Gloeotrichia cf. natans (Cyanobacteria) in the Continuous Permafrost Zone of Buotama River, Lena Pillars Nature Park, in Yakutia (Russia)
, Denis Davydov 2,3, Anna Vilnet 2, Sergey Sidelev 4,5, Ekaterina Chernova 6, Sophia Barinova 7,*, Olga Gabysheva 1 and Zoya Zhakovskaya 6Round 1
Reviewer 1 Report
The discovery of potentially toxigenic Gloeotrichia cf. natans 2 (Cyanobacteria) in the continuous permafrost zone of Buotama 3 River, Lena Pillars Nature Park in Yakutia, was made through 4 the observation of epilithic macrocolonies 5
This title is not in accordance with the findings presented in the manuscript.
I suggest it be modified to-
Gloeotrichia cf. natans 2 (Cyanobacteria) in the continuous permafrost zone of Buotama 3 River, Lena Pillars Nature Park in Yakutia (Russia)
It was completely consistent with the description of Gloeotrichia…
Which reference did you use to confirm the species?
Line 286
are not containing
Replace with “do not contain”
Line 286
Replace “evidence” with “ascertain”
Lines 383-395 reads more like the introduction. Please try to compare the information presented here with your findings, so that it will be a true discussion.
The text should be revised thoroughly to correct minor errors.
Author Response
Dear Editor,
Thank you and the Reviewer 1 for comments. Please find responses to each comment below.
With best regards,
Prof Sophia Barinova,
Corresponding author
Reviewer 1
Dear Reviewer!
We are thankful for the positive evaluation of our study.
1) This title is not in accordance with the findings presented in the manuscript.
I suggest it be modified to “Gloeotrichia cf. natans (Cyanobacteria) in the continuous permafrost zone of Buotama River, Lena Pillars Nature Park in Yakutia (Russia)”
Answer: We accept your proposal and change the title of the manuscript to the proposed one, which is more appropriate for the study.
2) It was completely consistent with the description of Gloeotrichia…
Which reference did you use to confirm the species?
Answer: The reference was added.
3) Line 286 “are not containing” replace with “do not contain”
Answer: Text has been rewritten.
4) Line 286
Replace “evidence” with “ascertain”
Answer: Text has been rewritten.
5)Lines 383-395 reads more like the introduction. Please try to compare the information presented here with your findings, so that it will be a true discussion.
Answer: The text was corrected according with your suggestion.
Editing of English language was made by native speaker of English.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The article by Gabyshev et al. describes the discovery of a cyanobacterium identified as Gleotrichia cf. natans in the permafrost zone in Yakutia. Identification was made using both morphological and genetic criteria. Comprehensive information about the hydrochemical conditions of habitat is also given in the article. Cyanobacteria have also been studied for the possibility of synthesizing cyanotoxins. The work is of interest from a biogeographical and ecological point of view, and can be recommended for publication, but still the manuscript requires some processing and changes.
First of all, in the title of the manuscript Gloeotrichia cf. natans has been characterized as potentially toxigenic. However, the authors showed that the described organism doesn't contain either the toxins or the genes responsible for the synthesis of toxins. Thus, the definition of this organism as potentially toxigenic is incorrect. This could be said if, for example, toxins were not found, but the genes encoding them were present. In the absence of both, one can say that this organism is safe. And clearly, "potentially toxigenic" should be removed from the title. If there is a possibility of the synthesis of some other toxins that havn't been experimentally tested, but can be potentially synthesized by this and/or related taxa of cyanobacteria according to the literature data, then attention should be focused on this in more detail. This problem is relevant to the entire text. Maybe it's better not to put cyanotoxins at the forefront? I would recommend paying more attention to ecology, along the way discussing cyanotoxins and showing that your isolate does not produce them.
Fig. 1. Please sign, on the figure "from" the Lena and Buotama rivers, if possible.
2.4. Water chemistry analysis. Please list the instruments used to measure water color, pH, ions, etc.
2.7. DNA extraction, purification and PCR. Please indicate the primer pairs used for the 16S-23S ITS sequence. References are for 16S gene sequence analysis only.
2.8. Molecular Analyses. Please indicate (here or further in the results) the length of the obtained sequences. Also specify NCBI Accession number if sequence is deposited.
3.3. Microscopic survey. Lines 247-248 repeat information from lines 235-236. Please leave this information in one place only.
3.4. Phylogeny. What is the percentage of similarity between the 16S of your isolate with neighboring species? Is there any reason to assume that the new organism is a new genus? There are also no data on 16S-23S ITS sequences, which were declared in the methods section.
4. Discussion. Link to fig. 3 in line 311 is apparently not correct. Please check
Extensive editing of English language required. Please contact a native speaker or a professional translator to correct the text.
Author Response
Dear Editor,
Thank you and the Reviewer 2 for comments. Please find responses to each comment below.
With best regards,
Prof Sophia Barinova,
Corresponding author
Reviewer 2
Dear Reviewer!
We are thankful for the positive evaluation and your valuable recommendations.
First of all, in the title of the manuscript Gloeotrichia cf. natans has been characterized as potentially toxigenic. However, the authors showed that the described organism doesn't contain either the toxins or the genes responsible for the synthesis of toxins. Thus, the definition of this organism as potentially toxigenic is incorrect. This could be said if, for example, toxins were not found, but the genes encoding them were present. In the absence of both, one can say that this organism is safe. And clearly, "potentially toxigenic" should be removed from the title. If there is a possibility of the synthesis of some other toxins that havn't been experimentally tested, but can be potentially synthesized by this and/or related taxa of cyanobacteria according to the literature data, then attention should be focused on this in more detail. This problem is relevant to the entire text. Maybe it's better not to put cyanotoxins at the forefront? I would recommend paying more attention to ecology, along the way discussing cyanotoxins and showing that your isolate does not produce them.
Answer: Paragraph describing Gloeotrichia cf. natans as a potentially toxigenic species was excluded from the manuscript. The title of the manuscript was corrected in accordance with your recommendation.
- Please sign, on the figure "from" the Lena and Buotama rivers, if possible.
Answer: The Lena River sign was added to the map (Fig. 1c), and the caption to the figure was corrected. Unfortunately, it is not possible to add the Buotama River and its sign due to the scale of the map.
2.4. Water chemistry analysis. Please list the instruments used to measure water color, pH, ions, etc.
Answer: We added the information about instruments used for chemical analysis.
2.7. DNA extraction, purification and PCR. Please indicate the primer pairs used for the 16S-23S ITS sequence. References are for 16S gene sequence analysis only.
Answer: The primer site map in the 16S-23S rRNA operon that was used in this study was shown at https://doi.org/10.1093/oxfordjournals.molbev.a003877. The forward primer 2 started at the 16S gene, and PCR with reverse primer 1 started at the 16S-23S rRNA operon. We also used internal primer 3.
2.8. Molecular Analyses. Please indicate (here or further in the results) the length of the obtained sequences. Also specify NCBI Accession number if sequence is deposited.
Answer: All necessary information was added to the text in subsection 2.8.
3.3. Microscopic survey. Lines 247-248 repeat information from lines 235-236. Please leave this information in one place only.
Answer: The paragraph was removed from section 3.3.
3.4. Phylogeny. What is the percentage of similarity between the 16S of your isolate with neighboring species? Is there any reason to assume that the new organism is a new genus? There are also no data on 16S-23S ITS sequences, which were declared in the methods section.
Answer: We added Table 2 with data of similarity between the 16S of our isolate with neighboring species. The similarity of our strain and other Gloeotrichia species were not less than 95%, so we assumed that this was a species of Gloeotrichia.
- Discussion. Link to fig. 3 in line 311 is apparently not correct. Please check
Answer: Thank you, the link was corrected.
Editing of English language was made by native speaker of English.
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
Dear authors!
Thanks for your work on the article. Your responses to my comments and the corrections made have completely satisfied me. I think that the article its present form can be recommended for publication.
