Response of Soil Physicochemical Properties and Microbial Community Composition in Larix olgensis Plantations to Disturbance by a Large Outbreak of Bark Beetle

Round 1

Reviewer 1 Report

I had now time reading your manuscript entitled, "Response of soil physicochemical properties and microbial community composition in Larix olgensis Plantation to disturbance by a large outbreak of bark beetle", which you submitted to the Forest.

I acknowledge that the author showed the nice manuscript. However, I regret that your manuscript still does not reach the standard as an original article published in Forest.

The descriptions of data are unclear and very long sentences. This is the main barrier to understanding throughout the manuscript, especially the introduction and discussion.

The abstract is still not detailed enough, the author should be shown the highlight for this research.

Introduction is unclear in some sentences (see the attached file).

Materials and methods! Several major comments are found in the part. The author might be shown as a map for the study site. It is easy to understand. Italic names are checked.

Results. Although it is clear, some minor corrections are found (see the attached file).

Discussion! Most problematic is the part, it might be used a very long sentence to present. It is also a barrier to understanding. The author should be clear between your data and the previous evidence.

Important information such as the compared data of plant, soil content and microbial data are clear.

As said in the nice academic data with you, I hope my comments in the annotated manuscript are helpful, despite being incomplete and not covering all problems.

Comments for author File: Comments.pdf

Author Response

Response to Reviewer 1 Comments

Thank you very much for your suggestions, they have helped us a lot with the article and we have made the changes as requested with a detailed description of the changes.

Point 1: pls changes: Soil physicochemical properties including soil moisture content, ....... were measured.

Response 1: The abstract has been revised as requested.

Point 2: more explain for these data from Refs 10-12.

Response 2: More explanation has been provided in accordance with the contents of the references.

Point 3: specific year?

Response 3: Changes to the content have been made as requested.

Point 4: These sentences are very confusing, the author should be revised as an easy understand to reader.

Response 4: Sentences have been rewritten as required.

Point 5: The author should be not wrote as this pattern. It is very confusing, Pls re-write!

Response 5: This part of the sentence was rewritten as requested.

Point 6: References are required. Pls add the situation about the serious harm as number or quatity.it is showed that economic data of Larix olgensis. This is the good data to support for your manuscript.

Response 6: The relevant descriptive data has been added as requested.

Point 7: Very hard to understand ! pls separate between the ojective and benefits from this reserach!. This is very long sentense. It is not easy to undertand.

Response 7: I have rewritten this section as requested.

Point 8: Is this experimental condition ? or the field study ?

If this study is focused on the field study, it should be used "study area, filed area or others".

Response 8: I have revised and added a schematic of the study area.

Point 9: Iitalic name?

Response 9: The writing of the Iitalic name of the plant has been corrected.

Point 10: Is this your previous data ? or other data, pls correct !

Response 10: Cited references have been added.

Point 11: This part should be moved in the introduction, not materials and methods.

Response 11: Changes have been made to the content.

Point 12: The criteria is showed as the previous evidence or not! or Is this the cretiria from the authors?

Response 12: The classification of different disturbance intensities was made by referring to the relevant literature and combining with the actual mortality situation in the experimental samples.

Point 13: pls specific where? is this the standad methods? It yes, pls add the references.

Response 13: Relevant references have been added.

Point 14: The authors should be clearly correlated between plant, soil properites and microorganism. Pls re-check! How to approve for this idea?

Response 14: Relevant references have been added.

Point 15: High resolution is required for Fig.1

Response 15: Picture clarity has been enhanced as requested.

Point 16: pls expain in all abbrevations such as CK, LI....v

Response 16: Abbreviations have been explained below the pictures as requested.

Point 17: pls expain in all abbrevations such as CK, LI....v

Response 17: Abbreviations have been explained below the pictures as requested.

Point 18: I am very confusing that these are the previous data or your present research. If this is the previous data, the author should not showed. Your topic data is the first and expain how why ?

Response 18: The discussion has been revised and the results of this study have been added.

Point 19: full stop after "different."

Response 19: The sentence it was originally in has been modified and the original sentence has been deleted.

Point 20: Evidence?

Response 20: Relevant references have been added.

Point 21: specific what are similar to your data!.

Response 21: Specific data descriptions have been added.

Point 22: your data ?

Response 22: Yes, through the results of the study, it was found that the bacteria were more sensitive to the response of different disturbance intensities.

Point 23: very long sentense! please rewrite!

Response 23: These sentences I reworked.

Point 24: what ? "Wu".

Response 24: Amended

Reviewer 2 Report

L09: The abstract has the components expected in a scientific abstract but is imbalanced, with too little emphasis on the introduction and objectives and too much detail in the results. This imbalance hampers its effectiveness in succinctly conveying the study's essence. Improving the abstract would involve expanding the introduction to more clearly define the study's significance, succinctly stating the research objectives, condensing the results to highlight only the most pivotal findings, and refining the conclusion to better encapsulate the study's contributions to the field.

L16 and further: PH → pH, many times in the text

L32: Keywords closely mirror the title, thus they may not fully leverage the potential to capture a wider array of related concepts that could draw in readers from adjacent fields or with varied interests. Effective keywords should ideally complement the title by incorporating broader or slightly different aspects of the research that are not already specified in the title. This approach can help in reaching a broader audience. For instance, if the title already includes "Larix olgensis," "soil physicochemical properties," and "microbial community structure," the keywords could benefit from including related but broader or more specific terms. Potential alternatives could involve underlying processes, specific methods or techniques used (e.g., "high-throughput sequencing"), broader themes (e.g., "forest ecosystem health"), or specific aspects of the disturbance (e.g., "ecological impact of pest outbreaks").

L37: Lower index for 2

L42-L46: The text should be divided into multiple sentences for clarity. Using periods instead of commas can improve readability and help delineate separate but related points. Applicable for the whole manuscript.

L53 till the end of sentence: The text's complex structure and lengthy sentences obscure clarity, necessitating division into shorter sentences. Additionally, some passages lack coherence, potentially confusing readers about the studies' conclusions. Simplifying and separating distinct findings would enhance readability and comprehension.

L55: Authors forgot to include this as the citation.

L80: “biomass Plant-microbe-soil” – Period is missing?

L83: Larch refers to a genus (Larix) rather than a single species, comprising several species adapted to cool climates. The introduction of "larch" without specifying the species, especially when referring to Larix olgensis later, might confuse readers unfamiliar with the context. A clearer introduction to the specific species studied from the outset would improve clarity and accuracy.

L86: The first appearance in the text, so full Latin name including the author of description is missing here.

L86-L97: The sentence is grammatically and stylistically cumbersome due to its length, awkward use of "cooperated," and somewhat disjointed structure. The repetition of "environmental changes" suggests a potential editing oversight. Consider revising for clarity, conciseness, and more precise language: E.g.: "In this study, we investigated the surface soil physicochemical properties and microbial community structural changes in a typical Larix olgensis plantation in eastern northeastern China using high-throughput sequencing. We explored the correlation between soil properties and microbial communities, assessing their responses to significant bark beetle disturbances amid environmental changes. This research aims to provide a scientific basis for understanding the long-term impacts of bark beetle outbreaks on soil and microorganisms, to develop effective forest restoration methods post-infestation, and to inform management strategies for deadwood."

L104-L106: The text unnecessarily repeats information about the site's elevation and terrain characteristics. To improve clarity and eliminate redundancy, it could be succinctly revised.

L109-L111: Some trees with the author of the description, some with the author of the description even when it's not the first mention, some without the author even when it is the first mention, and most are not in italics - need revision.

L113-L121: The phrase "plants.hm-2" is unusual and may confuse readers unfamiliar with this notation. It's commonly used to express plants per square hectometer (10,000 square meters) but could be clarified as "plants per hectare" for broader understanding. The passage "Larix olgensis plantation … were set up with control" is grammatically awkward. The plural subject ("plantations") and verb agreement ("were") are mismatched due to the singular noun "plantation." Also, the structure is slightly confusing because it initially sounds like the plantations were established as part of the study setup rather than being naturally affected and then studied.

L123-L133: The description of sample collection lacks clarity regarding the total number of samples collected. While the method of collecting soil samples and combining five soil samples from each site into a composite sample repeated three times is detailed, the exact number of sampling sites or plots within each disturbance category is not specified. This omission makes it difficult to ascertain the overall number of unique composite samples analyzed in the study. If each level of disturbance was represented by only one site from which three samples were taken, this study faces a classic issue of pseudoreplication. Such a sampling strategy raises significant concerns about the reliability and generalizability of the results. Treating samples from a single site as independent replicates does not account for spatial variability and can inflate the type I error rate, leading to misleading conclusions about the effects of disturbance on soil and microbial communities. A more robust experimental design, with true replication across multiple independently disturbed sites for each disturbance level, is essential to validate the findings.

L143: The text should specify which regions were amplified using universal primers.

L157-L165: The description of the statistical analysis mentions the use of one-way ANOVA for analyzing soil physicochemical properties and the application of the least significant difference method for testing differences between comparison parameters. However, there is no information on statistical analyses for microbial community data: The text describes the use of high-throughput sequencing and subsequent analyses like community barplot analysis, correlation heatmap, etc., yet it fails to mention the statistical tests used to analyze these complex microbial data. The mention of α diversity calculation does not come with an explanation of how these diversity indices were statistically compared across different disturbance levels. The narrative around the relative abundance of microbial populations suggests statistical comparisons were made, but details like how comparisons were tested and adjusted for multiplicity, are missing. Moreover, the mention of non-significant increases without providing context or specifics (e.g., what statistical test led to this conclusion) further obscures the analysis. In summary, while the text outlines some components of the statistical analysis, it falls short of providing a comprehensive and transparent account necessary for replicability and thorough scientific scrutiny. More detailed information on the statistical methods, precise results including effect sizes and confidence intervals, and a clearer linkage between the results presented and the statistical analyses conducted are needed for a robust evaluation of the study's findings.

L161: Without more detail, it remains unclear which exact metrics of alpha diversity are being assessed.

L166 and further: The description of results lacks detailed statistical outcomes (e.g., exact p-values, F-values, degrees of freedom) that would allow for a critical assessment of the data's statistical significance and effect sizes. When specific statistical results are mentioned as "significant," it's essential for transparency and reproducibility to include exact p-values and additional statistics. The absence of these details, especially if not presented elsewhere in the manuscript or in a table, may leave readers questioning the robustness and precision of the statistical analysis.

L169: Coefficient of variation? Should be specified at first appearance

L172: The phrase "and there was no difference" is ambiguously placed and does not clearly state what it refers to. This makes it confusing whether it suggests there was no difference in soil moisture content between the low-intensity and high-intensity disturbance groups, or if it refers to something else. For clarity and accuracy, the sentence should explicitly state between which groups or conditions "there was no difference" was observed.

L172 till the end of the sentence: The text could benefit from improved clarity and conciseness. The repeated use of "and" at the beginning of several clauses makes the text somewhat cumbersome. Additionally, the flow of information about different soil properties could be more organized to enhance readability.

Figure 1: The column charts, while informative, would greatly benefit from the addition of information regarding the number of samples compared in each group alongside the error bars. Overall, The manuscript fails to clarify whether the variability indicated in the figures represents standard deviation (SD) or standard error (SE).

L197-L198: The presentation of numerical values (319.3197, 325.5, 332.5, 329.3) without explicit clarification regarding their meaning or the specific disturbance levels they correspond to is a significant oversight. This lack of clarity hinders the reader's ability to fully understand the impact of disturbance on soil microbial community biodiversity.

L198: Why semicolon is so overused in the manuscript?

L198 till the end of the sentence: The sentence is cumbersome and challenging to comprehend due to its excessive length, convoluted structure, and repetitive phrases. The attempt to convey detailed information about the relative abundance of various microbial populations and their comparison across different disturbance levels is commendable. However, the execution significantly hampers readability and understanding. Issues include improper punctuation and a lack of clarity in presenting statistical findings and their implications. A more structured approach, possibly breaking down the information into shorter, more focused sentences and specifying comparisons and findings, would greatly enhance clarity and reader comprehension.

L213 and L224: The inclusion of fungal components, specifically an unclassified fungus and Botrytis cinerea, in a study primarily focused on bacterial populations raises several concerns, particularly when it's stated that "universal primers" were used without specifying which. If these primers were designed primarily for bacterial communities, their ability to amplify fungal DNA could indicate cross-reactivity or a broader spectrum of amplification than initially disclosed. This scenario could suggest issues with primer specificity. Given the focus on bacteria, the authors should clarify the scope of their primers and consider whether fungal data should be analyzed separately or potentially excluded if their inclusion wasn't intentional. Further, any unexpected findings, like the presence of significant fungal sequences, warrant a discussion regarding their origin, implications for the study's findings, and any measures taken to ensure data integrity, such as re-evaluating the primer choice.

L215-L216: Should be in italics

L216-L222: The sentence suffers from unnecessary repetition, particularly in its mentions of the relative abundance of Tenericutes being significantly higher in the medium-intensity group compared to the control group.

L232: Is this the result of a PERMANOVA analysis? I infer this based on the context provided. Authors are required to explicitly detail all statistical analyses employed, including this one, in the data analysis section. Furthermore, mentioning only the R2 value is insufficient; the inclusion of p-values is crucial for a comprehensive understanding and evaluation of the statistical significance of the findings.

Figure 2: The ACE Index is not the sole index for measuring alpha diversity; there are several indices, such as Shannon, Simpson, and Chao1, among others, used to assess species richness and evenness within communities. The late mention of the ACE (Abundance-based Coverage Estimator) Index in a single figure, without earlier specification in the data analysis section, is inadequate for comprehensive understanding. Clarification and proper abbreviation usage (ACE, not Ace) should be provided early in the methodology to ensure readers are fully informed about the metrics of alpha diversity being assessed. This oversight complicates the interpretation of what the authors refer to as "alpha diversity" throughout the manuscript.

Regarding Figure 2, there are several critical issues: 1) The figure is mislabeled as a histogram when it is, in fact, a bar chart. 2) It lacks error bars and an explanation of what they represent, which is essential for assessing data variability. 3) There is no information on the number of samples compared, which is crucial for evaluating the statistical significance and robustness of the findings. 4) The figure caption does not include a legend explaining what is being displayed; figures should be interpretable independently of the text. These oversights significantly hinder the reader's ability to understand and evaluate the presented data accurately.

Figure 4: There are numerous issues with the presentation and analysis. Firstly, the Kruskal-Wallis test is only mentioned in a figure title, with no corresponding description in the data analysis section. Furthermore, inserting titles directly into figures should be avoided unless necessary; here, such information could have been effectively conveyed in a caption. The rationale for selecting specific groups (two bacterial phyla and unclassified fungi) for presentation lacks clarity. Referring to these groups as "Species with significant differences at different levels of disturbance" without explaining the selection criteria or why unclassified fungi were included despite being an apparent misfit is confusing. Besides, these are not species.

Additionally, the manuscript fails to adequately describe the abbreviations used in the legend, the nature of the error bars, and the number of samples analyzed, all of which are essential for the figure to stand alone independently of the text. Moreover, the is inconsistency in denoting statistically significant differences—switching from letter annotations to a different style. These issues collectively detract from the plot's clarity, coherence, and reliability.

Figure 5: Not species, again.

L245 and further: The discussion section uncritically accepts the premise that varying levels of bark beetle-induced damage cause changes in soil and microbiota, without considering the possibility that soil composition and consequent microbiota changes at a micro-scale might influence tree susceptibility to beetle infestation. This lack of critical reflection on potential reverse causality overlooks the complex interplay between soil conditions, microbial communities, and tree health.

L277: Something is missing here

L285 and further: Across the whole discussion section: The sentence structure, length, and grammar have several issues that impact readability and clarity: The sentences are excessively long and complex, covering multiple distinct points to discuss. They attempt to convey too much information in a single run-on sentence, making it difficult for readers to follow. The frequent use of "and" contributes to the sentence's run-on nature. Breaking down this content into separate sentences or paragraphs would significantly improve clarity. The sentence structure could be improved for better grammatical correctness, particularly in terms of punctuation and the separation of ideas.

L334: of levels of?

L336: species?

L337: Given the use of probably bacterial-specific primers, it's expected that the majority of identified species groups would be bacterial.

L359: that that?

L377: Should be in italics

L385: Citation is missing for Wu

Many sentences in the manuscript are overly complex and lengthy, which may hinder comprehension. I suggest breaking these into shorter, more straightforward sentences. Some sections lack precision in language, especially regarding methodological descriptions and results interpretation. I have found instances of omitted words, phrases, or even whole parts of sentences that inappropriately repeat as if someone forgot to delete them. A thorough review for grammatical correctness is advisable to enhance the manuscript's professionalism and readability. For specific comments, please see the review.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 3 Report

The research topic of the authors is relevant. The scope of work is large. However, there are some comments on the presented material.

The abstract is too long: The abstract should be a total of about 200 words maximum. The first sentence of the abstract is very long. It is advisable to edit it or divide it into several. The full scientific name of Larix olgensis and other species should be indicated only at the first mention in the abstract and introduction. Further, it is enough to write L. olgensis. Latin names of genera and species must be in italics, for example in lines 109–111.

Keywords are needed to enhance the capabilities of search systems. Almost all keywords are present in the title; it is advisable to indicate keywords that are not repeated in the title to increase access to interested readers.

It would be necessary to indicate the species of bark beetle.....

Most of the papers mentioned in the Introduction concern evergreen coniferous species, only poplars are deciduous. Larch is a conifer species, but the foliage falls every year. Therefore, an unexpectedly large addition of litter, like that of pine after tree mortality, is not expected in larch forests. This should be noted.

Lines 86-97 formulate the aim and content of the work. However, the sentence is very long. The aim must be formulated clearly, and the tasks must be separately identified: shifts in soil physical and chemical properties, in the structure changes of microbial communities, and their interaction.....  

line 114: affected by climate change in 2020 – The climate has been changing for many years, and there was an outbreak of bark beetles in 2020.

Add a reference to bark beetle outbreaks on larch in the region. If there are no publications on the outbreak, who and when (what months?) surveyed the tree mortality?

It is necessary to indicate whether the site and stand characteristics were the same in the sample plots with different tree mortality, particularly, a slope, monocultures or mixed plantings, age, tree density, ....., what months were tree mortality records taken? Was precipitation and soil moisture the same in these areas? were samples taken within the crown projection?

Samples were taken in October 2021 after tree mortality in 2020. It would be advisable to repeat the analyses in subsequent years. The reaction may be short-lived.

Results. Figure 1. To add the explanation of axes titles. Figure 1. Soil physical properties and total carbon (TC), total nitrogen (TN), total phosphorus (TP), and total potassium (TK) contents in forested areas with different levels of disturbance.

Increase the font size on the x-axis. The title of the x-axis is “Intensity of tree mortality”. The names of the columns – Control (without tree mortality?), Low, Moderate, High.

lines 197–198: is it alfa-diversity? mention it!

lines 198–216. Too large a sentence. Divide it.

suggested: Figure 2. Alpha diversity index for different levels of disturbance. Explain the names of the columns. Better to give the same title to the x-axis for Figures 2, 3, 4. The legend is not necessary, all columns may be the same color.

Figure 3. Relative abundance of soil microorganisms on Phyllum level at different levels of disturbance

Suggested title for y-axis: Community abundance, %.

Figure 4. Species with significant differences at different levels of disturbance. The legend is not necessary, all columns may be the same color.

Figure 5. Heatmap analysis of the correlation between soil environmental factors and species with different mortality intensities.

Explain PH, SM, TC, TN, TP, and TK like in Figure 1.

Discussion. 4.1. Larch is a deciduous tree. Effects on litter and soil may differ from coniferous species. An indirect influence of larch mortality is also possible, for example, an increase in the proportion of other species in the plantation, which also affects soil properties and microorganism species composition. I think this part can be described more briefly. Compare results for other regions for larch, deciduous, and coniferous tree species.

4.2. Larch foliage falls off every year. Trees infested with bark beetles do not die immediately, and the needles may remain on the branches until the end of the season. Therefore, it would be interesting to evaluate the seasonal dynamics of larch needle litter in bark beetle outbreaks to find out how this affects the properties of the soil.

Some notes in the Manuscript are marked in yellow.

Comments for author File: Comments.pdf

Some notes in the Manuscript are marked in yellow.

Some sentences are very long.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Overall, I appreciate the effort made to revise the manuscript according to my comments. My initial critique was highlighting the need for a comprehensive overwriting to address issues of cumbersome sentences, awkward structure of sentences, and repetitive phrasing that significantly affect readability and comprehension across the document. While there has been an attempt to rewrite the manuscript, it appears that it still suffers from similar, widespread problems that were initially pointed out. The core issues—excessive sentence length, lack of clear punctuation, and grammatical issues — remain prevalent, detracting from the manuscript's overall quality and reader's ability to fully grasp the research findings. Authors should include breaking down complex information into shorter, more digestible sentences and ensuring that punctuation and structure facilitate rather than hinder understanding. I strongly recommend a thorough review of the manuscript to address these persistent issues comprehensively (maybe also use the check from a native speaker). See next section.

Others:

L90: The first appearance in the text, so full Latin name including the author of description is missing here.

L125: Figure 1: This is not a sufficient map, the study plots should be marked on map.

 L135: Is this part of the sentence still valid, if the authors add the information about tree sample plots above?

 L241: Authors should clarify why no fungi were classified despite the use of specific fungal primers focused on fungal analysis. This might indicate a mismatch with the reference database used for sequence classification. It's essential to disclose which databases were employed for both fungal and bacterial sequence comparisons, such as Silva or GreenGenes, as the choice significantly impacts microbial community analysis outcomes.

 L248: Why PERMANOVA is still not described in methods? The authors addressed the simpler issue (insert name of analysis and p-value) but forgot to mention this analysis also in the methods section.

Examples of awkward sentences (but only several, the whole manuscript is a problem):

L45-L46: This sentence lacks a proper subject-verb agreement, making it a fragment rather than a complete sentence. Moreover, the verb "affecting" is used incorrectly. Since the sentence is meant to convey a consequence of the previous action, it requires a verb in a form that can function as the main verb of a sentence.

L92-L94: This sentence is overly complex and combines multiple ideas without clear connections, making it difficult to follow. It tries to introduce the bark beetle as a pest, its effects, a historical note, and the consequences within one run-on sentence. The phrase "a large area of L. olgensis plantation forests are infested" has an agreement error. The use of commas does not effectively separate the ideas, leading to confusion.

L113-L114: The word "dominated" is used twice in different parts of the sentence, which might be stylistically inconsistent. Instead, it would be advisable to use synonyms or rephrase the sentence to eliminate this repetition. The list of tree species and their Latin names is not entirely clear due to the way they are enumerated, and because of the use of commas and conjunctions.

L171-L173: The sentence starts with the imperative verb "Calculate" but then shifts to an indicative statement. This creates a mismatch in sentence structure, suggesting it begins as a command but concludes as a descriptive statement.

L177-L180: The sentence could be refined for clarity and coherence.The phrase "the diversity Alpha diversity of a single sample" is repetitive and somewhat confusing. It seems like "Alpha diversity" is mistakenly introduced twice in a row.

L188-L192: The sentence shifts from past tense ("fluctuated", "was") to present tense ("is"), which disrupts the consistency of the narrative. The statement crams multiple observations into one long sentence, separated by commas, which makes it difficult to follow. Proper separation into distinct sentences can enhance clarity. The phrase "soil moisture content" is repeated multiple times, which is redundant. Using pronouns or rephrasing can make the sentence more readable.

L220: Diferent → different

L299-L336: This sentence is extremely long and covers multiple distinct ideas. It would be more digestible if broken into several sentences focused on specific points. The phrase "resulting in" is used multiple times in close succession, which can be repetitive and lead to a cumbersome read. Varying sentence structure and phrasing can improve readability. The sentence jumps between discussing soil moisture content, soil pH changes, carbon content, and the impact of pests, which could confuse readers. It's better to segment information into thematic paragraphs or sentences.

L369-L376: The sentence tries to convey multiple pieces of information about the role of Bacteroidetes and Tenericutes in the context of tree mortality and decomposition processes. Breaking it into shorter sentences would enhance clarity. The phrase "which may be the pathogens responsible for the death of trees under the influence of bark beetle" could be simplified to avoid redundancy, as the context of bark beetle disturbance is already established.

Author Response

Response to Reviewer 2 Comments

Thank you very much for your suggestions, they have helped us a lot with the article and we have made the changes as requested with a detailed description of the changes.

Point1:L90: The first appearance in the text, so full Latin name including the author of description is missing here.

Response1: The Latin in the text has been modified.

Point2:L125: Figure 1: This is not a sufficient map, the study plots should be marked on map.

Response2: The study area map has been revised.

Point3: L135: Is this part of the sentence still valid, if the authors add the information about tree sample plots above?

Response3: In order to represent the information of the sample plots more clearly.

Point4: L241: Authors should clarify why no fungi were classified despite the use of specific fungal primers focused on fungal analysis. This might indicate a mismatch with the reference database used for sequence classification. It's essential to disclose which databases were employed for both fungal and bacterial sequence comparisons, such as Silva or GreenGenes, as the choice significantly impacts microbial community analysis outcomes.

Response4: A comparative database of microbial communities has been added to the text.

Point5: L248: Why PERMANOVA is still not described in methods? The authors addressed the simpler issue (insert name of analysis and p-value) but forgot to mention this analysis also in the methods section.

Response5: The methodology has been described in the text: According to the PERMANOVA (Permutational Multivariate Analysis Of Variance) method, the total variance was decomposed using a half-metric or metric distance matrix to analyze the degree of explanation of the sample variance by the different environmental factors, with a larger R² indicating that the environmental factor indicator explains a higher degree of variance.

Comments on the Quality of English Language

Response6: An English language professional has been asked to revise and correct the writing throughout the text.

Examples of awkward sentences (but only several, the whole manuscript is a problem):

L45-L46: This sentence lacks a proper subject-verb agreement, making it a fragment rather than a complete sentence. Moreover, the verb "affecting" is used incorrectly. Since the sentence is meant to convey a consequence of the previous action, it requires a verb in a form that can function as the main verb of a sentence.

L92-L94: This sentence is overly complex and combines multiple ideas without clear connections, making it difficult to follow. It tries to introduce the bark beetle as a pest, its effects, a historical note, and the consequences within one run-on sentence. The phrase "a large area of L. olgensis plantation forests are infested" has an agreement error. The use of commas does not effectively separate the ideas, leading to confusion.

L113-L114: The word "dominated" is used twice in different parts of the sentence, which might be stylistically inconsistent. Instead, it would be advisable to use synonyms or rephrase the sentence to eliminate this repetition. The list of tree species and their Latin names is not entirely clear due to the way they are enumerated, and because of the use of commas and conjunctions.

L171-L173: The sentence starts with the imperative verb "Calculate" but then shifts to an indicative statement. This creates a mismatch in sentence structure, suggesting it begins as a command but concludes as a descriptive statement.

L177-L180: The sentence could be refined for clarity and coherence.The phrase "the diversity Alpha diversity of a single sample" is repetitive and somewhat confusing. It seems like "Alpha diversity" is mistakenly introduced twice in a row.

L188-L192: The sentence shifts from past tense ("fluctuated", "was") to present tense ("is"), which disrupts the consistency of the narrative. The statement crams multiple observations into one long sentence, separated by commas, which makes it difficult to follow. Proper separation into distinct sentences can enhance clarity. The phrase "soil moisture content" is repeated multiple times, which is redundant. Using pronouns or rephrasing can make the sentence more readable.

L220: Diferent → different

L299-L336: This sentence is extremely long and covers multiple distinct ideas. It would be more digestible if broken into several sentences focused on specific points. The phrase "resulting in" is used multiple times in close succession, which can be repetitive and lead to a cumbersome read. Varying sentence structure and phrasing can improve readability. The sentence jumps between discussing soil moisture content, soil pH changes, carbon content, and the impact of pests, which could confuse readers. It's better to segment information into thematic paragraphs or sentences.

L369-L376: The sentence tries to convey multiple pieces of information about the role of Bacteroidetes and Tenericutes in the context of tree mortality and decomposition processes. Breaking it into shorter sentences would enhance clarity. The phrase "which may be the pathogens responsible for the death of trees under the influence of bark beetle" could be simplified to avoid redundancy, as the context of bark beetle disturbance is already established.

Author Response File: Author Response.pdf

Reviewer 3 Report

Some notes in the Manuscript are marked in yellow.

Table 1. The 1^st column – Damage intensity

Planting year is the same and may be mentioned in the title of the table as well as the initial stand density.

Columns 3 and 4 have the same headings.

Some phrases are very long and difficult to understand. English needs correction.

Comments for author File: Comments.pdf

English needs correction.

Author Response

Response to Reviewer 3 Comments

Thank you very much for your suggestions, they have helped us a lot with the article and we have made the changes as requested with a detailed description of the changes.

Point1:Table 1. The 1^st column – Damage intensity

Response1: Changes have been made as requested.

Point2:Planting year is the same and may be mentioned in the title of the table as well as the initial stand density.

Response2: Changes have been made as requested.

Point3:Columns 3 and 4 have the same headings.

Response3: Changes have been made to the table contents

Point4:Some phrases are very long and difficult to understand. English needs correction.

Response4: An English language professional has been asked to revise and correct the writing throughout the text.

The annotations in the document have been amended as required.