Genetic Diversity and Structure of Pinus densiflora Siebold & Zucc. Populations in Republic of Korea Based on Microsatellite Markers
Round 1
Reviewer 1 Report
This paper aims at defining the genetic diversity, differentiation and structuration of Pinus densiflora, an important component of Republic of Korea forests.
It used an extanded sampling, covering the whole country, both in regions with high and low Pinus densiflora density.
The microsatellites method to retrieve the different genetic parameters is an efficient method for this level of diversity (intra-specific). Number of microsatelite used is appropriate.
Methods were, as far as I can judge, well conducted. All information such as primer sequences, motives, temperature etc, were given for each marker.
The authors found a high diversity and low differentiation within the more than 60 populations studied, as well as weak structuration.
This is in line with the life history of this species and breeding system.
I am not surprise to see the high bigger genetic differentiation with population of Mt Halla as other studies has shown similar pattern, for example in Zelkova serrata (Naciri et al. 2019), some populations of Jeju island are more connected genetically to Mainland Japan populations than Korean ones, for chloroplast only.
The authors discuss their results comparing with other gymnosperm species or the regions and in the context of forest management and conservation.
They report the interesting case of some populations with inbreeding depression that could be correlated to history of logging, fire and disease in the regions where the populations are growing.
This study is, from my point of view, interesting because it give new information for an important component of Republic of Korea and Eastern Asia flora, in the context of climate change and degradation of natural populations. I could be use as reference in futur studies.
I appreciated reading this study and recommand the publication in the present form.
Author Response
Response to Reviewer 1
Dear Reviewer,
We appreciate your kind opinion about manuscript ID Forests-1196885.
Sincerely yours,
Ahn Ji-Young
Author Response File: 
Author Response.docx
Reviewer 2 Report
The manuscript describes a very interesting and important study. The introduction outlines the nature of the problem, sets goals. The methods have been selected in accordance with modern requirements, described in detail, understandable and traceable. The obtained results and discussion confirm the defined conclusions.
Author Response
Response to Reviewer 2
Dear Reviewer,
We appreciate your kind opinion about manuscript ID Forests-1196885.
Sincerely yours,
Ahn Ji-Young
Author Response File: Author Response.docx
Reviewer 3 Report
Dear authors,
I've read your manuscript with great interest. In general, I like the manuscript. Please find attached my comments and suggestions, which I hope you will find useful and constructive.
Kind regrads
Comments for author File: 
Comments.pdf
Author Response
Response to Reviewer 3 Comments
Dear Reviewer
We appreciate your constructive comments and suggestions regarding manuscript ID Forests-1196885.
We tried our best to address your comments as much as possible in the revised manuscript.
The revised manuscript is appended alongside our responses to this letter.
I have left all track changes made in response to your comments and highlighted them in yellow.
Since your comments are written directly in the manuscript, we have corrected them by indicating page and paragraph numbers based on the manuscript.
However, in the process of revision, some pages and paragraph numbers have changed
Point 1: Abstract section, Page 1 paragraph 16 Reviewer comment that change words
Response 1: Page 1 paragraph 16, We revised “other” species to “conifer” species.
Point 2: Introduction section, Page 1 paragraph 28 Reviewer comment that add specify common names
Response 2: Page 1 paragraph 28, We revised “It is most widely” to “Japanese red pine is most widely….”
Point 3: Introduction section, Page 2 paragraph 44-45 Reviewer comment that add specify scientific names
Response 3: Page 2 paragraph 45–46, We revised “with pine gall midges or pine wilt disease.” to “with pine gall midges (Thecodiplosis japonensis) or pine wilt disease caused by pine-wood nematode (Bursaphelenchus xylophilus) [10].”
Point 4: Introduction section, Page 2 paragraph 54 Reviewer comment that change words
Response 4: Page 2 paragraph 55, We revised “conifer characteristics” to “needle and cone characteristics.”
Point 5: Material and Methods section, Page 2 paragraph 78 Reviewer comment that change word and Question
Response 5: Page 2 paragraph 80, We revised “Leaves were collected” to “Needles were collected.”
Question : Is there any other way?
Answer: If you are asking if there are other sampling methods, the method described is most commonly used. We tried to ensure that all trees that were sampled were at least 50 m apart.
Point 6: Result section, Page 6 paragraph 174-175, Table 2, Reviewer comment that add more information about microsatellite markers ‘calculate and present the number of alleles per locus (Na) and the polymorphism information content (PIC).’
Response 6: Page 6 paragraph 186–187, we calculated and presented Na and PIC in Table 2. These results are indicated in blue text. We also added the description of these calculations to the Materials and Methods (Page 4, paragraph 125–127)
Point 7: Discussion section, Page 9, paragraph 243-252, Table 4. Reviewer comment that ‘The results should not be presented in the Discussion section. Why did you used IAM model? Please specify in the Material and Methods section. Bottleneck should be tested for all populations. Table with the results can be added in the Supplementary material.’
Response 7: We tested all populations and presented the results in Supplementary materials ‘Table S1’. We deleted Table 4 and revised the Discussion (Page 9 paragraph 254–255). Moreover, we added a detailed description of the Bottleneck test in the Materials and Methods section (Page 5 Paragraph 148–154). We also added a description in the Results section (Page 6, paragraph 184–185)
Question: Why did you used IAM model?
Answer: We selected both the infinite allele mutation model (IAM) and the two-phase mutation model (TPM) and set the option to test according to a previously published analysis of P. densiflora in Japan [34]. The two models were selected because IAM has greater statistical power than the step mutation model (SMM) when using highly polymorphic markers. TPM is suitable when allele size is not equally distributed.
Point 8: Reviewer kindly comments that all species Scientific full name and recommend revised
Response 8: We have added the full scientific names for each species when they are first mentioned.
Point 9: Reviewer comment “English language and style are fine/minor spell check required”
Response 9: We rechecked the manuscript and re-edited it with the help of an English language expert.
We add one more reference and some references were reordered during revision. We changed the corresponding reference numbers in the manuscript.
Sincerely yours,
Ahn Ji-Young
Author Response File: Author Response.docx
Round 2
Reviewer 3 Report
Dear authors,
Thank you for addressing all the concerns/questions raised. The paper is well written and interesting to read. Additional corrections to minor methodological errors and text editing, and few comments and suggestions are marked in the attached document.
Kind regards
Comments for author File: Comments.pdf
Author Response
Dear Reviewer
Please see the attachment
Author Response File: Author Response.docx
