Analysis of the Alternative Splicing Events of Exogenous δ-Aminolevulinic Acid under NaCl Stress in Wild Jujube Seedlings

Round 1

Reviewer 1 Report

I have provided detailed and specific comments in the attached pdf file.

Comments for author File: Comments.pdf

Author Response

Please see the attachment

Author Response File: Author Response.docx

Reviewer 2 Report

The article is written with great care. In this work, Zhu et al. analyzed effect, especially alternative splicing events, of exogenous ALA under NaCl stress in wild jujube seedlings. They hypothesized that ALA treatment can alleviate the effects of salt stress on chlorophyll by altering the alternative splicing of genes related to photosynthesis, cystoid, chlorophyll metabolism and energy metabolism in wild jujube. In addition, the chlorophyll-related metabolites and genes of enzymes were verified in wild jujube.

Specific comments:

1.       Add the reference for the treated method and concentrations of NaCl and ALA (lines 107-108).

2.       The sampling time are confusing to me, sample at the sixth day of treatment is for RNA-sequencing? Why choose 24 h, 48 h, and 72 h for verification? (lines 113-114, 119)?

3.       How many biological replicates (line 116)?

4.       Add list panels with a, b, … in Figure 1, 2 and 4, together with figure legends. The figure legends are not sufficiently detailed.

5.       Uniform the four treatments in figures: CK or denoted as A; ALA or denoted as B; NaCl or denoted as C; NaCl + ALA, or denoted as D.

6.       Wrong description of Figure 2 and Table 5.

7.       Some values of listed enriched metabolic pathways were over P-value ≤ 0.05.

8.       In 3.5, authors mainly analyzed comparison groups of other treatments with CK, while comparison ALA vs. NaCl + ALA and NaCl vs. NaCl + ALA could be researched.

Author Response

Please see the attachment

Author Response File: Author Response.docx

Reviewer 3 Report

Overall, the manuscript is well written. The topic of this study is suitable for the journal and the results are of potential interest for scientific community. Methodology followed is clearly described. Obtained results have been nicely discussed in light of the recent publications in the area. However, there are also some flaws should be improved.

Some comments:

1. The authors should combine 2.1 and 2.2.

2. Please provide the model of RNA-seq platform.

3. In the 2.5.1 part, the author should add the Ref. of jujube genome.

4. Please confirm the reliability of internal reference gene.

5. Please state the biological and technical replicates in materials and methods part.

6. In 3.2, please provide the mapping rate of transcriptome and genome data.

7. Where the results of the RT-qPCR ?

8. Fig.1 and Fig. 4, SD or SE ? Please correct.

Author Response

Please see the attachment

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Thank you so much for your patience and perseverance in addressing my comments. I suggest the manuscript can be accepted in its present form.

Author Response

Thank you very much for your patient and scientifically rigorous guidance, it has been very beneficial and finally thank you for your recognition.

Reviewer 3 Report

The manuscript can be accepted in this version.

Author Response

Thank you very much for your patient and scientifically rigorous guidance, it has been very beneficial and finally thank you for your recognition.