Ecotoxicology and Environmental Safety 260 (2023) 115075

Contents lists available at ScienceDirect

Ecotoxicology and Environmental Safety

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

Combinations of waste seaweed liquid fertilizer and biochar on tomato

(Solanum lycopersicum L.) seedling growth in an acid-affected soil of
Jiaodong Peninsula, China
Xian-chao Shang a, 1, Manman Zhang b, 1, Yuqin Zhang c, Yiqiang Li d, Xin Hou a, *, Long Yang a, *
a
College of Plant Protection and Agricultural Big-Data Research Center, Shandong Agricultural University, Taian 271018, China
b
Citrus Research Institute, Southwest University, Chongqing 400712, China
c
Weihai Academy of Agricultural Sciences, Weihai 264200, China
d
Marine Agriculture Research Center, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, China

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

Edited by Dr Muhammad Zia-ur-Rehman Biochar application is an effective strategy for improving soil degradation and productivity. However, the effects
of the combination of biochar and other fertilizers to improve seedling growth in abiotic stress-affected soils
Keywords: remains unknown. We investigate the effect of biochar derived from reed straw (RBC) and waste seaweed liquid
Biochar fertilizer (SLF) on tomato (Solanum lycopersicum L.) seedling growth in an acid-affected soil of Jiaodong
Waste seaweed liquid fertilizer
Peninsula, China. The results revealed RBC, SLF, and the combination of RBC with SLF (RBC+SLF) significantly
Acid-affected soil
elevated the dry weight of tomatoes by 23.33 %, 29.93 %, and 63.66 %, respectively. The malondialdehyde
Microbial community
Metabolic pathway content in the tomato seedling roots, stems, and leaves was significantly lower in the RBC+SLF treatment, which
might be related to the enhanced contents of proline, soluble sugar, and soluble protein. The synthesis and
accumulation of zeatin riboside, indole-3-acetic acid, and gibberellic acid 3 in tomato under RBC+SLF amend­
ment may be attributed to the enhanced plant growth. Moreover, RBC, SLF, and RBC+SLF improved the soil
status (including ammonium nitrogen, nitrate nitrogen, laccase, and urease) in the acid-affected soil. Biochar and
waste seaweed liquid fertilizer significantly increased the relative abundance of Pseudomonas and Azospira
(beneficial bacteria) in tomato rhizosphere. The microbial amino acid metabolism was associated with changes
in soil properties and enzyme activities. Consequently, biochar and waste seaweed liquid fertilizer are viable soil
conditioners for acid-affected soil.

1. Introduction accumulates ions, and exacerbates soil hardening and acidification,

resulting in reduced tomato yield and quality (Song et al., 2022). Among
Tomato (Solanum lycopersicum L.) is one of the most important veg­ them, soil acidification is particularly prominent (Wang et al., 2021).
etables around the world, with over 7500 varieties grown globally and Refusing to complete statistics, the global acidic soil area reaches 3.95
an annual yield of over 175 million tons. As a high-value vegetable, billion hectares, accounting for about one-third of the land area.
tomatoes are usually grown in greenhouses in northern China (Wang Therefore, improving acid-affected soils caused by continuous cropping
et al., 2018b; Chen et al., 2022). Driven by high profits and guided by the and other environmental factors, especially the soil microbial environ­
market, the production of greenhouse tomatoes is highly intense with a ment, is of high importance.
high single-crop multiple cropping index (Mohammad et al., 2019; Chen The application of organic fertilizers aids in the remediation of acid-
et al., 2022). This intensive and unitary production mode has caused affected soil, as it effectively improves fertility and soil structure, and
many problems for sustainable production (Liu et al., 2019; Liang et al., increases soil chemical conservation characteristics (Chen et al., 2018).
2020) with multiple soil continuous cropping obstacles (Wang et al., Additionally, organic fertilizers can effectively regulate the microor­
2022; Tang et al., 2021; Wang et al., 2021). Continuous cropping de­ ganisms in the soil and release CO2 to promote photosynthesis (Rosa
creases the soil fertility, imbalances the soil microbial community, et al., 2018). Cooper et al. (2020) observed that the organic carbon

* Corresponding authors.
E-mail addresses: houxin@sdau.edu.cn (X. Hou), lyang@sdau.edu.cn (L. Yang).
1
These authors contributed equally to this paper.

https://doi.org/10.1016/j.ecoenv.2023.115075
Received 2 March 2023; Received in revised form 25 April 2023; Accepted 24 May 2023
Available online 1 June 2023
0147-6513/© 2023 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

content, pH, microbial biomass carbon, and cation exchange capacity of biochar or waste seaweed liquid fertilizer added (Control); 2) 3 % (w/w)
the treated soil significantly increased six years after fertilizer applica­ biochar was added to the acid-affected soil and evenly mixed (RBC) (Sun
tion. The soil microbial structure and diversity were gradually and et al., 2022; He et al., 2019); 3) 4 % (w/w) waste seaweed liquid fer­
significantly improved by organic fertilizers (Yan et al., 2021; Zhang tilizer was evenly added to the soil (SLF) (Deepika et al., 2020); and 4)
et al., 2019; Luo et al., 2022). Therefore, certain varieties and pro­ 1.5 % biochar and 2 % waste seaweed liquid fertilizer were combined
portions of organic fertilizer should be incorporated into acid-affected with the soil and mixed evenly (RBC+SLF). Then, we selected visibly
soil to alleviate the effects of acid stress on plants. similar tomato seedlings with five leaves, which were transplanted to
On the shores of China, which is over 32,000 km, various seaweeds the pots containing the different treatments. The pot experiments were
(kelp, sargassum, and bladder-wrack, etc.) grow naturally (i.e., in the conducted in a greenhouse with appropriate temperature (24–26 ◦ C)
Sea of Yellow, Bohai, and East China). The massive accumulation of and humidity (60 %− 70 %).
seaweed along the coastline causes environmental problems. To reduce After 45 days of growth, tomato seedling samples were collected for
the impacts, a large volume of seaweed is buried as waste (Yoruklu et al., further analysis. Following Zheng et al. (2021), we collected rhizosphere
2022). However, waste seaweeds may provide a natural bioresource for samples which were tightly adhered to the roots of tomato plants. The
organic fertilizers (Zhang and Sun, 2017; Yoruklu et al., 2022). Aban­ tomato rhizosphere soil samples were used to analyze the physico­
doned seaweeds are rich in organic compounds such as proteins, amino chemical properties, microbial community compositions, and enzymatic
acids, laminarin, alginate, and special sulfated polysaccharides (Khan activities.
et al., 2009; Gibilisco et al., 2022; Sousa et al., 2022). The plant growth
can be promoted by the fertilizers based on waste seaweeds which also
2.2. Tomato seedling analysis
improve plant stress resilience (Heo et al., 2005; Wang et al., 2016;
Wang et al., 2018a). Moreover, a variety of microorganisms have been
Fresh root, stem, and leaf samples of the tomato seedlings were
proved to exist in seaweed. For example, seaweed was mainly composed
collected from each pot to determine their biochemical characteristics.
of Bacteroidetes and Alphaproteobacteria, which was conducive to the
After adequately cleaning each tomato seedling, the stem height, root
evolution of microbial communities in soil (Egan et al., 2013; Tujula
length, and fresh/dry weight were measured. The contents of soluble
et al., 2010). Consequently, fertilizer based on waste seaweed can
sugar, soluble protein, malondialdehyde (MDA), and proline in the to­
effectively influence microbial community succession in the
mato (fresh roots, leaves, and stems) were analyzed using the corre­
acid-affected soil and enhance soil cultivability.
sponding test kits (Jiangsu Kete Biotechnology Co., Ltd., Nanjing City,
Recently, biochar made from various thermochemicals has attracted
China). In addition, the phytohormone (zeatin riboside (ZR), gibberellic
attention as a conditioner for biotic and abiotic stress-affected soils (Sun
acid3 (GA3), indole-3-acetic acid (IAA), and abscisic acid (ABA)) con­
et al., 2020). Biochar effectively alleviated abiotic stress in plants by
tents of the tomato seedlings were measured using the different enzyme-
increasing the nutrient availability in soil (Mahmoud et al., 2020; Sai­
linked immunosorbent assay (ELISA) kits (BYabscience, Nanjing City,
fullah et al., 2018). Biochar produced from corn straw decreases the
China, Jiangsu Kete Biotechnology Co., Ltd., Nanjing City, China),
concentration of Na+ in saline-alkali soil and enhanced the content and
respectively. Following the methodology described previously (Shang
availability of N, K, and P (Zhao et al., 2020). The plant growth in an
et al., 2021), the polyphenol oxidase (PPO), catalase (CAT), peroxidase
abiotic stress-affected soil was enhanced through the improvement of
(POD), and superoxide dismutase (SOD) were extracted and measured
soil physicochemical properties by biochar (Burrell et al., 2016). In
from the tomato samples using activity test kits (Beijing Solarbio Science
addition, biochar also conspicuously enhanced soil microbial diversity
& Technology Co., Ltd., Beijing city, China).
and shifted the microbial community to indirectly improve host growth
in stress-affected soils (Yan et al., 2021).
Although biochar and seaweed liquid fertilizers have been applied to 2.3. Analysis of the soil physicochemical properties and enzyme activities
the agricultural production, the effects of the combination of biochar
and waste seaweed fertilizer on abiotic stress-impacted soil are un­ The electrical conductivity (EC) and pH of the rhizosphere soil were
known. In this study, biochar and seaweed liquid fertilizer made from all determined at a water/soil mass ratio of 5:1. The amount of total
waste materials were investigated to determine their potential to nitrogen (TN) was determined by using standard method (Cooper et al.,
improve acid-affected soil of Jiaodong Peninsula, China. The main ob­ 2020), and the soil ammonium nitrogen (AN), and nitrate nitrogen (NN)
jectives were to examine the effect of waste seaweed liquid fertilizer and contents were detected in a Skalar Continuous Flow Analyzer 5000
biochar on the acid-affected soil, namely: (1) evaluate the effect of (Skalar Analytical BV, Breda, Netherlands). The available potassium
biochar and waste seaweed liquid fertilizer on tomato seedling growth (AK), and available phosphorus (AP) contents in the soil samples were
in an acid-affected soil; (2) investigate the effect of waste seaweed liquid estimated using the quinolinium phosphomolybdate and ammonium
fertilizer and biochar on the properties of acid-affected soil; (3) explore acetate-flame photometer methods, respectively. In addition, the po­
the relationships between the alterations in soil properties and soil mi­ tassium dichromate heating oxidation-volumetric method determined
crobial communities; and (4) predict the tomato rhizosphere microbial the levels of organic matter (OM). The organic carbon (OC) was
metabolic pathways after adding biochar and seaweed liquid fertilizer. analyzed using a Vario EL III element analyzer.
The activities of the soil laccase (SLA), catalase (SCAT), acid phos­
2. Material and methods phatase (SAP), sucrase (SSU), and urease (SUR) were determined using
enzyme activity test kits (Jiangsu Kete Biotechnology Co. Ltd., Nanjing
2.1. Microcosmic experiment design and sample collection City, China). The unit of soil enzyme activity (SLA, SCAT, SAP, SSU, and
SUR) was defined as follows, respectively. The amount of enzyme
The waste seaweed liquid fertilizer used in this study was obtained required to oxidize 1 nmol substrate 2,2′ -azino-bis (3-ethyl­
from the Weihai Academy of Agricultural Sciences (Weihai City, China), benzothiazoline-6-sulfonic acid) (ABTS) 1 min 1 g of soil at 37 ◦ C to
produced by biological methods from waste seaweed (bladder-wrack). provide one unit of laccase activity. One unit of catalase activity
Biochar made of reed straw was prepared following the procedure required to catalyze 1 µmol H2O2 degradation of air-dried soil (1 g) at
outlined in our laboratory (Sun et al., 2022). The acid-affected soil (pH 37 ◦ C every day. The amount of acid phosphatase required to release 1
＜6.21) was collected from a greenhouse with long-term vegetable nmol phenol of soil (1 g) one day at 37 ◦ C was one unit. One unit of
production in Jiaodong Peninsula, China. The microcosmic experiment sucrase activity required to produce 1 mg of reducing sugar 1 g of soil at
involved collecting an acid-affected soil and uniformly mixing it before 37 ◦ C. One unit of urease activity required to produce NH3-N (1 µg) per
dividing the soil between four treatments of: 1) the control with no gram of soil at 37 ◦ C.

2
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

Fig. 1. Stem height (a), root length (b), fresh weight (c), and dry weight (d) of tomato seedlings in an acid-affected soil. Control: acid-affected soil; RBC: acid-affected
soil+ 3 % biochar; SLF: acid-affected soil+ 4 % waste seaweed liquid fertilizer; RBC+SLF: acid-affected soil+ 1.5 % biochar+ 2 % waste seaweed liquid fertilizer.
Different letters indicate significant difference at P＜0.05.

2.4. Rhizosphere soil DNA extraction, PCR amplification, and 16S rRNA 97 %, operational taxonomic unit (OTU) clustering of non-repetitive
gene amplicon sequencing sequences was obtained using UPARSE (version 7.0.1090). The α-di­
versity (Chao 1 and Shannon index) was calculated using Mothur soft­
The FastDNA Spin Kit for Soil (MP Biomedicals, CA, United States) ware. To visualize variations in microbial compositions, non-metric
extracted the genomic DNA from the rhizosphere soil samples. multidimensional scaling (NMDS), and principal component analysis
Following the manufacturer’s instructions, 50 μL of DNA was extracted (PCA) of the treatment samples were applied using QIIME and R soft­
from rhizosphere soil (300 mg) and stored at − 20 ◦ C until required for ware (v. 3.5.3). Redundancy analysis (RDA) screened the environmental
further analysis. The primers 799 F/1193 R (reverse:5′ -ACG TCA TCC factors using variance inflation factor (VIF) analysis. Additionally, the
CCA CCT TCC-3′ ) amplified the V5 - V7 regions of the 16 S rRNA genes of Kyoto Encyclopedia of Genes and Genomes (KEGG) database (http://
the extracted DNA under the following conditions: 95 ◦ C (3 min), fol­ www.genome.jp/kegg/) was used to investigate the metabolic path­
lowed by 13 cycles of 95 ◦ C (30 s), 55 ◦ C (30 s), and 72 ◦ C (45 s), an ways of microorganisms in the soil using PICRUSt software.
extension at 72 ◦ C (10 min) (Bulgarelli et al., 2015). The PCR reactions
were performed in a 20 μL mixture containing 1 μL DNA template (10 ng 2.6. Statistical analysis
of DNA), 4 μL of 5 × FastPfu Buffer (TransGen Biotech, China), 0.8 μL of
each primer (5 μM total), 0.4 μL of FastPfu Polymerase (TransGen Data processing and figure creations used SPSS Statistics 20.0 (Chi­
Biotech, China), 2 μL dNTPs (2.5 mm, TransGen Biotech, China), and cago, USA) and Origin 8.5 (Northampton, USA), respectively. One-way
0.2 μL BSA (TransGen Biotech, China). After PCR amplification, the ANOVA and post-hoc Tukey’s tests analyzed the significance of the
bands were excised and purified with 1.2 % agarose gel using a PCR variations in the data. Different letters in the Figures and Tables indicate
Purification Kit. The purified PCR products from all samples were significant differences (P < 0.05).
collected, and paired-end sequencing was performed on the Illumina
MiSeq sequencing platform (Shanghai Majorbio Bio-pharm Technology 3. Results and discussion
Co., Ltd., Shanghai city, China).
3.1. Effect of biochar and waste seaweed liquid fertilizer on tomato
2.5. Amplicon sequence processing and bioinformatics analysis seedlings

The Quantitative Insights into Microbial Ecology pipeline analyzed 3.1.1. Tomato seedling growth
the 16S rRNA gene sequence data. Restricting the data to similarity of ≥ Recently, biochar has been proven to be effective in ameliorating

3
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

Fig. 2. Proline (a), MDA (b), soluble sugar (c), and soluble protein (d) contents of tomato seedlings in an acid-affected soil. Control: acid-affected soil; RBC: acid-
affected soil+ 3 % biochar; SLF: acid-affected soil+ 4 % waste seaweed liquid fertilizer; RBC+SLF: acid-affected soil+ 1.5 % biochar+ 2 % waste seaweed
liquid fertilizer.

plant growth in unfavorable soil environments (Zhao et al., 2020; Sun stresses, reflecting oxidative damage in plants (Li et al., 2017). Biochar
et al., 2020). The synergistic effects of biochar and waste seaweed liquid and waste seaweed liquid fertilizer led to a reduction in MDA level in the
fertilizer on the tomato seedlings growth profile were verified (Fig. 1 tomato seedlings, indicating that the biochar and waste seaweed liquid
and Fig. S1). Seedlings treated with RBC, SLF, and RBC+SLF had fertilizer can alleviate the oxidative damage (She et al., 2018). This may
significantly elevated stem heights by 28.11 %, 30.19 %, and 48.11 %, be due to the biochar and waste seaweed liquid fertilizer promoting
respectively (Fig. 1a). The RBC, SLF, and RBC+SLF treatments induced proline synthesis because ROS can be effectively removed by proline,
longer roots than the control (Fig. 1b). The fresh and dry weights of alleviating oxidative damage (Rady et al., 2019). The soluble sugar
tomato seedlings treated with biochar and waste seaweed liquid fertil­ content in the tomato seedling leaves grown with RBC, SLF, and
izer are provided in Fig. 1c,d. The RBC+SLF possessed the highest fresh RBC+SLF was significantly increased by 34.02 %, 48.03 %, and 49.92 %,
weights of tomato seedlings. In addition, the tomato seedling dry respectively (Fig. 2c). The soluble protein content in the tomato seedling
weights were dramatically increased by 23.33 % (RBC), 29.93 % (SLF), roots, stems, and leaves was significantly increased by the addition of
and 63.66 % (RBC+SLF). Effective soil conditioning might be the prin­ biochar and waste seaweed liquid fertilizer to the soil (Fig. 2d). Soluble
cipal reason for the enhanced growth of tomato seedlings (Yin et al., sugars and soluble proteins in plants are closely associated with host
2022). plant resistance to stressful environments (Du et al., 2020; Rady et al.,
2019). The significant increase in their contents reveals biochar and
3.1.2. Tomato seedling proline, MDA, soluble sugar, and soluble protein waste seaweed liquid fertilizer effectively improved tomato seedling
contents growth in an acid-affected soil.
RBC+SLF significantly increased the proline content in the roots,
leaves, and stems of tomato seedlings by 33.83 %, 108.70 %, and 112.33 3.1.3. Tomato seedling phytohormone content
%, respectively. However, RBC had no significant effect on any specific The effective response to environmental stresses in plant depends on
tomato part (Fig. 2a). The MDA content in the tomato seedling roots, the production and transport of phytohormones (Hao et al., 2020). In
stems, and leaves decreased in the RBC, and RBC+SLF treatments. particular, IAA plays a vital role in promoting the elongation of plant
Additionally, the MDA content in the tomato seedling leaves in the roots, and stems (Li et al., 2019). The IAA content of tomato seedling
control group was two times higher than that in the RBC+SLF treatment roots, stems, and leaves was significantly increased by SLF, and
(Fig. 2b). The accumulation of MDA in plant tissues is mainly caused by RBC+SLF treatments in this study (Fig. 3a). Biochar and waste seaweed
reactive oxygen species which can be induced by various adverse liquid fertilizer dramatically enhanced ZR content in the tomato

4
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

Fig. 3. IAA (a), ZR (b), GA3 (c), and ABA (d) contents of tomato seedlings in an acid-affected soil. Control: acid-affected soil; RBC: acid-affected soil+ 3 % biochar;
SLF: acid-affected soil+ 4 % waste seaweed liquid fertilizer; RBC+SLF: acid-affected soil+ 1.5 % biochar+ 2 % waste seaweed liquid fertilizer.

seedling stems by 32.68–64.52 % (Fig. 3b). The generation and accu­ RBC+SLF treatments were significantly increased (Fig. 4a,b,c). SOD and
mulation of ZR (a major endogenous cytokinin in plants) can promote POD can transform O-2 into an H2O2, reducing the toxicity to plant cells
cell division to enhance plant self-repair after exogenous insults (Hao (Shang et al., 2021). CAT can remove H2O2 and cooperates with SOD to
et al., 2018). In addition, SLF, and RBC+SLF positively affected the GA3 reduce oxidative damage to the cell membrane systems (Peng et al.,
levels in the tomato seedling roots and leaves. GA3 is a vital phytohor­ 2019). Biochar and waste seaweed liquid fertilizer significantly
mone in promoting root and stem elongation (Hao et al., 2020), in the enhanced the PPO activity in the tomato seedling roots by 57.14 %
tomato seedling the GA3 content was significantly promoted by 117.69 (Fig. 4d). PPO is related to the synthesis of quinone and lignin which are
% (in the roots), 19.66 % (in the stems), and 42.86 % (in the leaves) with beneficial for plants under abiotic stresses (Shang et al., 2021; Peng
the RBC+SLF treatment (Fig. 3c). The elongation of the roots and stems, et al., 2019). These results revealed that biochar and waste seaweed
and the increase in the biomass (Fig. 1) of the tomato seedlings grown liquid fertilizer can enhance multiple protective enzymes in tomato
with biochar and waste seaweed liquid fertilizer might be associated seedlings, which might be the mechanism used by tomato seedlings to
with various syntheses including IAA, ZR, and GA3. ABA is a good in­ grow in an acid-affected soil.
dicator of adverse soil stress and is used to evaluate the soil health status
(Liu et al., 2005). For example, host plants produce more ABA under 3.2. Effect of biochar and waste seaweed liquid fertilizer on soil
abiotic stress-affected soil (Duan et al., 2013). Uniformly, the biochar physicochemical properties
and waste seaweed liquid fertilizer lowered the ABA content in the to­
mato seedling roots by 23.26–64.75 %, but did not significantly affect Soil physicochemical properties are essential indicators that influ­
the content of ABA in the tomato seedling stems (Fig. 3d). The decreased ence nutrient supply and microbial activity for crops (Tang et al., 2022).
ABA content under the RBC+SLF treatment might be attributed to an The physicochemical properties of the acid-affected soil with the addi­
improved soil acidity. tion of biochar and waste seaweed liquid fertilizer are provided in
Table 1. RBC, SLF, and RBC+SLF significantly increased the pH of the
3.1.4. Protective enzyme activities in tomato seedlings soil samples by 18.40 %, 21.17 %, and 22.80 %, respectively. Therefore,
Plants fail to adapt and grow in stress-affected soils without using biochar and waste seaweed liquid fertilizer can reduce the soil acidifi­
protective enzyme systems, including SOD, POD, CAT, and PPO (Shang cation caused by continuous cropping (Song et al., 2021). The EC values
et al., 2021). Compared with the control group, the SOD, POD, and CAT reflect the ability of the soil to absorb, retain, and exchange ions (i.e., Cl-,
activities in tomato seedling roots and leaves under the SLF and Ca2+, Mg2+, and K+), and soils with higher EC values have less nutrient

5
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

Fig. 4. SOD (a), POD (b), CAT (c), and PPO (d) activities of tomato seedlings in an acid-affected soil. Control: acid-affected soil; RBC: acid-affected soil+ 3 % biochar;
SLF: acid-affected soil+ 4 % waste seaweed liquid fertilizer; RBC+SLF: acid-affected soil+ 1.5 % biochar+ 2 % waste seaweed liquid fertilizer.

leaching (Yu et al., 2019).

Table 1
Furthermore, biochar directly provides nutrients and also improves
Effect of waste seaweed liquid fertilizer and biochar on the soil properties of
soil nutrient availability for the host plants (Agegnehu et al., 2017;
tomato rhizosphere.
Zhang et al., 2021). The addition of RBC+SLF significantly improved the
Treatment Control RBC SLF RBC+SLF soil TN from 1.36 mg.g− 1 to 2.12 mg.g− 1. Treatments RBC, SLF, and
pH 6.14 ± 0.11c 7.27 ± 0.12b 7.44 7.54 ± 0.07a RBC+SLF all significantly increased the soil OM and OC. Moreover, all
± 0.13ab treatments remarkably elevated the soil NN content by 24.57 % (RBC),
EC (μS/cm) 321.68 355.59 363.89 408.04
35.90 % (SLF), and 90.38 % (RBC+SLF) when compared to the control.
± 12.00c ± 22.79bc ± 17.9b ± 22.25a
TN (mg.g− 1 1.36 ± 0.10b 1.48 ± 0.14b 1.56 ± 0.19b 2.12 ± 0.13a The AN content in the soil was significantly increased by 49.31 % (RBC),
soil) 74.38 % (SLF), and 149.31 % (RBC+SLF). Our findings supported pre­
AP (μg.g− 1 41.42 75.91 ± 2.54b 79.89 111.78 vious research demonstrating biochar promoted the decomposition of
soil) ± 3.57c ± 1.56b ± 2.16a soil organic matter and enhanced the abundance of nitrogen-fixing mi­
AK (μg.g− 1 196.74 219.02 230.12 311.78
soil) ± 8.90c ± 17.55bc ± 13.07b ± 14.75a
croorganisms, which was conducive to the availability of soil NN and AN
NN (μg.g− 1 4.68 ± 0.10d 5.83 ± 0.13c 6.36 ± 0.13b 8.91 ± 0.11a (Luo et al., 2017). The availability of K and P is crucial for the growth of
soil) plant in the acid-affected soil (Saifullah et al., 2018). The AP and AK
AN (μg.g− 1 3.63 ± 0.36c 5.42 ± 0.56b 6.33 ± 0.49b 9.05 ± 0.68a contents in rhizosphere soil were significantly increased by 169.87 %
soil)
and 58.47 % with biochar and waste seaweed liquid fertilizer, respec­
OM (mg.g− 1 8.81 ± 0.59d 10.02 ± 0.33c 13.38 16.16
soil) ± 0.60b ± 0.91a tively. This improvement might be related to the increased abundance of
OC (mg.g− 1 2.45 ± 0.16d 3.72 ± 0.19c 5.12 ± 0.14b 6.27 ± 0.30a rhizosphere bacteria (potassium-solubilizing, and phosphate
soil) -solubilizing) in the tomato and their inherent nutrients (Liu et al.,
EC: Electrical conductivity, TN: Total nitrogen, AP: Available phosphorus, AK: 2017). The property changes reflected that biochar and waste seaweed
Available potassium, NN: Nitrate nitrogen, AN: Ammonium nitrogen, OM: liquid fertilizer promoted tomato growth in an acid-affected soil by
Organic matter, OC: Organic carbon. Control: acid-affected soil; RBC: acid- improving the soil nutrient status.
affected soil+ 3 % biochar; SLF: acid-affected soil+ 4 % waste seaweed liquid
fertilizer; RBC+SLF: acid-affected soil+ 1.5 % biochar+ 2 % waste seaweed
liquid fertilizer.

6
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

Table 2 %, 42.29 %, and 67.73 % under RBC, SLF, and RBC+SLF, respectively.
Effect of biochar and waste seaweed liquid fertilizer on the enzyme activities of Zhao et al. (2021) reported that laccase activity in stress-affected soil
tomato rhizosphere soil. decreases significantly, which is consistent with our results. In addition,
Enzyme activities Control RBC SLF RBC+SLF biochar and waste seaweed liquid fertilizer significantly increased the
SLA 50.30 70.87 71.57 84.37
activities of SCAT and SUR, which may be due to the increased bacterial
(nmol⋅min− 1⋅g− 1) ± 3.68c ± 2.87b ± 2.85b ± 3.15a abundance at the genus level (Lee et al., 2021). The SAP and SSU in the
SCAT 9.30 18.60 32.03 42.33 SLF and RBC+SLF groups were significantly enhanced. The RBC treat­
(μmol⋅d− 1⋅g− 1) ± 1.28d ± 2.15c ± 3.21b ± 1.94a ment had no positive effects on the activities of SAP and SSU. Soil
SAP (μmol⋅d− 1⋅g− 1) 10.80 13.13 20.57 26.43
alkalization can activate acid phosphatase activity, which is consistent
± 2.49c ± 1.99c ± 1.96b ± 2.32a
SSU (U⋅g− 1) 5.70 6.80 23.67 35.97 with soil pH changes (Arteaga et al., 2022).
± 1.23c ± 1.75c ± 2.15b ± 2.75a
SUR (U⋅g− 1) 1636 3559 ± 95c 5668 ± 56b 6810
± 74d ± 122a 3.4. Response of soil microbial community to biochar and waste seaweed
SLA: soil laccase; SCAT: soil catalase; SAP: soil acid phosphatase; SSU: soil su­
liquid fertilizer amendments
crase; SUR: soil urease. Control: an acid-affected soil; RBC: an acid-affected
soil+ 3 % biochar; SLF: an acid-affected soil+ 4 % waste seaweed liquid fertil­ The 16S rRNA gene amplicon sequencing provided 708,400 clean
izer; RBC+SLF: an acid-affected soil+ 1.5 % biochar+ 2 % waste seaweed liquid reads. The rarefaction curves of all the soil samples reached the satu­
fertilizer. ration plateau, indicating that sequencing depth in this study was
enough to represent the microbial communities in each sample (Fig. S2).
3.3. Effect of biochar and waste seaweed liquid fertilizer on soil enzyme The Venn diagram (Fig. S3) shows the sequences were grouped into
activities 2547 bacterial OTUs. In addition, 870 bacterial OTUs were shared
among the four treatment groups, while 515 specific OTUs were unique
The effects of biochar and waste seaweed liquid fertilizer on the to RBC+SLF, 90 to SLF, 33 to RBC, and 63 were specific to the control
enzymatic activities of the tomato soil are provided in Table 2. SLA can group.
estimate soil status, a higher SLA activity is beneficial for plant growth Rhizosphere bacteria can be treated as the key evaluation indicators
(Zhao et al., 2021). The activity of SLA was dramatically higher by 40.89 in the abiotic stress-affected soil by playing important roles in nutrient
conversion, and organic matter degradation of soil (Feng et al., 2022; Liu

Fig. 5. The Chao1 (a), and Shannon (b) diversity indices and PCA (c), and NMDS (d) analysis of the bacterial communities of tomato rhizosphere soil. Control: acid-
affected soil; RBC: acid-affected soil+ 3 % biochar; SLF: acid-affected soil+ 4 % waste seaweed liquid fertilizer; RBC+SLF: acid-affected soil+ 1.5 % biochar+ 2 %
waste seaweed liquid fertilizer.

7
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

Fig. 6. The relative abundance and composition of bacterial communities at the phylum level (a), the significance comparisons of bacterial communities at the
phylum level (b), the heat map of the top 30 bacterial genera (c), and the significance comparisons of bacterial genera communities (d). Control: acid-affected soil;
RBC: acid-affected soil+ 3 % biochar; SLF: acid-affected soil+ 4 % waste seaweed liquid fertilizer; RBC+SLF: acid-affected soil+ 1.5 % biochar+ 2 % waste seaweed
liquid fertilizer.

et al., 2020). The biochar and waste seaweed liquid fertilizer mix At the phylum level, the dominant bacterial communities in all
(RBC+SLF) significantly increased the tomato soil bacterial Chao 1 and treatments were Proteobacteria, Actinobacteria, Firmicutes, and Bac­
Shannon indices (Fig. 5a,b). Therefore, the bacterial diversity, and teroidetes (Fig. 6a). The same composition of dominant bacterial phyla
abundance of rhizosphere soil were significantly increased under has been reported in other rhizosphere soils (Zheng et al., 2021; Mendes
RBC+SLF treatment. This significant increase in the relative abundance et al., 2018). Compared with the control group, the relative abundance
of soil bacteria might be due to the presence of a variety of active en­ of Proteobacteria under RBC+SLF treatment was significantly lower.
zymes and substances, such as protease, cellulase, ACC deaminase, IAA, Conversely, Firmicutes, Actinobacteria, Bacteroidetes, and Gemmati­
and siderophores (Shang et al., 2021; Guo et al., 2020). With a sufficient monadota were higher in RBC+SLF (Fig. 6b). Many Firmicutes strains
supply of these enzymes, the growth and reproduction of rhizosphere can produce proteases, lipases, cellulases, and other extracellular en­
bacteria are adequately promoted (Puri et al., 2020). Adding exogenous zymes, and metabolize a variety of soil substances (Lim et al., 2014). In
conditioners to the soil effectively increases the abundance of functional addition, Actinobacteria are involved in the metabolism of nitrogen and
bacterial communities (de Andrade et al., 2019; Shang et al., 2021). phosphorus in soil and assist in the decomposition of organic phos­
To further explore the variation in bacterial communities, we per­ phorus, and nitrogen (Kristensen et al., 2021).
formed PCA and NMDS analyses at the OTU level. The rhizosphere The relative abundances of the top 30 bacterial genera were observed
bacterial communities under Control, RBC, SLF, and RBC+SLF treat­ in all treatments (Fig. 6c). The dominant genera of rhizosphere soil
ments were distributed in different quadrants (Fig. 5c,d). The PC1 and samples were Acidovorax (4.61–14.20 %), Pseudomonas (4.13–7.71 %),
PC2 axis separation of the bacterial communities were 46.15 % and and Simplicispira (1.70–6.48 %). In soil, Pseudomonas can easily transfer
18.74 %, respectively. These findings demonstrate that the bacterial to the plant rhizosphere to form biofilms and antagonize pathogens
communities in the Control and RBC+SLF treatment were significantly (Pour et al., 2019; Shang et al., 2021). Moreover, Pseudomonas can
different. decompose cellulose in the rhizosphere environment by producing

8
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

Fig. 7. RDA of bacterial communities and properties (a), enzyme activities (b) in rhizosphere soil, and correlation heat maps of soil properties (c), soil enzyme
activities (d) and the dominant genera of bacteria. EC: Electrical conductivity; TN: Total nitrogen; AP: Available phosphorus; AK: Available potassium; NN: Nitrate
nitrogen; and AN: Ammonium nitrogen in (a,c). SLA: soil laccase; SCAT: soil catalase; SAP: soil acid phosphatase; SSU: soil sucrase; and SUR: soil urease in (b,d).
Control: acid-affected soil; RBC: acid-affected soil+ 3 % biochar; SLF: acid-affected soil+ 4 % waste seaweed liquid fertilizer; RBC+SLF: acid-affected soil+ 1.5 %
biochar+ 2 % waste seaweed liquid fertilizer.

xylanase and cellulose-degrading enzymes (Li et al., 2022). The addition 3.5. Correlations between soil properties, soil enzyme activities and
of RBC, SLF, and RBC+SLF significantly decreased the relative abun­ dominant soil microorganisms
dance of Methylophilus and Devosia. The RBC+SLF treatment signifi­
cantly enriched Ideonella, Ramlibacter, and Sphingomonas (Fig. 6d). Some As previously reported (Zhu et al., 2019; Zhou et al., 2019), the
bacterial isolates from the Sphingomonas genus can decompose organic application of biochar significantly influenced the conversion of nutri­
materials in the soil environment, providing necessary nutrients for ents in the soil and positively altered the soil enzyme activity enabling
plant growth (Song et al., 2021). the evolution of the microbial community. We used RDA to evaluate the
correlations between soil properties, soil enzyme activities, and
soil-dominant microorganisms (Fig. 7a,b). Of the soil nutrient in­
dicators, pH was the most affected by the soil bacterial communities,
followed by AP and NN. TN, AN, and NN contents were closer to

9
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

RBC+SLF amendment, indicating the combination of biochar and waste studies including the regulatory network of plant genes and soil
seaweed liquid fertilizer had the most significant impacts on the metabolite factors to be measured, our findings can improve the un­
ammonium nitrogen and organic matter conversion in the soil (Zheng derstanding of the association between soil conditioner and plant
et al., 2021). Moreover, TN was significantly positively correlated with growth in the context of abiotic pollution and help develop promising
five bacterial genera: Bacillus, Ideonella, Pseudomonas, Ramlibacter, and management strategies for soil pollution.
Sphingomonas. Intriguingly, TN had a significant positive correlation
with Pseudomonas (which contains multiple beneficial bacterial species), CRediT authorship contribution statement
which is consistent with the previous researches (Shang et al., 2021;
Zheng et al., 2021). Xian-chao Shang, and Manman Zhang: Resources, Conceptuali­
RDA revealed the SLA was positively correlated with SCAT, SAP, zation, Methodology, Writing-original draft. Yu-qin Zhang, and
SSU, and SUR. The RBC+SLF treatment had the most significant effect Yiqiang Li: Methodology, Data curation, Writing-review & editing.
on the five enzyme activities investigated in the acid-affected soil Long Yang: Resources, Writing-review & editing. Xin Hou: Funding
(Fig. 7b). This might be attributed to the evolution of the bacterial acquisition, Conceptualization, Writing-original draft.
community being induced by biochar and waste seaweed liquid fertilizer
(Feng et al., 2019). Additionally, SCAT was significantly positively Declaration of Competing Interest
correlated with Azospira which contains multiple nitrogen fixing bac­
terial species (Shang et al., 2021). However, the relative abundance of The authors declare that they have no known competing financial
Pseudomonas which is a beneficial rhizobacterium (Dimkić et al., 2022), interests or personal relationships that could have appeared to influence
showed no significant correlation with the soil enzyme in the soil the work reported in this paper.
(Fig. 7d).
Data availability
3.6. Change prediction analyses of the microbial metabolic pathways
No data was used for the research described in the article.
The KEGG pathways (level 1) most closely associated with the
rhizosphere soil microorganisms were divided into five categories: Acknowledgments
organismal systems (the relative abundances of the functional genes:
0.84–0.88 %), cellular processes (5.65–5.82 %), genetic information The authors are grateful for the financial support from the National
processing (14.83–15.64 %), environmental information processing Natural Science Foundation of China (32101289), Shandong Province
(15.40–16.89 %), and metabolism (46.46–47.34 %) (Fig. S4a). Inter­ Modern Agricultural Technology System (SDAIT-25-02), and Taishan
estingly, Ma et al. (2022) and Li et al. (2020) reported similar results for brand cigarette high-quality core raw material development and appli­
the functional genes of KEGG level 1 pathways. In addition, KEGG level cation in Shandong (202102004).
2 pathways were analyzed. The key microbial metabolic pathways were
membrane transport (12.36–13.90 %), amino acid metabolism Appendix A. Supporting information
(9.74–10.04 %), carbohydrate metabolism (8.73–9.00 %), replication
and repair (6.43–6.82 %) (Fig. S4b). Compared with the control group, Supplementary data associated with this article can be found in the
the RBC+SLF treatment decreased the membrane transport but online version at doi:10.1016/j.ecoenv.2023.115075.
increased the amino acid metabolism, replication and repair, and energy
metabolism. Significant changes in the soil microbial metabolism have References
been observed after soil inoculation with Bacillus subtilis (Li et al., 2021).
The RDA further demonstrated the cysteine and methionine metabolism Agegnehu, G., Srivastava, A.K., Bird, M.I., 2017. The role of biochar and biochar-compost
(ko00270), phenylalanine, tyrosine, and tryptophan biosynthesis in improving soil quality and crop performance: a review. Appl. Soil Ecol. 119,
156–170. https://doi.org/10.1016/j.apsoil.2017.06.008.
(ko00400), valine, leucine, and isoleucine biosynthesis (ko00290), and
Arteaga, J.F.M., Gluhar, S., Kaurin, A., Lestan, D., 2022. Simultaneous removal of arsenic
histidine metabolism (ko00340) were mainly affected by the RBC+SLF and toxic metals from contaminated soil: laboratory development and pilot scale
treatment (Fig. S4c). The increase in soil TN, NN, and AN may be due to demonstration. Environ. Pollut. 294, 118656 https://doi.org/10.1016/j.
envpol.2021.118656.
the enhanced amino acid and nitrogen metabolism genes (Huang et al.,
Bulgarelli, D., Garrido-Oter, R., Münch, P.C., Weiman, A., Dröge, J., Pan, Y., McHardy, A.
2021). C., Schulze-Lefert, P., 2015. Structure and function of the bacterial root microbiota
in wild and domesticated barley. Cell. Host. Microbe 17, 392–403. https://doi.org/
4. Conclusions 10.1016/j.chom.2015.01.011.
Burrell, L.D., Zehetner, F., Rampazzo, N., Wimmer, B., Soja, G., 2016. Long-term effects
of biochar on soil physical properties. Geoderma 282, 96–102. https://doi.org/
The combination of biochar and waste seaweed liquid fertilizer 10.1016/j.geoderma.2016.07.019.
dramatically promoted tomato seedling growth in an acid-affected soil Chen, L., Xie, X., Kang, H., Liu, R., Shi, Y., Li, L., Xie, J., Li, B., Chai, A., 2022. Efficiency
of calcium cyanamide on the control of tomato soil-borne disease and their impacts
of Jiaodong Peninsula, China. The RBC+SLF showed the highest on the soil microbial community. Appl. Soil Ecol. 176, 104522 https://doi.org/
improvement with significant promotion effects when compared to RBC 10.1016/j.apsoil.2022.104522.
or SLF in isolation. RBC, SLF, and RBC+SLF improved the soil proper­ Chen, W., Teng, Y., Li, Z., Liu, W., Ren, W., Luo, Y., Christie, P., 2018. Mechanisms by
which organic fertilizer and effective microbes mitigate peanut continuous cropping
ties, soil enzymatic activities, and enhanced the physiological status of yield constraints in a red soil of south China. Appl. Soil Ecol. 128, 23–34. https://doi.
the tomato seedlings. RBC+SLF enhanced proline, MDA, soluble sugar, org/10.1016/j.apsoil.2018.03.018.
soluble protein, and phytohormone contents. The beneficial bacteria (i. Cooper, J., Greenberg, I., Ludwig, B., Hippich, L., Fischer, D., Glaser, B., Kaiser, M., 2020.
Effect of biochar and compost on soil properties and organic matter in aggregate size
e., Pseudomonas and Azospira) abundance was also enhanced in fractions under field conditions. Agr. Ecosyst. Environ. 295, 106882 https://doi.org/
RBC+SLF, effectively improving an acid-affected soil quality and tomato 10.1016/j.agee.2020.106882.
seedling growth. In addition, significant correlations between soil de Andrade, F.M., de Assis Pereira, T., Souza, T.P., Guimarães, P.H.S., Martins, A.D.,
Schwan, R.F., Pasqual, M., Dória, J., 2019. Beneficial effects of inoculation of
nutrient indicators (TN, NN, and NN), bacterial communities, and soil
growth-promoting bacteria in strawberry. Microbiol. Res. 223–225, 120–128.
microbial amino acid metabolism were identified by either correlation https://doi.org/10.1016/j.micres.2019.04.005.
and prediction analyses. Thus, the combination of biochar and waste Deepika, P., MubarakAli, D., 2020. Production and assessment of microalgal liquid
seaweed liquid fertilizer is a potential soil conditioner for an acid- fertilizer for the enhanced growth of four crop plants. Biocatal. Agric. Biotechnol. 28,
101701 https://doi.org/10.1016/j.bcab.2020.101701.
affected soil for improved tomato vitality. Although the actual molec­ Dimkić, I., Janakiev, T., Petrović, M., Degrassi, G., Fira, D., 2022. Plant-associated
ular mechanism behind the presented conclusions requires further Bacillus and Pseudomonas antimicrobial activities in plant disease suppression via

10
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

biological control mechanisms - a review. Physiol. Mol. Plant P. 117, 101754. Liu, F., Jensen, C.R., Shahanzari, A., Andersen, M.N., Jacobsen, S.-E., 2005. ABA
https://doi.org/10.1016/j.pmpp.2021.101754. regulated stomatal control and photosynthetic water use efficiency of potato
Du, Y., Zhao, Q., Chen, L., Yao, X., Zhang, W., Zhang, B., Xie, F., 2020. Effect of drought (Solanum tuberosum L.) during progressive soil drying. Plant Sci. 168, 831–836.
stress on sugar metabolism in leaves and roots of soybean seedlings. Plant Physiol. https://doi.org/10.1016/j.plantsci.2004.10.016.
Biochem. 146, 1–12. https://doi.org/10.1016/j.plaphy.2019.11.003. Liu, H., Li, H., Ning, H., Zhang, X., Li, S., Pang, J., Wang, G., Sun, J., 2019. Optimizing
Duan, L., Dietrich, D., Ng, C.H., Chan, P.M.Y., Bhalerao, R., Bennett, M.J., Dinneny, J.R., irrigation frequency and amount to balance yield, fruit quality and water use
2013. Endodermal ABA signaling promotes lateral root quiescence during salt stress efficiency of greenhouse tomato. Agr. Water Manag. 226, 105787 https://doi.org/
in arabidopsis seedlings. Plant Cell 25, 324–341. https://doi.org/10.1105/ 10.1016/j.agwat.2019.105787.
tpc.112.107227. Liu, S., Tang, W., Yang, F., Meng, J., Chen, W., Li, X., 2017. Influence of biochar
Egan, S., Harder, T., Burke, C., Steinberg, P., Kjelleberg, S., Thomas, T., 2013. The application on potassium-solubilizing Bacillus mucilaginosus as potential biofertilizer.
seaweed holobiont: understanding seaweed-bacteria interactions. FEMS Microbiol. Prep. Biochem. Biotechnol. 47, 32–37. https://doi.org/10.1080/
Rev. 37 (3), 462–476. https://doi.org/10.1111/1574-6976.12011. 10826068.2016.1155062.
Feng, Y., Hu, Y., Wu, J., Chen, J., Yrjälä, K., Yu, W., 2019. Change in microbial Liu, Z., Liu, J., Yu, Z., Yao, Q., Li, Y., Liang, A., Zhang, W., Mi, G., Jin, J., Liu, X.,
communities, soil enzyme and metabolic activity in a Torreya grandis plantation in Wang, G., 2020. Long-term continuous cropping of soybean is comparable to crop
response to root rot disease. For. Ecol. Manag. 432, 932–941. https://doi.org/ rotation in mediating microbial abundance, diversity and community composition.
10.1016/j.foreco.2018.10.028. Soil Till. Res. 197, 104503 https://doi.org/10.1016/j.still.2019.104503.
Feng, Y., Zhang, H., Song, X., Ge, T., Zhu, J., Zhou, C., Cobb, K., Yan, X., Ruan, R., Luo, X., Liu, G., Xia, Y., Chen, L., Jiang, Z., Zheng, H., Wang, Z., 2017. Use of biochar-
Cheng, P., 2022. Microalgae as a potential conditioner for continuous cropping compost to improve properties and productivity of the degraded coastal soil in the
obstacles for taro (Colocasia esculenta L. Schott) production. J. Clean. Prod. 369, Yellow River Delta, China. J. Soils Sediment. 17, 780–789. https://doi.org/10.1007/
133356 https://doi.org/10.1016/j.jclepro.2022.133356. s11368-016-1361-1.
Gibilisco, P.E., Negrin, V.L., Idaszkin, Y.L., 2022. Assessing the use of two halophytes Luo, Y., van Veelen, H.P.J., Chen, S., Sechi, V., ter Heijne, A., Veeken, A., Buisman, C.J.
species and seaweed composting in Cu-pollution remediation strategies. Mar. Pollut. N., Bezemer, T.M., 2022. Effects of sterilization and maturity of compost on soil
Bull. 176, 113413 https://doi.org/10.1016/j.marpolbul.2022.113413. bacterial and fungal communities and wheat growth. Geoderma 409, 115598.
Guo, Q., Shi, M., Chen, L., Zhou, J., Zhang, L., Li, Y., Xue, Q., Lai, H., 2020. The https://doi.org/10.1016/j.geoderma.2021.115598.
biocontrol agent Streptomyces pactum increases Pseudomonas koreensis populations in Ma, S., Qiao, L., Liu, X., Zhang, S., Zhang, L., Qiu, Z., Yu, C., 2022. Microbial community
the rhizosphere by enhancing chemotaxis and biofilm formation. Soil Biol. Biochem. succession in soils under long-term heavy metal stress from community diversity-
144, 107755 https://doi.org/10.1016/j.soilbio.2020.107755. structure to KEGG function pathways. Environ. Res. 214 (2), 113822 https://doi.
Hao, Y., Ma, C., Zhang, Z., Song, Y., Cao, W., Guo, J., Zhou, G., Rui, Y., Liu, L., Xing, B., org/10.1016/j.envres.2022.113822.
2018. Carbon nanomaterials alter plant physiology and soil bacterial community Mahmoud, E., Baroudy, A.E., Ali, N., Sleem, M., 2020. Spectroscopic studies on the
composition in a rice-soil-bacterial ecosystem. Environ. Pollut. 232, 123–136. phosphorus adsorption in salt-affected soils with or without nano-biochar additions.
https://doi.org/10.1016/j.envpol.2017.09.024. Environ. Res. 184, 109277 https://doi.org/10.1016/j.envres.2020.109277.
Hao, Y., Ma, C., White, J.C., Adeel, M., Jiang, R., Zhao, Z., Rao, Y., Chen, G., Rui, Y., Mendes, L.W., Mendes, R., Raaijmakers, J.M., Tsai, S.M., 2018. Breeding for soil-borne
Xing, B., 2020. Carbon-based nanomaterials alter the composition of the fungal pathogen resistance impacts active rhizosphere microbiome of common bean. ISME
endophyte community in rice (Oryza sativa L.). Environ. Sci.: Nano 7, 2047–2060. J. 12, 3038–3042. https://doi.org/10.1038/s41396-018-0234-6.
https://doi.org/10.1039/C9EN01400D. Mohammad, H., Mohammad, A., Saeed, B., Naser, Az.A., 2019. The effect of deficit
He, L., Zhong, H., Liu, G., Dai, Z., Philip, C.B., Xu, J., 2019. Remediation of heavy metal irrigation on yield and yield components of greenhouse tomato (Solanum
contaminated soils by biochar: Mechanisms, potential risks and applications in lycopersicum) in hydroponic culture in Ahvaz region. Iran. Sci. Hortic. 254, 84–90.
China. Environ. Pollut. 252, 846–855. https://doi.org/10.1016/j. https://doi.org/10.1016/j.scienta.2019.04.084.
envpol.2019.05.151. Peng, G., Zhao, X., Li, Y., Wang, R., Qi, G., 2019. Engineering Bacillus velezensis with high
Heo, S.J., Park, E.J., Lee, K.W., Jeon, Y.J., 2005. Antioxidant activities of enzymatic production of acetoin primes strong induced systemic resistance in Arabidopsis
extracts from brown seaweeds. Bioresour. Technol. 96 (14), 1613–1623. https://doi. thaliana. Microbiol. Res. 227, 126297 https://doi.org/10.1016/j.
org/10.1016/j.biortech.2004.07.013. micres.2019.126297.
Huang, B., Jia, H., Han, X., Gou, J., Huang, C., Wang, J., Wei, J., Wang, J., Zhang, C., Pour, M.M., Saberi-Riseh, R., Mohammadinejad, R., Hosseini, A., 2019. Investigating the
2021. Effects of biocontrol Bacillus and fermentation bacteria additions on the formulation of alginate- gelatin encapsulated Pseudomonas fluorescens (VUPF5 and
microbial community, functions and antibiotic resistance genes of prickly ash seed T17-4 strains) for controlling Fusarium solani on potato. Int. J. Biol. Macromol. 133,
oil meal-biochar compost. Bioresour. Technol. 340, 125668 https://doi.org/ 603–613. https://doi.org/10.1016/j.ijbiomac.2019.04.071.
10.1016/j.biortech.2021.125668. Puri, A., Padda, K.P., Chanway, C.P., 2020. In vitro and in vivo analyses of plant-growth-
Khan, W., Rayirath, U.P., Subramanian, S., Jithesh, M.N., Rayorath, P., Hodges, D.M., promoting potential of bacteria naturally associated with spruce trees growing on
Critchley, A.T., Craigie, J.S., Norrie, J., Prithiviraj, B., 2009. Seaweed extracts as nutrient-poor soils. Appl. Soil Ecol. 149, 103538 https://doi.org/10.1016/j.
biostimulants of plant growth and development. J. Plant Growth Regul. 28 (4), apsoil.2020.103538.
386–399. https://doi.org/10.1007/s00344-009-9103-x. Rady, M.M., Elrys, A.S., El-Maati, M.F.A., Desoky, E.M., 2019. Interplaying roles of
Kristensen, J.M., Singleton, C., Clegg, L.-A., Petriglieri, F., Nielsen, P.H., 2021. High silicon and proline effectively improve salt and cadmium stress tolerance in
diversity and functional potential of undescribed “acidobacteriota” in danish Phaseolus vulgaris plant. Plant Physiol. Biochem. 139, 558–568. https://doi.org/
wastewater treatment plants. Front. Microbiol. 12, 643950 https://doi.org/ 10.1016/j.plaphy.2019.04.025.
10.3389/fmicb.2021.643950. Rosa, D.D., Rowlings, D.W., Biala, J., Scheer, C., Basso, B., Grace, P.R., 2018. N2O and
Lee, J.K., Park, H.J., Cha, S.J., Kwon, S.J., Park, J.H., 2021. Effect of pyroligneous acid CO2 emissions following repeated application of organic and mineral N fertiliser
on soil urease, amidase, and nitrogen use efficiency by Chinese cabbage (Brassica from a vegetable crop rotation. Sci. Total Environ. 637–638, 813–824. https://doi.
campestris var. Pekinensis). Environ. Pollut. 291, 118132 https://doi.org/10.1016/j. org/10.1016/j.scitotenv.2018.05.046.
envpol.2021.118132. Saifullah, Dahlawi S., Naeem, A., Rengel, Z., Naidu, R., 2018. Biochar application for the
Li, H., Lei, P., Pang, X., Li, S., Xu, H., Xu, Z., Feng, X., 2017. Enhanced tolerance to salt remediation of salt-affected soils: challenges and opportunities. Sci. Total Environ.
stress in canola (Brassica napus L.) seedlings inoculated with the halotolerant 625, 320–335. https://doi.org/10.1016/j.scitotenv.2017.12.257.
Enterobacter cloacae HSNJ4. Appl. Soil Ecol. 119, 26–34. https://doi.org/10.1016/j. Shang, X.-C., Cai, X., Zhou, Y., Han, X., Zhang, C.-S., Ilyas, N., Li, Y., Zheng, Y., 2021.
apsoil.2017.05.033. Pseudomonas inoculation stimulates endophytic Azospira population and induces
Li, H., Xu, X., Zhang, M., Zhang, Y., Zhao, Y., Jiang, X., Xin, X., Zhang, Z., Zhang, R., systemic resistance to bacterial wilt. Front. Plant Sci. 12, 738611 https://doi.org/
Gui, Z., 2022. Accelerated degradation of cellulose in silkworm excrement by the 10.3389/fpls.2021.738611.
interaction of housefly larvae and cellulose-degrading bacteria. J. Environ. Manag. She, D., Sun, X., Gamareldawla, A.H.D., Nazar, E.A., Hu, W., Edith, K., Yu, S., 2018.
323, 116295 https://doi.org/10.1016/j.jenvman.2022.116295. Benefits of soil biochar amendments to tomato growth under saline water irrigation.
Li, J., Guan, Y., Yuan, L., Hou, J., Wang, C., Liu, F., Yang, Y., Lu, Z., Chen, G., Zhu, S., Sci. Rep. 8, 14743. https://doi.org/10.1038/s41598-018-33040-7.
2019. Effects of exogenous IAA in regulating photosynthetic capacity, carbohydrate Song, L., Niu, X., Zhang, N., Li, T., 2021. Effect of biochar-immobilized Sphingomonas sp.
metabolism and yield of Zizania latifolia. Sci. Hortic. 253, 276–285. https://doi.org/ PJ2 on bioremediation of PAHs and bacterial community composition in saline soil.
10.1016/j.scienta.2019.04.058. Chemosphere 279, 130427. https://doi.org/10.1016/j.chemosphere.2021.130427.
Li, Q., You, P., Hu, Q., Leng, B., Wang, J., Chen, J., Wan, S., Wang, B., Yuan, C., Zhou, R., Song, Q., Fu, H., Shi, Q., Shan, X., Wang, Z., Sun, Z., Li, T., 2022. Overfertilization
Ouyang, K., 2020. Effects of co-contamination of heavy metals and total petroleum reduces tomato yield under long-term continuous cropping system via regulation of
hydrocarbons on soil bacterial community and function network reconstitution. soil microbial community composition. Front. Microbiol. 13, 952021.
Ecotox. Environ. Saf. 204, 111083 https://doi.org/10.1016/j.ecoenv.2020.111083. Sousa, F., Martins, M., Sousa, B., Cristiano, S., Manuel, A., Ruth, P., Fernanda, F., 2022.
Li, Q., Xing, Y., Fu, X., Ji, L., Li, T., Wang, J., Chen, G., Qi, Z., Zhang, Q., 2021. Wrack composed by Fucus spp, Ascophyllum nodosum and Pelvetia canaliculata limits
Biochemical mechanisms of rhizospheric Bacillus subtilis-facilitated phytoextraction metal uptake and restores the redox homeostasis of barley plants grown in Cu-
by alfalfa under cadmium stress – Microbial diversity and metabolomics analyses. contaminated soils. J. Plant Growth Regul. 41, 3544–3555. https://doi.org/
Ecotox. Environ. Saf. 212, 112016 https://doi.org/10.1016/j.ecoenv.2021.112016. 10.1007/s00344-021-10532-x.
Liang, H., Chen, Q., Liang, B., Hu, K., 2020. Modeling the effects of long-term reduced N Sun, K., Han, L., Yang, Y., Xia, X., Yang, Z., Wu, F., Li, F., Feng, Y., Xing, B., 2020.
application on soil N losses and yield in a greenhouse tomato production system. Application of hydrochar altered soil microbial community composition and the
Agr. Syst. 185, 102951 https://doi.org/10.1016/j.agsy.2020.102951. molecular structure of native soil organic carbon in a Paddy soil. Environ. Sci.
Lim, J.W., Chiam, J.A., Wang, J.-Y., 2014. Microbial community structure reveals how Technol. 54, 2715–2725. https://doi.org/10.1021/acs.est.9b05864.
microaeration improves fermentation during anaerobic co-digestion of brown water Sun, R., Zheng, H., Yin, S., Zhang, X., You, X., Wu, H., Suo, F., Han, K., Cheng, Y.,
and food waste. Bioresour. Technol. 171, 132–138. https://doi.org/10.1016/j. Zhang, C., Li, Y., 2022. Comparative study of pyrochar and hydrochar on peanut
biortech.2014.08.050.

11
X.-c. Shang et al. Ecotoxicology and Environmental Safety 260 (2023) 115075

seedling growth in a coastal salt-affected soil of Yellow River Delta, China. Sci. Total acidified soil. Chemosphere 298, 134191. https://doi.org/10.1016/j.
Environ. 833, 155183 https://doi.org/10.1016/j.scitotenv.2022.155183. chemosphere.2022.134191.
Tang, H., Chen, M., Wu, P., Faheem, M., Feng, Q., Lee, X., Wang, S., Wang, B., 2022. Yoruklu, H.C., Ozkaya, B., Demir, A., 2022. Optimization of liquid fertilizer production
Engineered biochar effects on soil physicochemical properties and biota from waste seaweed: a design of experiment based statistical approach.
communities: a critical review. Chemosphere, 137025. https://doi.org/10.1016/j. Chemosphere 286, 131885. https://doi.org/10.1016/j.chemosphere.2021.131885.
chemosphere.2022.137025. Yu, H., Zou, W., Chen, J., Chen, H., Yu, Z., Huang, J., Tang, H., Wei, X., Gao, B., 2019.
Tang, L., Hamid, Y., Chen, Z., Lin, Q., Shohag, M.J.I., He, Z., Yang, X., 2021. Biochar amendment improves crop production in problem soils: a review. J. Environ.
A phytoremediation coupled with agro-production mode suppresses Fusarium wilt Manag 232, 8–21. https://doi.org/10.1016/j.jenvman.2018.10.117.
disease and alleviates cadmium phytotoxicity of cucumber (Cucumis sativus L.) in Zhang, L., Sun, X., 2017. Addition of seaweed and bentonite accelerates the two-stage
continuous cropping greenhouse soil. Chemosphere 270, 128634. https://doi.org/ composting of green waste. Bioresour. Technol. 243, 154–162. https://doi.org/
10.1016/j.chemosphere.2020.128634. 10.1016/j.biortech.2017.06.099.
Tujula, N.A., Crocetti, G.R., Burke, C., Thomas, T., Holmstrom, C., Kjelleberg, S., 2010. Zhang, X., Zhong, Z., Bian, F., Yang, C., 2019. Effects of composted bamboo residue
Variability and abundance of the epiphytic bacterial community associated with a amendments on soil microbial communities in an intensively managed bamboo
green marine Ulvacean alga. ISME J. 4 (2), 301–311. https://doi.org/10.1038/ (Phyllostachys praecox) plantation. Appl. Soil Ecol. 136, 178–183. https://doi.org/
ismej.2009.107. 10.1016/j.apsoil.2018.12.025.
Wang, F., Wang, X., Song, N., 2021. Biochar and vermicompost improve the soil Zhang, Y., Wang, J., Feng, Y., 2021. The effects of biochar addition on soil
properties and the yield and quality of cucumber (Cucumis sativus L.) grown in plastic physicochemical properties: a review. Catena 202, 105284. https://doi.org/
shed soil continuously cropped for different years. Agr. Ecosyst. Environ. 315, 10.1016/j.catena.2021.105284.
107425 https://doi.org/10.1016/j.agee.2021.107425. Zhao, W., Zhou, Q., Tian, Z., Cui, Y., Liang, Y., Wang, H., 2020. Apply biochar to
Wang, H., Zhu, Y., Xu, M., Cai, X.Y., Tian, F., 2022. Co-application of spent mushroom ameliorate soda saline-alkali land, improve soil function and increase corn nutrient
substrate and PGPR alleviates tomato continuous cropping obstacle by regulating availability in the Songnen Plain. Sci. Total Environ. 722, 137428 https://doi.org/
soil microbial properties. Rhizosphere 23, 100563. https://doi.org/10.1016/j. 10.1016/j.scitotenv.2020.137428.
rhisph.2022.100563. Zhao, Z., Xia, L., Qin, Z., Cao, J., Mohammed, A.A.O., Toland, H., 2021. The
Wang, M.P., Chen, L., Li, Y.T., Chen, L., Liu, Z.Y., Wang, X.J., Yan, P.S., Qin, S., 2018a. environmental fate of phenanthrene in paddy field system and microbial responses
Responses of soil microbial communities to a short-term application of seaweed in rhizosphere interface: effect of water-saving patterns. Chemosphere 269, 128774.
fertilizer revealed by deep amplicon sequencing. Appl. Soil Ecol. 125, 288–296. https://doi.org/10.1016/j.chemosphere.2020.128774.
https://doi.org/10.1016/j.apsoil.2018.02.013. Zheng, Y., Han, X., Zhao, D., Wei, K., Zhang, C.S., 2021. Exploring biocontrol agents from
Wang, Y., Zhang, Y., Gao, Z., Yang, W., 2018b. Breeding for resistance to tomato microbial keystone taxa associated to suppressive soil: a new attempt for a biocontrol
bacterial diseases in China: challenges and prospects. Hortic. Plant J. 4 (5), 193–207. strategy. Front. Plant Sci. 12, 655673 https://doi.org/10.3389/fpls.2021.655673.
https://doi.org/10.1016/j.hpj.2018.08.004. Zhou, G., Qiu, X., Zhang, J., Tao, C., 2019. Effects of seaweed fertilizer on enzyme
Wang, Y.F., Fu, F.Y., Li, J.J., Wang, G.S., Wu, M.M., Zhan, J., Chen, X.S., Mao, Z.Q., activities, metabolic characteristics, and bacterial communities during maize straw
2016. Effects of seaweed fertilizer on the growth of Malus hupehensis Rehd. seedlings, composting. Bioresour. Technol. 286, 121375 https://doi.org/10.1016/j.
soil enzyme activities and fungal communities under replant condition. Eur. J. Soil biortech.2019.121375.
Biol. 75, 1–7. https://doi.org/10.1016/j.ejsobi.2016.04.003. Zhu, J., Peng, H., Ji, X., Li, C., Li, S., 2019. Effects of reduced inorganic fertilization and
Yan, T., Xue, J., Zhou, Z., Wu, Y., 2021. Biochar-based fertilizer amendments improve rice straw recovery on soil enzyme activities and bacterial community in double-rice
the soil microbial community structure in a karst mountainous area. Sci. Total paddy soils. Eur. J. Soil Biol. 94, 103116 https://doi.org/10.1016/j.
Environ. 794, 148757 https://doi.org/10.1016/j.scitotenv.2021.148757. ejsobi.2019.103116.
Yin, S., Zhang, X., Suo, F., You, X., Yuan, Y., Cheng, Y., Zhang, C., Li, Y., 2022. Effect of
biochar and hydrochar from cow manure and reed straw on lettuce growth in an

12