Assessment of Variability: Chloroplast Microsatellite DNA, Defoliation, and Regeneration Potential of Old Pine Stands of Different Origins in the Context of Assisted Genotype Migration
Round 1
Reviewer 1 Report
The author studied the genetic variation and natural regenerations among transferable and non-transferable (local) populations. This research is interesting, and I have some questions.
1. Figure 1 has a low resolution, and some information is not clear, such as longitude and latitude.
2. In section 2.3, please add the producers of the software used in the study.
3. Line 199, the level of genetic diversity is expressed as a percentage (98%). Please consider whether this is appropriate.
4. Line 211, the genetic differentiation (Fst) between MIE and PLE should be 1.6% or 1.577%.
5. The Fst values for the KOC ranged from 4% to 12%, which distinguished it from the local populations, and I suppose that may be caused by the different origins. However, according to the results of genetic differentiation (Fst) and PCoA, stand GAC (local) showed an obvious genetic differentiation from the other local populations (MIE and PLE), and it has a comparatively close relationship to the transferable population (KOC). I just want to know the reasons, and it should be mentioned in the manuscript.
Author Response
The author studied the genetic variation and natural regenerations among transferable and non-transferable (local) populations. This research is interesting, and I have some questions.
Dear Reviewer
Thank you for your appreciation of my work on analysing the effects of seed transfer. I know that the original text required some minor adjustments, which I have made. I will describe the details in relation to the questions posed. The text also underwent a language revision MDPI 52334.
Figure 1 has a low resolution, and some information is not clear, such aslongitude and latitude.
I have the drawing in high resolution pdf however the doc. format limits its posting. The quality of Figure 1. will be improved as the text is assembled.
In section 2.3, please add theproducers of thesoftware used in the study.
- Haplotype: References 24; Eliades, N.-G.; Eliades, D.G. Haplotype Analysis: Software for Analysis of Haplotypes Data. Distributed by the Authors; Forest Genetics and Forest Tree Breeding; Georg-Augst University: Goettingen, Germany, 2009.
- ARLEQUIN version 3.0: References 25; Excoffier, L.; Lischer, H.E.L. Arlequin suite ver. 3.5: A new series of programs to perform population genetics analyses under Linux and Windows. Mol. Ecol. Resour. 2010, 10, 564–567.
- GenALEx 6.5 References 26; Peakall, R.; Smouse, P. Genealex 6.5: Genetic Analysis in Excel. Population genetic software for teaching and research. Mol. Ecol. Notes 2006, 6, 288–295.
Line 199, the level of genetic diversity is expressed as a percentage (98%). Please consider whether this is appropriate.
Line 211, the genetic differentiation (Fst) between MIE and PLE should be 1.6% or 1.577%.
The value has been corrected. Thank you.
The Fst valuesfor the KOC ranged from 4% to 12%, which distinguished it from the localpopulations, and I suppose that may be caused by the different origins. However, according to the results of genetic differentiation (Fst) and PCoA, stand GAC (local) showed an obvious genetic differentiation from the other local populations (MIE and PLE), and it has a comparatively close relationship to the transferable population (KOC). I just want to know the reasons, and it should be mentioned in the manuscript.
Thank you for reading the text and drawing attention to the phenomenon of the local GAC population. The GAC stand was 100% defoliated by insects several times between 1990 and 1995, resulting in natural selection and the death of some trees. It is likely that the genetic distance from local populations is due to selection, but this is only a logical conclusion without empirical evidence. In the absence of sufficient empirical data (e.g., number of trees withered due to defoliation), the GAC population was not analysed in the text of the manuscript.
Best Regards
Paweł Przybylski
Author Response File: 
Author Response.docx
Reviewer 2 Report
This paper focuses on the differences in the effects of cpSSR on the health status and regeneration potential of stands from exotic or native seeds. The assessment of stand stability traits and their regeneration potential determines the sustainability of the ecosystem and can address potential selection for unsuitable genotypes in both parents and progeny. The overall work is satisfying, but several problems should be resolved at first before its acceptance:
1. The language of this manuscript must be modified since many language flaws could be identified which makes it difficult to understand.
2. Why SSR instead of SNP or other molecular markers was selected in this study?
3. L51-54: this part should be stated before L41-43.
4. L66: the meaning of this sentence is confusing.
5. Is it too general to conclude that KOC has a higher number of haplotypes (A), haplotype richness (Rh), and genetic diversity (He) than other trees grown from exotic seeds, and that it lacks a selection mechanism?
6. The information in fig2 is too concise, and I’m wondering if the forest type could cause an effect on the outcome of the deciduous part of the leaf canopy.
Author Response
This paper focuses on the differences in the effects of cpSSR on the health status and regeneration potential of stands from exotic or native seeds. The assessment of stand stability traits and their regeneration potential determines the sustainability of the ecosystem and can address potential selection for unsuitable genotypes in both parents and progeny. The overall work is satisfying, but several problems should be resolved at first before its acceptance:
Dear Reviewer
Thank you for your appreciation of my work on analysing the effects of seed transfer. I know that the original text required some major adjustments, which I have made. I will describe the details in relation to the questions posed. The text also underwent a language revision MDPI 52334.
The language of this manuscript must be modified since many language flaws could be identified which makes it difficult to understand.
The text of the manuscript was linguistically revised under number 52334.
Why SSR instead of SNP or other molecular markers was selected in this study?
SSR markers are a useful molecular biology tool. They are an old type of markers, but the results obtained with them are valid and prove biological phenomena. One example is the effect of seed transfer on the gene pool analysed in the manuscript. I have planned SSR analyses as part of the research project (which is subject to constraints, including economic constraints), but I hope that the results obtained will allow application to other projects focusing on KOC using newer analytical tools such as SNPs.
L51-54: this part should be stated before L41-43.
The sentences list, in chronological order, the events that confirm the negative effects of spontaneous seeds transfer. Please check that the current version of the text is correct.
L66: the meaning of this sentence is confusing.
The sentence has been corrected.
Is it too general to conclude that KOC has a higher number of haplotypes (A), haplotype richness (Rh), and genetic diversity (He) than other trees grown from exotic seeds, and that it lacks a selection mechanism?
The sentence has been corrected, and clarified. The original version was general.
The information in fig2 is too concise, and I’m wondering if the forest type could cause an effect on the outcome of the deciduous part of the leaf canopy.
The information in figure 4 indicates differences in defoliation between the stands studied. The results did not allow us to prove significant differences in defoliation between the stands.
Best Regards
Paweł Przybylski
Author Response File: Author Response.docx
Round 2
Reviewer 2 Report
The author has well addressed all my concerned issues, and for me it's satisfying to accept it in its current form.
Author Response
Dear Reviewer
Thank you for your support of the manuscript.
Best Regards
Paweł Przybylski
