Effect of Karst Microhabitats on the Structure and Function of the Rhizosphere Soil Microbial Community of Rhododendron pudingense

Round 1

Reviewer 1 Report (Previous Reviewer 3)

The manuscript has been resubmitted with significant improvement, and all comments have been addressed. The minor errors need to be corrected. As an example, "soil microbial" should be revised to "Soil microbes" in the Abstract.

There is a need to check for minor errors in English.

Author Response

Review 1:

• The manuscript has been resubmitted with significant improvement, and all comments have been addressed. The minor errors need to be corrected. As an example, "soil microbial" should be revised to "Soil microbes" in the Abstract.

Reply: We sincerely thank the reviewer for careful reading. As suggested by the reviewer, we have corrected the “soil microbial” into “Soil microbes”.

 

• There is a need to check for minor errors in English.

Reply: We have polished the content of the manuscript, and we hope the revised manuscript will be accepted by you.

Author Response File: Author Response.docx

Reviewer 2 Report (Previous Reviewer 2)

The materials and methods section for soil physical and chemical properties is completely missing, this is a whole addition of data not shown in the previous version of the manuscript that directly appears in the results.

The data analysis section should be more detailed. The process of analysis are often better explained in the results section than in the materials and methods ones, please add all the details about each analysis performed and state one by one which software was used to perform it.

Several mistypes and format issue are present in the manuscript I encourage you to proofread the paper to fix them.

L128 first of all I encourage you to pay attention to the fact that the author is spelled Riley and the citation format is incorrect. You choose a very outdated reference, nowadays the golden standard method to sample rhizospheric soil is to shake off loosely attached soil, following excised roots are placed in sterile water or saline solution or phosphate buffer, shaken and finally centrifuged to recover the rhizospheric soil. I understand that you followed a different method but please at least follow the reference you picked accordingly. In fact, Riley et al. states “The rhizosphere and rhizoplane soil samples came from distances of approximately 1-4 mm and 0-2 mm from the root surface, respectively. Portions of roots plus rhizoplane soil, rhizosphere soil and non rhizosphere soil were immediately extracted for 5 min with distilled water at a water:material ratio of 2:1.”  therefore considering you didn’t sample roots portion but only shaken soil you haven’t sampled “Soil layer (0 ~ 4 mm) on the root surface” as you state but according to Riley if anything the 1-4mm. 

L 133 "mixed with about 10 g" in the previous version of the manuscript you stated 20 g… which version is the correct ones? 

L138 please state what data you are reporting. are those means ± standard deviation? Same for table 3

L161 the first time you use an acronym you should write it in the extended form with the acronym between brackets and only following on with the acronym alone.

L217 OTU not OUT

 

The quality of English language has been slightly improved; however it must still be improved to assure a good readability and to fix the abundant grammar mistakes.

Author Response

Review 2:

(1) The materials and methods section for soil physical and chemical properties is completely missing, this is a whole addition of data not shown in the previous version of the manuscript that directly appears in the results.

Reply:We feel sorry for our carelessness. In 2.4 of Materials and Methods, we added the content of the determination of physical and chemical properties of soil. Thanks for your correction.

 

(2) The data analysis section should be more detailed. The process of analysis are often better explained in the results section than in the materials and methods ones, please add all the details about each analysis performed and state one by one which software was used to perform it.

Reply: According to your suggestion, we gave a more detailed description of this part, especially the description of operating software.

 

(3) Several mistypes and format issue are present in the manuscript I encourage you to proofread the paper to fix them.

Reply: We were really sorry for our careless mistakes. Here are some changes:

L 84, L 141, L 143---- "R. Pudingense" had been replaced by "R. pudingense";

L 114, L 118----"-" had been replaced by "to";

L 291, L 315----"OUT" had been replaced by "OTU";

Thank you for your reminder.

 

(4) L128 first of all I encourage you to pay attention to the fact that the author is spelled Riley and the citation format is incorrect. You choose a very outdated reference, nowadays the golden standard method to sample rhizospheric soil is to shake off loosely attached soil, following excised roots are placed in sterile water or saline solution or phosphate buffer, shaken and finally centrifuged to recover the rhizospheric soil. I understand that you followed a different method but please at least follow the reference you picked accordingly. In fact, Riley et al. states “The rhizosphere and rhizoplane soil samples came from distances of approximately 1-4 mm and 0-2 mm from the root surface, respectively. Portions of roots plus rhizoplane soil, rhizosphere soil and non rhizosphere soil were immediately extracted for 5 min with distilled water at a water:material ratio of 2:1.”  therefore considering you didn’t sample roots portion but only shaken soil you haven’t sampled “Soil layer (0 ~ 4 mm) on the root surface” as you state but according to Riley if anything the 1-4mm.

Reply: The description of rhizosphere soil sampling method in the manuscript is not very clear, and the author's name is incorrectly quoted. I apologize for this. Thank you for your suggestions. The content of this study is only a part of the total experimental design. In our earlier experimental design, the roots of Rhododendron pudingense were sampled, but the experimental content of roots was not involved in this study, so we omitted this part. Since the root environment is relatively dry and the soil viscosity is not strong, it is easy to brush the 0-4mm soil on the root surface after the root is taken out. Perhaps due to careless literature review, the rhizosphere soil was not found to be 1-4 mm soil. We have corrected this mistake.

 

(5) L 133 "mixed with about 10 g" in the previous version of the manuscript you stated 20 g… which version is the correct ones?

Reply: In the process of revising the manuscript, we rewrote this paragraph and carelessly changed “20 g” into “10 g”. Now it has been revised into 20 g (L 148). Thank you for your reminding.

 

(6) L 138 please state what data you are reporting. are those means ± standard deviation? Same for table 3.

Reply: We modify in L 246, and the mean ± variance expressed refers to Table 3 and Table 5.

 

(7) L161 the first time you use an acronym you should write it in the extended form with the acronym between brackets and only following on with the acronym alone.

Reply: The whole process has been replenished. The green background is highlighted in the manuscript (L 193).

 

(8) L217 OTU not OUT

Reply: We sincerely thank the reviewer for careful reading. "OUT" has been changed to "OTU".

 

(9) Comments on the Quality of English Language

The quality of English language has been slightly improved; however it must still be improved to assure a good readability and to fix the abundant grammar mistakes.

Reply: Thanks for your suggestion. We have tried our best to polish the language in the revised manuscript.

Reviewer 3 Report (New Reviewer)

The current manuscript entitled “Effect of karst microhabitats on the structure and function of  rhizosphere soil microbial community of Rhododendron pudingense” by Yuan et al. deals with the detection of the composition, quantity and functional types of rhizosphere soil microbial community in Rhododendron pudingense under three kinds of karst microhabitats (soil surface, rock gully, and rock surface) in Wangmo Country (WM), Zhenning Country (ZN) and Qinglong Country (QL).  After a careful reading, I found this manuscript interesting and suitable for publication in the Sustainability journal after minor changes. My specific comments are:

1.      Please do not repeat the keywords which already appeared in the title.

2.      Line 50: Please add “Therefore” before “It is of great…”.

3.      Line 74: Please replace “articles” with “studies”.

4.      Line 77: Please correct: Rhododendron Pudingense with Rhododendron pudingense.

5.      Line 95: Please replace “-“ to “to”.

6.      Correct scientific name at lines 123 and 126 and other parts of the whole manuscript.

7.      Line 537: if the effect was significant the p should be < 0.05 and not >.

Scientific names should be corrected in whole manuscript. Minor syntax and grammatical mistakes carefully.

Author Response

Review 3:

The current manuscript entitled “Effect of karst microhabitats on the structure and function of  rhizosphere soil microbial community of Rhododendron pudingense” by Yuan et al. deals with the detection of the composition, quantity and functional types of rhizosphere soil microbial community in Rhododendron pudingense under three kinds of karst microhabitats (soil surface, rock gully, and rock surface) in Wangmo Country (WM), Zhenning Country (ZN) and Qinglong Country (QL).  After a careful reading, I found this manuscript interesting and suitable for publication in the Sustainability journal after minor changes.

Reply: Many thanks for reviewer’s valuable comments.

 

• Please do not repeat the keywords which already appeared in the title.

Reply: Thanks for your suggestion, the keywords have been modified. —L 35

 

• Line 50: Please add “Therefore” before “It is of great…”.

Reply: "Therefore" has been added.—L 53

 

• Line 74: Please replace “articles” with “studies”.

Reply: According to your suggestion ,"articles" has been replaced with "studies".

 

• Line 77: Please correct: Rhododendron Pudingense with Rhododendron pudingense.

Reply: The word error here has been fixed.

 

• Line 95: Please replace “-“ to “to”.

Reply: According to your suggestion, we have changed "-" into "to".

 

• Correct scientific name at lines 123 and 126 and other parts of the whole manuscript.

Reply: The scientific name of the whole manuscript has been corrected correctly.

 

• Line 537: if the effect was significant the p should be < 0.05 and not >.

回复：根据您的建议，我们将“>”改为“<”。

 

• 学名应更正全文。仔细处理轻微的语法和语法错误。

回复：感谢您的仔细检查。我们对我们的粗心大意感到抱歉。根据您的意见，我们进行了更正。

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report (Previous Reviewer 2)

The manuscript in the present form has really been improved from the original version, yet the English language should be further improved.

Following few aspect that still should be fixed:

-section 2.4 please provide references for the used methods since no descriptions of the protocols are provided...

- (6) L 138 please state what data you are reporting. are those means ± standard deviation? Same for table 3. Reply: We modify in L 246, and the mean ± variance expressed refers to Table 3 and Table 5.

I don't see this modifications in the resubmitted manuscript please add this information in each table caption!

The quality of English language should be further improved

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

1. Well written article. However, there was no significant differences among three habitats (Soil Surface, Rock Gully and Rock Surface) with respect to microbial diversity. Depth of soil, soil texture, soil structure, organic matter and moisture content will definitely play a role on microbial diversity. Kindly look into this. 

Reviewer 2 Report

In the manuscript entitled “Effect of karst microhabitats on the structure and function of rhizosphere soil microbial community of Rhododendron pudingense” submitted to sustainability, the authors deal with the study of the rhizospheric microbial community of Rhododendron pudingense in several karst microhabitats. The submitted manuscript is generally interesting, and present conceptually well-organized tables and figures. However the manuscript in the current form present several major flaws that should be addressed and fixed.

 

Major issues

I recommend you check out the definition of rhizospheric soil and current sampling techniques for it. According to your sampling paragrapher the soil you isolated it's not the true rhizospheric fraction. The 2.3 section should be rewritten to made it clearer and more readable.

Several parts of the manuscript should undergo a linguistic revision to make it more comprehensible and readable.

in several points of the manuscript the significance of the results is overgeneralized drawing conclusions not fully supported by data.

The resolution of figures is poor and sometimes it makes hard to read the labels and the data reported (e.g. figure 2) I suggest to improve it to fully valorize the presented figures.

 

Minor issues

Throughout the manuscript there are many mistyping, grammar mistakes and improper use of terminology, I warmly suggest you to proofread it and improve the readability.

OUT is incorrect the correct acronym is OTU and should be reported in the not shortened form the first time it appears.

PCoA should be reported in statistical analysis.  

L225 “flora of fungi” is a serious conceptual mistake.

Reviewer 3 Report

The manuscript entitled “Effect of karst microhabitats on the structure and function of rhizosphere soil microbial community of Rhododendron pudingense” establishes the theoretical basis for explaining Rhododendron pudingense's adaptation to karst environments. The methods are well described, and the results are clearly interpreted. The figures and tables are well presented. Overall, the paper is well written and meets the journal's publication standard. Here are some suggestions I would like to make:

 

Major comments:

Introduction

It is important to state scientific questions or hypotheses, but these are not included in this introduction.

Materials and Methods

1. L116-133: The information will be easily accessed if the information of karst microhabitats are included in the forms. Please consider it.

2. L134: Important data, such as the age, height, and ground diameter of plants are not provided. The corresponding information needs to be supplemented.

3. L170: The data analysis is not detailed enough. Pleases describe the process of analysis according to the section of results.

Results

1. L181: The “sequencing data” maybe not necessary in the manuscript, but they could be displayed in forms of supplementary materials. Please consider it.

2. L217: OTUs may be incorporated into the section on microbial diversity. Figure 4 should be considered to add the significant analysis. Please consider it.

3. L277-286: Genus-level data are not included in the manuscript. Please verify it.

Discussion

1. The results are only slightly described but not explained deeply. Moreover, the physical and chemical properties of soils can show some differences in various karst microhabitats. Thus, the properties of soils, the important indexes measured, should be estimated due to their relationships and soil microbes. Currently, the relevant information cannot be incorporated into the study.

2. Limitation and Prospect, a vital section, maybe considered to incorporated into the manuscript.

Conclusion

This section may be summarized methodically, and several implications in habitat protection and exploitation may be discussed.

Minor comments:

L13, L35, L47: the word “microbial” is adjective, and please verify it. Please consider “microbes”. Please check the entire manuscript for similar errors.

L246: Please notice that the “gate level” is not exact. Please correct it.

L399: The format of references should be utilized correctly. Please check the entire manuscript for similar errors.

Note: The grammars and languages in the manuscript could be improved with the help of native experts if possible.