Exploring Polyploidization in Nigella sativa L.: An Applicable Strategy Towards Crop Improvement

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript“Exploring Polyploidization in Black Cumin: Comparison of the Morphology, Cytology, Yield Characteristics, and Thymoquinone Content in Diploid versus Tetraploid Plants” has strong practicality and I agree to publish it after modification.  Several aspects need to be modified.
1. the abstract content needs to add some data support and description of the results, not just ordinary textual descriptions.
2. there is repetition in the writing content, with 2.2 and 2.6 being completely identical.
3. Figure 5 should indicate the peak time of Thymoquinone in Figures a, b, and c.
4.it is suggested to adjust the writing logic of the manuscript. Firstly, the difference in ploidy between diploid and tetraploid should be demonstrated through cytology and flow cytometry. Then, the differences in germination rate, morphology, and yield between diploid and tetraploid should be introduced, followed by the difference in Thymoquinone content. This way, the logic of the manuscript is more reasonable, and the corresponding materials, methods, and discussion order are more in line with logic.

Author Response

Response to Reviewer # 1

Comments and Suggestions for Authors

The manuscript “Exploring Polyploidization in Black Cumin: Comparison of the Morphology, Cytology, Yield Characteristics, and Thymoquinone Content in Diploid versus Tetraploid Plants” has strong practicality and I agree to publish it after modification.  Several aspects need to be modified.

Response: Thank you for your encouragement words, also many thanks for your valuable comments which aimed to improve the MS. All changes or corrections you asked us to do are highlighted in yellow in the manuscript.  
1. The abstract content needs to add some data support and description of the results, not just ordinary textual descriptions.

Response: Ok, done
2. There is a repetition in the writing content, with 2.2 and 2.6 being completely identical.

Response: Thank you for the valuable notification, I deleted 2.6., and the Statistical analyses became 2.6.  then became 2.7. due to the logic rearrangement of titles in M and M section as you suggested to do  
3. Figure 5 should indicate the peak time of Thymoquinone in Figures a, b, and c.

Response: I added the Retention time (RT) of Thymoquinone in Figures a, b, and c.

4. It is suggested to adjust the writing logic of the manuscript. Firstly, the difference in ploidy between diploid and tetraploid should be demonstrated through cytology and flow cytometry. Then, the differences in germination rate, morphology, and yield between diploid and tetraploid should be introduced, followed by the difference in Thymoquinone content. This way, the logic of the manuscript is more reasonable, and the corresponding materials, methods, and discussion order are more in line with logic.

Response: I am totally agree with you, done

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript is a research article on the effect of polyploidization on morphology, cytology, yield Characteristics, and thymoquinone content of Black cumin. The manuscript is interesting and publishable, and could contribute to the effective breeding of Nigela sativa.   I have some concerns/ comments for the authors.

1. The title should be revised. It is ambiguous and unnecessarily long.
2. The scientific name of black cumin should be used in the title instead of English. Black cumin could be something in some parts of the world.
3. Abstract: The needs revision. I have provided an elaborate comment in the annotated manuscript.
4. The introduction is well written and provides a good rationale and objective for this study.
5. Methodology: Information on how the seeds were sourced and their accession number should be added to the methodology. This is impratnt for the reproducibility of this study.
6. Use the formula function in Microsoft Word to write the formulas appropriately.
7. Tables 1 and 2 are not necessary. Summarise how you carried out the study succinctly in the text. Table 2 could be useful only if you relate your data to varying environmental conditions
8. Results: Line 281: What do you mean by previous parameters? May be measured parameters?
9. Discussion: Line 386-387; provide the reference for the statement.
10. The discussion needs to be improved. The discussion should focus on your own results and not discussing already published works.

The conclusion needs to be improved. Authors should focus on drawing a conclusion from their results and the implications of this study to the black cumin breeding program.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

The manuscript should be edited appropriately, and scientific names mentioned in the text should be properly written with their authority. 

Author Response

Response to Reviewer # 2

Comments and Suggestions for Authors

The manuscript is a research article on the effect of polyploidization on morphology, cytology, yield Characteristics, and thymoquinone content of Black cumin. The manuscript is interesting and publishable, and could contribute to the effective breeding of Nigella sativa.   I have some concerns/ comments for the authors.

Response: Thank you for your supporting words, also many thanks for your valuable comments which aimed to improve the MS. All changes or corrections you asked us to do are highlighted in yellow in the manuscript.  

1. The title should be revised. It is ambiguous and unnecessarily long.

Response: Thank you for your comment. I changed the title to be “Exploring Polyploidization in Nigella sativa L.: An Applicable Strategy towards Crop Improvement”           I hope it is accepted

2. The scientific name of black cumin should be used in the title instead of English. Black cumin could be something in some parts of the world.

Response: You are right. Ok done, the scientific name of black cumin is included in the modified title

3. Abstract: The needs revision. I have provided an elaborate comment in the annotated manuscript.

Response: we revised the abstract according to your comments in the pdf file

4. The introduction is well written and provides a good rationale and objective for this study.

Response: Thank you for your encouragement words

5. Methodology: Information on how the seeds were sourced and their accession number should be added to the methodology. This is important for the reproducibility of this study.

Response: The source of the seeds (the breeder line of tetraploid plants induced using 2,4-dinitroaniline 10 mg L^-1 for 24 h), and the selected line of diploid plant which are originated from the breeding program at Faculty of Agriculture, Kafrelsheikh University, Egypt, according to El-Mahrouk et al. 2015). These lines are produced by the breeder (one of the authors), on 2015 so, there is no registration number that usually used for the lines obtained from gene banks 

 

6. Use the formula function in Microsoft Word to write the formulas appropriately.

Response: Thank you for this comment, I have learned something new

Ok done for all equations (formulas) mentioned in the study      

.

7. 7. Tables 1 and 2 are not necessary. Summarize how you carried out the study succinctly in the text. Table 2 could be useful only if you relate your data to varying environmental conditions

Response: Tables 1 and 2 has been deleted,  

8. Results: Line 281: What do you mean by previous parameters? May be measured parameters?

Response: Yes, measured parameters I changed in the text

9. Discussion: Line 386-387; provide the reference for the statement.

Response: Ok, done   I added reference for that statement

10. The discussion needs to be improved. The discussion should focus on your own results and not discussing already published works.

Response: Ok, We modified the discussion

11. The conclusion needs to be improved. Authors should focus on drawing a conclusion from their results and the implications of this study to the black cumin breeding program.

Response:
12. Comments on the Quality of English Language

The manuscript should be edited appropriately, and scientific names mentioned in the text should be properly written with their authority. 

Response:  We revised English, and we checked all scientific names and added their authority

 

Responses to the comments in the pdf file

Comment	Response
Lines 21-22 Does it only apply to tetraploid? Please rephrase	These markers can support selecting the tetraploid and diploid plants during the seedling stage by the plant breeders.
Lines 24-25 This methodology is shallow. The methodology should be captured succinctly.	Field experiments on tetraploids and diploids were carried out to evaluate the morphological and yield traits of both plants . Also, some cytological studies and GC analyses were conducted to achieve these goals.
Lines 25-27 How?	The cotyledon leaf of a diploid is taller and more pointed than the shorter tetraploid which has rounded leaf tops
Lines 27-29 This is not supposed to be. It should be part of the conclusion	Ok, it is omitted and added to conclusion
Lines 29-31 This looks superficial. Superior in what way. This quantitative study. You need to quite figures you have analyzed	Ok, this part rephrased again
Lines 31-33 This is not clear. Please present your results and  the implication on your objectives	Ok, this part is rephrased for more clarification
Line 41 What is the meaning of this? Not everybody will know that you are talking about chromosome number and the plant family. Please write clearly	Ok done, the needed information were added to the text
Line 72 Add authority to the scientific name	we checked all scientific names and added their authority
Line 76 Authority? Please add authority to all the scientific names mentioned in the text.
Line 133 Add how the seeds used in this study were source and their accessions for reproducibility purpose	The source of the seeds (the breeder line of tetraploid plants induced using 2,4-dinitroaniline 10 mg L-1 for 24 h), and the selected line of diploid plant which are originated from the breeding program at Faculty of Agriculture, Kafrelsheikh University, Egypt, according to El-Mahrouk et al. 2015). These lines are produced by the breeder (one of the authors), on 2015 so, there is no registration number that usually used for the lines obtained from gene banks.  We added this information in the text
Lines 162-168 Use formula function to write the formulas appropriately	Ok done, We used Microsoft Word to write the formulas mentioned in the study
Lines 171-172 Please delete table 1	Ok, done
Lines 175-176 Please delete this table or summarize the information and add to the methodology	Ok, done
Line 281 previous or measured parameters	Ok, measured
Line 312 Make sure all the values in the table are in the same decimal points	Ok, done as I could understand the comment
Line 387 Provide reference or this accession	Ok, done
Line 397 Add authority	Ok, done
Lines 519-523 This is not needed. This should be part of introduction at least	Ok, I deleted it

 

 

 

Reviewer 3 Report

Comments and Suggestions for Authors

In this manuscript, El-Mahrouk et al investigated morphological, cytological, and yield-related differences between diploid and tetraploid black cumin plants. This is a valuable contribution, as polyploidization is a widely used strategy in crop improvement, but comprehensive trait evaluations are relatively limited in Nigella sativa. The study is well-structured, with a clear methodology and relevant results. However, there are some issues in the manuscript that need to be addressed for better conciseness and accuracy. Please find my comments below:

 

In addition to the germination rate, I wonder whether the authors have noticed any difference in terms of the timing of germination, in the context of different ploidy levels.

 

I wonder whether the authors could supplement the p values and sample size information in Figure 1, 4, and 5d, keeping the consistence across the whole manuscript (for example, in Table 4, p values are listed).

 

What is the scale bar size in Fig. 2c-2f?

 

Fig. 3e, 6a, and 6c need a scale bar in the picture.

 

Typos:
In line 285: ‘diplopod’

In line 530: ‘Furthemore’

Author Response

Response to Reviewer 3

Comments and Suggestions for Authors

In this manuscript, El-Mahrouk et al investigated morphological, cytological, and yield-related differences between diploid and tetraploid black cumin plants. This is a valuable contribution, as polyploidization is a widely used strategy in crop improvement, but comprehensive trait evaluations are relatively limited in Nigella sativa. The study is well-structured, with a clear methodology and relevant results. However, there are some issues in the manuscript that need to be addressed for better conciseness and accuracy. Please find my comments below:

Response: Thank you for your encouragement words, also many thanks for your valuable comments which aimed to improve the MS. All changes or corrections you asked us to do are highlighted in yellow in the manuscript.  

In addition to the germination rate, I wonder whether the authors have noticed any difference in terms of the timing of germination, in the context of different ploidy levels.

 Response: We added the needed information in the text

I wonder whether the authors could supplement the p values and sample size information in Figure 1, 4, and 5d, keeping the consistence across the whole manuscript (for example, in Table 4, p values are listed).

Response:  Ok, done

 What is the scale bar size in Fig. 2c-2f?

 Response: we added the scale bar for Fig. 2c-2f but it changed to 3c-3f

Fig. 3e, 6a, and 6c need a scale bar in the picture.

 Response: Ok done (magnification power of 15×) for 3e which became 4e

Also, for 6a and 6c   is  1000×  and they became 1a  and 1c

Typos:
In line 285: ‘diplopod’     Ok, done  diploid

In line 530: ‘Furthemore’      these lines are deleted

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The authors have significantly improved the manuscript and addressed my comments. I therefore recommend the manuscript for publication.