Ectomycorrhizospheric Microbiome Assembly Rules of Quercus mongolica in the Habitat of SongRong (Tricholoma matsutake) and the Effect of Neighboring Plants

Round 1

Reviewer 1 Report

 

Dear authors,

 

I have enjoyed the manuscript titled “Host ectomycorrhizospheric microbiome assembly rules of SongRong (Tricholoma matsutake) and the Neighboring plants’ effects on them.” This manuscript provides valuable data on the microbiome associated with the root of Quercus mongolica and its relationship with the surrounding vegetation. Yet, please find the comments below that would help improve the quality of the manuscript.

 

My major points of revision are as follows:

 

I doubt that the title of this manuscript is appropriate for the result, as a large part of the study focuses on ectomycorrhizospheric microbiome of Q. mongolica, rather than T. matsutake and its relationship with nearby organisms. There are several research studies on the relationship between T. matsutake and other microbes (mycelial growth, seedling tests, ... etc.). Please consider adding more discussions on T. matsutake to keep the content aligned with the title.

 

Furthermore, from my experience, PICRUST2 uses only a small portion (less than 1/3) of fungal microbiome data for the prediction of sample functional abundance due to a small database. The fungal OTU/ASV references utilized in the PICRUST2 analysis were mostly those from agriculture or engineering studies, which may have a bias. What percentage of fungal sequences was used in PICRUST2 in this manuscript, and what are the taxonomic identities of these sequences?

 

Materials and Methods 2.4: How did you filter ectomycorrhizal fungi? Please state the database/reference used for the process. Also, describe the criteria used for the quality filtering of sequences during the QC process.

 

L144: Please describe a clear definition of “relative abundance” and how it was calculated in detail. However, be aware that the concept of relative abundance using ITS and 16S might not be appropriate for a statistical test because of a PCR bias and different copy numbers of rRNA sets in microorganisms. Please consider adding a statistical test presenting only the presence/absence data of OTU/ASV.

 

L275: I think these paragraphs are about Fig. 4, but I could not find a relevant figure or a table here. Furthermore, the order of Fig. 4 and 5 are reversed in the manuscript. Please check the paragraph and the figures.

 

L345: The authors mentioned the microbiome of a habitat rhizospheric soil in the M&M. Please explain how/whether you processed the habitat rhizospheric soil samples or used the data in previous studies.

 

L372: Are there any previous studies focused on the role of up-regulated pathways in the mycorrhizal microbiome? I recommend discussing the roles of these pathways (and enzymes) in the microbiome in this paragraph.

 

And these are some minor points:

 

Materials and Methods 2.3: Please describe the DNA extraction method and PCR protocol in detail.

 

Fig. 1e: Please add a Simpson index plot. Both plots show Shannon index.

 

Table 2: Which statistical test did you use to find differences in physicochemical properties and climatic factors? Please add the details in Materials and Methods.

 

There were several typos in the manuscript and references. In addition, there were many grammatical errors in the manuscript. Please revise the overall manuscript before submission.

 

L16: In order to clarified → in order to clarify

L25: affact → affect

L69: microbiom → microbiome

L334: gether → gather

 

Legends of Fig. 1, Table 1, Fig. 4: Italicize the species names.

 

Reference No. 7: Please check the names of the authors (to lower case) and species name (to italic)

Reference No. 16, 67: Please check the names of the authors (to lower case)

Reference No. 17, 81: Please check the journal title (to lower case)

Reference No. 18, 19, 21, 24, 28, 30, 32, 50, 65: Please check the species name (to italic)

Reference No. 29: Please check the species name (to italic, and edit the first letters of the scientific names to capital)

Additional notes: Some of the journal titles are in abbreviation, while the others are listed in full names. Please check those references and unify the format according to the journal guideline.

 

 

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

Title: Improve the title because it does not represent the work done

Abstract: The objective is not well defined

                    Missed the conclusion of the work in the abstract

Keywords: The words in this topic must not be in the title

Introduction: It is important to cite some values ​​in the examples cited in the literature regarding symbiosis.

Material and methods

Line 91 exchange ml for mL

Neighboring plants were not identified?

Conclusion

It can be more objective. As this seems more discussion then conclusion.

 

Author Response

Please see the attachment

Author Response File: Author Response.docx