Enhancing Yield and Improving Grain Quality in Japonica Rice: Targeted EHD1 Editing via CRISPR-Cas9 in Low-Latitude Adaptation

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript with the title “Enhancing Yield and Improving Grain Quality in Japonica Rice: Targeted EHD1 Editing via CRISPR-Cas9 in Low Latitude Adaptation”, studied the two high-yield northeast Japonica genotypes (Jiyuanxiang 1 and Yinongxiang 12) with the purpose to edit the EHD1 gene, with goal of achieving higher adaptability across latitudes. The manuscript proposes these preliminary results could be helpful in obtaining elite rice genotypes faster, overcoming time-consuming traditional breeding methods that so far limited the expansion of japonica rice crops into the south. The results are novel.

 

Title: replace Enhangcing with Enhancing

 

Abstract provides a good summary of the manuscript and main finding.

 

Introduction:

Line 31, when scientific names of subspecies are given, these have to be preceded by scientific name of species: Oryza sativa.

Lines 65-70 present both the conclusion and recommendation. I suggest to only present at the end of the introduction the aim and objectives, and move these lines at the end of the manuscript as concluding remark.

 

 

Figure 4, components A and D – please mention in the caption the statistical test used to assign the letter case significance. Figure 4 I have not found cited anywhere in the text. Check also figure 3 and all the tables and figure to be cited in the text at their right place.

 

Material and Method – 4.8. for which data set was used the t-test? I find it mentioned first time here. Authors chose Duncan, a parametric test. Was the data distributed normally to ensure the appropriate test was used? If data was normally distributed a parametric ANOVA is applied first, which indicates if a post-hoc test (such as Duncan is needed), hence for traits from table 2, an ANOVA would indicate if needs to be explored further with a Duncan or not.

At Line 184, “traits did not show significant differences among these lines”, I suppose the authors found this based p values following ANOVA, and not directly with Duncan. Because p value would have indicated that is not worth applying a post-hoc test such as Duncan for those traits. Why two different test were used (t and Duncan)?

 

Given the novelty of the findings I suggest to introduce the p-values following ANOVA for each trait so that we know how far for significance ehd1 mutants were regarding agronomic productivity parameters.

 

Letter case assignment – when authors assigned the letter case significance, in Figure 2 (based on Duncan test) they assigned “a” to the lowest values, while at Figure 4A-B they assigned “a” to highest value (unknown test). Table 2, letter assignment gives highest value “a” still following Duncan test, different than the approach at figure 2 using same test. I strongly recommend to use the same system in all the manuscript and all the data – when assigning letters. A robust statistic is essential for the credibility of the findings.

 

Best regards.

Comments on the Quality of English Language

English grammar, syntax and style need moderate revision.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript authored by Song et al. describes gene editing in rice using the CRISPR/CAS9 system. The mutant plants, with Ehd1 edited, displayed enhanced agronomic phenotypes. The study is well-executed, and the data is excellent. However, the manuscript needs a revision due to the lack of details in most parts.

  In sum,   Line 33 and throughout the manuscript- ponica(space)[2].   Northeast and northeast. It should be consistent throughout the manuscript.   Line 77- remove "(LD)" as it is already introduced before.   Line 92- These variances suggest differing editing efficiencies across genetic backgrounds and rice varieties- nothing new    Except Table 1, Tables and Figures with statistical analysis need to provide the number of samples tested (n=?).   (WT) appeared three times throughout the MS. You can remove it Other than the initial introduction.   Figure 3 is missing a caption   Inconsistency of italic or not italic for mutant plant. Ex- ehd1 line 186   The discussion part is weak. The authors should discuss why the flowering time differed greatly in the mutant while grain quality went up. Do you have any thoughts or guesses? Why did gRNA1 and 2 show great differences in gene-edited rates? How can you improve this?    Plasmid construction- can you provide the final binary sequence with a map?  Comments on the Quality of English Language

Go over the manuscript a few times.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Dear authors,

most of the issues have been addressed in this version of the manuscript.

Best regards.

Comments on the Quality of English Language

some typing errors have been fixed by the authors.