Heterologous Grafting Improves Cold Tolerance of Eggplant
, Qian Li 1, Xin Wang 1, Xuefeng Li 1, Feng Liu 2 and Jianguo Yang 1,*
Round 1
Reviewer 1 Report
-The results of the MS Sustainability -1853205 are new for the understanding the molecular mechnism of improvement of cold tolerance in grafted eggplant, in particular and in heterologous grafted plants, in general. They are useful for further study of molecular mechanism underlying heterlogous grafted plants enhanced cold tolerance.
-The English of the MS is appropriate and understandable. However, there are many limitations in grammar, sentences and words as highlighted in reviewed MS.
-The cited referances are up to date and relevant. However, there are some references in the list (highlighted in reviewed MS) were not cited in the text. They need to be removed or cited in the text.
-The MS can be accepted for publication after revision based on reviwer's comments and sugestions
Comments for author File: 
Comments.pdf
Author Response
please see in the attchment
Author Response File: Author Response.pdf
Reviewer 2 Report
General comments:
The manuscript titled“ Heterologous Grafting Improves Cold Tolerance of Eggplant”， and investigate four grafting combinations of eggplant from cold-sensitive line J55 and cold-resistant line J65 were subjected to morpho-physiological experiments and transcriptome sequencing to compare their responses to cold stress. And all results suggest that grafting improves the cold tolerance of eggplant caused by material by increasing the enzyme activity and maintaining the stability of the plant. The paper were well structured but must be updated before publish.
I have some suggestions, as shown below:
1, line102 method 2.6， please indicate the method for cDNA and all the methods must be as detailed as possible.
2, In the result part, please keep consistent, Fig. or Figure?
3, what happened with table 1 and Figure 7.
4 so many minor mistakes in the manuscript and messed up, for example sometimes B6 and sometimes b6.
5 The figure legend is very short and not clear, please updated as much as you can
6. English writing must be updated, and the reference needs to be checked one by one.
Minor revision:
Line 8: However，there is
Line15: the sentence start from “Speculated that” seems incomplete sentence.
Line 19: B6
Line 21: extra space between through and increasing
Line22： The heterografte ialone also
Line 28: However, it is sensitive to low temperature
Line 45: Arabidopsis thalian should be italic
Line 49: what is CS/CS and CS/CF means here, write the full name when it is first appears
Line 52: what is SA stand for
Line 142: 5,230 and 6,222 genes, please keep consistent for the whole manuscript.
Line 156: Who speculated what?
Line162: Figure3 the labels ABCD can be on the top left, as same with other figures
Line 186: it is wired to start a sentence with numbers, can be there are 76 DEGs
Line 190: Aligned the figure well, same with figure 5, 6
Line 340, while bold here
Author Response
please see in the attachment
Author Response File: Author Response.pdf
Reviewer 3 Report
The article “Heterologous Grafting Improves Cold Tolerance of Eggplant” by Wang and colleagues describes the RNAseq analysis of grafting experiments between cold tolerant and susceptible eggplant lines.
I am sure the authors put a lot of effort in the generation of the RNAseq data, however, the article is incomprehensible in the state it is. I would seriously recommend the authors to counter-read the article, use some English editing, restructure everything and make new figure outlines before submitting it anywhere.
Major comments:
Introduction: There is not really a structure, basically the authors put everything in which is somehow remotely related to “cold tolerance”. The first paragraph is alright, maybe expand this a bit in a reasonable frame. The 2nd and 3rd paragraph look more like a discussion (maybe transfer most of it to there)?
Material and methods: You need a better description. What is normal growth temperature of eggplant? What is a 50 well petri-dish? What is the substrate you used?
The n number is missing for most experiments.
The RNAseq description I not good enough.
Why would anyone pick randome genes for qPCR confirmation? You are supposed to pick genes where you know that they are regulated by cold stress.
Results:
Root stock and scion of line J65 improve the chilling tolerance (which sounds very strange to me). Since the general idea of the grafting is adjustement of the cold tolerance by a root stock we can stop at this point.
Since it is unclear if line J55 has a higher yield/ is performing better or whatever, the project should have stopped there. The best performing line based on the given data would just be J65, and the grafting is unnecessary.
For the transcriptome, what is the sequencing depth and the genome coverage. How often did you repeat the rna seq? (based on a graph 3 times each?)
Identification of DEGs: It is tiresome to read, what kind of information does the reader get when you just list the number of Up/down DEGs? Please prepare the data in an appropriate way.
GO and KEGG analysis: You just listed the most accumulated DEGs and put them even in just one figure, same for the KEGG analysis. What is the meaning of this? Please prepare the data in an appropriate way.
The same is true for the metabolic pathways and transcription factors. Without the interpretation of the authors, the data are worthless.
The qPCR confirmation does not make sense with randomly picked genes. Please use proper genes for control.
Discussion:
I am not sure what you did there. You randomly make remarks about the regulation in other plants, without linking it properly to your research.
Author Response
please see in the attachment
Author Response File: Author Response.pdf
Round 2
Reviewer 3 Report
In the revision of the article “Heterologous Grafting Improved Cold Tolerance of Eggplant” Wang et al. describe their results of an RNAseq analysis of homo- and hetero-grafts of eggplants to identify the molecular mechanism behind cold tolerance.
The authors took great effort in rewriting the manuscript and changing the figures.
The general story line is now much better. Most of my previous points were answered in an acceptable form.
However, there are still a few points which need to be addressed.
1. The English, specifically in the newly added parts is in a non-acceptable form. The authors mentioned that they will use an English editing service after this revision round?
When this is the case it would be acceptable.
2. You changed the figures quite a lot, but they do not get better.
Don’t get me wrong, Figure 8 and 9 are great, good effort!
Figures 2, 3, 4, 5 need larger axis titles
Figures 6 and 7 are still not readable (Try to read the writing, its impossible). Especially the headers would be important
3. Yes, I understand your purpose of picking random genes to confirm your RNA seq data.
Still, that is not a good style. It is always prefereable to confirm it with some genes where you know the response. In your case some genes that are inducible by cold (either increase expression, or repression).
Here a list with some minor issues (not exhaustive, there is too much):
Line 13 -> What are specifical DEGs
Line 16 -18 -> So you listed all GO clusters of the DEGs, but why?
Line 23-23 -> Ah so the hormones are important, that is nice. I suppose they are in relationship to the KEGG pathway? When yes you should state that.
Line 25 -> self not sef , grafted not grated
Line 27-29 -> I do not understand the sentence.
Line 41-43 -> Please adjust the grammar.
Line 85 -> on large scale applications I suppose not “and”
Line 88-89 -> grammar
Line 93 -> what is figleaf gourd ?
Line 119 -> ”(“ is missing
Line 295 -> was not were
Line 296 -> need space after fullstop
Line 206 -> Differential expressed genes (DEGs)
Line 220/221 -> “significantly seriously” is not a thing
Line 221, Line 225 and other-> Beware “,” spacing rules
Line 281 ->grafted not grafed
Line 304 -> analyzed not anlysised
Line 328 -> That is not a complete sentence.
Line 334 -> yet another form of analyzed “analysede”
Line 592 -> wrong form of “analysed”
Figure 2: X and Y axis are too small to read. No size scale in panel A.
Figure 3: Axis descriptions are too small.
Figure 4: Axis descriptions are to small. In the legend you write something about 8 tissues. Not clear what you mean by that.
Figure 5: “Statistical of the differentially expressed genes” -> what do you mean by that?
Figure 6: Well now you have a different representation of the same data, which is as unreadable as the figure before. It is impossible to read, let alone to infer something from it.
Actually the figure (when someone would take the effort to understand it) carrys even less information than the previous figure.
Figure 6: Also not readable.
Figure 8 and 9: Much better!
Author Response
please see in the attachment
Author Response File: Author Response.pdf
