation, signal transduction and replication and repair.
level of the soil sample under the different pressure of BC. A total of 10 bac- The KEGG database identified the differences in the functional metabolic
terial from the bacterial communities were identified, including Nitrospirae, pathways of soil microorganisms and obtained a difference map of func-
Latescibacteria, Rokubacteria, Actinobacteria, Firmicutes, Chloroflexi, tional metabolic pathways at the three levels (Fig. 5b). The results showed

Fig. 4. Bacterial composition and abundance at phylum level.

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Fig. 5. Difference diagram of soil microbial functional pathway at two (a) and three (b) levels with different treatments.

that the metabolic pathways, biosynthesis of secondary metabolites, bio- metabolic pathways of microorganisms. The composition of dominant bac-
synthesis of antibiotics, biosynthesis of amino acids and ribosome of C1, teria phylum in soil under different treatments was similar, while the abun-
C2 and C3 were more abundant than those in K2. In this process, BC was dance of soil microbial bacteria phylum varied. This study was helpful for
utilized by cells to promote metabolism and thus change the abundance understanding the environmental risks of different sources of BC.
and diversity of microbial communities. From Fig. S4, the two-component
systems (TCSs) pathways for functional metabolism of microorganisms in CRediT authorship contribution statement
C2 and C3 were more abundant than that in C1. TCSs could sense and re-
spond to various physical and chemical biological stimuli, regulate gene ex- Changlu Hu: Writing & review, Editing, Data analysis.
pression and influence metabolism (Lazar and Tabor, 2021), and the Fadan Lei: Writing & review, Editing, Data analysis.
microorganisms in C2 and C3 biofilm made better use of the characteristics Xue Zhang: Data analysis.
of TCSs to adapt to soil environmental conditions. The ATP-binding cassette Jiabao Shi: Data analysis.
(ABC) transporters pathway was more abundant than that of C2 in the func- Jing Li: Review.
tional metabolic pathway of C3 microorganisms, and the ABC transporters Xin Zuo: Editing.
protein was involved in the assembly of exopolysaccharides, which was of Shuxing Li: Editing.
great significance for biofilm formation and the dehydration tolerance of Zhuozhi Ouyang: Writing & review, Data analysis, Investigation, Meth-
microorganisms (Vanderlinde et al., 2010). The higher ABC transporters odology, Conceptualization, Project administration, Funding acquisition.
in C3 biofilm indicated that there were more pathways to promote biofilm Xuetao Guo: Conceptualization & Review.
formation and respond to external stresses, which meant that C3 biofilm
was denser and more stable than C2 biofilm (Huang et al., 2022). The addi- Data availability
tion of BC affected the structure and metabolic function of soil microbial
community by changing the content of soil organic carbon. The composi- Data will be made available on request.
tion of dominant bacteria phylum in soil was similar, while the abundance
of soil microbial bacteria phylum caused by different BC varied. The pres- Declaration of competing interest
ence of BC changed the biodiversity in soil. The 1:4 BC had a longer-term
positive impact on soil microbial community than other proportions of BC. The authors declare that they have no known competing financial inter-
ests or personal relationships that could have appeared to influence the
4. Conclusion work reported in this paper.

In this study, polystyrene MPs (PS) and maize straw with different mass Acknowledgements
ratios were used as raw materials to prepare different BC by pyrolysis. The
presence of PS had little effect on the crystal structure of BC. Nevertheless, The study was supported by the National Natural Science Foundation of
the greatly changed C content and oxygen-containing functional groups of China (No. 42207279), the Young Talent Fund of Association for Science
BC occurred and the value of O/C and content of N element of BC decreased and Technology in Shaanxi, China (No. 20220706), the Natural Science
with more PS. The order of functional groups of BC formed by co-pyrolysis Foundation of Shaanxi Province, China (No. 2021JQ-167), China Postdoc-
was: C=C > C-O > C-H > Si-O-Si. Besides, the main component of leaching toral Science Foundation (No. 2021M692652), Chinese Universities Scien-
from three kinds of BC was humic-like and fulvic-like acid. The input of ex- tific Fund (No. 2452020225) and the Open Research Fund Program of
ogenous BC into the soil affected the abundance, composition and Cultivation Project of Double First-Class Disciplines of Environmental

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C. Hu et al. Science of the Total Environment 881 (2023) 163398

Science and Engineering, Beijing Technology and Business University 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.
(BTBU) (No. ESE2022YB08). J. Hazard. Mater. 429, 128353.
Li, C., Zhang, C., Gholizadeh, M., Hu, X., 2020a. Different reaction behaviours of light or
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