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Chemical and Biological Technologies in Agriculture
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Effects of peanut shell biochar and fermented cow manure on plant growth and metabolism of tomato

Chemical and Biological Technologies in Agriculture volume 11, Article number: 113 (2024) Cite this article

Abstract

Background

This experimental study used peanut shell biochar and fermented cow manure as the main raw materials forming a substrate for tomato plants.

Results

Substrates were created from peanut shell biochar, fermented cow manure, slag, and vermiculite mixed in volume ratios of 6:0:1:2, 5:1:1:2, 4:2:1:2, and 3:3:1:2, respectively. Comparisons were made to a control substrate composed of peat, slag, and vermiculite in a volume ratio of 6:1:2, respectively. As the proportion of biochar in the substrate increased, the bulk density showed a downward trend while the total porosity, aeration porosity, and water holding capacity showed upward trends. As the proportion of cow manure increased, the total N, available K, Ca, and Mg in the substrate increased. Tomatoes demonstrated similar or better growth than the control at experimental substrate composition ratios of 6:0:1:2 and 5:1:1:2. This was reflected in seedling strength index, seedling growth, chlorophyll content, root growth, plant carbohydrates, purine metabolism, caffeine metabolism, galactose metabolism, and starch and sucrose metabolism. The results of this study indicate the experimental substrate composition ratios of 6:0:1:2 and 5:1:1:2 were the most beneficial in terms of supporting the growth of tomato plants.

Conclusions

The study confirms biochar in composite substrate promotes plant growth by improving the root environment and plant metabolism. This investigation provides new information to moderate the use of peat and support efforts to achieve carbon neutrality through the creative utilization of agricultural waste.

Graphical abstract

Background

Tomatoes are one of the most widely planted crops worldwide. Substrate plays a crucial role during the growth process of tomatoes. Soilless cultivation is a new method that can provide good conditions for plant growth and reproduction. Peat is an ideal soilless cultivation substrate, with stable physical properties and chemical properties that create good moisture, fertilizer, air, and heat conditions for plants [1]. However, peat resources are limited and non-renewable, and excessive exploitation of peat can cause damage to wetland systems [2, 3]. In recent years, extensive research has been conducted on the use of various agricultural waste materials, such as rice husks [4], coconut husks [5], wheat straw [6], and sugarcane bagasse [7], as raw materials for composting or for use in combinations as alternative substrates to peat.

Biochar is produced by the pyrolysis of organic waste materials under anaerobic and high-temperature conditions [8]. The world is facing the effects of climate change, which are mainly attributed to rising levels of greenhouse gases. Biochar has seen a surge in popularity due to its potential in terms of increasing carbon storage and mitigating greenhouse emissions [9]. Biochar has a porous structure, which improves nutrient retention and utilization efficiency, reduces nutrient leaching, and promotes nutrient cycling in plants [10, 11]. Applying biochar as a soil amendment increases phosphorus (P) content and microbial activity [12]. The surface of biochar contains abundant functional groups that effectively enhance the cation exchange capacity of the soil. The addition of biochar is an effective means to change the fertility of sandy soil. In addition, the numerous active sites on the surface of biochar help it to better adsorb nutrients [11]. Biochar applied to soil can adsorb heavy metals, resulting in improved tomato yield and quality [13]. Furthermore, biochar reduces the damage of saline alkali soil to wheat, grape, and peonies by improving soil physicochemical properties, increasing soil enzyme activity, and enriching microbial rhizosphere communities [14,15,16].

Animal waste (feces) is the main waste in animal husbandry. Improper handling of animal waste can cause a series of environmental problems, such as soil, water, and air pollution. The rapid growth in related operations resulting in the large-scale production of animal waste raises concerns during waste treatment. In recent years, animal waste has been mostly treated through composting, a process of nutrient recycling in which microorganisms decompose waste and convert it into fertilizer [17, 18]. The applying compost to the soil can promote plants growth and yield by improving soil fertility and the physical and chemical properties of the soil [19, 20]. Moreover, addition of compost as an amendment to soil alters the rhizosphere microbial community of tomato and provides beneficial compounds for their growth [21]. Replacing chemical fertilizer with fermented cow manure has also been shown to change the metabolic products of tea seedlings, ultimately improving the quality of tea trees [22].

Some preliminary studies have been conducted on biochar and animal manure compost as seedling substrates. Mixing 5% chicken manure and 60 or 70% biochar with commercial substrates can replace peat as a substrate for tomato seedlings [23]. The ratio of 70% biochar to 30% earthworm manure as a seedling substrate was shown to be superior to other manure and biochar ratios [24]. However, these experiments mostly focused on the physicochemical properties of the composite substrate and the physiological conditions of plant growth, so questions about how the composite substrate affects plant growth and metabolism remain to be addressed.

This paper describes an experiment that used peanut shell biochar, fermented cow manure, slag, and vermiculite as raw materials to study the effect of the ratio of biochar to animal manure on the physicochemical properties of the substrate and tomato growth metabolism. This experiment analyzed the physicochemical properties of the substrate and the absorption of nutrients by the plants. In addition, metabolomics was used to analyze the differential plant metabolites, revealing the regulatory effects of different substrates on plants in order to select the optimal substrate formula suitable for tomato growth. The results provide a new perspective for the utilization of agricultural waste as a renewable resource.

Materials and methods

Plant materials

The tomato cultivar ‘Kang Bing 011’ was used in this study. The plants were divided into five groups of three replicates, with the seeds sown in a 50 well tray and placed in a 28 ℃ incubator to induce germination. Plants were subject to normal growth management for 35 days without fertilization.

Substrate preparation

The biochar was produced from peanut shell which was pyrolyzed at 350–500 °C for 2 h with carbonization furnace (Longkang machinery Equipment Co. Ltd, Henan, China). Before being pyrolyzed, the peanut shell was impregnated in 2% H3PO4 solutions for 24 h, and then dried at 60 °C. The impregnation volume ratio (acid solution:biochar) was 1:3. Fermented cow manure was provided by Tianjin Biotechnology Co. Ltd. (Henan, China). The properties of the peanut shell biochar and fermented cow manure are shown in Table 1.

Table 1 Properties of peanut shell biochar and cow manure
Full size table

Five cultivation substrates were assessed: peat:slag:vermiculite (6:1:2, CK), peanut shell biochar:slag:vermiculite (6:1:2, T1), peanut shell biochar:fermented cow manure:slag:vermiculite (5:1:1:2, T2), peanut shell biochar:cow manure:slag:vermiculite (4:2:1:2, T3), and peanut shell biochar:cow manure:slag:vermiculite (3:3:1:2, T4). The substrate ratios are all volume ratios.

Substrate physicochemical properties analysis

Triplicate substrate samples were assessed on day 0 (sowing date) with respect to pH, electrical conductivity (EC), bulk density, total porosity, aeration porosity, holding porosity, and air–water ratio according to methods published by Guo [25].

Determination of nutrient content

Total nitrogen (N), total P, total potassium (K), alkali N, available P, available K, calcium (Ca), and magnesium (Mg) content were measured using the method described by Bao [26]. In brief, samples of substrate and plant were air dried and ground through a 100 mesh sieve. After H2SO4–H2O2 digestion, total N content was determined by the Kjeldahl N determination method and total P was determined by the molybdenum antimony colorimetric method. After HNO3 digestion, total K, Ca, and Mg contents were determined by atomic absorption spectroscopy. Substrate alkali N was determined by the alkaline diffusion method, available P was determined by the NaHCO3 (Olsen) method, and available K was extracted by the NH4OAC leaching method.

Determination of plant growth indicators

The emergence rate of plants was measured on day 10 and plant samples were collected on day 35. Plant samples were measured in terms of plant height, stem diameter, dry weight, leaf area, chlorophyll content, roots, seedling strength index, and G value.

A root scanner (LA2400, Regent Instruments, Quebec, CA) was used to analyze root morphology, including root volume, root surface area, and root length. Leaf area analysis was conducted using ImageJ software.

To determine dry weight, plants were harvested and cut open along the root base to separate the aboveground and underground parts. Plants were washed then dried at 120 ℃ for 20 min and then at 80 ℃ to a constant weight. The dry weight of each part was determined and used to calculate the G value [27] and seedling strength index [28] as follows:

$$G\; value=\frac{whole\; plant\; dry\; weight}{seedling\; days},$$
$$Seedling\; strength\; index=\left(\frac{stem\; diameter}{plant\; height}+\frac{root\; dry\; weight}{shoot\; dry\; weight}\right)\times whole\; plant\; dry\; weight.$$

Determination of root enzyme activity and root activity

The determination of nitrate reductase (NiR) activity was carried out using the sulfonamide colorimetric method. ATPase and acid phosphatase activity were measured using a kit (Yike Biotechnology, Shanghai, China). Root vitality was determined using the red tetrazolium method (TTC) [29].

Determination of chlorophyll content and plant carbohydrates

Chlorophyll content was extracted using a 95% ethanol extraction method. Soluble total sugar was determined using the anthrone colorimetric method, with the sample extracted using boiled distilled water. Cellulose was extracted using sulfuric acid digestion and measured using the anthrone colorimetric method. Starch content was determined by adding calcium acetate chloride and boiling for extraction and dissolution, and then using a spectrophotometer to measure absorbance [29].

Determination of metabolome

Extraction of metabolites

Tomato leaf samples (25 mg) were placed in an Eppendorf (EP) tube to which homogenization beads and 500 μL of extraction solution (methanol:acetonitrile:water in a volume ratio of 2:2:1) were added. The mixture was vortexed for 30 s, placed in a homogenizer at 35 Hz for 4 min, then transferred to an ice water bath with ultrasound for 5 min. The homogenization steps were repeated three times.

LC–MS/MS analysis

Samples were frozen at − 40 ℃ for 1 h, after which they were placed in a 4 ℃ centrifuge at 12,000 rpm for 15 min. To reduce any deviations in analytical results and avoid errors caused by the instrument itself, an equal amount of supernatant from all samples was taken and mixed into quality control (QC) samples for machine testing before, during, and after liquid chromatography with tandem mass spectrometry (LC–MS/MS) analysis of the test sample. Non-targeted metabolite analysis was performed using Vanquish (Thermo Fisher Scientific) ultra-high performance LC. The instrument was connected to an ACQUITY UPLC BEH Amide column (2.1 mm × 50 mm, 1.7 μm; Waters Ltd.) for chromatographic separation of target compounds [30]. The A phase for the LC analysis contained 25 mmol/L ammonium acetate and 25 mmol/L ammonia in water, while the B phase was acetonitrile. Samples (2 μL) were injected at a temperature of 4 ℃. Metabolomics data analysis was performed using the method proposed by Wu et al. [31].

Statistical analysis

Data were analyzed using DPS 9.01, and the least significant difference (LSD) method was used for significance testing (p < 0.05). Microsoft Excel 2020 was used for data organization and chart drawing. All data represent the mean ± standard deviation of at least three replicates. Metabolite data plotting involves clustering analysis of differential metabolite abundance values using Pheatmap (V1.0.12) from R and drawing a heatmap. KEGG enrichment functional analysis was performed on differential metabolites using ClusterProfiler (V4.6.0).

Results

Physical properties of substrate

Compared to CK, the pH significantly decreased under the T1 and T2 treatments and significantly increased under the T3 and T4 treatments (Table 2). None of the treatments had holding porosities that differed from CK. EC and bulk density significantly increased under all treatments. Total porosity and air–water ratio significantly decreased under the T2, T3, and T4 treatments. Aeration porosity was significantly lower in all treatments than CK.

Table 2 Physical properties of different composite substrates
Full size table

Substrate nutrient content

As the proportion of cow manure in the substrate increased, the values of total K increased while alkali N climbed and then declined. Total N, available K, and Ca were significantly higher under the T4 treatment but lower under the T1 treatment compared to CK (Table 3). Total P and available P were significantly higher in all treatments than the control, with the largest increase noted for the T1 treatment. The T4 treatment had the highest Mg content and the T2 treatment the lowest.

Table 3 Chemical properties of different composite substrates
Full size table

Plant growth status

Under the T2 treatment, the emergence rate was 100%; however, the emergence rate under the T3 and T4 treatments was significantly lower, at 83 and 47% compared to CK, respectively. The G value for CK was significantly higher than under any of the four treatments. The seedling strength index was 34.8 and 14.7% higher under the T1 and T2 treatments, respectively, than for CK (p < 0.05). However, the seedling strength index was significantly lower under the T4 treatment compared to CK. The CK treatment had the highest shoot dry weight, while the T1 treatment had the highest root dry weight (Table 4).

Table 4 Plant growth indicators under different substrate treatments
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Compared to CK, the T1 treatment increased the stem diameter but significantly decreased plant height (Fig. 1). The CK treatment had the highest plant height and the smallest stem diameter, and there was no significant difference in leaf area compared to other treatments. The T1 treatment significantly increased the root surface area of tomato plants and the T4 treatment decreased root volume compared to CK.

Fig. 1
figure 1

The growth of tomato plants under different treatments: A plant height; B stem diameter; C leaf area; D root length; E root surface area; and F root volume. Columns with different letters represent significant differences between treatments (p < 0.05) based on the LSD method

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Root system nutrient absorption capacity

Compared to CK, root activity was higher under the T1, T3 and T4 treatment (p < 0.05). T3 had the highest value of root activity, which was significantly higher than for T1 and T4 (Fig. 2). Acid phosphatase activity was higher under the CK, T2, T3, and T4 treatments than under the T1 treatment. T2 and T4 had higher NiR activity and ATPase activity values than plants grown under other treatments (p < 0.05). The CK and T1 treatments had the lowest NiR and ATPase activities, respectively.

Fig. 2
figure 2

The absorption of nutrients by roots under different treatments: A root activity; B NiR activity; C acid phosphatase activity; and D ATPase activity. Columns with different letters represent significant differences between treatments (p < 0.05) based on the LSD method

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Plant nutrient content

The CK treatment had the lowest total N, total P, and total K content and the highest Ca and Mg content (Table 5). The T3 treatment had the highest total N and total P content, at 2.7 and 5.3 times higher, respectively, compared to CK (p < 0.05). The T4 treatment resulted in higher total K values and lower Ca and Mg values than the CK treatment (p < 0.05). Under all four treatments, the values of total N, total P, and total K were all higher while the values of Ca and Mg were all lower relative to CK (p < 0.05).

Table 5 Total N, total P, total K, Ca, and Mg content of tomato plants
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Chlorophyll content in leaves

Compared to the CK treatment, the various treatments significantly promoted Chl a, Chl b, total chlorophyll (Chl a + b), and carotenoid (Car) content (Fig. 3). All chlorophyll indicators increased the most under the T1 treatment (p < 0.05). The T2 treatment showed the smallest increase in the Chl a and total chlorophyll content while the T4 treatment showed the smallest increase in the Chl b and carotenoid content (p < 0.05).

Fig. 3
figure 3

Chlorophyll content under different treatments: A Chl a; B Chl b; C Chl a + b; and D Car. Columns with different letters represent significant differences between treatments (p < 0.05) based on the LSD method

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Carbohydrates in plants

Total soluble sugar content significantly increased under the T2 treatment, but decreased under the T1, T3, and T4 treatments, compared to CK. Moreover, starch content significantly increased under the T1 (28.8%) and T2 (15.7%) treatments but decreased under the T3 (− 24%) and T4 (− 45.2%) treatments compared to CK. The different treatments also had different effects on the cellulose content. Compared to CK, the T1 treatment had a significantly higher (7.0%) cellulose content while the T2 (− 57.2%), T3 (− 40.9%), and T4 (− 54.0%) treatments had significantly reduced cellulose content (Fig. 4) compared to CK.

Fig. 4
figure 4

Effect under different treatments on carbohydrates of tomato plants: A total soluble sugar content; B starch content; and C cellulose content. Columns with different letters represent significant differences between treatments (p < 0.05) based on the LSD method

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Cluster analysis of tomato metabolites under different substrate treatments

Clustering the same treatment group into one category indicates the differences within each group are relatively small. Based on the relative content of differential metabolites, the first step was to cluster within the same treatment group, then to cluster T1 with T2 and T3 with T4, then to cluster CK with the T1 and T2 group, and then to cluster the T3 and T4 group with the T1, T2, and CK group (Fig. 5).

Fig. 5
figure 5

Cluster heatmap analysis of differential metabolites under different substrate treatments. The relative content of metabolites is indicated by the different colors in the graph, with red and blue indicating higher and lower expression levels, respectively

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Cluster analysis and KEGG enrichment analysis of differential metabolic pathways

To further clarify the metabolic pathways of differential substances, enrichment analysis was conducted using the KEGG database (Fig. 6). The results show differential metabolites are mainly enriched in caffeine metabolism, galactose metabolism, starch and sucrose metabolism, and purine metabolism.

Fig. 6
figure 6

KEGG enrichment analysis of the top 20 differential metabolites

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In the pathways of galactose, starch, and sucrose metabolism, the metabolites melibiose, sucrose, trehalose, and maltose were significantly downregulated under the T1, T3, and T4 treatments and significantly upregulated under the T2 treatment compared to CK (Fig. 7). α-D-Galactose-1P and D-glucose-6P were significantly upregulated under the T2 treatment and significantly downregulated under the T3 and T4 treatments compared to CK. UDP-glucose and UDP-galactose were significantly downregulated under the T4 treatment compared to CK.

Fig. 7
figure 7

Changes in the metabolic pathway network of galactose, starch, and sucrose in tomato leaves under different substrate treatments. Both upward and downward adjustments are based on CK as a reference

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In purine and caffeine metabolism, all treatment groups had significantly upregulated dGmp, AMP, and 7- and 1-methyluric acid compared to CK (Fig. 8). Compared to CK, the T1 and T2 treatments were significantly upregulated in 3ʹ-AMP and downregulated in guanosine. Compared to CK, the T2, T3, and T4 treatments were significantly upregulated in adenine and downregulated in L-glutamine, inosine, and caffeine. The T3 and T4 treatments were significantly upregulated in adenosine and downregulated in 1,3,7-trimethyl-uric acid. The T1 treatment was significantly downregulated in 7-, 3-, and 1-methylxanthine.

Fig. 8
figure 8

Changes in the metabolic pathway network of caffeine and purine in tomato leaves under different substrate treatments. Both upward and downward adjustments are based on CK as a reference

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Discussion

The peanut shell biochar and cow manure composite promotes plant growth

The strong seedling index of the T1 and T2 treatments was significantly higher than that of CK. Furthermore, stem diameter and root surface area of plants under the T1 treatment were significantly higher than those under CK. Meanwhile, T1 and T2 showed a significantly increase in root dry weight. As such, the T1 and T2 treatments promoted the growth of tomato plants during the seedling stage. The results of this study indicate biochar and fermented cow manure are feasible alternatives to peat. The use of peanut shell biochar fixes carbon thus reducing carbon dioxide emissions [32], while the use of cow manure effectively replaces agricultural fertilizers. This proposed recycling and reuse of peanut shells and cow manure in agriculture is therefore in line with the requirements of sustainable agricultural development [33].

The peanut shell biochar and cow manure composite promotes plant growth by improving substrate properties

Root is related to the substrate structure and physical properties of the substrate [34]. The composite substrate of T1 and T2 improved root growth while the root was inhibited under T4 compared to CK. The T1 treatment resulted in physical properties that were the most similar to the CK treatment. The total porosity, aeration porosity, and air–water ratio of the T4 substrate were significantly lower, while the bulk density was significantly higher, than for the CK substrate. This results in poor ventilation in the T4 substrate. When the permeability of the substrate is poor, the root system cannot obtain sufficient oxygen, which forces the root system to stop growing or even die [35]. Good porosity enhances the permeability of the substrate, providing space for root growth and allowing the roots to absorb sufficient water and nutrients. The T1 and T2 treatment have a high biochar content. Biochar has a large number of pores and micropores into which roots can grow [36, 37]. This supports the observation that plants under the T1 and T2 treatments had more developed root systems than those under the T4 treatment.

The nutritional components in the substrate determine the nutritional content of the plant. As the ratio of cow manure increased, the N and K content in the substrate increased and the total K content in the plants increased, while the total N and total P content in the plants first increased and then decreased. N, P, and K are components of substances in cells and are involved in metabolism. Moderate nutrient content in the substrate can promote photosynthetic capacity, plant growth, yield and quality [38, 39]. Moreover, N has a synergistic effect with both P and K, and appropriate proportions can promote the absorption of other elements by plants, thereby promoting plant growth [40]. Thus, the good growth in the T1 and T2 treatments is partially attributed to the presence of appropriate nutrients. On the contrary, the T4 substrate was the most nutrient rich yet some measures of plant growth were lower compared to plants grown in the CK substrate. Excessive nutrients in the substrate can inhibit plant absorption of nutrients and lead to salt accumulation, thereby inhibiting root growth [41, 42]. A higher proportion of biochar in the T1, T2, and T3 treatments mitigated the downside caused by the nutrient richness of cow manure in the T4 composite substrate.

The peanut shell biochar and cow manure composite promotes plant growth by altering plant metabolites

Metabolomics results show different substrate treatments had a significant impact on the caffeine, purine, sucrose and starch, and galactose metabolism pathways of tomatoes, indicating these substances are related to how the different substrates affect tomato growth.

Sugar plays an irreplaceable role in plant growth and development [43, 44]. Sucrose and maltose were upregulated under the T2 treatment compared to the other treatments. The upregulation of sucrose is caused by the upregulation of α-D-galactose-1P, while the upregulation of maltose is caused by the upregulation of trehalose, D-glucose-6P, and sucrose. The T2 treatment resulted in the highest soluble sugar content; this is attributed to the stronger sugar metabolism of plants under the T2 treatment that thus results in the accumulation of more soluble sugars. Sugar provides energy for plants, enabling them to better absorb and utilize nutrients and thereby promoting plant growth and development. This observation also aligns with the significantly higher seedling strength index in the T2 treatment than in the control. The starch and cellulose contents were significantly higher under the T1 treatment than the other treatments, indicating the sugar in the T1 treatment was stored in the form of these insoluble sugars. Starch, as a product of photosynthesis, can be converted into soluble sugars under the action of related enzymes for plant respiration. At the same time, starch provides energy for plant growth and development through hydrolysis. This explains why the seedling strength index of the T1 treatment was significantly higher than the control. Increases in starch content and chlorophyll content in plants are closely related, and is one reason for the observation that the T1 treatment had both high chlorophyll content and high starch content [45, 46].

Caffeine content is higher in young leaves than in old leaves, so a higher caffeine content means plant vitality is stronger [47]. This may be one of the reasons why plants from the T1 and CK treatments were robust. Caffeine originates from the 7-methylxanthine pathway of purine metabolism. Although the 7-methylxanthine content decreased under the T1 treatment, caffeine catabolites (3- and 1-methylxanthine) showed downregulation. The synchronous decrease of synthesis and decomposition maintained the caffeine content at CK levels. Interestingly, dGmp, AMP, and 3ʹ-AMP in purine metabolism increased under the T1 treatment. The decrease in 7-methylxanthine may be attributed to the accumulation of 7-methyuric acid catalyzed by xanthine dehydrogenase. More detailed information about caffeine metabolism is needed in further studies.

Conclusions

The results of this study indicate 6:0:1:2 and 5:1:1:2 ratios of peanut shell biochar:fermented cow manure:slag:vermiculite were the most beneficial in terms of supporting the growth of tomato plants. These substrate compositions improve the absorption of nutrients by plants by changing the physicochemical properties of the substrate, leading to changes in plant metabolic activity and ultimately promoting plant growth.

Availability of data and materials

No datasets were generated or analyzed during the current study.

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Acknowledgements

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Funding

This work was supported financially by the Special Fund for Henan Agriculture Research System, China (HARS-22–07-G4).

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  1. College of Horticulture, Henan Agricultural University, Zhengzhou, 450046, Henan, China

    Yanying Zhu, Qianqian Di, Meng Li, Qingjie Du & Huaijuan Xiao

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  2. Qianqian Di
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  3. Meng Li
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Contributions

Yanying Zhu and Qingjie Du designed the experiment. Yanying Zhu, Qianqian Di, Meng Li, and Qingjie Du performed the experiment and analyzed data. Yanying Zhu wrote the paper. Qingjie Du and Huaijuan Xiao reviewed and checked all the details. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Qingjie Du or Huaijuan Xiao.

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Supplementary Information

40538_2024_638_MOESM1_ESM.xlsx

Additional file 1: Table S1. Differential metabolites identified after comparing four groups (CK vs. T1, CK vs. T2, CK vs. T3, CK vs. T4) in both positive and negative ion modes. Table S2. Differential metabolites identified after multiple comparisons (CK vs. T1 vs. T2 vs. T3 vs. T4) in both positive and negative ion modes. Table S3. After comparing multiple groups (CK vs. T1 vs. T2 vs. T3 vs. T4) under positive and negative ion modes, the top 20 most significant differential metabolic pathways were identified, and functional analysis and classification annotation were performed.

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Zhu, Y., Di, Q., Li, M. et al. Effects of peanut shell biochar and fermented cow manure on plant growth and metabolism of tomato. Chem. Biol. Technol. Agric. 11, 113 (2024). https://doi.org/10.1186/s40538-024-00638-1

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