Identification and Fine Mapping of Osdsm3, a Drought-Sensitive Gene in Rice (Oryza sativa L.)

Round 1

Reviewer 1 Report

In their manuscript, Deng et al. characterized the drought-sensitive mutant, dsm3, using EMS. The authors successfully mapped the location of the Osdsm3 gene and provided some data on its regulatory role in the drought stress mechanism. The clarity of the data and the quality of writing are commendable. My main concern centers around whether introducing Osdsm3 into the dsm3 mutant could restore its drought-sensitive phenotype or influence the expression of marker genes. This portion of the data is pivotal in supporting the central claim of the manuscript.

 

Specific Comments:

 

Line 14-16:  It is imperative to provide the full names of EMS, MDA, Pn, POD, and SOD for clarity.

 

Line 25: "Arabidopsis thaliana" should be italicized for proper formatting.

 

Line 31: "Oryza sativa" should be italicized as well.

 

Line 189: Consider adding a description detailing how the dsm3 mutant was identified through EMS treatment.

 

Line 191, Fig 1A:  What is the phenotype of the dsm3 mutant under normal growth conditions? Are there any developmental differences between the dsm3 mutant and WT plants?

 

Line 200:  The statement "These findings suggest that the withered leaf-tip in dsm3 has a negative impact on its growth, development, and rice yield" is inaccurate. The results only demonstrate reductions in various agronomic traits for the dsm3 mutant; there is no evidence connecting these reductions to the withered leaf-tip.

 

Fig 2C-E:  Statistical information for these panels needs to be included.

 

Fig 2F-I:  The titles of these panels are in mg/g or U/mg, while the Y-axis represents percentages. The titles should be revised for consistency.

 

Line 220-222: Consider rephrasing the sentence as follows: "This suggests that differences in root water use efficiency between ZH8015 and dsm3 may have contributed to the observed significant decrease in maximum root length and root-shoot ratio in dsm3."

 

Fig 3C:  Providing length and width values for both WT and dsm3 would allow for a better assessment of cell size differences between the two.

 

Line 255: Is there a reference for this statement? Please provide a source if applicable.

 

Line 262: For Figure 4E-H, the statistical significance needs to be indicated in the figure or accompanying text.

 

Line 265: Suggest revising to: "while SOD activity also exhibited a notable reduction."

 

Line 294: Could information about the domain of the GIL1 protein and any conserved domains that might hint at its function be provided?

 

Line 302:  Should be "No. of normal plants".

 

Line 319-321:   A clearer phrasing could be: "Based on these findings, we speculate that genes related to drought resistance are significantly more induced in response to drought stress in dsm3 than in the WT."

 

Fig 6C: The figure legend for the " tissue-specific expression of Osdsm3" should clarify which gene was used as the control and set to a relative expression value of 1. This information is essential for interpreting the data accurately.

 

Line 340:"Arabidopsis thaliana" should be italicized.

 

Line 347: Suggest revising to: "the fluorescence signal of GFP."

 

Line 348-351: The claim that DSM3-GFP localizes to the cytoplasm while free GFP is distributed throughout the cell is not evident in Figure 7B. Both DSM3-GFP and free GFP appear to be located on the plasma membrane. Additionally, the quality of Figure 7B does not provide a clear conclusion. A higher quality figure is needed for clarity.

 

The manuscript's English language quality is acceptable, and I have no difficulty comprehending the authors' main points.

Author Response

Dear Reviewer,

We would like to express our sincere gratitude to you for your thoughtful and constructive comments on our manuscript entitled "Identification and Fine Mapping of Osdsm3, a Drought-Sensitive Gene in Rice (Oryza sativa L.)". We have carefully considered all the comments and made significant revisions to the manuscript accordingly. Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Deng et al. report "Identification and Fine Mapping of Osdsm3,,,," In their report they  discovered a drought-sensitive mutant, designated as dsm3, arising from the progenies of indica rice variety Zhonghui8015 treated with EMS. Under drought stress conditions, dsm3 exhibited increased levels of MDA and H2O2, reduced Pn, and decreased activity of POD and SOD. Genetic analysis revealed a single recessive gene, designated as Osdsm3. Sequencing analysis unveiled a mutation 20 (1275G→A) in the exon of the candidate gene (LOC_Os01g10680), leading to premature translation termination. Phylogenetic analyses indicated that Osdsm3 shares evolutionary homology with UNE1, an intracellular transport protein found in Arabidopsis thaliana.

 

Major

1.     L103-105. The molecular mechanisms and genetics in relation to abiotic stress have been broadened. The authors draw an abrupt conclusion against tons of biblical reports in the area.  

2.     L104-108 Some of genetic evidence is provided? Currently, knock-out technology is available.  Those evidence is critical to draw the hypothesis of the authors.

3.     L220-228. The Fig 2F is not described. In addition, The graphs, Fig 2G -2I are not really described.

4.     Table2. How were the mutants measured among F2 population? Was the conclusion drawn based on either sequence observation or phenotype count?  

 

 

Minor

L14. What is MDA?

L56. Ref 12 and 13 represent 44 drought genes?

L103-105. The molecular mechanisms and genetics in relation to abiotic stress have been broadened. The authors draw an abrupt conclusion against tons of biblical reports in the area.  

L114. Provide the pedigree of 02428. At least even the parental cultivars of the line.

L282. If the gene is recessive, the symbol has to be small characterized.

Author Response

Dear Reviewer,

We would like to express our sincere gratitude to you for your thoughtful and constructive comments on our manuscript entitled "Identification and Fine Mapping of Osdsm3, a Drought-Sensitive Gene in Rice (Oryza sativa L.)". We have carefully considered all the comments and made significant revisions to the manuscript accordingly. Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

I reviewed the study titled "Identification and Fine Mapping of Osdsm3, a Drought-Sensitive Gene in Rice (Oryza sativa L.)". The authors have done a good job, but it still needs some refinement before publication.

 

All my observation is highlighted directly in the text.

 

I wish the authors success.

Comments for author File: Comments.pdf

Make minor changes, following the journal's guidelines.

Author Response

Dear Reviewer,

We would like to express our sincere gratitude to you for your thoughtful and constructive comments on our manuscript entitled "Identification and Fine Mapping of Osdsm3, a Drought-Sensitive Gene in Rice (Oryza sativa L.)". We greatly appreciate your valuable suggestions. We have meticulously incorporated your recommendations, including improvements in writing format, icon colors, and the addition of references. Please refer to the revised manuscript for a detailed overview of these changes.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

The report has a limitation in proving the function of the gene and it has to be addressed. 

Author Response

Dear Reviewer,

We highly appreciate your suggestion. Indeed, conducting gene function validation would significantly enhance the reliability of our results. We plan to undertake gene function validation in future work. As this submission primarily focuses on the identification and fine mapping of the drought-sensitive gene Osdsm3 in rice, and has not yet reached the cloning stage, we regret that we may not be able to provide gene function validation in this study.