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Article

Effect of Co-Application of Chinese Milk Vetch and Iron-Modified Biochar on Rice in Antimony-Polluted Soil

by
Yejie Hu
1,†,
Xinglong Xiang
2,†,
Wenjie Jiang
1,
Guiyuan Meng
1,
Jing Zhou
1,
Zhenzhen Guo
1,
Jinxiu Zhou
1,
Haiying Tang
1,*,†,
Jianqun Miao
3,*,† and
Kareem Morsy
4
1
School of Agriculture and Biotechnology, Hunan University of Humanities, Science and Technology, Loudi 417000, China
2
Hunan Qingyang Lake Forestry Technology Co-Limited, Ningxiang 410600, China
3
School of Computer Information and Engineering, Jiangxi Agricultural University, Nanchang 330045, China
4
Biology Department, College of Science, King Khalid University, Abha 61413, Saudi Arabia
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Agronomy 2024, 14(9), 1887; https://doi.org/10.3390/agronomy14091887
Submission received: 26 July 2024 / Revised: 12 August 2024 / Accepted: 20 August 2024 / Published: 23 August 2024
(This article belongs to the Section Agricultural Biosystem and Biological Engineering)
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Abstract

:
Antimony (Sb) toxicity is a serious concern across the globe due to its hazardous impacts on plants and living organisms. The co-application of Chinese milk vetch (CMV) and biochar (BC) is a common agricultural practice, however, the effects of combined CMV and BC in mitigating Sb toxicity and bio-availability remain unclear. Therefore, this study investigated the impacts of CMV, rape straw biochar (RBC), and iron-modified biochar (FMB) and their combinations on rice productivity, physiological, and biochemical functioning of rice and Sb availability. Antimony toxicity caused a marked reduction in rice growth and productivity by decreasing chlorophyll, and anthocyanin synthesis, leaf water contents, osmolyte synthesis and antioxidant activities while, increasing hydrogen peroxide (H2O2), electrolyte leakage (EL), and malondialdehyde (MDA) production and Sb accumulation. Co-application of CMV and FMB increased biomass (29.50%) and grain yield (51.07%) of rice by increasing chlorophyll, and anthocyanin synthesis, leaf water contents, osmolyte synthesis, antioxidant activities, and decreasing production of H2O2, EL, and MDA and Sb accumulation in roots (90.41%) and shoots (96.38%). Furthermore, the combined addition of CMV and FMB also reduced the soil available Sb by 75.57% which resulted in less accumulation of Sb in plant parts and improved growth and yield. Given these facts, these findings indicate that co-application of CMV and FMB is a promising approach to remediate Sb-polluted soils and improve sustainable and safer rice productivity.

1. Introduction

Antimony (Sb) is a toxic and non-essential metalloid for humans and its excessive intake can cause heart disease, liver damage, cancer, and respiratory issues [1,2]. In recent years, anthropogenic activities like mining and smelting substantially increased the Sb concentration in soil and water [3,4]. The concentration of Sb in contaminated areas has reached up to thousands of mgkg−1 [5,6] which is greater than the permissible limit of 36 mg·kg−1 [7]. Antimony is a non-essential metalloid, however, plants readily uptake Sb which negatively affects their growth and human health by entering the human food chain [6,8]. Plants grown in Sb-polluted soils can accumulate high levels of Sb which induces chlorosis and decreases leaf water contents, chlorophyll synthesis, photosynthetic efficiency, and nutrient uptake leading to significant growth losses [9,10]. The excessive concentration of Sb also promotes the production of reactive oxygen species (ROS) which causes oxidation of cellular structure and even leads to plant death [2]. Plants grown in Sb-polluted soils are a major source of Sb entry into humans [11,12], thus, it is imperious to adopt appropriate measures to prevent the absorption of Sb by plants and subsequent impacts on humans and animals.
Antimony is present in four different states (−3, 0, +3, and +5), and among these states, +3 (Sb-III) and +5 (Sb-V) are the most prevalent states in the environment [13,14]. Sb-III is considered to be 10 times more toxic to plants as compared to Sb-V [15]. Plants exhibit significant differences in their efficiency in absorbing and transporting Sb-III and Sb-V [16]. Generally, plant roots easily absorb Sb-III as compared to Sb-V [17,18]. Rice is an important food crop in China and other developing nations and it is often grown in Sb-polluted soils due to its economic demands [19]. Rice plants can accumulate Sb in roots, stems, leaves, and grains, therefore, rice could be a main source of Sb intake in humans by eating Sb-contaminated rice [19].
Different strategies including biological remediation and in immobilization are being used globally to remediate contaminated soils [20,21]. Biochar (BC) is a carbon-rich material known as black gold and it has received appreciable attention to remediate polluted soils due to its bio-compatibility and eco-friendly properties [22,23]. It is well-documented that BC decreases the availability, mobility, and toxicity of heavy metals [24]. The application of BC can decrease the mobilization of Sb in soils by forming the complex between human acid and organic matter of BC and Sb, thereby reducing the availability of Sb to plants [25,26]. Nonetheless, the effect of BC to remediate Sb pollution could be limited, therefore different methods including acidification, alkalization, and oxidation are being used to improve the capacity of BC to remediate polluted soils [27]. Among them, the application of iron-modified biochar (FMB) has emerged as an effective approach to remediate polluted soils owing to its lower cost, environmentally friendly nature, and appreciable ability to adsorb Sb [28]. Different studies documented the remarkable efficiency of FMB to remediate Sb- and As-polluted soils due to its appreciable specific surface area and oxygen functional groups [29,30,31].
Chinese milk vetch (Astragalus sinicus L.: CMV) is a green manure crop that is typically applied to rice fields to improve crop yield and soil productivity [32]. Recently, it has been documented that CMV could also be used to remediate heavy metals in polluted soils [33]. Recent studies have reported that CMV decreased the bioavailability of Cd by changing the soil properties and modulating the microbial community [34,35]. Another study reported that CMV decreased the solubility of copper (Cu) and its uptakes by rice plants [36]. Nonetheless, there could be some constraints to using CMV, for instance, lower remediation efficacy and long-term treatment [36].
The role of FMB and CMV in mitigating the toxic effects of heavy metals in rice is well-reported in the literature. However, no study reported the co-incorporation effects of FMB and CMV in reducing Sb toxicity and accumulation in rice plants. Thus, it is important to elucidate the impacts of FMB and CMV in mitigating the Sb toxicity and mobility, and the mechanism involved in this process. Herein, we hypothesized that the co-incorporation of FMB and CMV can improve rice growth and productivity by improving physiological functioning, antioxidant activities, and immobilizing Sb. Thus this study was amid with the following objectives:(1) determine the effects of RBC, FMB, and CMV on the growth, physiological, and biochemical activities of rice; (2) determine the effects of RBC, FMB, and CMV on Sb availability in plant parts and rice productivity.

2. Materials and Methods

2.1. Soil Collection and Material Preparations

The present pot study was performed at Hunan University of Humanities, Science and Technology (28°46′ N, 115°36′ E). The study site has a sub-tropical monsoon climate with a cold winter and wet summer. The soil for filling the pots was collected from the experiment field which is used for rice and rapeseed production. The collected soil was sieved and pots having a capacity of 10 kg were filled with soil. A sub-sample of soil from collected soil was taken to determine the different soil properties. The soil was identified as silt loam with acidic pH (5.42), organic matter content of 44.4 g·kg−1, total nitrogen content of 2.53 g·kg−1, available phosphorus of 17.44 mg·kg−1, and available potassium of 177.60 mg·kg−1. The rapeseed was collected dried and pyrolyzed (600 °C) for 8 h to prepare the BC. For preparation of FMB, rapeseed straws were ground and soaked for 3 h in FeSO4·7 H2O solution. Then, these residues were dried (110 °C) for 24 h and pyrolyzed to prepare the FMB. The straws of CMV were also collected, dried, and ground and then applied to pots according to treatments. The information about properties of different materials are given in Table 1.

2.2. Experimental Treatments

The study was comprised of different treatments: control (no CMV and no BC), CMV, RBC, FMB, CMV + RBC, and CMV + FMB. All the amendments including CMV, RBC, and FMB were applied at the rate of 2.5%. The study was performed in a completely randomized design and each treatment has three replications. Moreover, an antimony level of 250 mg·kg−1 was maintained in each pot before applying the treatments. The pots with a capacity of 10 kg were filled with soil and Sb concentration of 250 mg·kg−1 was obtained by using C8H4K2O12Sb2 (analytical grade). The antimony in the form of an aqueous solution of C8H4K2O12Sb2 was added to soil. Then, pots were placed in the dark for four weeks, thereafter, pots were taken and the soil of each pot was placed on a plastic sheet. RBC, CMV, and FMB were thoroughly mixed with soil and each pot was filled again. Five rice seedlings were grown in each pot and were regularly irrigated and all other practices were kept constant to obtain a good seeding establishment.

2.3. Chlorophyll Contents and Leaf Water Contents

Next, 0.5 g fresh leaves of rice were taken and chopped in 80% acetone solution to obtain the extract. Therefore, the obtained extract was centrifuged and absorbance was noted at 663, 645, and 470 nm to determine the concentration of chlorophyll a, chlorophyll b, and carotenoids. For determining relative water contents (RWC), top fresh leaves were taken and weighed (FW), and then these leaves were placed in water for 24 h and weighed again (TW). Following, these leaves were removed from the water and oven-dried and weighed (DW) again, and finally, RWC was measured with the following equation: RWC = (FW − DW)/(TW − DW) × 100. To determine electrolyte leakage (EL), 0.5 g fresh rice leaves were collected and placed in water for 24 h, and EC1 was measured. Then, these leaf samples were autoclaved for 120 min and a second EC2 was taken, and finally, EL was measured with the following equation: EL% = (EC1 − EC2) × 100.

2.4. Determination of Oxidative Stress Markers, Potential Osmolyte, and Antioxidant Activities

We used 0.5 g fresh rice leaves and ground them in 5% tri-chloroacetic acid (TCA) solution to obtain the extract. After that, this extract was centrifuged for five minutes, then 1 mL extract and 100 µL of potassium phosphate buffer (PPB) were mixed and placed in a water bath. Later, the absorbance was noted at 390 nm to determine the concentration of hydrogen peroxide (H2O2) [37]. In the case of malondialdehyde (MDA), again 0.5 g fresh rice leaves were ground using TCA solution and centrifuged (12,000 rpm) for 10 min and then this mixture was heated (100 °C) for 30 min; absorbance was noted to determine MDA concentration [38].
For the determination of free amino acids (FAA) and total soluble proteins (TSP), 0.5 g fresh leaves were ground by using PPB. To determine o FAA, 1 mL extract was taken and centrifuged (15,000 rpm) for 15 min. Thereafter, 1 mL of each nanhydrin (2%) and pyridine (10%) was added in tubes containing plant extract and heated for 30 min. Then, volumes of tubes were increased to 15 mL by adding water and absorbance was noted (570 nm) [39]. To determine TSP, 1 mL extract was added with 2 mL of Bradford reagent, and the solution was left for 15 min at room conditions and absorbance was noted (590 nm: [40]). To determine soluble sugars (SS), 1–2 mL extract was collected and placed on the prism of the refractometer and the concentration of brix was measured. In the case of proline, fresh leaves (0.5 g) were taken and ground with 10 mL of sulpho-salicylic acid (3%), then they were centrifuged (10,000 rpm) for 10 min. Thereafter, acid nin-hydrin was added to the extract and boiled for 30 min at 90 °C and absorbance was noted at 520 nm [41].
To determine the concentration of ascorbate peroxidase (APX), catalase (CAT), and peroxidase (POD), 0.5 g fresh leaves were collected and ground by using the PPB. To determine the concentration of APX, the mixture containing 100 μL of enzyme extract, ascorbate (7.5 mM), H2O2 (300 mM), and PPB (25 mM) was prepared and absorbance was noted at 290 nm to determine APX activity [42]. Moreover, to determine CAT, 100 μL extract was mixed with 100 μL H2O2 and 1000 μL PPB, and absorbance was noted at 240 nm to determine CAT [43]. To determine POD activity, the 100 μL leaf extract was taken and centrifuged (12,000 rpm) for 15 min and absorbance was noted at 470 nM [44]. Lastly, to determine superoxide dismutase (SOD) activity, the mixture containing 400 µL H2O2, 25 µL buffer, 100 µL Triton, 50 µL nitro blue tetrazolium chloride (NBT), 50 µL sample, and 50 µL riboflavin was prepared and later absorbance (560 nm) was noted [44].

2.5. Determination of Growth and Yield Traits

All plants were harvested from each plot to determine different growth and yield traits. All the harvested plants were weighed to determine biomass yield, then kernels were separated and weighed to determine kernel yield. Moreover, tillers from each pot were manually counted and the root length of five plants was measured and averaged.

2.6. Determination of Plant and Soil Sb Concentration

The plant samples (roots and shoots) were collected, dried, and ground to make powder. Then, 100 mg of each root and shoot samples were placed in tubes and 2 mL of nitric acid (HNO3) were added into tubes. Thereafter, samples were digested (150 °C) for two hours, the digested mixture was allowed to cool, and then these tubes were placed in a water bath to obtain a clear solution. Lastly, the volume of solution was made to 50 mL by adding 1% HNO3 solution, filtered, and later analyzed by using ICP-MS to determine the concentration of Sb in plant samples. For determination of soil Sb concentration, 500 mg soil was collected and added with 2 mL of HNO3 and 6 mL of hydrochloric acid (HCl) in a Teflon bomb and then this mixture was placed in a water bath (95 °C). Afterward, the solution was allowed to cool and volume was made up to 50 mL and Sb concentration was recorded with atomic absorbance spectrophotometry.

2.7. Statistical Analysis

The data were verified for normal distribution and homogeneity of variance. The data collected on different traits were analyzed by one-way analysis of variance (ANOVA) by using Statistix 8.1. The difference among means were separated by using Tukey’s honestly significant difference (HSD) test (p ≤ 0.05). The figures used in the experiment were prepared by using Sigma-plot-8 software.

3. Results

3.1. Growth and Yield Traits

The application of different amendments showed a significant impact on the growth and yield traits of rice grown in Sb-polluted soil (Table 2). The maximum root length (31.43 cm) was observed with the combined use of CMV and FMB, followed by application of FMB (29.63 cm) and CMV application (28.33 cm), and the lowest root length (21.43 cm) was observed in control (Table 2). Further, taller plants (84.10 cm) with more tillers (77/pot) were also observed with integrated use of CMV and FMB that remained the same with the application of FMB and shorter plants (61 cm) with minimum tillers (46/pot) were observed in control (Table 2). Lastly, maximum biomass (187.79 g) and grain yield (139.93 g/pot) were also noted with the combined addition of CMV and FMB followed by FMB and CMV + RBC, and the lowest biomass and grain yield was recorded in control (Table 2).

3.2. Photosynthetic Pigments and Leaf Relative Water Contents

A significant reduction in photosynthetic pigments and RWC was observed with Sb toxicity (Table 3). However, the application of different amendments showed promising results and significantly improved the synthesis of photosynthetic pigments and RWC (Table 3). The combined application of CMV and FMB increased chlorophyll-a and chlorophyll-b contents by 45.45% and 84% as compared to control (Table 3). Likewise, diverse amendments also showed promising results and increased concentrations of carotenoid and anthocyanin contents (Table 3). The maximum concentration of carotenoid (4.61 mg/g FW) and anthocyanin (10.37 mg/g FW) was noted with the application of CMV and FMB followed by FMB and CMV + RBC, and the lowest carotenoid (3.52 mg/g FW) and anthocyanin (6.96 mg/g FW) was recorded in control (Table 3). Leaf RWC also showed a significant reduction with Sb toxicity, however, the application of CMV + FMB significantly maintained the higher RWC as compared to amendments (Table 3).

3.3. Oxidative Stress Markers and Osmolytes

The results indicate that Sb stress significantly increased the production of EL, MDA, and H2O2 (Figure 1). However, the application of CMV, FMB, and RBC significantly reduced the production of the aforementioned oxidative stress markers (Figure 1). Notably, integrated CMV decreased the EL, MDA, and H2O2 production by 125.49%, 80.63%, and 135.48% respectively as compared to the control. On the other hand, RMB decreased EL, MDA, and H2O2 production by 88.52%, 71%, and 113.15% while combined CMV and RBC decreased EL, MDA, and H2O2 production by 37.60%, 51%, and 90.43% (Figure 1).
The results evidenced a significant increase in proline, TSP, FAA, and SS concentration with the application of different amendments (Figure 2). The maximum concentration of proline (0.93 mg/g FW) and TSP (10.99 mg/g FW) was recorded from pots supplied with combined CMV and FMB, followed by FBM and combined CMV and RBC. The lowest proline (0.58 mg/g FW) and TSP (7.55 mg/g FW) was observed in control (Figure 2).
Maximum FAA (9.73 mg/g FW) and SS (18.91 mg/g FW) were recorded from plants treated with combined CMV and FMB, and the lowest concentration of FAA (6.15 mg/g FW) and SS (10.31 mg/g FW) was observed in control (Figure 2). In addition, maximum proline and SS concentration was also observed with the combined use of CMV and FMB and the lowest proline and SS concentration was observed in control (Figure 2).

3.4. Antioxidant Activities

The dynamics of the change in antioxidants with different treatments are presented in Figure 3. The activities of all the studied antioxidants were increased with the application of different amendments (Figure 3). The maximum activity of APX, CAT, POD, and SOD was seen with the combined use of CMV and FMB followed by the application of FMB, CMV, and RBC and the lowest antioxidant activities were observed in control (Figure 3). Therefore, it can be sensed that the application of all the amendments appreciably increased antioxidant activities which helped to counter the toxic effects of Sb (Figure 3).

3.5. Antimony Concentration in Soil and Plant

The application of different amendments significantly reduced the Sb accumulation in rice roots and shoots (Table 4). The maximum root (56.23 mg·kg−1) and shoot Sb (16.83 mg·kg−1) concentration was found in the control as compared to the control (Table 4). The co-application of CMV and RMB appreciably reduced the root and shoot Sb concentration by 91.34% and 96.38%, respectively, as compared to the control (Table 4). Likewise, FMB, RBC, and CMV also reduced the root concentration by 51.27%, 23.77%, and 47.31%, respectively (Table 4). Moreover, the same treatments also resulted in a reduction of 80%, 26.82%, and 15.51% in shoot Sb concentration (Table 4). The application of different amendments also reduced the soil Sb concentration, minimum soil Sb concentration (129.67 mg·kg−1) was observed with the combined addition of CMV and FMB, and maximum soil Sb concentration was observed in control (227.67 mg·kg−1).

4. Discussion

Antimony is a non-essential metalloid for plants and its excessive concentration induces toxic effects on plants [2]. In the present study, antimony toxicity reduced the growth and yield of rice plants (Table 2). This is consistent with previous studies where authors also found a marked reduction in growth and yield traits of different crops like rice, maize, and wheat with Sb toxicity [45,46]. The results indicate that rice plants accumulated a significant quantity of Sb which reduced the plant growth by inducing oxidative stress and disturbing the plant functioning [47,48]. Apart from this, Sb also increased ROS that damaged the cellular structures and increased the production of MDA and EL causing significant growth and yield losses. Chinese milk vetch and FMB significantly improved the growth and yield of rice which aligns with earlier findings where authors noted a significant increase and growth and biomass production with the application of FMB.
The application of FMB facilitates the transformation of an accessible form of Sb to a less accessible form which in turn decreases Sb toxicity and subsequent growth reduction. Moreover, the application of CMV and FMB might have increased the nutrient absorption, and increased synthesis of chlorophyll, anthocyanin, osmolyte, antioxidant activities, and reduced Sb accumulation, resulting in better growth and yield (Table 2). The application of CMV and FMB also decreased the Sb accumulation in shoots and reduced the Sb translocation efficiency from roots to shoots which could also be a reason for improved plant growth [49]. It is also documented that biochar-induced changes in soil properties also affect the mobility and speciation of heavy metals [50]. The improved soil porosity and permeability with biochar application leads to the oxidation of Sb-III into Sb-V which plays an important role in mitigating Sb toxicity. Interestingly, the application of CMB and FMB appreciably increased the growth and yield of rice which might be because CMV and FMB changed the oxidation state of Sb and ensured the better availability of nutrients.
Chlorophyll is an important pigment needed for photosynthesis and it plays a critical role in light absorption and transformation during photosynthesis. The results indicate that chlorophyll and carotenoids also exhibited a marked reduction following Sb treatment (Table 3). Antimony enters the plant cells and binds with the sulfhydryl group of chloroplast proteins, which leads to disruption of the chloroplast structure and function. Also, Sb increases the activity of chlorophyll-degrading enzymes which reduces the chlorophyll synthesis and leads to a reduction in growth and biomass production [51,52]. The application of CMV and FMB considerably increased the chlorophyll and anthocyanin concentration (Table 3) and maintained the better leaf RWC. The application of CMV and FMB restrained the Sb uptake by rice plants which might have protected the photosynthetic apparatus and ensured better nutrient uptake (Fe and Mg) thereby resulting in better chlorophyll synthesis [53]. The synthesis of abscisic acid (ABA) is significantly increased under stress conditions which increases the activity of chlorophyllase which causes a reduction in chlorophyll synthesis [54]. The application of CMV and FMB might have protected the photosynthetic apparatus and decreased the ABA synthesis and subsequent chlorophyllase activity, therefore, resulting in better chlorophyll synthesis [55,56]. Additionally, biochar application might also increase the uptake and absorption of Fe and Mg by plants which ensures a significant increase in chlorophyll synthesis [57]. In the present study, Sb toxicity caused a marked reduction in leaf RWC which could be due to poor root growth and restricted water uptake. Contrarily, CMV and FMB effectively improved root growth (Table 2) and restricted the Sb uptake which ensured better water uptake thus resulting in better RWC.
Anthocyanin regulates the accumulation of ROS and maintains the plant’s photosynthetic efficiency by protecting the photosynthetic apparatus [58]. The application of CMV and FMB increased the concentration of anthocyanin which might improve the photosynthetic efficiency of rice plants by decreasing ROS production and protecting the photosynthetic apparatus. The impaired plant growth under heavy metal toxicity is linked with excessive ROS production [59]. H2O2 is a major form of ROS that induces lipid peroxidation by increasing MDA production which affects the cellular membranes and ion transport [60]. The results indicate that levels of MDA, EL, and H2O2 were significantly increased in rice plants after exposure to Sb stress which is similar to previous studies [61,62]. To counter heavy metals-induced oxidative stress, plants have excellent antioxidant defense systems [63]. The results indicate that rice plants showed an increase in antioxidant activities (APX, CAT, POD, and SOD) under Sb toxicity which was further increased with the co-application of CMV and FMB. Therefore, co-application of CMV and FMB significantly reduced H2O2 production due to improved antioxidant activities which was evidenced in terms of lower MDA and EL (Figure 1). Catalase helps to mitigate the toxic effects of peroxides in plants, while SOD decomposes superoxide radicals into H2O2 [63]. Peroxidase is also an important antioxidant that lowers the heavy metals-induced oxidative damage by changing the phenols into quinines [64,65]. Ascorbate peroxidase is also a key enzyme that scavenge H2O2 under heavy metals stress [66]. Taken together, the co-application of CMV and FMB increased the antioxidant activities which quenched the ROS production and resulted in lower MDA and H2O2 production. Plants also produce different osmo-regulatory substances to maintain osmotic potential and mitigate ROS production [67,68]. The results indicate that rice plants showed an increase in the synthesis of osmoregulatory substances (proline, TSP, FAA, and SS) with the co-application of CMV and FMB under Sb toxicity (Figure 2). This could be ascribed to increased Sb accumulation in plant parts and N uptake which play a major role in the synthesis of proteins and amino acids.
Rice is a non-hyper-accumulator Sb plant and it absorbs most of Sb in roots and avoids its transport to aerial plant parts. This mechanism is very essential to eliminate the toxic effects of Sb in aerial plant parts [2,57]. In this study, rice plant roots accumulated more Sb as compared to shoots. The application of all the amendments, particularly co-application of CMV and FMB, significantly reduced the Sb accumulation in roots and shoot and soil Sb concentration. The co-application of CMV and FMB might decrease Sb mobility by forming complexes between organic matter and humic acid of biochar and Sb. Therefore, the reduction in mobility of Sb following the co-application of CMV and FMB led to a significant decrease in Sb accumulation in plant parts [26]. Likewise, Wang et al. [69] also reported that the co-application of CMV, biochar, and rice straw can reduce the availability of heavy metals. The application of organic amendments can also cause redistribution of heavy metals within different fractions [70]. This indicates that co-application of CMV and FMB might convert the more accessible Sb form into less accessible forms therefore resulting in less availability of Sb.

5. Conclusions

The co-application of CMV and FMB improved rice productivity by improving photosynthetic pigments and leaf water status and reducing Sb-induced oxidative damage through an increase in antioxidant activities and osmolyte accumulation. The co-incorporation of CMV and FMB also decreased the Sb bio-availability which was linked with a substantial reduction in soil Sb concentration. This study highlights that a combination of CMV and FMB can be the best option to mitigate Sb toxicity and bio-availability. Therefore, the findings of this study hold immense potential to improve crop production in Sb-polluted areas. However, this study was conducted in controlled conditions which may not fully capture the complexities of plant–soil interactions in natural environments. Hence, field studies are needed to validate the effectiveness of CMV and RMB in reducing Sb toxicity and remediating Sb-polluted soils. This study also contained only one level of Sb stress, thereby, studies are needed to explore the effects of varying concentrations of Sb and different types of biochar in mitigating the toxic effects of Sb on plants. It is also suggested that in-depth biochemical and molecular studies must be conducted to provide better insights regarding the role of CMV and RMB in mitigating Sb toxicity. Further, the role of CMV and RMB on non-enzymatic antioxidants should also be explored. The present study only tested the concentration of Sb in plants, therefore, in future studies, authors must include the quantification of macro and micro nutrients in plant tissues. This will provide a better overview of potential elements and nutrients that may be synergistic or antagonistic to Sb-induced effects.

Author Contributions

Conceptualization, J.Z. (Jinxiu Zhou), X.X. and H.T.; writing—original draft preparation, J.Z. (Jinxiu Zhou), X.X., H.T. and J.M.; data curation, W.J., G.M., J.Z. (Jinxiu Zhou), Z.G. and Y.H., writing—review and editing, W.J., G.M., J.Z. (Jing Zhou), Z.G., Y.H. and K.M., visualization, K.M., supervision, H.T. and J.M. All authors have read and agreed to the published version of the manuscript.

Funding

This work was supported and funded by the National Natural Science Foundation of China (32060318); Hunan Provincial Natural Science Foundation of China (2023JJ50473, 2023JJ50474, 2023JJ50476); 2023 Industry-University Collaborative Education Project of the Higher Education Department of the Ministry of Education (230801720230445); 2023 Hunan University Students Innovation and Entrepreneurship Training Program (S202310553008); 2024 Hunan College Students ’ Innovative Training Program ‘Study on the Effect of Combined Application of Chinese Milk Vetch and Biochar on Alleviating Antimony Toxicity to Rice’ (234242204); 2024 Ministry of Education Supply and Demand Docking Employment Education Project: Research on employment-oriented practical teaching model reform of plant production courses (2024011153277); Science Project of Hunan Provincial Department of Education (21B0786); and 2023 Open Projects of Key Laboratory of Crop Physiology, Ecology and Genetic Breeding, Ministry of Education, Jiangxi Agricultural University (202302).

Data Availability Statement

Data are contained within the article and further inquiries can be directed to the corresponding author.

Acknowledgments

The authors are thankful to Muhammad Umair Hassan for his suggestions that improved the quality of our work. The authors extend their appreciation to the Deanship of Research and Graduate Studies at King Khalid University for supporting this work through a large groups project under grant number R.G.P.2/15/45.

Conflicts of Interest

Author Xinglong Xiang was employed by the company Hunan Qingyang Lake Forestry Technology Co-Limited. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Agronomy 14 01887 g001
Figure 1. Effect of CMV straw, rape biochar, and iron-modified biochar on EL (A), MDA (B), and H2O2 (C) concentration of rice grown in antimony-polluted soil. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
Figure 1. Effect of CMV straw, rape biochar, and iron-modified biochar on EL (A), MDA (B), and H2O2 (C) concentration of rice grown in antimony-polluted soil. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
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Figure 2. Effect of CMV straw, rape biochar, and iron-modified biochar on proline (A), TSP (B), FAA (C), and soluble sugar (D) concentration of rice grown in antimony-polluted soil. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
Figure 2. Effect of CMV straw, rape biochar, and iron-modified biochar on proline (A), TSP (B), FAA (C), and soluble sugar (D) concentration of rice grown in antimony-polluted soil. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
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Figure 3. Effect of CMV straw, rape biochar, and iron-modified biochar on APX (A), CAT (B), POD (C), and SOD (D) activity of rice plants grown in antimony-polluted soil. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
Figure 3. Effect of CMV straw, rape biochar, and iron-modified biochar on APX (A), CAT (B), POD (C), and SOD (D) activity of rice plants grown in antimony-polluted soil. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
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Table 1. Properties of different materials used in the study.
Table 1. Properties of different materials used in the study.
PropertiesRapeseed BiocharIron-Modified BiocharChinese Milk Vetch
pH 9.569.766.5
Carbon contents612 g·kg−1622 g·kg−146.31 g·kg−1
Nitrogen contents 4.33 g·kg−14.61 g·kg−122.3 g·kg−1
Table 2. Effect of CMV straw, rape biochar, and iron-modified biochar on growth and yield traits of rice grown in antimony-polluted soil.
Table 2. Effect of CMV straw, rape biochar, and iron-modified biochar on growth and yield traits of rice grown in antimony-polluted soil.
TreatmentsRoot Length (cm)Plant Height (cm)Tillers/PotBiological Yield/Pot (g)Grain Yield/Pot (g)
Control21.43 ± 5.16 b61.00 ± 6.61 c46.00 ± 5.79 d145.01 ± 7.36 bc92.62 ± 2.12 d
CMV 28.33 ± 2.46 ab79.50 ± 2.21 ab60.67 ± 5.12 c146.73 ± 1.07 a105.92 ± 4.15 c
RBC27.77 ± 4.97 ab78.40 ± 3.37 ab64.00 ± 5.66 c174.66 ± 7.13 ab125.50 ± 4.96 b
FMB29.63 ± 1.15 a74.90 ± 4.44 ab74.00 ± 0.71 ab179.53 ± 3.47 bc132.58 ± 4.66 ab
CMV + RBC27.60 ± 1.39 ab73.70 ± 8.26 b65.67 ± 4.55 bc139.66 ± 3.14 c103.35 ± 5.41 c
CMV+ FMB31.43 ± 4.18 a84.10 ± 4.15 a77.00 ± 3.94 a187.79 ± 6.73 a139.93 ± 2.80 a
Control: no CMV and no biochar addition. CMV: Chinese milk vetch; RBC: rape biochar; FMB: iron-modified biochar. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
Table 3. Effect of CMV straw, rape biochar, and iron-modified biochar on photosynthetic pigments and leaf water contents of rice grown in antimony-polluted soil.
Table 3. Effect of CMV straw, rape biochar, and iron-modified biochar on photosynthetic pigments and leaf water contents of rice grown in antimony-polluted soil.
TreatmentsChl-a mg/g FWChl-b mg/g FWCart. mg/g FWAnth. mg/g FWRWC (%)
Control0.44 ± 0.017 c0.25 ± 0.013 d3.52 ± 0.11 e6.96 ± 0.44 e59.00 ± 0.77 e
CMV 0.48 ± 0.028 c0.28 ± 0.019 d3.71 ± 0.21 d7.81 ± 0.33 d65.36 ± 0.57 d
RBC0.58 ± 0.020 ab0.37 ± 0.033 bc4.14 ± 0.33 c9.13 ± 0.16 c77.69 ± 2.05 b
FMB0.61 ± 0.031 ab0.40 ± 0.041 ab4.34 ± 0.11 b9.74 ± 0.46 b82.32 ± 1.99 ab
CMV + RBC0.55 ± 0.029 b0.34 ± 0.012 c4.02 ± 0.22 c8.93 ± 0.52 c72.33 ± 2.04 c
CMV + FMB0.64 ± 0.026 a0.46 ± 0.017 a4.61 ± 0.44 a10.37 ± 0.39 a85.88 ± 3.51 a
Control: no CMV and no biochar addition. CMV: Chinese milk vetch; RBC: rape biochar; FMB: iron-modified biochar. Chl: chlorophyll, Cart: carotenoid, Anth.: anthocyanin, RWC: relative water content, FW: fresh weight. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
Table 4. Effect of CMV straw, rape biochar, and iron-modified biochar on root, shoot, and soil antimony concentration.
Table 4. Effect of CMV straw, rape biochar, and iron-modified biochar on root, shoot, and soil antimony concentration.
TreatmentsRoot Sb Concentration (mg·kg−1)Shoot Sb Concentration (mg·kg−1)Soil Sb Concentration (mg·kg−1)
Control 56.23 + 5.99 a16.83 + 1.42 a227.67 + 15.50 a
CMV 51.43 + 9.08 ab15.83 + 6.08 ab142.33 + 11.03 d
RBC 45.43 + 4.39 ab13.27 + 3.26 abc184.00 + 13.43 b
FMB 37.17 + 9.33 bc 9.35 + 3.58 bc138.43 + 11.41 d
CMV + RBC 38.17 + 6.00 bc14.57 + 0.55 abc158.07 + 12.10 c
CMV+ FMB29.53 + 5.16 c8.57 + 4.04 c129.67 + 14.50 e
Control: no CMV and no biochar addition. CMV: Chinese milk vetch; RBC: rape biochar; FMB: iron-modified biochar. Sb: antimony. The data are the means of three replications with ±SE and different letters indicating significance among means at p ≤ 0.05.
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Hu, Y.; Xiang, X.; Jiang, W.; Meng, G.; Zhou, J.; Guo, Z.; Zhou, J.; Tang, H.; Miao, J.; Morsy, K. Effect of Co-Application of Chinese Milk Vetch and Iron-Modified Biochar on Rice in Antimony-Polluted Soil. Agronomy 2024, 14, 1887. https://doi.org/10.3390/agronomy14091887

AMA Style

Hu Y, Xiang X, Jiang W, Meng G, Zhou J, Guo Z, Zhou J, Tang H, Miao J, Morsy K. Effect of Co-Application of Chinese Milk Vetch and Iron-Modified Biochar on Rice in Antimony-Polluted Soil. Agronomy. 2024; 14(9):1887. https://doi.org/10.3390/agronomy14091887

Chicago/Turabian Style

Hu, Yejie, Xinglong Xiang, Wenjie Jiang, Guiyuan Meng, Jing Zhou, Zhenzhen Guo, Jinxiu Zhou, Haiying Tang, Jianqun Miao, and Kareem Morsy. 2024. "Effect of Co-Application of Chinese Milk Vetch and Iron-Modified Biochar on Rice in Antimony-Polluted Soil" Agronomy 14, no. 9: 1887. https://doi.org/10.3390/agronomy14091887

APA Style

Hu, Y., Xiang, X., Jiang, W., Meng, G., Zhou, J., Guo, Z., Zhou, J., Tang, H., Miao, J., & Morsy, K. (2024). Effect of Co-Application of Chinese Milk Vetch and Iron-Modified Biochar on Rice in Antimony-Polluted Soil. Agronomy, 14(9), 1887. https://doi.org/10.3390/agronomy14091887

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