Microbial Composition of Natural, Agricultural, and Technogenic Soils of Both Forest and Forest-Tundra of the Russian North

Round 1

Reviewer 1 Report

I have read the article titled “Microbiome of Natural, Agricultural and Technogenic Soils of Forest and Forest‐tundra of the Russian North”. Your article is written well and is at a level that will contribute to the literature. But, some grammatical misusages and some incomprehensible sentences have been detected in the article. In the material and method section, there are deficiencies in taking soil samples and which method is used. Also, some additional needs in the M&M part. I added some minor suggestions in the PDF file.

 Title

The title is clear and shows the manuscript.

Abstract

Some grammatical misusages have been detected in this part.

 

Introduction

There is also a grammatical problem here. Some important sentences should be cited from more recent studies. I think a better literature review is needed in this part.

Material and Method

There are also some grammatical problems in this part. The methodology needs to be rewritten well and understandable. There is no information about the climate, topography, and vegetation characteristics of the research areas. This lack of information needs to be corrected.

 

Results

There is also a grammatical problem here. This section of the manuscript is clear, relevant, and presented well.

Discussion

This section has been written clearly, is relevant, and presented well.

Conclusion

This section has been written clearly, is relevant, and presented well.

The figures/tables/images/schemes are appropriate and they are properly showing the data.

 

 

 

Comments for author File: Comments.pdf

Your article is written well and is at a level that will contribute to the literature. But, some grammatical misusages and some incomprehensible sentences have been detected in the article. 

Author Response

Reply for 1^rd reviewer comments

 

I have read the article titled “Microbiome of Natural, Agricultural and Technogenic Soils of Forest and Forest‐tundra of the Russian North”. Your article is written well and is at a level that will contribute to the literature. But, some grammatical misusages and some incomprehensible sentences have been detected in the article. In the material and method section, there are deficiencies in taking soil samples and which method is used. Also, some additional needs in the M&M part. I added some minor suggestions in the PDF file.

 Title

The title is clear and shows the manuscript.

Abstract

Some grammatical misusages have been detected in this part.

Reply - Grammatical issues has been improved

 

Introduction

There is also a grammatical problem here. Some important sentences should be cited from more recent studies. I think a better literature review is needed in this part.

Reply - Grammatical issues has been improved

Material and Method

There are also some grammatical problems in this part. The methodology needs to be rewritten well and understandable. There is no information about the climate, topography, and vegetation characteristics of the research areas. This lack of information needs to be corrected.

 

Reply – data on vegetation cover and climate provided.

 

Results

There is also a grammatical problem here. This section of the manuscript is clear, relevant, and presented well.

Discussion

This section has been written clearly, is relevant, and presented well.

Conclusion

This section has been written clearly, is relevant, and presented well.

The figures/tables/images/schemes are appropriate and they are properly showing the data.

 

Dear reviewer, thank you for all suggestions in the text, all changes are tracked by red color in updated text.

Sincerely yours, corresponding author,

Evgeny Abakumov

Reviewer 2 Report

The manuscript tiled "Microbiome of Natural, Agricultural and Technogenic Soils of Forest and Forest-tundra of the Russian North".  The idea of comparison between different soil via their biodiversity and their chemical and physical properties is very good. Also, presence of the phyla such as; Protobacteria, Actinobateriota, Acidobateriota, Bacteroidota, Chroloflexi, 20 Planctomycetota, Verrucomicrobiota and Firmicutes in the examined soils revealed that most of the examined soils are fertile and it seems healthy, especially the natural and forest soils.

Title: should be changed into:  Bacteribiome of Natural, Agricultural and Technogenic Soils of both Forest and Forest-tundra of the Russian North.

Materials and methods;

Accordingly, studying the physical and chemical properties of the examined soils should be included in this study and should be tabulated. Biodiversity based on the 16S rRNA was indicated only for the bacterial not for the fungi. I think the studying of the fungi based on the 18S rRNA will help more to obtain a good recommendation and for the examined soils. The microbial analysis was performed but the authors did not reported the types of the bacteria was obtained and the types of media which used. The number of bacterial species obtained by sequencing when compared with those obtained by plate microbiology. The living and nonliving bacteria, how could the authors would be sure based on the metabiome. The soil DNA concentration and purity did not mentioned in the materials and methods part.  The image of the 16S rRNA amplicone on the agrose gel should be included to show the quality and the DNA integrity. The intact DNA could be indicator for the living bacteria for what extent.  The website/source for the programs used in sequence analysis should be included.

Results and discussion:

 Figure 2 should be improved because the names of bacterial phyla is not appeared in good manner.

Citation is not enough and quite poor, so, more references should be added to make the discussion is more acceptable.

Conclusion:

Is too long and should be summarized.

 

Author Response

Reply for 2nd reviewer comments

 

Dear reviewer, thank you for all suggestions in the text, all changes are tracked by red color in updated text.

 

 

The manuscript tiled "Microbiome of Natural, Agricultural and Technogenic Soils of Forest and Forest-tundra of the Russian North".  The idea of comparison between different soil via their biodiversity and their chemical and physical properties is very good. Also, presence of the phyla such as; Protobacteria, Actinobateriota, Acidobateriota, Bacteroidota, Chroloflexi, 20 Planctomycetota, Verrucomicrobiota and Firmicutes in the examined soils revealed that most of the examined soils are fertile and it seems healthy, especially the natural and forest soils.

Title: should be changed into:  Bacteribiome of Natural, Agricultural and Technogenic Soils of both Forest and Forest-tundra of the Russian North.

Reply – we are agree, the title has been changed in current version.

Materials and methods;

Accordingly, studying the physical and chemical properties of the examined soils should be included in this study and should be tabulated

Reply – there is a table 1 with agrochemical data in paper text.

Biodiversity based on the 16S rRNA was indicated only for the bacterial not for the fungi. I think the studying of the fungi based on the 18S rRNA will help more to obtain a good recommendation and for the examined soils.

Reply – we are agree, that this additional data may improve paper, but, we have only data, based on 16rRNA, thus, we also changed title of the manuscript as it was suggested.

 The microbial analysis was performed but the authors did not reported the types of the bacteria was obtained and the types of media which used. The number of bacterial species obtained by sequencing when compared with those obtained by plate microbiology. The living and nonliving bacteria, how could the authors would be sure based on the metabiome.

Reply – we have not compared living and non living bacteria and we have not used any types of media in this research.

The soil DNA concentration and purity did not mentioned in the materials and methods part. 

Reply - DNA was purified and diluted according to Illumina sequencing requirements (200-400 ng per sample). This information has been added to text.

 

 The image of the 16S rRNA amplicone on the agrose gel should be included to show the quality and the DNA integrity. The intact DNA could be indicator for the living bacteria for what extent.  The website/source for the programs used in sequence analysis should be included.

Reply – we have to clarify if it is necessary to add this data massive to Supplementary? If yes, it should be about 10 separate pages. Let us know, please.

 

Results and discussion:

 Figure 2 should be improved because the names of bacterial phyla is not appeared in good manner.

Reply – figure quality has been improved.

Citation is not enough and quite poor, so, more references should be added to make the discussion is more acceptable.

Reply – some references added.

Conclusion:

Is too long and should be summarized

Reply – corrected

Sincerely yours, corresponding author,

Evgeny Abakumov

Reviewer 3 Report

General comment:

The manuscript discusses the Microbiome of Soils in Northern Russian regions. The contents are interesting and worth reading. However, there are several professional comments/suggestions that the authors should consider.

 

Title:

The present title the authors provided is quite descriptive and conveys the main focus of the manuscript, which appears to be about the microbiome in different types of soils (natural, agricultural, and technogenic) found in specific regions (forest and forest-tundra) of the Russian North. However, it is long and could be improved for better readability and clarity. I suggest the authors consider using a shorter title.

 

Abstract:

Elaborate on the types of disturbances or agricultural practices that have been studied and their potential implications for soil microbiota. Mention the specific techniques and methodologies used for metagenomic sequencing (e.g., library preparation, sequencing platforms) to give readers a clearer understanding of the study's technical aspects. Clarify the rationale for selecting the particular regions in Russia for soil sampling. Were there specific factors or characteristics that led to their inclusion in the study? It's important to provide more context on the significance of the results and how they contribute to the existing body of knowledge. How does this study fill the knowledge gap mentioned earlier, and how can this new understanding of soil microbiomes be practically applied for soil reclamation and amelioration in severe climatic conditions? The authors should consider adding some quantitative information about the taxonomic diversity of the microbiomes in the various ecosystems studied. Mention any significant findings, such as differences in diversity or abundance of specific microbial groups. Please provide a clearer description of the "depression of microbiome communities" mentioned in the abstract. Elaborate on what this means and how it relates to the formation of new microbial communities in quarries. Ensure consistency in the use of terms; for instance, the abstract mentions "agrogenic" and "agro-soils," which seem to refer to the same concept. Stick to one term to avoid confusion. The authors should consider rephrasing the sentence: "Thus, data on soil microbiome composition may be used for further driving of soil properties for reclamation and amelioration practices in severe climatic conditions of northern Russia." This sentence is crucial, but it could be more straightforward in conveying how the findings can practically impact soil management strategies.

 

Introduction:

The authors can add more context on the current state of soil microbiology research and how it relates to the need for a global understanding of the soil microbiome (mentioned in lines 29-31). Thus, highlight the gaps in knowledge that your study aims to address. Expand on the importance of the soils in Russia's boreal forests and polar belt as an agricultural resource (mentioned in lines 32-34). Provide a clearer transition between the discussion on agricultural impacts in different regions (lines 38-48) and the importance of soil microbiome research in these areas (lines 49-53). Elaborate on the role of the soil microbiome in soil profile development and soil-biochemical processes (mentioned in lines 54-62). Explain how understanding the taxonomic and functional diversity of the soil microbiome can contribute to managing soil formation processes. To enhance the introduction's flow, consider reorganizing the paragraph starting from line 62 to better connect the discussion on soil microbiome diversity in natural conditions with the importance of studying disturbed soils (mentioned in lines 68-71). Further, elaborate on the knowledge gaps regarding the soil microbiome in Russia (mentioned in lines 79-91). Provide a clearer rationale for why studying the taxonomic composition of the soil microbiome in specific regions is essential and how your study will contribute to filling these gaps. The authors should strongly consider separating the aim of the study (line 92) from the specific objectives (lines 95-98) to make them more distinct and easier to follow. Next, provide a brief overview of the research methodology in the introduction. This can help readers understand how the study was conducted and what methods were used to investigate the taxonomic composition and biodiversity parameters of the soil microbiome. Finally, the authors should consider adding a more concise and impactful closing statement for the introduction, summarizing the importance of the study and its potential contributions to soil microbiology research in Russia and beyond.

 

Methodology:

Provide a brief rationale for the selection of the specific study sites and their significance in representing different natural zones and levels of anthropogenic disturbance. Providing more details about the specific locations of the study sites (e.g., coordinates, geographical features) to enhance the reproducibility and comparability of the study. Elaborate on the reason for selecting the topsoil depth of 0-10 cm for sampling and why this depth range was considered representative of the objectives of the study. Also, provide references/citations to support this reason.  Add information on how the triplicate soil samples from each location were combined to create an average sample weighing 500 g for routine soil analyses (lines 131-132). Explain the rationale behind using 2g samples for microbiological analysis and the number of replications. In the methodology section for chemical and microbiological analysis, provide more information on the specific steps and protocols used in the soil analyses (e.g., the exact method used for pH measurement, the equations or references used to calculate the content of available forms of ammonium nitrogen and nitrate nitrogen, the Kirsanov method for determining free potassium and phosphorus). Readers need to know this information for the repeatability of experiments in future.  Consider providing more details on the quality control process for the isolated total soil DNA (line 147) to ensure the reliability of the subsequent sequencing results. Add more information on the sample preparation process for sequencing the V4 variable region of the 16S rRNA gene. Include details on the PCR conditions and cycling parameters used for amplification. Elaborate on the general processing of the sequences using the dada2 and phyloseq packages (lines 152-160). Explain the steps involved in trimming, quality filtering, and chimera removal, as well as any specific parameters or thresholds used. Provide more information on how the taxonomic annotation of the 16S rDNA amplicon sequences was performed using the naive Bayesian classifier (lines 158-160). Mention any relevant settings or cutoffs used for taxonomic classification.

 

Results & discussion:

The authors should consider grouping the Results and Discussion into subsections to significantly improve the clarity and organization of the manuscript. Provide a clear introduction to the results section, highlighting the main objectives of the study and the key findings obtained from the analysis of soil chemical characteristics and soil microbiome composition. When discussing the results of soil chemical characteristics (lines 168-179), provide more context on why certain trends were observed, such as the higher soil organic carbon content in agrosoils compared to primary soils of mines, and the variation in pH values across different sampling locations. The section discussing the alpha diversity metrics (lines 198-203), provides more context on why certain soils, such as those of mines, demonstrated higher biodiversity deviation than others. Explain the potential implications of these findings for soil ecosystem stability and resilience. When discussing the beta diversity data (lines 214-257), provide more insights into the factors that may be driving the differences in taxon representation between disturbed and undisturbed soils. Explain how these findings align with previous research in the field. Consider discussing the potential implications of the observed changes in microbial diversity for soil ecosystem functioning and nutrient cycling (lines 258-266). The discussion of the wide diversity of soils in Northern European Russia and Western Siberia (lines 283-290), provides more context on how the combination of natural factors and various types of anthropogenic activity may influence the soil microbiome and ecosystem dynamics. Elaborate on the significance of using different-aged technogenic and agrogenic disturbed landscapes for modelling the formation of climax microbial communities (lines 290-294). Explain how this approach can contribute to a better understanding of soil microbiome dynamics in response to different disturbances. In the final paragraph of the discussion (lines 301-304), provide more specific examples of the ecological and physiological activity of the soil microbiome and how these activities relate to soil ecosystem functions and services.

 

Conclusion:

Reiterate the significance of the study in expanding knowledge of the soil microbiome in the taiga-forest and forest-tundra regions of Northern Eurasia (lines 307-310). Highlight the contribution of the research to filling gaps in geographical coverage and understanding microbial diversity in these ecosystems. Emphasize the key dominant phyla found in the soil microbiome across the studied regions (lines 311-313). Explain the potential implications of increased microbial diversity for soil recovery processes and the establishment of new soil types. Consider providing potential future research directions to expand the understanding of soil microbial biodiversity in the polar-boreal ecotone (lines 321 and 327).

 

References:

Consider adding recent papers from 2020 to 2023 that are relevant to the study's topic, such as those focusing on the soil microbiome in similar ecosystems or regions. The authors should ensure that the references cited are relevant to the research and complement the findings, helping readers gain a broader understanding of soil microbiome studies in the regions of interest.

 

Figures & tables:

The current figures and tables are appropriate for presenting the results and supporting the discussions. No additional figures or tables are suggested as they are already sufficient for the content presented, except if other reviewers suggest so.

 

The quality of the English Language written by the authors is clear and coherent. 

Author Response

Dear reviewer!

Thank you for all comment,

Below you can find point by point reply for your comments,

All changes are red color tracked in text.

 

General comment:

The manuscript discusses the Microbiome of Soils in Northern Russian regions. The contents are interesting and worth reading. However, there are several professional comments/suggestions that the authors should consider.

 

Title:

The present title the authors provided is quite descriptive and conveys the main focus of the manuscript, which appears to be about the microbiome in different types of soils (natural, agricultural, and technogenic) found in specific regions (forest and forest-tundra) of the Russian North. However, it is long and could be improved for better readability and clarity. I suggest the authors consider using a shorter title.

Reply – now the title is shortened and changed.

 

 

Abstract:

Elaborate on the types of disturbances or agricultural practices that have been studied and their potential implications for soil microbiota. Mention the specific techniques and methodologies used for metagenomic sequencing (e.g., library preparation, sequencing platforms) to give readers a clearer understanding of the study's technical aspects.

Reply - corrected

 

Clarify the rationale for selecting the particular regions in Russia for soil sampling. Were there specific factors or characteristics that led to their inclusion in the study? It's important to provide more context on the significance of the results and how they contribute to the existing body of knowledge. How does this study fill the knowledge gap mentioned earlier, and how can this new understanding of soil microbiomes be practically applied for soil reclamation and amelioration in severe climatic conditions? The authors should consider adding some quantitative information about the taxonomic diversity of the microbiomes in the various ecosystems studied. Mention any significant findings, such as differences in diversity or abundance of specific microbial groups. Please provide a clearer description of the "depression of microbiome communities" mentioned in the abstract. Elaborate on what this means and how it relates to the formation of new microbial communities in quarries. Ensure consistency in the use of terms; for instance, the abstract mentions "agrogenic" and "agro-soils," which seem to refer to the same concept. Stick to one term to avoid confusion.

 

Reply – done, corrected

The authors should consider rephrasing the sentence: "Thus, data on soil microbiome composition may be used for further driving of soil properties for reclamation and amelioration practices in severe climatic conditions of northern Russia." This sentence is crucial, but it could be more straightforward in conveying how the findings can practically impact soil management strategies.

Reply – the sentence is rephrased 

 

Introduction:

The authors can add more context on the current state of soil microbiology research and how it relates to the need for a global understanding of the soil microbiome (mentioned in lines 29-31).

Reply – added, corrected

 

Thus, highlight the gaps in knowledge that your study aims to address. Expand on the importance of the soils in Russia's boreal forests and polar belt as an agricultural resource (mentioned in lines 32-34).

Reply - corrected

Provide a clearer transition between the discussion on agricultural impacts in different regions (lines 38-48) and the importance of soil microbiome research in these areas (lines 49-53). Elaborate on the role of the soil microbiome in soil profile development and soil-biochemical processes (mentioned in lines 54-62). Explain how understanding the taxonomic and functional diversity of the soil microbiome can contribute to managing soil formation processes.

Reply - corrected

 

To enhance the introduction's flow, consider reorganizing the paragraph starting from line 62 to better connect the discussion on soil microbiome diversity in natural conditions with the importance of studying disturbed soils (mentioned in lines 68-71).

Reply - corrected

 

Further, elaborate on the knowledge gaps regarding the soil microbiome in Russia (mentioned in lines 79-91). Provide a clearer rationale for why studying the taxonomic composition of the soil microbiome in specific regions is essential and how your study will contribute to filling these gaps. The authors should strongly consider separating the aim of the study (line 92) from the specific objectives (lines 95-98) to make them more distinct and easier to follow. Next, provide a brief overview of the research methodology in the introduction. This can help readers understand how the study was conducted and what methods were used to investigate the taxonomic composition and biodiversity parameters of the soil microbiome. Finally, the authors should consider adding a more concise and impactful closing statement for the introduction, summarizing the importance of the study and its potential contributions to soil microbiology research in Russia and beyond.

 

Reply – corrected, improved.

 

Methodology:

Provide a brief rationale for the selection of the specific study sites and their significance in representing different natural zones and levels of anthropogenic disturbance. Providing more details about the specific locations of the study sites (e.g., coordinates, geographical features) to enhance the reproducibility and comparability of the study.

Reply – corrected, additional information is provided in text.

 

Elaborate on the reason for selecting the topsoil depth of 0-10 cm for sampling and why this depth range was considered representative of the objectives of the study. Also, provide references/citations to support this reason.  Add information on how the triplicate soil samples from each location were combined to create an average sample weighing 500 g for routine soil analyses (lines 131-132).

Reply – corrected, clarified

 

 Explain the rationale behind using 2g samples for microbiological analysis and the number of replications.

Reply - corrected

 

In the methodology section for chemical and microbiological analysis, provide more information on the specific steps and protocols used in the soil analyses (e.g., the exact method used for pH measurement, the equations or references used to calculate the content of available forms of ammonium nitrogen and nitrate nitrogen, the Kirsanov method for determining free potassium and phosphorus). Readers need to know this information for the repeatability of experiments in future.

Reply – required information has been added.

 

  Consider providing more details on the quality control process for the isolated total soil DNA (line 147) to ensure the reliability of the subsequent sequencing results. Add more information on the sample preparation process for sequencing the V4 variable region of the 16S rRNA gene. Include details on the PCR conditions and cycling parameters used for amplification.

Reply – details on quality control can be find on link https://github.com/a-zverev/16s-amplicon-processing

Elaborate on the general processing of the sequences using the dada2 and phyloseq packages (lines 152-160). Explain the steps involved in trimming, quality filtering, and chimera removal, as well as any specific parameters or thresholds used. Provide more information on how the taxonomic annotation of the 16S rDNA amplicon sequences was performed using the naive Bayesian classifier (lines 158-160). Mention any relevant settings or cutoffs used for taxonomic classification.

 

Reply – required information is added. 

 

Results & discussion:

The authors should consider grouping the Results and Discussion into subsections to significantly improve the clarity and organization of the manuscript. Provide a clear introduction to the results section, highlighting the main objectives of the study and the key findings obtained from the analysis of soil chemical characteristics and soil microbiome composition. When discussing the results of soil chemical characteristics (lines 168-179), provide more context on why certain trends were observed, such as the higher soil organic carbon content in agrosoils compared to primary soils of mines, and the variation in pH values across different sampling locations. The section discussing the alpha diversity metrics (lines 198-203), provides more context on why certain soils, such as those of mines, demonstrated higher biodiversity deviation than others.

 

Reply – corrected, more detalized.

 Explain the potential implications of these findings for soil ecosystem stability and resilience. When discussing the beta diversity data (lines 214-257), provide more insights into the factors that may be driving the differences in taxon representation between disturbed and undisturbed soils. Explain how these findings align with previous research in the field.

Reply - corrected

 

Consider discussing the potential implications of the observed changes in microbial diversity for soil ecosystem functioning and nutrient cycling (lines 258-266). The discussion of the wide diversity of soils in Northern European Russia and Western Siberia (lines 283-290), provides more context on how the combination of natural factors and various types of anthropogenic activity may influence the soil microbiome and ecosystem dynamics. Elaborate on the significance of using different-aged technogenic and agrogenic disturbed landscapes for modelling the formation of climax microbial communities (lines 290-294). Explain how this approach can contribute to a better understanding of soil microbiome dynamics in response to different disturbances. In the final paragraph of the discussion (lines 301-304), provide more specific examples of the ecological and physiological activity of the soil microbiome and how these activities relate to soil ecosystem functions and services.

 Reply – corrected.

Conclusion:

Reiterate the significance of the study in expanding knowledge of the soil microbiome in the taiga-forest and forest-tundra regions of Northern Eurasia (lines 307-310). Highlight the contribution of the research to filling gaps in geographical coverage and understanding microbial diversity in these ecosystems. Emphasize the key dominant phyla found in the soil microbiome across the studied regions (lines 311-313). Explain the potential implications of increased microbial diversity for soil recovery processes and the establishment of new soil types. Consider providing potential future research directions to expand the understanding of soil microbial biodiversity in the polar-boreal ecotone (lines 321 and 327).

 

Reply – the conclusion is rewritten and reformulated.

 

References:

Consider adding recent papers from 2020 to 2023 that are relevant to the study's topic, such as those focusing on the soil microbiome in similar ecosystems or regions. The authors should ensure that the references cited are relevant to the research and complement the findings, helping readers gain a broader understanding of soil microbiome studies in the regions of interest.

Reply  - a number of recent references has been added.

 

 

Figures & tables:

The current figures and tables are appropriate for presenting the results and supporting the discussions. No additional figures or tables are suggested as they are already sufficient for the content presented, except if other reviewers suggest so.

 Reply – thank you, the quality of table 2 is improved.

Reviewer 4 Report

This manuscript presents a detailed analysis of the taxonomic diversity of overripe and anthropogenically disturbed soils in various ecosystems of Russia. Overall, this manuscript is an interesting study , well-structured with essential literature review. However, the presentation in the manuscript as currently constituted is inadequate there is a need for extensive and major revision to improve the presentation of the study. Additionally, the manuscript shows a lack of attention to detail and formatting issues as set out in my comments as follows:

-Title: the current title is meaningful and brief

- The general standard of English requires attention with careful proof checking to correct grammatical issues

-The abstract requires revision and improvement to better summarise the study. The main results is not clear, to represent just important results. The reader of an abstract needs to its recommended! Tell them please, in as few sentences as possible, what they will find, and key details just.

-In Introduction, clarify the novelty of your work and your contribution to the body of knowledge, and identify a gap in the previous knowledge and provide sufficient evidence to support the direction of your research. Furthermore, clarify the research component in your work.

- Please depended modern references is made to Introduction relating to direction of your research.

-Material and Methods with sub-section, are fine,

-There is a need for clearer discussion relating to the contribution to the body of knowledge with Results, text and figure captions were need to re-written, which should be improved.I

-Improve the Figure quality 2.

-Conclusion is need re-write and it is too long , a research paper should end with a well-constructed conclusion. You can usually do this in one paragraph, you can raise some open questions and set the scene for the next study, this is a good place to register your thoughts about possible future work, try to explain to your readers what more could be done? What do you think are the next steps to take? What other questions warrant further investigation? Remember, the conclusion is the last part of the essay that your reader will see, so spend some time to re-writing the conclusion so that you can end on a high note.

 It’s advised to have English native speaker to  review paper.

 

Author Response

Dear reviewer, thank you for all suggestions in the text, all changes are tracked by red color in updated text.

 

This manuscript presents a detailed analysis of the taxonomic diversity of overripe and anthropogenically disturbed soils in various ecosystems of Russia. Overall, this manuscript is an interesting study , well-structured with essential literature review. However, the presentation in the manuscript as currently constituted is inadequate there is a need for extensive and major revision to improve the presentation of the study. Additionally, the manuscript shows a lack of attention to detail and formatting issues as set out in my comments as follows:

-Title: the current title is meaningful and brief

The general standard of English requires attention with careful proof checking to correct grammatical issue

• reply -the quality of the English language has been improved

-The abstract requires revision and improvement to better summarise the study. The main results is not clear, to represent just important results. The reader of an abstract needs to its recommended! Tell them please, in as few sentences as possible, what they will find, and key details just.

Reply - the abstract has been rewritten.

-In Introduction, clarify the novelty of your work and your contribution to the body of knowledge, and identify a gap in the previous knowledge and provide sufficient evidence to support the direction of your research. Furthermore, clarify the research component in your work.

Reply - corrected

Please depended modern references is made to Introduction relating to direction of your research.

Reply - new references has been added

-Material and Methods with sub-section, are fine,

-There is a need for clearer discussion relating to the contribution to the body of knowledge with Results, text and figure captions were need to re-written, which should be improved.I

Reply - corrected.

-Improve the Figure quality 2.

Reply - corrected.

-Conclusion is need re-write and it is too long , a research paper should end with a well-constructed conclusion. You can usually do this in one paragraph, you can raise some open questions and set the scene for the next study, this is a good place to register your thoughts about possible future work, try to explain to your readers what more could be done? What do you think are the next steps to take? What other questions warrant further investigation? Remember, the conclusion is the last part of the essay that your reader will see, so spend some time to re-writing the conclusion so that you can end on a high note.

Reply - Conclusion has been shortened.

Sincerely yours, corresponding author,

Evgeny Abakumov