CsAFS2 Gene from the Tea Plant Intercropped with Chinese Chestnut Plays an Important Role in Insect Resistance and Cold Resistance
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe proposed article is undoubtedly interesting from the point of view of studying the mechanisms of plant resistance to various stress influences. The results of the study are presented consistently and supported by figures and tables.
However, the performance can be improved.
You will find specific comments below:
1. The abstract does not mention an important result about the role of trichomes and their interaction with the α-gene responsible for farnesene.
2. In the introduction, the information given on lines 46-53 is unnecessary.
3. In scientific articles, it is customary to use abbreviations for compounds presented for the first time. Therefore, the names of hormones should be deciphered.
4. The list of references is very carelessly designed. Authors should clarify all references listed in the list of cited literature. As an example, I note that the text does not mention works 1, 2,3, 76,7,9, etc.
Author Response
Thank you very much for taking the time to review this manuscript. We greatly appreciate your valuable feedback and constructive comments. These insights have significantly contributed to the improvement of our manuscript. We have carefully considered each suggestion and have made substantial revisions to address the concerns raised. In this revised version, changes to our manuscript were all highlighted by yellow colored text in the manuscript, and the incorrect English expressions in the manuscript have been corrected the modified parts have been highlighted by blue.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsAbstract. Section "stress regulation mechanisms in tea plants" which species does this part of the "Abstract" refer to? It is unclear which specific species the name of the tea plants refers to, whether Castanea mollissima or Camellia sinensis; specify which genus and species you are referring to.
Introduction. α-Farnesene is produced in tea plants in response to different herbivore attacks; In a study with Camellia sinensis (DOI: 10.3390/ijms20174151), α-farnesene synthase (CsAFS) was functionally characterized, but not in Castanea mollissima.
Expand the background of insect resistance of tea plants, with recent publications:
1) Nie, X., Zhao, S., Hao, Y. et al. Transcriptome analysis reveals key genes involved in the resistance to Cryphonectria parasítica during early disease development in Chinese chestnut. BMC Plant Biol 23, 79 (2023). https://doi.org/10.1186/s12870-023-04072-7
2. Lombardero MJ, Ayres MP, Álvarez-Álvarez P and Castedo-Dorado F (2022) Defensive patterns of chestnut genotypes (Castanea spp.) against the gall wasp, Dryocosmus kuriphilus. Front. For. Glob. Change 5:1046606. doi: 10.3389/ffgc.2022.1046606
Results. The results of the vital role of CsAFS2 in improving cold and insect resistance are well shown in figures and tables.
Discussion. The compound α-Farnesene and α-farnesene synthase (CsAFS) are not included in the Discussion.
References. Update References
Additional comments:
1. What is the main question addressed by the research?
The main question addressed by the research is to know the regulatory mechanisms that affect stress in tea plants by growing stress-resistant tea plants.
2.1. What parts do you consider original or relevant for the field?
I consider that the original part of the study is to know the mode of expression of the AFS genes in tea plants interplanted with Chinese chestnut.
2.2. What specific gap in the field does the paper address?
The specific gap in the field that the article addresses is that the study was carried out in tea plantation intercropped with Chinese chestnut
3. What does it add to the subject area compared with other published material?
The study contributes little to the topic compared to other published articles because there is a report (doi: 10.3389/ffgc.2022.1046606) indicating that genetic resistance to chestnut wasp of chestnut genotypes is not related to terpenes (α-farnesene) as indicated in this study.
4. What specific improvements should the authors consider regarding the methodology?
In the methodology, the content of the sesquiterpene (α-farnesene) that is related to plant defense is not characterized in the experimental treatments carried out.
4.1 What further controls should be considered?
Under growth conditions, the culture media are barely described, there is no data on what MS, BA and NAA are, although they are known in plant tissue culture, they must be described.
5.1. Please describe how the conclusions are or are not consistent with the evidence and arguments presented.
The conclusions have little consistency with the evidence and arguments presented, since more studies on the increase in resistance to cold and insects in tea plants are required to understand these interactions in tea plantations intercropped with Chinese chestnut.
5.2 Please also indicate if all main questions posed were addressed and by which specific experiments.
The main questions raised were addressed through the specific experiments
6. Are the references appropriate?
No, the references must be updated
7. Please include any additional comments on the tables and figures and quality of the data.
In Figures 7 and 8 the letters are small and blurry, which makes them difficult to read, the others are fine along with the Tables.
Author Response
Thank you very much for taking the time to review this manuscript. We greatly appreciate your valuable feedback and constructive comments. These insights have significantly contributed to the improvement of our manuscript. We have carefully considered each suggestion and have made substantial revisions to address the concerns raised. In this revised version, changes to our manuscript were all highlighted by yellow colored text in the manuscript, and the incorrect English expressions in the manuscript have been corrected the modified parts have been highlighted by blue.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for Authors Dear Author,I have read the manuscript. Following are my suggestions.
1. What is the difference between deforestation and exploiting. Change this sentence.
2. research that show that the AFS. Change this sentence.
3. I can see the SAS result and it is missing in 1d. Where is standard error bar ?
4. What is the significance of Figure 2. If the gene was already selected based in the previous studies or literature, than it can be avoided. Gene Cloning and Sequence analysis not needed.
5. Don't start sentence with numeric. 20 T1 tobacco.
6. What is the transgenic efficiency ?
7. Figure 4, no SAS and SE
8. empty vector the control tobacco plant, means ?
9. Figure 6 can be given in bigger size. Scale is not visible.
10. ( i ) empty vector the control tobacco plant; ( ii-iv ) transgenic tobacco lines. ?
11. Figure 7 and 8, why SAS was performed separately between 25 C and 4 C.
12. obvious blue discolouration ? Change to blue staining.
13. Table 1. Amount -> Number
14. Log2FC, why no SE ?
15. Tender leaves ? Any numbers like 3rd leaves from top.
16. What is followed by gel cutting and recovery ?
17. Change Twenty 4-year-old wingless to Twenty numbers of 4-year-old wingless.
18. Which test was used in SPSS. ANOVA, p-value ?
All the best!
Comments on the Quality of English Language
It needs extensive English language correction.
Author Response
Thank you very much for taking the time to review this manuscript. We greatly appreciate your valuable feedback and constructive comments. These insights have significantly contributed to the improvement of our manuscript. We have carefully considered each suggestion and have made substantial revisions to address the concerns raised. In this revised version, changes to our manuscript were all highlighted by yellow colored text in the manuscript, and the incorrect English expressions in the manuscript have been corrected the modified parts have been highlighted by blue.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsReview
Article «CsAFS2 Gene from the Tea Plant Intercropped with Chinese Chestnut Plays an
All my comments have been taken into account. Corresponding changes have been made to the text.
Reviewer 2 Report
Comments and Suggestions for AuthorsAbstract. Section "stress regulation mechanisms in tea plants" which species does this part of the "Abstract" refer to? It is unclear which specific species the name of the tea plants refers to, whether Castanea mollissima or Camellia sinensis; specify which genus and species you are referring to.
Answer. tea plants (Camellia sinensis) intercropped with Chinese chestnut (Castanea mollissima)
Introduction. α-Farnesene is produced in tea plants in response to different herbivore attacks; In a study with Camellia sinensis (DOI: 10.3390/ijms20174151), α-farnesene synthase (CsAFS) was functionally characterized, but not in Castanea mollissima.
Answer. Additionally, α-farnesene was accumulation rose notably in tea plants infested by various pests (tea tapeworm, tetranychus kanzawai, tea looper, or tea green leafhopper) [5-7]
Discussion. The compound α-Farnesene and α-farnesene synthase (CsAFS) are not included in the Discussion.
Answer. 3.4 CsAFS2 Might Enhance Insects and Cold Resistance in Plants by Releasing α-Farnesene.
References.References were updated with suggested citations
