Isolation of FLOWERING LOCUS C and Preliminary Characterization in the Floral Transition of Xinjiang Precocious Walnut
Round 1
Reviewer 1 Report
Dear Authors,
Horticulturae-2398086
The brief report entitled „Isolation of FLOWERING LOCUS C and preliminary characterization in the floral transition of Xinjiang precocious walnut“ represents an important news on isolation, sequence alignment, and expression analysis of FLC gene homolog in precocious walnut (Juglans regia) native to Xinjiang region in western China. The results of the expression analyses using JrFLC overexpression in a heterologous system of A. thaliana revealed an analogous role of FLC in walnut to FLC in A. thaliana revealing delayed flowering and decreased expression of flowering-related genes such as SOC1 and LFY.
I can recommend this short report for publication in Horticulturae.
However, I have a few important comments on the present manuscript:
In Figure 2A-F and Figure 3B showing various micro-photos at various scales, appropriate scale bars have to be added.
In Figure 3C,D,E, asterisks indicating significant differences have to be adequately explained in the figure legend, i.e., the kind of statistical test used and the level of statistical significance have to be given. In addition, a short section on the data statistical evaluation has to be added to Materials and methods section providing basic information on the statistical software used, statistical method, i.e., kind of statistical test and level of statistical signifikance used for the data evaluation.
Final recommendation: Accept after a minor revision.
Comments for author File: 
Comments.docx
Author Response
#R1
The brief report entitled „Isolation of FLOWERING LOCUS C and preliminary characterization in the floral transition of Xinjiang precocious walnut“ represents an important news on isolation, sequence alignment, and expression analysis of FLC gene homolog in precocious walnut (Juglans regia) native to Xinjiang region in western China. The results of the expression analyses using JrFLC overexpression in a heterologous systém of A. thaliana revealed an analogous role of FLC in walnut to FLC in A. thaliana revealing delayed flowering and decreased expression of flowering-related genes such as SOC1 and LFY.
I can recommend this short report for publication in Horticulturae.
Response: We sincerely thank the reviewer for the supportive comments.
However, I have a few important comments on the present manuscript:
In Figure 2A-F and Figure 3B showing various micro-photos at various scales, appropriate scale bars have to be added.
Response: We have fixed it. Please see these newly added bars in new revised Figures, and descriptions in the revised figure legends (highlighted in Red).
In Figure 3C,D,E, asterisks indicating significant differences have to be adequately explained in the figure legend, i.e., the kind of statistical test used and the level of statistical significance have to be given. In addition, a short section on the data statistical evaluation has to be added to Materials and methods section providing basic information on the statistical software used, statistical method, i.e., kind of statistical test and level of statistical signifikance used for the data evaluation.
Response: We have revised it accordingly. Please see it in the revised descriptions related to Figure 3C,D,E. Moreover, we have also added these descriptions in the Materials and Methods (see Line 112-115).
Final recommendation: Accept after a minor revision.
Response: We again thank the reviewer for above supportive comments.
Author Response File: Author Response.docx
Reviewer 2 Report
In this brief report the key flowering-time gene FLOWERING LOCUS C (FLC) was isolated in two precocious Xinjiang walnuts varieties. Furthermore, a preliminary characterization of the floral transition is provided. More specifically, previous findings along with the new achieved results showed that FLC in the considered varieties plays an important role in regulating the floral transition. Therefore, the work is very interesting leading to the individuation of a strong candidate gene for the genetic improvement of flowering time in walnut, even though further studies are needed to confirm this preliminary result.
The activities developed in this study are well detailed.
However, a clearer description of the overall experimental design and the adopted methodological approach would be appreciated.
Furthermore, an integration with the results achieved through the previous RNA sequencing work carried out by the same laboratory is suggested.
Overall, the work deserves to be published after minor revisions.
Minor editing of English language required
Author Response
#R2
In this brief report the key flowering-time gene FLOWERING LOCUS C (FLC) was isolated in two precocious Xinjiang walnuts varieties. Furthermore, a preliminary characterization of the floral transition is provided. More specifically, previous findings along with the new achieved results showed that FLC in the considered varieties plays an important role in regulating the floral transition. Therefore, the work is very interesting leading to the individuation of a strong candidate gene for the genetic improvement of flowering time in walnut, even though further studies are needed to confirm this preliminary result.
The activities developed in this study are well detailed.
Response: We sincerely thank the reviewer for the supportive comments.
However, a clearer description of the overall experimental design and the adopted methodological approach would be appreciated.
Response: We have fixed it. Please see the revision listed below (also see Line 56-58):
In this Brief Report, we isolated the key floral transition gene FLC and measured its dynamic expression levels at multiple growth stages. Furthermore, we confirmed its molecular function via transgenic expression in wild type (Col-0) of A. thaliana.
Furthermore, an integration with the results achieved through the previous RNA sequencing work carried out by the same laboratory is suggested.
Response: We have added discussion for the integration of previous RNA-seq and this study. Please see this addition in new revised mansucript. Please see Line 193-200.
Overall, the work deserves to be published after minor revisions.
Response: We again thank the reviewer for above supportive comments.
Author Response File: Author Response.docx
Reviewer 3 Report
The report "Isolation of FLOWERING LOCUS C and Preliminary Characterization in the Floral Transition of Xinjiang Precocious Walnut" discusses the identification and characterization of a crucial gene involved in the regulation of flowering time in precocious walnuts. The authors show that this gene, called JrFLC, plays an important role in regulating the transition from vegetative growth to reproductive growth in walnut trees.
The article is well-written and provides a clear and concise summary of the research findings. The authors use a variety of techniques, including sequence alignment, gene expression analysis, and overexpression studies, to demonstrate the function of JrFLC in regulating flowering time in walnuts. The results of the study suggest that JrFLC acts as a floral repressor, delaying the onset of flowering in precocious walnuts and increasing the number of leaves produced.
The study has important implications for the breeding of precocious walnuts, as it identifies a potential target for genetic improvement of flowering time. The authors suggest that further study of JrFLC is needed to fully understand its function and potential for genetic manipulation.
Overall, the article provides valuable insights into the molecular mechanisms underlying flowering time regulation in walnut trees and highlights the importance of precocious germplasms for enhancing breeding efficiency. The study adds to our understanding of plant development and has important implications for the agricultural industry.
I only have a few minor suggestions regarding the text.
1) Please, provide the initials of the author of the species’ name when mentioning it for the first time (e.g. Juglans regia L.).
2) The English style is good, although minor punctuation corrections are needed.
3) In the Abstract, please explain what Xinjiang is (a variety, a region?)
4) Names of varieties should be written in ‘’.
5) Keywords should be written alphabetically and not repeat words from the title.
6) In the materials and methods, web pages should be presented as references, according to the MDPI formatting style.
7) Line 102: do not start a sentence with an abbreviation.
8) Photographs lack scale bars.
9) In the graphs, it is unclear if we see standard errors or standard deviations.
10) DOI numbers are missing in the Reference list.
After making all the necessary changes, the article can be accepted for publication.
The English style is good, although minor punctuation corrections are needed.
Author Response
#R3
The report "Isolation of FLOWERING LOCUS C and Preliminary Characterization in the Floral Transition of Xinjiang Precocious Walnut" discusses the identification and characterization of a crucial gene involved in the regulation of flowering time in precocious walnuts. The authors show that this gene, called JrFLC, plays an important role in regulating the transition from vegetative growth to reproductive growth in walnut trees.
The article is well-written and provides a clear and concise summary of the research findings. The authors use a variety of techniques, including sequence alignment, gene expression analysis, and overexpression studies, to demonstrate the function of JrFLC in regulating flowering time in walnuts. The results of the study suggest that JrFLC acts as a floral repressor, delaying the onset of flowering in precocious walnuts and increasing the number of leaves produced.
The study has important implications for the breeding of precocious walnuts, as it identifies a potential target for genetic improvement of flowering time. The authors suggest that further study of JrFLC is needed to fully understand its function and potential for genetic manipulation.
Overall, the article provides valuable insights into the molecular mechanisms underlying flowering time regulation in walnut trees and highlights the importance of precocious germplasms for enhancing breeding efficiency. The study adds to our understanding of plant development and has important implications for the agricultural industry.
Response: We sincerely thank the reviewer for the supportive comments.
I only have a few minor suggestions regarding the text.
1) Please, provide the initials of the author of the species’ name when mentioning it for the first time (e.g. Juglans regia L.).
Response: Done. Thanks!
2) The English style is good, although minor punctuation corrections are needed.
Response: We have improved the language of our manuscript followed by a languge service, with a twice improvement. Thus, for other types of minor corrections, we believe that, it can be fixed during the proof stage.
3) In the Abstract, please explain what Xinjiang is (a variety, a region?)
Response: We have fixed it. Now it was ‘the precocious walnuts of Xinjiang region’.
4) Names of varieties should be written in ‘’.
Response: We agree with your scientific rigorism, but actually all variety names have been written in ‘’ already in our initial version. Thanks!
5) Keywords should be written alphabetically and not repeat words from the title.
Response: We agree with your scientific rigorism. Done. Many thanks!
6) In the materials and methods, web pages should be presented as references, according to the MDPI formatting style.
Response: Thanks. we have added two refs [15], [17], and [18].
7) Line 102: do not start a sentence with an abbreviation.
Response: We have fixed it. Now it was ‘Primers of RT-qPCR...’
8) Photographs lack scale bars.
Response: Similar to comment by #R1, we have fixed it.
9) In the graphs, it is unclear if we see standard errors or standard deviations.
Response: We have fixed it. Now in the figure legends, we have added ‘bars indicated the standard deviations’.
10) DOI numbers are missing in the Reference list.
Response: Done.
After making all the necessary changes, the article can be accepted for publication.
Response: We again thank the reviewer for above supportive comments.
