Genome-Wide Identification of DREB Transcription Factor Family and Functional Analysis of PaDREB1D Associated with Low-Temperature Stress in Phalaenopsis aphrodite

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

why to study only PaDREB1D  (the authors identify  31 genes, and choose only one, PaDREB1B, of them for further analysis.

The authors needs more justification in the results section that make a clear-cut idea why to follow only this gene, the argument I could not found and for me is not clear in the manuscript. Moreover beside PaDREB1B, in the phylogenetic-tree PaDREB1C is closely related to PaDREB1B, why the authors only use one gene? And it would very interesting to use other PaDREB homologue that did not result in the same phenotypes resulting from the PaDREB1B overexpression.

Without this experimental context for me is very difficult to understand the meaningful of the results.

Author Response

Dear reviewer,

Thank you for your valuable comments!

First of all, we need to make it clear: we can understand what you said about the close relationship between 1B and 1C genes in evolutionary development, but in this paper we have been studying the gene function of 1D.

Secondly, our explanation to your question is as follows:

As you can see, the structure of this paper is as follows: identification to Phalaenopsis DREB gene family members (31) → and then targeting to 4 DREB1 subgroup members → then try to overexpress DREB1 genes in PLBs and to test gene functional validation.

The reason why we only did the functional validation of PaDREB1D is as follows: we tried to overexpress 4 genes (in PLBs), but the only one that finally succeeded and could have enough samples for the functional validation is PaDREB1D. This has to do with the success rate of the genetic transformation system, and it is not that we didn't want to do it. So, starting with result 3.3, we have only released the results of PaDREB1D related studies.

As a plant lover, I hope that each of my transgenic plants can grow healthily; as a botanical experimenter, I also hope that my work can be perfect. But the reality is so cruel that only PaDREB1D of the transgenic work was successful during my working period, which I must say was surprising, but not perfect as you say.

The above is the whole situation, I hope you can understand.

We make the following additions: We explain this situation in the discussion section (line 422), the revised parts in the manuscript have been marked in red.

Thank you again for your correction and feedback!

Hope you have a nice day!

Your sincerely, 

authors

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Line 108: Replace qRT-PCR with qPCR

Line 105: Description of Real‑Time Quantitative PCR (RT‑qPCR) method should include details such as the concentration of template or qPCR validation data, according to "Minimum Information for Publication of Quantitative Real-Time PCR Experiments" published by Bustin in 2009.

Regarding phylogenetic analysis, why does the phylogenetic tree not show the bootstrap values? How many bootstrap replicates were conducted?

Author Response

Dear reviewers:

Thank you for your valuable comments.

In response to your comments, we have made the following reply (t he revised parts in the manuscript have been marked in red)

1     Line 108: Replace qRT-PCR with qPCR

We have changed it to qPCR at line 108.

2     Line 105: Description of Real‑Time Quantitative PCR (RT‑qPCR) method should include details such as the concentration of template or qPCR validation data

We apologize for overlooking this point and have made corresponding additions in Method 2.2 (line 105).

3     Regarding phylogenetic analysis, why does the phylogenetic tree not show the bootstrap values?  How many bootstrap replicates were conducted?

We apologize for missing this point when writing the manuscript, and we have made a correction on line 143.

 

Thank you again for your correction and feedback!

Hope you have a nice day!

Your sincerely,

authors

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Based on the corrections and comments from the authors, I accept the manuscript.