A Critical Review of the Effectiveness of Biochar Coupled with Arbuscular Mycorrhizal Fungi in Soil Cadmium Immobilization

Round 1

Reviewer 1 Report

Comments to the manuscript:

 A critical review of the effectiveness of biochar coupled with arbuscular mycorrhizal fungi in the soil cadmium immobilization.

General comments:

This study aims to identify the influencing factors of soil AMF community composition in the Hg mining area. The authors must clearly explain the statistical analyses used in this work as well as how they interpret the results of these analyses.

I suggest the following changes in the tables:

Table 1. The authors must remove; it is not necessary to be exhaustive with the description of impacts on humans.

Table 2. The authors must change the title as it shows both positive and negative effects.

Table 3. It is not understandable; there are too many acronyms without explanation.

I suggest the following changes in the figures:

Figure 2. There are many distractors that are not discussed in the text, for example, fishing.

Figure 4. Please put in the figure caption what PH and CEC means.

Figure 5. Please put in the figure caption what GRSP means.

 Figure 7. Pease explain what the results of the random forest Analysis means and how it is interpreted.

Line 375, please correct “extraarticular hyphae”.

Lines 381-382, please change “arbuscular” to “arbuscule”.

Line 401, write the scientific name in italics.

Line 441 and in advance, always write the scientific name in italics.

Lines 524-529, check what BCF means. Is it Bioaccumulation Factor or Bioconcentration Factor?

Lines 525 and 535, is TF for Transfer Factor or Translocation Factor?

Line 609, Random Forest Analysis is not explained before how it was performed (include in Research methodology).

Line 681, please change “arbuscular” to “arbuscule”.

I suggest the following changes in the tables:

Table 1. The authors must remove; it is not necessary to be exhaustive with the description of impacts on humans.

Table 2. The authors must change the title as it shows both positive and negative effects.

Table 3. It is not understandable; there are too many acronyms without explanation.

I suggest the following changes in the figures:

Figure 2. There are many distractors that are not discussed in the text, for example, fishing.

Figure 4. Please put in the figure caption what PH and CEC means.

Figure 5. Please put in the figure caption what GRSP means.

 Figure 7. Pease explain what the results of the random forest Analysis means and how it is interpreted.

Line 375, please correct “extraarticular hyphae”.

Lines 381-382, please change “arbuscular” to “arbuscule”.

Line 401, write the scientific name in italics.

Line 441 and in advance, always write the scientific name in italics.

Lines 524-529, check what BCF means. Is it Bioaccumulation Factor or Bioconcentration Factor?

Lines 525 and 535, is TF for Transfer Factor or Translocation Factor?

Line 609, Random Forest Analyss is not explained before how it was performed (include in Research methodology).

Line 681, please change “arbuscular” to “arbuscule”.

Author Response

Response to Reviewer #1

Thank you very much for your valuable comments and suggestions. We have carefully studied them and revised the manuscript completely. In addition, the English edit of this manuscript has been modified by a native English expert. The responses are given as follows.

Comment on the tables

Comment 1: Table 1. The authors must remove; it is not necessary to be exhaustive with the description of impacts on humans.

Response 1: We appreciate your suggestions and patient corrections to our manuscript. We have deleted Table 1. Please take a look at it.

Comment 2: Table 2. The authors must change the title as it shows both positive and negative effects.

Response 2: Thank you very much for your valuable comments. We have changed the title of Table 2, which has been changed to Table 1 in the revised version of manuscript.

Comment 3:  Table 3. It is not understandable; there are too many acronyms without explanation.

Response 3: Thank you for your valuable comments. We have elucidated the meanings of all acronyms of Table 3, which has been changed to Table 2 in the revised version of manuscript.

Comment on the figures

Comment 1: Figure 2. There are many distractors that are not discussed in the text, for example, fishing.

Response 1: We appreciate your valuable comment. However, we were requested by another reviewer to delete Figure 2 and provide an explanation solely focusing on the geochemical behavior of cadmium in the environment. Therefore, we have deleted it as it was requested.

Comment 2: Figure 4. Please put in the figure caption what PH and CEC means.

Response 2: Thank you very much for your valuable suggestion and comments. We have elucidated the acronyms of PH and CEC of Figure 4, which has been changed into Figure 2 in the revised version of manuscript.

Comment 3:  Figure 5. Please put in the figure caption what GRSP means.

Response 3: Thank you for valuable comment. We have added the full meaning of GRSP in Figure 5 caption, which has been changed into Figure 3 in the revised version of manuscript..

Comment 4: Figure 7. Please explain what the results of the random forest Analysis means and how it is interpreted.

Response 4: Thank you for your valuable comment. Typically, random Forest Analysis is a machine learning technique that is often used for predictive modeling and classification. In the context of our statement, it seems like you are considering the application of Random Forest Analysis to conduct a critical review of the effectiveness of biochar coupled with arbuscular mycorrhizal fungi in the soil cadmium immobilization. Ultimately, the Random Forest Analysis can serve as a data-driven approach to evaluate and understand the complex interactions between biochar, mycorrhizal fungi, and soil cadmium immobilization. It helps identify the most influential factors and their combined effects on the outcome of interest.

Here's how we applied Random Forest Analysis in this review paper:

1. Data collection: Gather data related to the effectiveness of biochar and arbuscular mycorrhizal fungi in immobilizing cadmium in the soil. This data may include experimental results, field studies, and other relevant information.
2. Feature selection: Identify key features or variables that are important in assessing the effectiveness of the mentioned soil amendments. These features could include biochar properties, mycorrhizal fungi presence, soil characteristics, and cadmium levels.
3. Training the Model: Use historical data to train the Random Forest model. The model will learn the patterns and relationships between the chosen features and the effectiveness of cadmium immobilization.
4. Prediction: Apply the trained model to new or unseen data to predict the effectiveness of biochar and arbuscular mycorrhizal fungi in soil cadmium immobilization. This can provide insights into how these factors contribute to the desired outcome.
5. Evaluation: Assess the accuracy and reliability of the model's predictions. This step is crucial for determining the validity of the Random Forest Analysis in predicting the effectiveness of biochar and mycorrhizal fungi in immobilizing cadmium.

Comment on the lines

Comment 1: Line 375, please correct “extraarticular hyphae”.

Response 1: We appreciate for your valuable comment. As shown in lines 358 and 362 , we have changed “extraarticular hyphae” into “extraradical hyphae”.

Comment 2: Lines 381-382, please change “arbuscular” to “arbuscule”.

Response 2: Thank you very much for your valuable comment. As shown in line 664, we have corrected that miswritten words.

Comment 3:  Line 401, write the scientific name in italics.

Response 3: Thank you very much. We have written those words in italics as indicated in lines 330-332.

Comment 4: Line 441 and in advance, always write the scientific name in italics.

Response 4: Thank you for your valuable comment. We have corrected these errors (lines 424 and 427)

Comment 5: Lines 524-529, check what BCF means. Is it Bioaccumulation Factor or Bioconcentration Factor?

Response 5: Thank you for your valuable comment. As shown in Line, 509 BCF stands for Bioconcentration Factor. Subsequently, we have meticulously rectified all identified errors within the manuscript to enhance its accuracy and overall quality.

Comment 6: Lines 525 and 535, is TF for Transfer Factor or Translocation Factor?

Response 6: Thank you for your valuable comment. As shown in line 510, TF refers translocation factor. Subsequently, we have meticulously rectified all identified errors within the manuscript to enhance its accuracy and overall quality.

Comment 7: Line 609, Random Forest Analysis is not explained before how it was performed (include in Research methodology).

Response 7: Thank you for your valuable comment. As shown in lines 101-103, we have included Random Forest Analysis in research methodology.

Comment 8: Line 681, please change “arbuscular” to “arbuscule”.

Response 8: Thank you for your valuable comment. We have changed “arbuscular” into “arbuscule” (line 664).

Author Response File: Author Response.pdf

Reviewer 2 Report

The work entitled "A critical review of the effectiveness of biochar coupled with arbuscular mycorrhizal fungi in the soil cadmium immobilization" presents an extensive review of the relationship between biochar and Cd, between AMF and this same contaminant and the synergistic effect of biochar-AMF. Bibiographical references are appropriate and the topic is well addressed and clearly expressed. However, I consider that certain Tables and Figures should be reviewed, corrected and/or eliminated from the manuscript, since they contain repeated information in the text without providing extra data. This would prevent the manuscript from appearing repetitive.

There are specific suggestions and corrections regarding the presentation of some data or corrections in the format in which the bibliographical references are presented, which I detail in the attached file.

Comments for author File: Comments.pdf

Author Response

Response to Reviewer #2

Thank you very much for your valuable comments and suggestions. We have carefully studied them and revised the manuscript completely. In addition, the English edit of this manuscript has been modified by a native English expert. The responses are given as follows.

General comments:

Comment 1: Different font size.

Response 1: We appreciate your suggestions and patient corrections to our manuscript. As shown in line 12-13, we have corrected these lines and ensured that all words are in the same font size.

Comment 2: I suggest rephrasing the sentence as: Biochar and arbuscular mycorrhizal fungi (AMF) can effectively immobilize cadmium in the soil in an environmentally friendly way.

Response 2: Thank you sincerely for your valuable comments aimed at enhancing our manuscript. We have made the necessary corrections to that sentence as per your suggestions (lines 14-15).

Comment 3:  I suggest removing the comma.

Response 3: Thank you for your valuable comments. We have remove that comma (line 46) and we have checked whole manuscript.

Comment 4: Given the length of the text and the number of figures, I suggest presenting Figure 1 and part of the information about the bibliographic compilation process (lines 101 to 122) as supplementary material.

Response 4: We appreciate your suggestions and patient corrections to our manuscript. We have presented Figure 1 as Figure S1 and part of the information about the bibliographic compilation process (lines 104 to 145) in supplementary materials.

Comment 5: I suggest eliminating Figure 2. The information presented here does not provide additional data to those already expressed in the text (lines 129 to 138). Although the caption of the figure says "Geochemical behavior of cadmium in the environment", nothing is found about the behavior of this heavy metal. Lastly, the quality of the figure does not seem appropriate.

Response 5: Thank you for your valuable suggestions and comments. As indicated in the manuscript, we have removed Figure 2, which was titled geochemical behavior of cadmium in the environment.

Comment 6:  This sentence (Line 142) must be rewritten for better understanding. Is there more current data on soil contamination in China?

Response 6: Thank you very much for your valuable comments. We have rewritten that sentence (see lines 116-117). The current used data are the one which are available in published papers and websites.

Comment 7: For line 148, I suggest adding more details on the differential bioavailability according to the chemical form of cadmium

Response 7: Thank you for your valuable comment and patient for our correction. We have added details on the differential bioavailability according to the chemical form of cadmium (see lines 118-128).

Comment 8: I suggest eliminating Table 1 since it is very extensive and is not related to the topic of this review work.

Response 8: Thank you for your valuable comment. We have eliminated Table 1 from our manuscript, which was titled “the potential effects of cadmium on different human body parts”.

Comment 9: For line 173, References to other acronyms and the bibliographic citation of the original work are missing

Response 9: Thank you for your valuable comment. As indicated in Lines 158-160 (Current title of figure 1), all acronyms have been fully defined

Comment 10: For lines 175-177, rewrite this sentence.

Response 10: Thank you for your valuable comment. As shown in lines 148-156, we have rewritten these sentences and whole paragraph was revised.

Comment 11: For line 187, I suggest rewriting as " and functional group complexation, among others".

Response 12: Thank you for your valuable comment. We have rewrite that sentence according to your suggestion (see line 164).

Comment 13: For line 216, SI and S2 are nomenclatures specific to Meng et al. work, so it is not appropriate to use them outside the context of that work. I suggest rewriting the sentences on lines 214 to 218.

Response 14: Thank you for your valuable comment. These lines have been rewritten (see line 196-198).

Comment 15: For line 308-315, correct the miswritten.

Response 15: Thank you for your valuable comment. These lines have been corrected (see line 288-295).

Comment 16: For line 117 (Figure 5), In the figure, which is indicating the yellow circle with reference "affect the host gene and the cell wall"? I suggest improving the edges of the box in the figure

Response 17: Thank you for your valuable comment. The edges of the figures have been enhanced, resulting in improved clarity. Figure 5 has been changed into Figure 3.

Comment 18: For lines 320-324, this paragraph needs context, what soil is it referring to? Even if there is a reference at the end, I would suggest being clearer for the reader of the review.

Response 18: Thank you for your valuable comment. We have amended that paragraph ( see lines 301-307).

Comment 20: For lines 348-349, I suggest writing the specific name in italics first and the abbreviation in parentheses: Funneliformis mosseae (FM), in this case and throughout the entire manuscript.

Response 20: Thank you for your valuable comment. We have changed accordingly (lines 330-332).

Comment 21: For lines 401, these abbreviations had already been presented, so it is not necessary to rewrite the specific name, however previously capital letters were used (FM, RI).

Response 21: Thank you for your valuable comment. We have changed accordingly (line 384).

Comment 22: For lines 479, Incorrect reference format.

Response 22: Thank you for your valuable comment. As shown in Line 460, incorrect format was corrected and whole manuscript was checked.

Comment 23: For lines 510, incorrect reference format.

Response 23: Thank you for your valuable comment. As shown in line 476, incorrect formatting has been corrected, and the entire manuscript has been thoroughly reviewed.

Comment 24: For lines 529-535, correct errors in this paragraph.

Response 24: Thank you for your valuable comments. All errors have been corrected, and the entire manuscript has been thoroughly reviewed ( see lines 508-525).

Comment 25: For lines 544, for Figure 6. Correct miswritten words.

Response 25: Thank you for your valuable comment. All errors have been corrected, and the entire manuscript has been thoroughly reviewed (see line 529).

Comment 26: For lines 583-592, correct reference format.

Response 26: Thank you for your valuable comment. All errors have been corrected, and the entire manuscript has been thoroughly reviewed (see lines 571 and 575).

Comment 27: For table 3, TP is not defined in the Table note.

Response 27: Thank you for your valuable comment. TP in Table 2 (former table 3) represents total phosphorus and the entire manuscript has been thoroughly checked.

Comment 28: For figure 7, TP was not defined in the Figure caption.

Response 28: Thank you for your valuable comment. TP represents total phosphorus and the entire manuscript has been thoroughly checked (see line 607).

Comment 29: For line 628, table 1 does not refer to this topic, but Table 2 does.

Response 29: Thank you for your valuable comment. We have deleted Table 1, means that table 2 has been changed to table 1 in the revised manuscript.

Author Response File: Author Response.pdf

Reviewer 3 Report

Review Comments for Manuscript ID jof-2854348

The manuscript provides a thorough evaluation of the synergistic effects of biochar and arbuscular mycorrhizal fungus (AMF) on cadmium immobilization in soil. The authors meticulously investigated numerous elements that influence the efficacy of this integrated method. The extensive examination of the mechanisms, such as biochar's adsorption capacities and AMF's improvement of nutrient uptake, provides vital insights into the combined application's many benefits.

After careful consideration, I would like to recommend this manuscript for publication in JoF (ISSN 2309-608X), provided the authors address major revisions. These suggestions are outlined below:

Abstract:

1.           There appears to be a font type inconsistency in lines 12–13 of the abstract.

Introduction:

2.        I suggest identifying the Sustainable Development Goals (SDGs) that intersect with the theme addressed in the manuscript. Including this information could enhance the significance and visibility of the authors' research.

In reference to the manuscript sections discussing:

3.  Cadmium mobility in soil and water, including aspects such as bioavailability, and leaching into groundwater (Lines 141-169).

4.  The potential adverse health effects resulting from anthropogenic activities leading to cadmium pollution (Lines 175-183).

5.   The inherent properties of biochar, derived from various raw materials, and their impact on effective cadmium sequestration, along with their implications in the cadmium sequestration process (Lines 639-645)

Given the critical nature of these topics, I would like to highlight the potential importance of incorporating Life Cycle Assessment (LCA) methodologies in the analysis. An LCA approach could provide a comprehensive understanding of the environmental impact associated with cadmium mobility, anthropogenic activities, and the use of biochar. Moreover, LCA can offer valuable insights into the overall sustainability of the proposed solutions.…

If the authors have not yet come across studies employing Life Cycle Assessment to address these specific mechanisms, it might be worth considering that the authors promote research in this area.

6.  The phrase "Biochar enhances plant biomass" could be changed to “Biochar increases both aboveground and belowground plant biomass.” (Lines 304-306)

7. In plant physiology, the research revealed that the combined treatment significantly decreased the malondialdehyde content across various cadmium levels by 69.1%, indicating a reduction in oxidative stress and cellular damage. Simultaneously, the treatment boosted antioxidant enzyme activity by 54.3%, 83.4%, and 52%, respectively. It would be beneficial for the authors to explicitly mention that the measurement of malondialdehyde serves as an indicator of oxidative stress and cellular damage, and thus, its decrease is a favorable outcome. (Lines 503-506). Similarly, it would be valuable for the authors to provide explanations for measurements such as Glutathione, Lipid Peroxidation, Ascorbate Peroxidase, Amino Acid Content, Glutathione Reductase, Glycine Betaine Content, and Total Phenols. Providing detailed insights into these parameters will enhance the overall understanding of the study and its implications. (Lines 511-516).

8.  In the manuscript, it would be beneficial to provide additional details on the conventional methods employed for biochar preparation, as well as information on the inoculation of arbuscular mycorrhizal fungus (AMF) in plants.

9.  The authors have cited a work that utilized random forest analysis to identify key factors, such as biochar type, pH, AMF type, and soil cadmium concentration. Employing a deep learning approach could provide valuable insights and serve as a guiding framework for exploring intricate mechanisms involved in the interactions observed. I encourage the authors to consider incorporating this suggestion into their discussion or future research directions."

Sincerely,

Comments for author File: Comments.pdf

Author Response

Response to Reviewer #3

Thank you very much for your valuable comments and suggestions. We have carefully studied them and revised the manuscript completely. In addition, the English edit of this manuscript has been modified by a native English expert. The responses are given as follows.

Abstract:

Comment 1: There appears to be a font type inconsistency in lines 12–13 of the abstract.

Response 1: Thank you for reviewing our manuscript. We appreciate your suggestions and patient corrections to our manuscript. We have made these sentences consistency and passed whole manuscript. Please see the first two lines in red color of the abstract.

 Introduction:

Comment 2: I suggest identifying the Sustainable Development Goals (SDGs) that intersect with the theme addressed in the manuscript. Including this information could enhance the significance and visibility of the authors' research.

Response 2: Thank you sincerely for your valuable comments aimed at enhancing our manuscript. Based on Sustainable Development Goals (SDGs) on cadmium, Cadmium is a non-essential and toxic element for humans mainly affecting kidneys and the skeleton. It is also a carcinogen by inhalation. Cadmium is accumulated in bone and may serve as a source of exposure later in life. Cadmium is used in batteries, paints, plastics, electroplating etc. It is released to the atmospheric environment from metals production, fossil fuel combustion. Phospherous fertilisers and sewage sludges are also a major source of environmental releases of cadmium. As shown in lines 38-42, examples of SDGs are evinced in the introduction, which shows that cadmium is headache issue worldwide. However, the information on the remediation of cadmium using biochar-AMF is a new area of study, which is why we have focused on conducting research in this field.

In reference to the manuscript sections discussing:

Comment 3:  Cadmium mobility in soil and water, including aspects such as bioavailability, and leaching into groundwater (Lines 141-169).

Response 3: Thank you for your valuable comments. As shown in lines 141-168, we have edited and improved the meaning of that part (see lines 116-139).

Comment 4: The potential adverse health effects resulting from anthropogenic activities leading to cadmium pollution (Lines 175-183).

Response 4: We appreciate your suggestions and patient corrections to our manuscript. As shown in lines 175-185, we have rewritten these sentences (see lines 146-151).

Comment 5: The inherent properties of biochar, derived from various raw materials, and their impact on effective cadmium sequestration, along with their implications in the cadmium sequestration process (Lines 639-645)

Given the critical nature of these topics, I would like to highlight the potential importance of incorporating Life Cycle Assessment (LCA) methodologies in the analysis. An LCA approach could provide a comprehensive understanding of the environmental impact associated with cadmium mobility, anthropogenic activities, and the use of biochar. Moreover, LCA can offer valuable insights into the overall sustainability of the proposed solutions.…

If the authors have not yet come across studies employing Life Cycle Assessment to address these specific mechanisms, it might be worth considering that the authors promote research in this area.

Response 5: Thank you very much for your valuable suggestion and comments. As shown in lines 638-651, we have revised that content accordingly. Based on the information provided below, our manuscript is fulfilled Life Cycle Assessment (LCA) methodologies. Life Cycle Assessment (LCA) methodologies for this topic involves a systematic and comprehensive analysis of the environmental impacts associated with the entire life cycle of the biochar-AMF system.

Here's a step-by-step guide:

1. Define the Scope and Objectives:
• Clearly define the objectives of your LCA, specifying what aspects of the biochar-AMF system you want to assess (e.g., cadmium immobilization effectiveness, environmental impacts).
• Identify the functional unit, which represents the quantifiable unit of the system's performance (e.g., per unit of cadmium immobilized, per unit of biochar produced).
2. System Boundary:
• Define the system boundary by determining the life cycle stages to be considered (e.g., biochar production, AMF cultivation, application, and end-of-life disposal).
• Consider the direct and indirect processes related to each life cycle stage.
3. Inventory Analysis:
• Collect data on inputs and outputs for each life cycle stage, including raw materials, energy consumption, emissions, and waste generation.
• Consider the specific characteristics of the biochar production process, AMF cultivation, and the overall application in the context of soil cadmium immobilization.
4. Impact Assessment:
• Use impact categories relevant to the objectives of this study, such as global warming potential, eutrophication, or toxicity.
• Evaluate the potential environmental impacts associated with each life cycle stage and express them in quantitative terms.
5. Interpretation:
• Interpret the results by analyzing the significance of identified impacts.
• Identify areas of improvement or hotspots within the life cycle stages.
• Consider trade-offs and synergies between different environmental impact categories.
6. Sensitivity Analysis:
• Perform sensitivity analyses to assess the influence of key parameters or uncertainties on the overall results.
• Understand how variations in factors like biochar production methods or AMF cultivation conditions affect the LCA outcomes.
7. Comparison and Benchmarking:
• Compare the LCA results with alternative approaches or technologies for soil cadmium immobilization.
• Benchmark against existing environmental standards or guidelines.
8. Reporting and Communication:
• Present the findings in a clear and accessible manner.
• Communicate the implications of the LCA results for decision-making and policy development.
9. Iterative Process:
• LCA is often an iterative process, and refinement may be needed based on feedback and further data collection.

Therefore, by applying these steps, we can use Life Cycle Assessment methodologies to assess the environmental performance of the biochar-AMF system in soil cadmium immobilization and gain insights into potential environmental improvements or trade-offs associated with its application.Top of Form

Comment 6:  The phrase "Biochar enhances plant biomass" could be changed to “Biochar increases both aboveground and belowground plant biomass.” (Lines 304-306).

Response 6: Thank you very much for your valuable comments. We have rewritten that sentence accordingly (Please see the lines 283-286).

Comment 7: In plant physiology, the research revealed that the combined treatment significantly decreased the malondialdehyde content across various cadmium levels by 69.1%, indicating a reduction in oxidative stress and cellular damage. Simultaneously, the treatment boosted antioxidant enzyme activity by 54.3%, 83.4%, and 52%, respectively. It would be beneficial for the authors to explicitly mention that the measurement of malondialdehyde serves as an indicator of oxidative stress and cellular damage, and thus, its decrease is a favorable outcome. (Lines 503-506). Similarly, it would be valuable for the authors to provide explanations for measurements such as Glutathione, Lipid Peroxidation, Ascorbate Peroxidase, Amino Acid Content, Glutathione Reductase, Glycine Betaine Content, and Total Phenols. Providing detailed insights into these parameters will enhance the overall understanding of the study and its implications. (Lines 511-516).

Response 7: Thank you for your valuable comment and suggestions. Typically, malondialdehyde (MDA) is a natural byproduct of lipid peroxidation, a process that occurs when reactive oxygen species (ROS) interact with polyunsaturated fatty acids in cell membranes. Reactive oxygen species are highly reactive molecules that can be generated during normal metabolic processes or in response to various stressors, such as exposure to pollutants, radiation, or toxins. When cells experience oxidative stress, the balance between the production of ROS and the cell's ability to neutralize them is disrupted. This imbalance can lead to the peroxidation of lipids in cell membranes, resulting in the formation of malondialdehyde. MDA is a reactive aldehyde and a breakdown product of lipid peroxidation, which can be measured as a marker of oxidative damage. The detection and quantification of malondialdehyde levels are commonly used as a biomarker to assess the extent of lipid peroxidation and, by extension, the degree of oxidative stress and cellular damage in biological systems. Elevated levels of malondialdehyde are indicative of increased oxidative stress, and a decrease in MDA levels is generally considered a positive outcome, suggesting a reduction in oxidative damage to cellular structures. This is an essential factors for AMF and biochar association. Its measurement is detailed in several published papers such as paper written y Guo et al “Effects of Biochar and AM Fungi on Growth, Mineral Elements and Cadmium Uptake of Mulberry under Cadmium Stress. IOP Conf. Ser. Earth Environ. Sci. 2021, 687, doi:10.1088/1755-1315/687/1/012021”. That is why we did not write its measurement processes. Same as for measurements such as Glutathione, Lipid Peroxidation, Ascorbate Peroxidase, Amino Acid Content, Glutathione Reductase, Glycine Betaine Content, and Total Phenols. They are all detailed in published papers. To provide the measurement it should be like repletion which is not necessary for a review paper.

Comment 8: In the manuscript, it would be beneficial to provide additional details on the conventional methods employed for biochar preparation, as well as information on the inoculation of arbuscular mycorrhizal fungus (AMF) in plants.

Response 8: Thank you for your valuable comment. The conventional methods employed for biochar preparation, as well as information on the inoculation of arbuscular mycorrhizal fungus (AMF) in plants are fruitful information, however, these methods were detailed in published papers (http://dx.doi.org/10.1007/s42773-024-00301-w) and (doi:10.1007/s11104-020-04678-y). Therefore, we think that to repeat information should not be better.

Comment 9: The authors have cited a work that utilized random forest analysis to identify key factors, such as biochar type, pH, AMF type, and soil cadmium concentration. Employing a deep learning approach could provide valuable insights and serve as a guiding framework for exploring intricate mechanisms involved in the interactions observed. I encourage the authors to consider incorporating this suggestion into their discussion or future research directions."

Response 9: Thank you for your valuable comment. We have incorporated the suggestion of the utilization of a deep learning approach in the discussion or future research directions. Please see the red color in the discussion or future research directions (see lines 656-662).

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

The review paper makes a valuable contribution to the field of soil remediation by exploring the synergistic potential of biochar and AMF in immobilizing cadmium. By addressing current knowledge gaps and highlighting practical implications, the paper offers insights that could inform future research efforts and environmental management practices.

The authors have adequately addressed the items outlined in the review.

Author Response

Response to the editor's and reviewers’ comment

Thank you very much for your valuable comments and suggestions. We have carefully studied them and revised the manuscript completely according to editors’ comment. The responses are given as follows.

Comment 1: L60: There are grammar mistakes. Remember "AMF" (arbuscular mycorrhizal fungi) are plural; “80%” is not accurate, we often use “more than 80%”. I suggest you correct as "AMF can form symbiosis with the majority of land plants"

Response 1: We appreciate your suggestions and patient corrections to our manuscript. As shown in line 60, we have corrected that sentence according to your suggestions.

Comment 2: L61: Refs [16,17] are not good references, which can be replaced by two reviews:
Shi, Z., Li, K., Zhu, X., & Wang, F. (2020). The worldwide leaf economic spectrum traits are closely linked with mycorrhizal traits. Fungal Ecology, 43, 100877.
Wang, F., RENGEL, Z. (2024). Disentangling the contributions of arbuscular mycorrhizal fungi to soil multifunctionality. Pedosphere. https://doi.org./10.1016/j.pedsph.2023.12.015

Response 2: We thank you for your suggestions. As shown in Line 61 and references lists (reference 16 & 17), we have replaced those references.

Comment 3: L61: correct “It” as “They” or “AMF”; Please check the whole manuscript to correct such mistakes.  

Response 3: We appreciate your patient corrections to our manuscript. As indicated in Line 61, we have replaced it by they.

Comment 4: L131-136: wrong citation order; [40], [41]…

Response 4: We appreciate your suggestions and patient corrections to our manuscript. As shown in Lines 132-134, we have corrected wrong citation order.

Comment 5: Figure 2: why use “Reduces” first but “Reduce” second?

Response 5: Very sorry for that mistake and we thank you for your valuable comment. Therefore, we have used “reduces” for both sentences. We have replaced that the 1^st figure with another corrected figure.

Comment 6: L177-178: you need not explain “pH”

Response 6: Thank you very much for your valuable comment. We have deleted the explanation of pH (see lines 176-177).

Comment 7: L320: Pan et al. [77] is wrong. Check all the citations and confirm they match the cited references.

Response 7: Thank you for your correction. We have checked the correctness of all references (see lines 319-320).

Comment 8: L776: Ref [31] is totally same as Ref [74] (L872-874); check all the references.

Response 8: We appreciate your suggestions and patient corrections to our manuscript. We have removed that repeated reference. In addition, we have checked whole manuscript (See reference list).

Comment 9: L304: “It” is not clear.

Response 9: We appreciate your suggestions and patient corrections to our manuscript. We have made it clear (see lines 303-304).

Comment 10: L314: correctly use subscript and superscript in “NO3−“

Response 10: We appreciate your suggestions and patient corrections to our manuscript. We have corrected it (see line 313).

Comment 11: L398: correct “AM fungi” as “AMF”

Response 11: We appreciate your suggestions and patient corrections to our manuscript. We have corrected “AM fungi” as “AMF” (see line 397).

Comment 12: L408: Where is “[91]”?

Response 12: We appreciate your suggestions and patient corrections to our manuscript. We have inserted that reference (see line 395). In addition, we have checked all references.

Comment 13: L422: replace “repair”; check the whole manuscript.

Response 13: We appreciate your suggestions and patient corrections to our manuscript. As shown in Line 421, we have revised that sub-title.

Comment 14: L428: Italicize “G.”

Response 14: Thank you for your valuable comment. As shown in Line 427, we have italicized “G”.

Comment 15: L439: It is wrong “abiotic impact”; Rephrase this sentence

Response 15: We appreciate your suggestions and patient corrections to our manuscript. As shown in Lines 438-439, we have rephrased that sentence.

Comment 16: L440: “can also impacted” is wrong. Please check the language and correct grammar mistakes.

Response 16: We appreciate your suggestions and patient corrections to our manuscript. We have corrected that sentence (see line 439).

Comment 17: L488: Guo et al. [109] should be correct as “Guo and Li [109]”; in addition, [109] has been introduced in L468; Is it necessary to repeat it again?

Response 17: We appreciate your suggestions and patient corrections to our manuscript. We have deleted that reference in L467. Means that it is better to use it only on L486.

Comment 18: L542: provide the number of this ref.

Response 18: We appreciate your suggestions and patient corrections to our manuscript. We have provided that reference number (see line 540).

Comment 19: Table 2: italicize the Latin names; there is a space between genus and species name;

Response 19: We appreciate your suggestions and patient corrections to our manuscript. We have corrected those errors (see table 2). In addition, there is a space between genus and species name was provided.

Comment 20: L757-760: check the title.

Response 20: We appreciate your suggestions and patient corrections to our manuscript. We have checked that references (See references list).

Comment 21: L833: check the title

Response 21: We appreciate your suggestions and patient corrections to our manuscript. We have corrected it (See references list).

Comment 22: L835-836: Wrong authors’ surnames; provide journal title. Check all the references carefully.

Response 22: We appreciate your suggestions and patient corrections to our manuscript. We have provided authors’ surnames (see references list)

Comment 23: L853: Italicize “Zea Mays”.

Response 23: We appreciate your suggestions and patient corrections to our manuscript. We have italicized it (see reference 124).

Comment 24: L861: Italicize “Triticum Aestivum”; check all the Latin names.

Response 24: We appreciate your suggestions and patient corrections to our manuscript. We have italicized it (see reference 70).

Comment 25: L918: check the authors. “W.F.F”?

Response 25: We appreciate your patient corrections to our manuscript. We have checked and corrected all references (See references list).

Author Response File: Author Response.docx