Micro-Environmental Variation in Soil Microbial Biodiversity in Forest Frontier Ecosystems—Implications for Sustainability Assessments

Round 1

Reviewer 1 Report

The present study afford very useful information on soil sampling method, as well as soil microbial communities changes with soil depths and spatial distance across the land-use type ( in forest, grassland and horticulture systems). The manuscript is well written and I have been enjoyed reading the ms. One issue I cared is the details of the community composition of soil microbes at varies depths and land-use type, particularly the functional microbes such as Glomeromycota. The authors need to be very careful for the conclusions based on this issue.  

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

This manuscript is a good start to understanding the spatial and soil-depth differences in soil sample microbiology for forest, grassland, and crops in northern Patagonia in Argentina. The co-authors need to make improvements to the manuscript in order for this to be suitable for publication in MDPI Sustainability. The majority of the edits involve “clarification” of what has already been done (exception being presentation of stats noted in edit 15 if these have not been run yet) below). I would be willing to review this manuscript after co-authors have made the following TWENTY-FIVE substantive edits:

    1)      The title needs to be improved so it is more concise and clear. MDPI Sustainability format for titles capitalizes all major words 

    2)      In general, the manuscript needs to do a better job at balancing the technicality of the writing with providing cross-disciplinary readers enough “context” so that the technicality is more easily understood. If this manuscript was submitted to a more discipline specific journal, then this likely would not need to be done. MDPI Sustainability is a cross-disciplinary journal that is read by people with broad academic backgrounds. 

    3)      When you use the abbreviations i.e. and e.g. you need to add a comma after these. The abbreviation i.e., means “in other words” as opposed to e.g., which means “for example.” 

    4)      Please add a new Figure 1 which is a map of the geographic area of Argentina with the different land use types (e.g., forest, crops, pasture, etc.) shown relative to the sampled area. Typically this includes a smaller inset map indicating where the study area is located with respect to Western Hemisphere. Most readers are not automatically familiar with northern Patagonia (e.g., that this is actually closer to the Pampas relative to Tierra del Fuego and in the mountains rather than closer to the coast). The geographic location of the study area is important for the reader to understand contextually. Please add latitude and longitude on the map border as well as a distance scale in kilometers as well as north arrow for all maps. 

    5)      The term “horticulture” throughout the manuscript is used when you have a picture in Figure 1 on L117 of what appears to be soybeans (if this is not, then please indicate what crop exactly this is). Please clarify since if commodity crops then these are classified as “commodity agriculture” or “commodity field crops” since horticulture is defined as “fruits, vegetables, flowers, or ornamental plants.” 

    6)      In sub-section 2.1., please indicate what types of crops are grown in the sampled area (include Latin name in italics). 

    7)      Do not abbreviate Fig. or Figs. in the writing and write out as Figure and Figures, respectively, throughout the manuscript. 

    8)      On L128 and anywhere else, use the × symbol and not * to indicate multiplication. 

    9)      For sub-section headers like 2.1. in italics, please capitalize all major words so for example, 2.1. Study Site and Soil Sampling Design. 

    10)  At the end of the paragraph on L201, please clarify for the reader the importance of using semi-variograms for helping correct for spatial autocorrelation (I assume this was done?). Meaning why this is important for spatial statistical models? Sustainability is a cross-disciplinary journal so you cannot assume that every reader knows this. 

    11)  Similarly in the Introduction section, please introduce for the reader the concepts of alpha, beta, and gamma diversities so there is contextual understanding when you present results on alpha and beta diversities. 

    12)  In the Methods section, add equations for alpha and beta diversity indicators used in the analysis. Add a blank row above and below the equation and all the way to the right you have (1) for example for equation 1. 

    13)  Add space on either side of signs like < so for example on L292, this should be p < 0.05. Make similar edits if need be throughout the manuscript. 

    14)  Paragraphs are a minimum of 3 sentences. So please for example move L294-296 to the end of the paragraph on L287. Make similar edits if need be throughout the manuscript (e.g., L471-473). 

    15)  For Table 1, why are statistical significant differences between land use types not presented as capital letters A, B, C (please present with lower case letter significant differences which I assume is for spatial distance a, b)? 

    16)  Delete L442-445 as this is from the instructions for authors Word template (never was deleted from the original template). 

    17)  In all figures, if there is a negative sign used such as for the axis labels, then this needs to use the longer endash symbol and not the shorter hyphen. The endash is consistently used throughout the manuscript which is correct. 

    18)  Figures and tables are stand alone in terms of understanding and the reader should not have to refer to the writing to understand what abbreviations stand for. Please write out EC, TN, TC, and CN in Figure 6 for example. 

    19)  For each major section of the manuscript (e.g., Introduction, Methods, Results, Discussion), please define abbreviations such as those in 17) above. Readers often do not read in direct order. 

    20)  In the paragraph on L474-484, please clarify the use of alpha and beta diversity. Assuming alpha diversity is the diversity within a particular sample and that beta diversity is the diversity between samples, why are samples that are taken by spatial distances treated as the same sample? Aren’t they different samples? Why is beta diversity which now accounts for soil depth of the sample categorized as beta diversity? Are the sub-samples by depth for any particular sampling location being defined as different samples? Please clarify this in the writing. The way that this is written implies that a sample is one soil depth across space (holding land use constant). Is that correct? 

    21)  There are areas where the writing needs to be broken up into shorter paragraphs (e.g., L485-515). Each paragraph has a topic sentence followed by supporting sentences. 

    22)  In the Discussion section, there are suggestions for sampling made and the implication is that there is broad generalization of research results. How specific are your results to the El Manso Valley? What is the justification for these generalizations (L497-515)? 

    23)  Do not write the Conclusions section in bullet points or numbered points. Please write in paragraph format. 

    24)  Please end the Conclusion section on what future research needs to do given the context of your results. Keep the Conclusion section to one paragraph (maximum of two paragraphs). 

    25)  The References formatting needs to be consistent with MDPI Sustainability. Please use journal names that are in italics and that are abbreviated (e.g., J. Plant Nutr. Soil Sci.) and for abbreviations look these up via web search by writing the journal name and then abbreviation in the search bar. Year needs to be in bold. Volume (issue) needs to be in italics. The DOI needs to be in https:// format.

In general, the writing is good. I will edit for line-number specific edits after the first round of revisions are made, but given the very good quality of the English writing, these will likely be limited in number.

Author Response

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Author Response File: Author Response.docx

Reviewer 3 Report

Micro-environmental variation of soil microbial biodiversity in forest frontier ecosystems – implications for field sampling design in sustainability assessments

Generally, this is an interesting data set and the description of the study mostly understandable and logical. However, the paper has to be improved with major modifications. Also, serious consideration should be given to the English grammar. Although it is understandable, it must be improved. Oxford punctuation should be applied in the whole paper.

Concerning Figure 2: please improve the quality of the figure. Choose fancy colors, avoid dark grey, and delete jitter points

Concerning Figures 4 and 5: delete letter "a" in the legend

Conclusion part: Bullets should be avoided.

Supplementary Materials are not available to review So please provide them with the next manuscript version

Moderate editing of English language required.

Author Response

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Author Response File: Author Response.docx

Reviewer 4 Report

Manuscript: sustainability-2761450-peer-review-v1

Micro-environmental variation of soil microbial biodiversity in 2 forest frontier ecosystems – implications for field sampling de-3 sign in sustainability assessments

Summary:

The manuscript presents a study about the evaluation of the changes in soil microbial communities with soil depth and spatial distance across contrasting land uses (forest, grassland and horticulture), in Northern Patagonia (Argentina). Soil chemical characterization, 16S rRNA gene sequencing and DNA quantification were determined and related to the micro-environmental variation in microbial biomass, α-β-diversity, and communities' assembly processes.

Overall opinion:

The subject addressed in the manuscript is interesting, as well as the achieved results, and meets the publishing objectives of the journal.

The manuscript is well written, structured and organised, and there is an easy flow between the methodology, results, discussion, and conclusion. The introduction gives the basic background of the research, the justification/relevance of the topic under discussion, and the objectives of the work. The methodology is well described, and the presentation of data and discussion are illustrated with appropriate tables and figures. The major findings are presented and discussed in an objective mode. So, I recommend the publication of the manuscript.

The text needs final editing to correct a few typos.

Author Response

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Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Sustainability

Manuscript Draft

Manuscript Number: 2761450

Title: Micro-environmental Variation of Soil Microbial Biodiversity in Forest Frontier Ecosystems – Implications for Sustainability Assessments

Article Type: Research article

This manuscript is a good start to understanding the spatial and soil-depth differences in soil sample microbiology for forest, grassland, and crops in northern Patagonia in Argentina. The co-authors have made the requested improvements to the manuscript in order for this to be suitable for publication in MDPI Sustainability. What remains are minor edits. I do NOT need to review this manuscript after co-authors have made the following THREE substantive edits as well as line-number specific edits:

     1)      Figure 1a lacks a distance scale in km. 

     2)      For Table 1, are the (47), (42), and (41) in the first column citations? If so, they need to be [47], [42], and [41]. Are they number of samples? Not clear.

     3)      The names of co-authors in Author Contributions starting on L814 need to be abbreviated (this was initially done but for some reason was changed to names being fully written out which is not correct)

Specific Line Number of Manuscript Figure/Table comments (note that requested change of word(s) in quotations where NO edits need to be made for writing before/after each “…”):

L226 – Do not indent since this is part of the paragraph that starts on L224

L243 – Do not indent since this is part of the paragraph that starts on L241

L249 – Do not indent since this is part of the paragraph that starts on L247

L371 – Write out as Figure (use Word Find & Replace to check of any other cases of Fig. to change to Figure)

L727 – Use e.g., and not i.e., (e.g., means for example; i.e., means in other words)

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

This manuscript version has been well enhanced compared with the previous one and several comments have been addressed. I provided a general appreciation of the manuscript and how it can be improved to achieve a good standard for publication. Here are other considerations and suggestions before acceptance.

L58 there is ...... should be paraphrased with "Evidence shows that"

and "there is" or "it is" should be avoided in the scientific writing. 

L89 same comment as L58: should be paraphrased

L404 and L409 you should take care of Oxford punctuation.

L659 and L674 same comment as L58 : should be paraphrased

The conclusion should be improved.

L745 it is ...... same comment as L58 : should be paraphrased

Minor editing of English language required.

Oxford punctuation should be taken into account in the whole paper.

Author Response

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Author Response File: Author Response.pdf