A Gene Encoding a SHINE1/WAX INDUCER1 Transcription Factor Controls Cuticular Wax in Barley
Round 1
Reviewer 1 Report
Title: the title is not convenient. The data provided here are not enough to support the claim by the authors. Propose a more suggestive title rather than a definitive one with less evidence. This also applies to the language used in the abstract.
Abstract
The abstract is poorly written and does not give the expected information to reflect the claim by the authors in the title given to this manuscript. Consider revising
Introduction
- The title of the MS indicates that the target gene has a TF activity and functions in Hv. Yet, not a single statement talks about wheat and why the authors are interested in this particular TF.
- In addition, the authors should add a paragraph introducing TF in general, and TF or genes encoding wax in Hv
- The use of “in this paper” is not appropriate (line 60)
- Lines 60–66: I could not see the hypothesis the authors tried to reject and accept. Add the aim of the study.
- The HvWIN1 is suddenly mentioned at the end of the introduction with no roots.
M&M
- This section should be improved. Indicate why these particular cultivars and mutants were selected for this study.
- More information are required about the growth conditions. Describe the omitted details (lines 69–77)
- Line 75: indicate the source of the compost
- Lines 79–94: indicate the weight of samples for cuticle integrity assay. Specify if the samples were processed immediately after being harvest or were stored (indicate the storage conditions if applicable) prior to be processed further.
- Lines 87–88: add the nm to specify the wavelength of the absorbance
- Lines 90–92: use appropriate mathematical signs for x and minus
- Lines 94: how the authors justify the use t-test where they have parents lines and mutants lines (more than two groups). In this case, changes in the mutants are compared with the parents or WT. t-test is not appropriate to test the hypothesis. I urge the authors to use an appropriate statistical analysis and re-write the results. In addition, indicate which experiment design was used.
- Lines 105–106: describe the PCR reaction and condition here. Indicate the sequencing company details.
- Line 109: specify the weight of samples used for gene expression
- Line 118: simply use HvACT7
- Lines 124–125: I think the logic here should be …samples were frozen in liquid nitrogen and stored at –80 °C. Specify the number of replications
- Line 131: can use as previously described instead of as in…
- Line 140: replace the full stop with a colon. Re-write lines 141–145 to give a better sense. To my understanding, plants ensemble database was used to BLAST the SHINE proteins…did you use a single reference protein sequence or many?
- Line 157: write MEGA in full
- Was the data in Figure 2C generated by the present study or retrieved from the databases? Kindly specify the source
- It is not clear why the authors measure the gene expression as well as other components only in Bowman and BW407 and how the data compared for their significance. The results are confusing and should be clarify. The identity of Bowman and BW407 should have been described in subsection 2.1, as of now it is not clear.
- How many alleles were of the mutants were used in the study?
Discussion
- This section should be improved after improving the results section.
- A conclusion should be drawn.
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
##Minor points:
Lin3 33, what are VLCFAs?
Lines 30-35 rewrite for clarity
Line 76 what is the light intensity of the supplement light?
Line 95 do not start a sentence with an abbreviation
Major points
The materials methods section is quite confusing and not well written:
- Based on the Supplementary table, you have 44 Germplasm but for some experiments are performed on a few germplasm without any explaination.
- We do not know in which germplasm for instance cuticule integrity measument and gene expression was done
- It is not clearly stated why data analysis (sequence retrieval and phylogeny, haplotype analysis) was done.
In sum, we do know understand your experimental design.
Author Response
Please see the attachment
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
The authors have improved the manuscript, and I appreciate their answers.
Author Response
Please see the attachment
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Line 98: Twenty-five instead of 25
Line 114: You should remove [44]
- Materials and Methods: 2.1. You should add in the main manuscript a sample table containing only the main germplasms used in this study for ease reading.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
