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Article

Biochar Herbicide Protection Pods for Mitigating Herbicide Sensitivity in Tomato Plants

by
Sandipan Sil
1,
Fernanda Reolon de Souza
1,
Bailey Bullard
2,
Todd Mlsna
2 and
Te-Ming Tseng
1,*
1
Department of Plant and Soil Sciences, Mississippi State University, 32 Creeman St., Starkville, MS 39762, USA
2
Department of Chemistry, Mississippi State University, 310 President Cir, Starkville, MS 39762, USA
*
Author to whom correspondence should be addressed.
Agronomy 2025, 15(5), 1188; https://doi.org/10.3390/agronomy15051188
Submission received: 8 April 2025 / Revised: 5 May 2025 / Accepted: 12 May 2025 / Published: 14 May 2025
(This article belongs to the Special Issue Advancing Weed Biology Through Innovation and Technology for Sustainable Agriculture)
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Abstract

:
Tomato is a major crop, and efforts are ongoing to enhance its resilience to biotic and abiotic stresses. Weed management remains a key challenge, prompting the search for sustainable alternatives to reduce the impact of excessive herbicide use. Biochar is a promising alternative, as it enriches the soil, improves its water retention capacity, promotes its regeneration and increased fertility, delays nutrient leaching, and improves fertilizer use efficiency. This study aimed to investigate the efficiency of biochar use in mitigating stress caused by different herbicides. Two different biochar materials, Douglas fir and rice husk, were used. Tomato seeds were sown in pots and arranged in a randomized design. At the 4V stage (28 days after sowing), the herbicides S-metolachlor, metribuzin, and halosulfuron were applied. Plant length, injury, antioxidant enzyme activity, ascorbate peroxidase (APX), catalase (CAT), guaiacol peroxidase (GPOD), glutathione reductase (GR), and hydrogen peroxide content (H2O2) were assessed 7 and 14 days after herbicide application. Plants treated with biochar and submitted to herbicide treatments showed significantly higher growth parameters and fewer injuries when compared to plants treated with herbicides without biochar. The antioxidant response of the plants followed the same trend; smaller plants with more injuries showed greater H2O2 accumulation and significantly higher antioxidant enzyme activity. These findings highlight the protective effect of biochar, particularly Douglas fir biochar, as it effectively mitigated herbicide-induced oxidative stress and helped maintain plant growth and structural integrity under treatment conditions.

1. Introduction

Tomato (Solanum lycopersicum L.) is a highly sought-after vegetable worldwide due to its nutritional value and abundance of antioxidant compounds [1]. Its cultivation in greenhouses has considerably increased its commercialization, which is attributed to the use of climate control methods and alternative soils, allowing to obtain higher yields per acre and improving the farmer’s ability to sustain production throughout the year. The vegetable production sector has gained significant importance, as reflected in the sharp rise in import value from 2.4% in 2021 to 5.7% in 2023, demonstrating its growing role in the industry [2].
Tomato production in greenhouses has important bottlenecks, such as extreme weather conditions, pest resistance, poor infrastructure, and post-harvest losses [3,4]. Among these factors, weed management is a potential problem, and although it impacts greenhouse production to a lesser extent, it is a potential problem in field production, as it causes reduced productivity due to competition for space, water, and even nutrient sources. Because it is susceptible, tomato production depends on the use of chemicals, which, in excess, makes production unsustainable. Species such as Palmer amaranth (Amaranthus palmeri), purple (Cyperus rotundus) and yellow (Cyperus esculentus) nutsedges, and large crabgrass (Digitaria sanguinalis) are major weeds in tomato fields, with nutsedges being the most troublesome and leading to a notable decline in fruit quality. In the southern United States, purple and yellow nutsedges are the most problematic, resulting in losses of up to 44% in tomato plants due to their high competitiveness [5].
Although herbicides are the most effective approach to weed control, their indiscriminate use can affect non-target plants and animals. Prolonged and high-dose applications lead to contamination and negatively impact soil health, causing significant yield losses [6]. This is exemplified by the case of organic tomato growers in Tupelo, Mississippi, who suffered substantial losses due to 2,4-D drift in their production fields [7].
In this sense, to support sustainable agriculture and reduce reliance on chemical herbicides, attention has turned to biochar—a carbon-rich material produced through pyrolysis of organic waste like wood and crop residues under low-oxygen conditions [8]. The type of biomass used determines the characteristics of the biochar—it can be derived from manure, wood (e.g., Douglas fir), or crop residues (e.g., rice husk) [9]. These properties allow biochar to retain water and nutrients and make them available to plants and microorganisms [10], helping self-replication and supporting plant growth due to its porous structure and chemical stability [11].
Studies have shown that adding biochar to soil can increase water holding capacity and available water capacity, particularly in soils with coarse texture and a high number of macropores [12,13]. Due to a high hydrogen/carbon ratio in biochar, the mineral pool in soils is effectively decomposed, leading to more nitrogen for plants to utilize [14]. Furthermore, biochar application has been found to enhance the nitrification process with autotrophic nitrifying bacteria, resulting in increased plant nutrient uptake, foliar gas exchange, and biomass production [15]. This is primarily attributed to biochar’s pore structure, low density, and large specific surface area, which alter soil physicochemical properties by increasing soil pH, helping reduce leaching, and reducing micronutrient oxidation by gas exchange and cation exchange. Previous research has also shown that biochar addition can decrease nitrogen fertilizer demand in greenhouse tomato cultivation [16] and has the potential to increase fruit yield and improve tomato quality by enhancing photosynthesis and nutrient uptake [17].
Biochar amendments are not new and have a history of application for absorbing and neutralizing herbicides and many other organic compounds in addition to being efficient in mitigating numerous abiotic stresses. Adding biochar helps mitigate some of the negative effects of salt stress and inadequate irrigation, as they slow the uptake of ions, thus mitigating ionic and osmotic stresses in tomatoes and benefiting the growth of seedlings under various stresses [18].
Herbicide sorption and desorption in soil are the basis for studying the environmental behavior and biotoxicity of pesticides. Thus, the ability of biochar to adsorb pesticides may be a key factor that can affect not only mobility and conversion processes, such as chemical transport, leaching, and bioavailability in soil, but also herbicide uptake and utilization by plants [19].
From this perspective, the addition of biochar together with chemical herbicides in weed management may be a promising alternative [20,21]. The hypothesis is that this approach will mitigate the effect of herbicide stress on tomato plants, preserving soil properties and maintaining sustainable and profitable production. This study aimed to demonstrate that the application of biochar can effectively alleviate the physiological stress and potential phytotoxic effects caused by the indiscriminate use of the herbicides S-metolachlor, metribuzin, and halosulfuron, which are widely utilized for weed management in the tomato crop.

2. Materials and Methods

2.1. Experiment Background

The experiment was conducted between June and July 2024 in a greenhouse at Mississippi State University, MS, USA, with a temperature minimum of 21 °C, average of 26 °C, and maximum of 32 °C with an average relative humidity of 45%, monitored with a WatchDog A150 Temp/RH recorder (Spectrum Technologies, Aurora, IL, USA) at the R.R. Foil Plant Science Research Center (43°04′33″ N, 89°25′27″ W).
The experimental unit consisted of 0.4825 L polyethylene pots filled with commercial substrate without fertilizers. The plant material used was tomato (Solanum lycopersicum L.-F1 hybrids-Harris Seeds).
For this study, rice husk and Douglas fir biochars were used. The rice husk biochar was produced by slow pyrolysis of rice (Oryza sativa L.) at 450–550 °C under low-oxygen conditions. It has an alkaline pH (8.5–10.0), high porosity, large surface area (50–200 m2 g−1), and high fixed carbon content. Douglas fir biochar was obtained as a byproduct from bio-syngas production and is commercially available (Black Owl Biochar Environment Ultra, Biochar Supreme Inc., Everson, WA, USA).
The treatments consisted of (i) control (C), i.e., no biochar treatment; (ii) rice husk biochar pods (RH); and (iii) Douglas fir biochar pods (DF) (Table 1). Twenty-eight days after sowing (V4 stage), the plants were sprayed with the herbicides S-metolachlor (6.72 kg ha−1), metribuzin (1.12 kg ha−1), and halosulfuron (0.044 kg ha−1) at twice the standard application rate used in the field, overestimating the stress condition (Table 2) (one plant kept aside as a control without any herbicide), inside a spray chamber (Generation III track sprayer, DeVries Manufacturing, Inc., Hollandale, MS, United States) equipped with an FC-GA110-02 nozzle (Teejet) calibrated to deliver 187 L ha−1 and calibrated for a spraying speed of 3.22 km h−1, covering a spray length of 1.83 m and a spray pressure of 276 kPa. The distance of the plants from the spray tip was approximately 53.3 cm, based on average plant height.
The experimental units were watered to field capacity prior to herbicide application, and watering was repeated for the remainder of the experiment as needed. The experiment was conducted twice in a randomized block design with four replicates per treatment, producing 48 experimental units.

2.2. Visual Injury Analysis

The visual injury was monitored and recorded 7 and 14 days after application of treatments (DAT), where 0% signifies an injury-free plant and 100% implies the death of the plant (Table 3).

2.3. Growth Parameters

The length and fresh weight of the tomato plants were taken (in cm and mg, respectively) 7 and 14 DAT, respectively, with the measurements carried out in different plants. The length of the plants was measured from the base of the soil to the tip of the topmost leaf.
P l a n t   l e n g t h   i n c r e a s e % = I n c r e a s e   i n   l e n g t h   b e t w e e n   t w o   t i m e   p o i n t s F i n a l   l e n g t h   o f   t h e   p l a n t   a t   l a s t   t i m e   p o i n t × 100

2.4. Enzymatic Activity Assays

Biochemical analyses were performed on plant material collected 7 and 14 DAT. To measure enzyme activities, 200 mg of leaf tissues were ground in liquid nitrogen with 5% PVPP and homogenized in phosphate buffer (pH 7.8) containing EDTA and sodium ascorbate. The mixture was centrifuged at 12,000× g for 20 min at 4 °C, and the resulting supernatant was used as the crude enzyme extract. Protein content was determined using bovine serum albumin as a standard [22].
CAT activity was obtained using the method described by Azevedo et al. [23]. The assay mixture consisted of 100 mM potassium phosphate buffer, pH 7.0, with 12.5 mM hydrogen peroxide and crude enzyme extract. CAT activity was measured as a decline in absorbance at 240 nm. APX activity was determined by a reaction mixture consisting of 100 mM potassium phosphate buffer, pH 7.4, 0.5 mM sodium ascorbate, 0.1 mM hydrogen peroxide, and an aliquot of the enzyme [24], and the rate of ascorbate oxidation was monitored at 290 nm. GR activity was determined according to Cakmak et al. [25] by following the decrease in absorbance at 340 nm due to NADPH oxidation. The reaction mixture consisted of 50 mM potassium phosphate buffer, pH 7.8, 1 mM oxidized glutathione (GSSG), 75 μM NADPH, and an enzyme aliquot. GPOD activity was assayed by monitoring tetraguayacol production by the reduction of hydrogen peroxide at 470 nm. The reaction consisted of 100 mM potassium phosphate buffer, pH 7.0, 0.1 μM EDTA, 5 mM guayacol, and 15 mM hydrogen peroxide [26].
To determine the H2O2 content, the leaves (200 mg) were ground in 0.1% (w:v) trichloroacetic acid (TCA). The homogenate was centrifuged (12,000× g, 4 °C, 20 min), and the supernatant was added to 10 mM potassium phosphate buffer, pH 7.0, and 1 M potassium iodide. The absorbance of the reaction was measured at 390 nm. The content of H2O2 was given on a standard curve prepared with known concentrations of H2O2 [27].

2.5. Data Collection and Analysis

The three-way analysis of variance (ANOVA) and the principal component analysis were performed using the JMP® Pro 18 (Statistical Discovery™, from SAS Institute Inc., Cary, NC, USA) software for the analysis of data, which were subjected to ANOVA, and significance levels were calculated using LS means differences. Student’s t-test analysis with Bonferroni correction (p < 0.05) was performed to distinguish the differences between herbicide treatment within each biochar treatment. The graphs were plotted using the GraphPad Prism8 software. Principal component analysis (PCA) was performed using the same software for multivariate analysis.

3. Results

3.1. Fresh Weight and Plant Length

The fresh weights of tomato plants treated only with biochar were comparatively higher than the control in all aspects in both periods evaluated, showing a significant difference between treatments. When compared to the control, there was an increase of 54% and 46% in the fresh weight of plants subjected to DF biochar and RH biochar, respectively. Plants not treated with biochar and subjected to treatment with different herbicides presented significantly lower fresh weight values when compared to the control. The same trend was observed for plant length. Those treated with biochar had the greatest increase in their length (Figure 1A,B).

3.2. Injury%

Severe injuries were observed on plants treated with herbicides, with metribuzin causing the highest level of damage at both 7 and 14 days after treatment (DAT). In contrast, plants treated with biochar but not exposed to herbicides exhibited the least injury. Across all time points, plants without biochar consistently showed greater injury compared to those treated with rice husk (RH) and Douglas fir (DF) biochars (Figure 2).

3.3. Stress Response

In plants treated only with biochar without herbicides, the activity of APX and CAT (Figure 3) was significantly lower. Within each treatment, the plants without biochar showed increased activity, followed by those with rice husk and Douglas fir, with a significant difference between them.
For APX activity, in rice husk biochar pods at 7 DAT (Figure 3A), we observed a 33% decrease in activity when no herbicides were applied, a 35% decrease when S-metolachlor was applied, a 44% decrease when metribuzin was considered, and a 37% reduction due to halosulfuron application compared to the control or no biochar plants. In Douglas fir biochar pods compared to no biochar plants at 7 DAT, we observed a reduction of 55% when no herbicides were applied and a 37%, 53%, and 39% reduction when S-metolachlor, metribuzin, and halosulfuron were applied, respectively. At 14 DAT (Figure 3B,D), there was a subtle increase in the activity of these enzymes in all treatments. However, they followed the same response trend. For the rice husk, there was a 19% reduction when no herbicides were applied, a 32% reduction when S-metolachlor was used, a 46% reduction in metribuzin application, and a 40% reduction when halosulfuron was applied. At 14 DAT, with no herbicide, there was a steep reduction of 54%, whereas when S-metolachlor, metribuzin, and halosulfuron were applied, we observed a reduction of 28%, 49%, and 42%, respectively (Figure 3B).
The CAT activity at 7 DAT (Figure 3A) with RH-treated biochar pods vs. those without biochar was 21% when no herbicide was involved, 44% with S-metolachlor, 45% with metribuzin, and 39% with halosulfuron. At 14 DAT, the decreasing trend was −13% without herbicides and 40%, 35%, and 38% for S-metolachlor, metribuzin, and halosulfuron, respectively. For DF-treated biochar pods, the reduction was even higher against no biochar at 7 and 14 DAT. A 49% reduction was observed without herbicide, and S-metolachlor, metribuzin, and halosulfuron were responsible for 54%, 57%, and 48%, respectively, at 7 DAT. At 14 DAT, the reduction was 52% without herbicide and 50%, 47%, and 42% under S-metolachlor, metribuzin, and halosulfuron, respectively (Figure 3D).
The activity of GPOD and GR enzymes was significantly higher when plants were subjected to herbicide treatment compared to plants without herbicide and with biochar alone (Figure 4). For GPOD in plants treated with RH at 7 DAT, there was a 40% reduction when no herbicide was present and a 14%, 36%, and 27% reduction when S-metolachlor, metribuzin, and halosulfuron were present, respectively (Figure 4A); at 14 DAT, there was a 29% reduction in the condition without herbicide, while the application of S-metolachlor, metribuzin, and halosulfuron resulted in a reduction rate of 44%, 33%, and 35%, respectively (Figure 4B). Biochar treated with DF compared to the control or no biochar also showed a reduction at 7 and 14 DAT. At 7 DAT (Figure 4A), there was a 58%, 18%, 40%, and 26% reduction with no herbicide, S-metolachlor, metribuzin, and halosulfuron, respectively. At 14 DAT, 46%, 41%, 44%, and 48% reductions were observed with no herbicide, S-metolachlor, metribuzin, and halosulfuron, respectively (Figure 4B).
The activity of GPOD and GR enzymes was significantly higher in plants subjected to herbicide treatment when compared to plants without herbicide and only with biochar (Figure 4). For GR in plants treated with RH at 7 DAT, there was a 40% reduction when no herbicide was present and a 14%, 36% and 27% reduction when S-metolachlor, metribuzin, and halosulfuron were present, respectively (Figure 4A); at 14 DAT, there was a 29% reduction in the condition without herbicide, while the application of S-metolachlor, metribuzin, and halosulfuron resulted in a reduction rate of 44%, 33%, and 35%, respectively (Figure 4B). Biochar treated with DF compared to the control or no biochar also showed a reduction at 7 and 14 DAT. At 7 DAT (Figure 4C), there was a 58%, 18%, 40%, and 26% reduction under no herbicide, S-metolachlor, metribuzin, and halosulfuron, respectively. At 14 DAT, reductions of 46%, 41%, 44%, and 48% were observed when no herbicide, S-metolachlor, metribuzin, and halosulfuron were used, respectively (Figure 4D).
At 7 DAT (Figure 5A), the comparison of RH-treated plants with pruned biochar against those without biochar showed a 16% reduction when no herbicide was present. At the same time, in the presence of S-metolachlor, metribuzin, and halosulfuron, there was a reduction of 30%, 26%, and 26%, respectively. At 14 DAT (Figure 5B), a 26% reduction was observed when no herbicide was present, and a 25%, 31%, and 25% reduction when S-metolachlor, metribuzin, and halosulfuron were added, respectively. For tomato plants with DF-pruned biochar, the % reduction was greater than plants with RH-pruned biochar when compared to the control at 7 and 14 DAT. At 7 DAT, there was a reduction of 33% for untreated and 32%, 27%, and 31% for S-metolachlor, metribuzin, and halosulfuron, respectively. At 14 DAT, the reduction rates were 43% for untreated and 29%, 37%, and 28% for S-metolachlor, metribuzin, and halosulfuron, respectively (Figure 5).

4. Discussion

Biochar increased the growth of plants under herbicide stress compared with no biochar plants, especially with DF biochar. Furthermore, plants treated with biochar showed less damage due to herbicide stress, demonstrating the protective effect of the treatment. Studies carried out by Zhao et al. [28] revealed that adding biochar improved the SPAD (leaf chlorophyll content index), soluble solids content, sugar/acid ratio, nutrient absorption, and dry matter production of tomato plants. This is directly related to its high porosity and aromatic structure that allocates better adsorption of chemical molecules when compared to soil organic matter [29]. As observed in this study, these properties positively influenced the initial plant growth parameters.
Biochar application has been shown to improve soil fertility by sequestering carbon, increasing water holding capacity, reducing contaminant uptake, and promoting soil aeration, nutrient cycling, and microbial activities [30]. In this study, plants with the lowest increase in their growth parameters also showed significant damage (Figure 2) because tomato plants are especially sensitive and susceptible to injury due to excessive herbicide use [31]. These injuries are physiological manifestations of plants subjected to herbicide stress triggered by oxidative stress, which also impairs their growth and development, triggering a delay in their length and weight [32,33,34].
Previous research in greenhouse tomato cultivation has also shown that biochar addition can positively influence growth parameters by decreasing the demand for nitrogen fertilizers [16]. Wang et al. [17] found that biochar has the potential to increase fruit yield and improve tomato quality by enhancing photosynthesis and nutrient uptake. Similarly, Akhtar et al. [35] observed that biochar addition resulted in increased yield and enhanced tomato quality under reduced irrigation conditions.
Kamyab et al. [36], in their review, explored the benefits of biochar in attenuating the effects caused by compounds such as atrazine, an ALS inhibitor, as well as the herbicide halosulfuron, which is widely used for tomato crops was used in the present study. There are a large number of studies on the impact of biochar application in agricultural soils on the sorption–desorption and degradation of herbicides. Qiu et al. [37] evaluated the role of wheat-derived biochar in the biodegradation of atrazine in soil and observed enhanced degradation of the herbicide due to stimulation of microbial growth by soluble nutrients in the biochar. Yu et al. [38] found that soil amended with biochar derived from the pyrolysis of red gum chips enhanced the sorption of diuron and increased the nonlinearity of the adsorption isotherm and the extent of sorption–desorption hysteresis.
The responses regarding the adsorption and desorption of herbicides vary according to the type of herbicide and biochar. In some cases, the adsorptive capacity of biochar is high, preventing the bioavailability of the herbicide and keeping it in the soil, as found in the study by Zhang et al. [39], where the incorporation of about 1% biochar into soils showed decreased biodegradation of benzonitrile due to enhanced sorption [39]. Yang et al. [40] found that the use of biochar promotes reduced microbial degradation of diuron with herbicidal efficacy in barnyard grass and decreased uptake of chlorpyrifos by Chinese chives and spring onions [41,42].
Herbicides such as S-metolachlor, metribuzin, and halosulfuron, widely used in tomato crops, without proper management can generate oxidative stress through the exacerbated production of reactive oxygen species (ROS) by plants, which damage plant cells [43] and alter their DNA [44], impairing their growth. Thus, the balance between the formation and elimination of ROS is necessary for the normal functioning of plants [45,46].
In the presence of stress factors, plants mobilize a complex protection system: their enzymatic system, involving enzymes such as ascorbate peroxidase (APX), catalase (CAT), and glutathione reductase (GR) that are present in different cellular compartments. Furthermore, H2O2 content is an essential non-enzymatic indication of oxidative stress that can quickly diffuse across membranes with the help of aquaporins located in the plasma membrane, reaching distant sites wherever damage or stress occurs [47]. Non-enzymatic antioxidants present in all cellular compartments can detoxify ROS and radicals directly or decrease the substrates for these antioxidant enzymes [48]. In this study, tomato plants without biochar showed a reduction in their growth parameters in an inversely proportional relationship to stress injury (Figure 1 and Figure 2). For these plants, the increased activity of APX and CAT (Figure 3) suggests an attempt to mitigate the oxidative damage resulting from H2O2 accumulation.
The main defensive role against oxidative stress is to neutralize toxic H2O2 into less harmful components. This cycle is a redox balance system composed of ascorbate peroxidase (APX) and glutathione reductase (GR) accessible in all cellular compartments [49]. In this process, the APX converts H2O2 and ascorbate into H2O and monodehydroascorbate (MDHA) by utilizing ascorbate as an electron donor [50]. As occurred in this study, the increased levels of these enzymes provide evidence of their activity. This effect is especially noticeable in plants without biochar treatment, which were more exposed to stress (Figure 3A,B).
Guaiacol peroxidase (GPOD), a heme-containing enzyme, plays a key role in mitigating oxidative stress. It catalyzes the oxidation of aromatic substrates such as guaiacol in the presence of hydrogen peroxide (H2O2). Through this process, GPOD helps plant cells manage reactive oxygen species (ROS) under stressful conditions [51].
This protective response is further supported by the observed increase in catalase (CAT) activity (Figure 3C,D), another heme-containing enzyme that decomposes H2O2 into water and oxygen, contributing to cellular detoxification [52].
Glutathione reductase does not directly decompose hydrogen peroxide like catalase, APX, and GPOD. However, it is essential to maintain the antioxidant system in balance, allowing other antioxidant enzymes to perform their functions effectively. In other words, GR is essential for regenerating the antioxidant compounds that protect cells against oxidative damage. The regeneration of reduced glutathione (GSH) from oxidized glutathione (GSSG), mediated by glutathione reductase (GR), is a crucial step for the maintenance of redox balance and is required to provide tolerance against stress conditions [53]. The increased GR activity (Figure 4C,D) in herbicide-stressed plants, especially in the absence of biochar, is another indication of the plant response and an attempt to keep the system in balance, but mainly supports the role of biochar in stress mitigation, which has already been demonstrated by its ability to reduce phytotoxicity and alleviate stress levels, playing a key role in soil remediation [54,55].

5. Conclusions

Based on our findings, the use of biochar pods represents a promising strategy to mitigate herbicide-induced stress in tomato cultivation, particularly during early plant development. Biochar, a simple and traditional carbon-based amendment, enhanced plant vigor, reduced susceptibility to damage, and improved antioxidant regulation. These benefits were consistent across all tested conditions, highlighting biochar’s potential as a complementary input to conventional herbicide-based weed management. While we refer to it as a cost-effective and sustainable alternative, this is relative to the increased resilience observed in treated plants, which may reduce the need for additional chemical inputs or replanting due to early damage. However, future studies assessing the economic viability and long-term field performance of biochar under varying agronomic conditions are essential to validate its broader applicability. To further explore the benefits of herbicide-protective pods (HPPs), we propose analyzing fruit nutritional quality, agronomic traits, and gene expression related to stress responses, aiming to better understand the mechanisms involved and support future implementation.

Author Contributions

Conceptualization, S.S., T.-M.T. and T.M.; methodology, S.S., F.R.d.S., T.-M.T., B.B. and T.M.; software, S.S. and F.R.d.S.; validation, S.S., F.R.d.S. and T.M.T; formal analysis, S.S., F.R.d.S. and T.M.T; investigation, S.S., T.-M.T., B.B. and T.M.; resources, T.-M.T., B.B. and T.M.; data curation, S.S. and F.R.d.S.; writing—original draft preparation, S.S., F.R.d.S. and T.-M.T.; writing—review and editing, S.S., F.R.d.S. and T.M.T; visualization, S.S., F.R.d.S. and T.-M.T.; supervision, F.R.d.S., T.-M.T. and T.M.; project administration, T.-M.T.; funding acquisition, T.-M.T. All authors have read and agreed to the published version of the manuscript.

Funding

This research was partially funded by the Southern Integrated Pest Management Center (S23-046) and the Mississippi Department of Agriculture and Commerce–Specialty Crop Block Grant Program (902297018). This material is based upon work supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture, Hatch project, under accession number 230100.

Data Availability Statement

Dataset available on request from the authors.

Conflicts of Interest

The authors declare no conflicts of interest.

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Figure 1. Fresh weight and height of tomato plants grown with and without biochar at 7 (A,C) and 14 (B,D) days after application of different herbicides. Error bars correspond to the 95% confidence interval. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
Figure 1. Fresh weight and height of tomato plants grown with and without biochar at 7 (A,C) and 14 (B,D) days after application of different herbicides. Error bars correspond to the 95% confidence interval. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
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Figure 2. Injury in tomato plants grown with and without biochar at 7 (A) and 14 (B) days after application of different herbicides. Error bars correspond to the 95% confidence interval. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
Figure 2. Injury in tomato plants grown with and without biochar at 7 (A) and 14 (B) days after application of different herbicides. Error bars correspond to the 95% confidence interval. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
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Figure 3. Ascorbate peroxidase (APX) and catalase (CAT) activity in tomato leaves grown with and without biochar at 7 (A,C) and 14 (B,D) days after application of different herbicides. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
Figure 3. Ascorbate peroxidase (APX) and catalase (CAT) activity in tomato leaves grown with and without biochar at 7 (A,C) and 14 (B,D) days after application of different herbicides. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
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Figure 4. Guaiacol peroxidase (GPOD) and glutathione reductase (GR) activity in tomato leaves grown with and without biochar at 7 (A,C) and 14 (B,D) days after application of different herbicides. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).H2O2 accumulation was significantly higher in plants without biochar, and the same trend was followed even when herbicides were applied (Figure 5). Among the biochars, RH-treated biochar had a higher hydrogen peroxide content than DF-treated biochar plants.
Figure 4. Guaiacol peroxidase (GPOD) and glutathione reductase (GR) activity in tomato leaves grown with and without biochar at 7 (A,C) and 14 (B,D) days after application of different herbicides. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).H2O2 accumulation was significantly higher in plants without biochar, and the same trend was followed even when herbicides were applied (Figure 5). Among the biochars, RH-treated biochar had a higher hydrogen peroxide content than DF-treated biochar plants.
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Figure 5. Hydrogen peroxide (H2O2) content in tomato leaves grown with and without biochar at 7 (A) and 14 (B) days after application of different herbicides. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
Figure 5. Hydrogen peroxide (H2O2) content in tomato leaves grown with and without biochar at 7 (A) and 14 (B) days after application of different herbicides. Error bars represent the 95% confidence interval. Different letters indicate statistically significant differences between herbicide treatments within each biochar condition, based on one-way ANOVA followed by pairwise Student’s t-tests with Bonferroni correction (p ≤ 0.05, n = 10).
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Table 1. Biochar types with herbicide application setup.
Table 1. Biochar types with herbicide application setup.
Types of Biochar Treatment with Herbicide Application
No BiocharRice Husk (RH) * HPPsDouglas Fir (DF) HPPs
Control (no herbicide)RH (no herbicide)DF (no herbicide)
Control + S-metolachlorRH + S-metolachlorDF + metolachlor
Control + metribuzinRH + metribuzinDF + metribuzin
Control + halosulfuronRH + halosulfuronDF + halosulfuron
* High-performance polymers.
Table 2. Lists of herbicides used and their respective rates.
Table 2. Lists of herbicides used and their respective rates.
HerbicideTrade NameRates Used (2× Standard Rates)
S-metolachlorDual II Magnum6.72 kg ha−1
MetribuzinGlory1.12 kg ha−1
HalosulfuronProfine0.044 kg ha−1
Table 3. Injury% and their respective symptomology.
Table 3. Injury% and their respective symptomology.
Injury%Symptomology
0–10No injuries were seen, and no reduction in growth or yellowing of leaves seen
11–30Little to moderate injury and yellowing of leaves
31–50Yellowing at the base; basal leaves are more yellowish, and leaf curling is seen; growth reduction is moderate
51–70Yellowing of leaves is seen, more with growth reduction; moderate to severe injuries are seen
71–95Severely injured, with almost all yellow leaves and no growth seen
96–100Almost dead or dead plant
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Sil, S.; Souza, F.R.d.; Bullard, B.; Mlsna, T.; Tseng, T.-M. Biochar Herbicide Protection Pods for Mitigating Herbicide Sensitivity in Tomato Plants. Agronomy 2025, 15, 1188. https://doi.org/10.3390/agronomy15051188

AMA Style

Sil S, Souza FRd, Bullard B, Mlsna T, Tseng T-M. Biochar Herbicide Protection Pods for Mitigating Herbicide Sensitivity in Tomato Plants. Agronomy. 2025; 15(5):1188. https://doi.org/10.3390/agronomy15051188

Chicago/Turabian Style

Sil, Sandipan, Fernanda Reolon de Souza, Bailey Bullard, Todd Mlsna, and Te-Ming Tseng. 2025. "Biochar Herbicide Protection Pods for Mitigating Herbicide Sensitivity in Tomato Plants" Agronomy 15, no. 5: 1188. https://doi.org/10.3390/agronomy15051188

APA Style

Sil, S., Souza, F. R. d., Bullard, B., Mlsna, T., & Tseng, T.-M. (2025). Biochar Herbicide Protection Pods for Mitigating Herbicide Sensitivity in Tomato Plants. Agronomy, 15(5), 1188. https://doi.org/10.3390/agronomy15051188

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