Brenneria nigrifluens Isolated from Aesculus hippocastanum L. Bark in Hungary
and Anita Végh 1Round 1
Reviewer 1 Report
The article titled "Brenneria nigrifluens isolated from Aesculus hippocastanum L. bark in Hungary" present the first notification of bacteria Brenneria nigrifluens on hungarian chestnuts. Authors highlight that this species, like other relates species of Brenneria, are increasing problem in this region, on many deciduous trees. From this point, any notification like that, describing new hosts and the potential threat is important.
Although linguistic changes are necessary. In some paragraphs the importance of the study is not highlighted enough and reader may lost the plot. The importance to the local ecosystems or urban green areas should be emphasized. In the results authors are describing their other observation of Brenneria in Hungary, so it will be worth to try to make a prediction of the future of this genera in local ecosystem.
Authors correctly made the tests of pathogenicity and correctly decribed, that single infection by Brenneria nigrifluens is not causing he same symptoms as were observed in the nature. Although studied species can cause necrosis of bark and is able to infect the chestnut.
There is a need to make also some stylistic changes in the manuscript:
- Citation needs to be changed according to the journal requirements.
- There is a lack of Conclusion part, which is required by journal.
More detailed information are included in the pdf attached below.
Comments for author File: 
Comments.pdf
Author Response
Manuscript 1551195
Response to Reviewer1
Dear Editor, Assistant Editor and Reviewer,
Thank you for giving us the opportunity to submit a revised draft of the manuscript „Brenneria nigrifluens isolated from Aesculus hippocastanum L. bark in Hungary” for publication in the Journal Forests. We appreciate the time and effort that you and the reviewers dedicated to providing feedback on our manuscript and are grateful for the insightful comments on and valuable improvements to our paper. We have incorporated most of the suggestions made by the reviewers. Those changes are highlighted within the manuscript. Please see below, in green, for a point-by-point response to the reviewers’ comments and concerns. All page numbers refer to the revised manuscript file with tracked changes.
Reviewers’ Comments to the Authors:
Reviewer 1
1. Comment from Reviewer 1: The article titled "Brenneria nigrifluens isolated from Aesculus hippocastanum L. bark in Hungary" present the first notification of bacteria Brenneria nigrifluens on hungarian chestnuts. Authors highlight that this species, like other relates species of Brenneria, are increasing problem in this region, on many deciduous trees. From this point, any notification like that, describing new hosts and the potential threat is important.
Response: Thank you.
Comment from Reviewer 1: Although linguistic changes are necessary. In some paragraphs the importance of the study is not highlighted enough and reader may lost the plot. The importance to the local ecosystems or urban green areas should be emphasized. In the results authors are describing their other observation of Brenneria in Hungary, so it will be worth to try to make a prediction of the future of this genera in local ecosystem.
Response: We thank the reviewer for pointing this out. We have added the suggested content to the manuscript. Please see page 10 of the revised manuscript, lines 4-11.
Comment from Reviewer 1: Authors correctly made the tests of pathogenicity and correctly described, that single infection by Brenneria nigrifluens is not causing the same symptoms as were observed in the nature. Although studied species can cause necrosis of bark and is able to infect the chestnut.
Response: Thank you.
Comment from Reviewer 1: There is a need to make also some stylistic changes in the manuscript: Citation needs to be changed according to the journal requirements.
Response: We thank the reviewer for pointing this out. We have made the change.
Comment from Reviewer 1: There is a lack of Conclusion part, which is required by journal.
Response: We apologize for the missing Section. We have added it to the manuscript. Please see page 11 of the revised manuscript, lines 22-30.
More detailed information are included in the pdf.
Responses to the comments in the pdf attached:
- Comment from Reviewer 1
Response: We apologize for our error. Please see page 1, 2 of the revised manuscript, line 36, 37 and lines 1 and 4.
- Comment from Reviewer 1
Response: Thank you for pointing this out. We have corrected the sentence. Please see page 2 of the revised manuscript, line 5.
- Comment from Reviewer 1
Response: We have fixed the error. Please see page 2 of the revised manuscript, line 6.
- Comment from Reviewer 1
Response: We have fixed the error. Please see page 2 of the revised manuscript, lines 5-7.
- Comment from Reviewer 1
Response: We thank the reviewer for pointing this out. We have added the suggested content to the manuscript. Please see page 1 of the revised manuscript, lines 28-32.
- Comment from Reviewer 1
Response: We have fixed the error. Please see page 4 of the revised manuscript, line 32.
- Comment from Reviewer 1
Response: We thank the reviewer for pointing this out. We have added the suggested content to the manuscript. Please see page 5 of the revised manuscript, lines 2-3.
- Comment from Reviewer 1
Response: This observation is correct. We have marked by rhombuses the strains which were used in our research. Please see page 9 of the revised manuscript.
- Comment from Reviewer 1
Response: We have fixed the error. Please see page 10 of the revised manuscript, line 10, 11.
- Comment from Reviewer 1
Response: We thank the reviewer for pointing this out. We have removed those sentences, because tissue samples we have already collected in other locations of Hungary are part of present research, but the investigations are still ongoing.
- Comment from Reviewer 1
Response: We have corrected the typo. Please see page 10 of the revised manuscript, line 16.
- Comment from Reviewer 1
Response: We have revised the text to address your concerns and hope that is now clearer. Please see page 10 and 11 of the revised manuscript, lines 4-17 and lines 1-20.
Sincerely yours,
Prof. Dr. László Palkovics
Author Response File: Author Response.pdf
Reviewer 2 Report
The manuscript entitled „Brenneria nigrifluens isolated from Aesculus hippocastanum L. bark in Hungary“ describes identification of causal agent of longitudinal cracks on the trunk and branches and intensive brown liquid oozing on Aesculus hippocastanum trees. Causal agent belong to the genus Brenneria ie the closest phylogenetic relation with Brenneria nigrifluens according to phylogenetic analysis of 16S rRNA gene region. Authors have adequately addressed the following points which include complete description of symptoms, the host, disease observation, the geographic locations where samples that were used in the study were collected, proof of strains pathogenicity, 16S rRNA strains sequencing, and re-isolation of bacteria from the inoculated plants, fulfilling Koch’s postulates.
I have a few comments:
- The technical aspects of the paper are not correct.
(1) References are not used as is stated in Instruction for Authors - “References must be numbered in order of appearance in the text (including citations in tables and legends) and listed individually at the end of the manuscript. In the text, reference numbers should be placed in square brackets [ ] and placed before the punctuation; for example [1], [1–3] or [1,3]. Similarly, authors should modify Section References;
(2) Section Conclusion is missing;
- In Table 1 authors should cited references for presence of Brenneria species per countries;
- Although the authors confirmed the presence of Brenneria onAesculus hippocastanum trees, it seems that phylogenetic analysis of the 16S rRNA gene region did not give clear species determination. Therefore, some other support for taxonomic identity of Hungarian strains is needed. I highly recommend sequencing more genes such as MLSA;
- The manuscript requests extensive editing before it is published. I will leave the decision to the editor.
Author Response
Manuscript 1551195
Response to Reviewer2
Dear Editor, Assistant Editor and Reviewers,
Thank you for giving us the opportunity to submit a revised draft of the manuscript „Brenneria nigrifluens isolated from Aesculus hippocastanum L. bark in Hungary” for publication in the Journal Forests. We appreciate the time and effort that you and the reviewers dedicated to providing feedback on our manuscript and are grateful for the insightful comments on and valuable improvements to our paper. We have incorporated most of the suggestions made by the reviewers. Those changes are highlighted within the manuscript. Please see below, in green, for a point-by-point response to the reviewers’ comments and concerns. All page numbers refer to the revised manuscript file with tracked changes.
Reviewers’ Comments to the Authors:
Reviewer 2
1. Comment from Reviewer 2: The manuscript entitled „Brenneria nigrifluens isolated from Aesculus hippocastanum L. bark in Hungary“ describes identification of causal agent of longitudinal cracks on the trunk and branches and intensive brown liquid oozing on Aesculus hippocastanum trees. Causal agent belong to the genus Brenneria ie the closest phylogenetic relation with Brenneria nigrifluens according to phylogenetic analysis of 16S rRNA gene region. Authors have adequately addressed the following points, which include complete description of symptoms, the host, disease observation, the geographic locations where samples that were used in the study were collected, proof of strains pathogenicity, 16S rRNA strains sequencing, and re-isolation of bacteria from the inoculated plants, fulfilling Koch’s postulates.
Response: Thank you.
I have a few comments:
Comment from Reviewer 2: The technical aspects of the paper are not correct.
(1) References are not used as is stated in Instruction for Authors - “References must be numbered in order of appearance in the text (including citations in tables and legends) and listed individually at the end of the manuscript. In the text, reference numbers should be placed in square brackets [ ] and placed before the punctuation; for example [1], [1–3] or [1,3]. Similarly, authors should modify Section References;
Response: We thank the reviewer for pointing this out. We have made the change.
Comment from Reviewer 2: (2) Section Conclusion is missing;
Response: We apologize for the missing Section. We have added it to the manuscript. Please see page 12 of the revised manuscript, lines 22-30.
Comment from Reviewer 2: In Table 1 authors should cited references for presence of Brenneria species per countries;
Response: We thank for the reviewer for pointing this out. We have made the change. Please see pages 2-4 of the revised manuscript.
Comment from Reviewer 2: Although the authors confirmed the presence of Brenneria on Aesculus hippocastanum trees, it seems that phylogenetic analysis of the 16S rRNA gene region did not give clear species determination. Therefore, some other support for taxonomic identity of Hungarian strains is needed. I highly recommend sequencing more genes such as MLSA;
Response: We appreciate the reviewer’s insightful suggestion and agree that it would be useful to perform an analysis of the proposed housekeeping genes. Our results confirmed that we identified Brenneria species from horse chestnut trees and are grouped to Brenneria nigrifluens isolates, with the type strain from Germany, a walnut strain from Serbia and the highest identity was with a walnut isolate from Hungarian, formed a single branch on Maximum Likelihood phylogenetic tree. Our study’s aim was to draw attention to the fact that another pathogen is present in Aesculus hippocastanum, which have already been classified by IUCN as ‘Vulnerable’ species. This can be important information for professionals conducting research on horse chestnuts.
Comment from Reviewer 2: The manuscript requests extensive editing before it is published. I will leave the decision to the editor.
Response: We have sent the manuscript for revision.
We would like to thank the referee again for taking time to review the manuscript.
We look forward to hearing from you regarding our submission and to respond to any further questions and comments may have.
Sincerely yours,
Prof. Dr. László Palkovics
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
Well edited paper. I do not have further suggestions .
