MtTGA1 Transcription Factor Enhances Salt Tolerance through Hormonal Regulation and Antioxidant Enzyme Activity in Medicago truncatula

Round 1

Reviewer 1 Report

Dear Editor

Researchers examined the regulation mechanism of MtTGA1 in response to salt stress in this study. The study topic is interesting and novel, but the following comments need to be corrected:

1. There is a need for authors to report the exact location of the promoter region on the genome, which chromosome, etc.

2. Summarize the legends of the y-charts in Figure 4

3. As part of the real-time PCR analysis, which gene is the housekeeper's gene?

4. I think that the abstract should be written more precisely, for example, the difference in gene expression should be included in the abstract

5. ​The discussion was not deep enough. Reorganize and rewrite

   Best Regards

 

Dear Editor

Researchers examined the regulation mechanism of MtTGA1 in response to salt stress in this study. The study topic is interesting and novel, but the following comments need to be corrected:

1. There is a need for authors to report the exact location of the promoter region on the genome, which chromosome, etc.

2. Summarize the legends of the y-charts in Figure 4

3. As part of the real-time PCR analysis, which gene is the housekeeper's gene?

4. I think that the abstract should be written more precisely, for example, the difference in gene expression should be included in the abstract

5. ​The discussion was not deep enough. Reorganize and rewrite

   Best Regards

 

Author Response

Response letter 1

Dear reviewer,

Thank you for your hard work and valuable comments. These comments are very helpful to improve the quality of the manuscript. We have studied all comments carefully and have made conscientious correction. Revised portion are marked with revision mode on the original manuscript. Point-by-point replies were listed as follows.

1. There is a need for authors to report the exact location of the promoter region on the genome, which chromosome, etc.

We added the exact location of the promoter to the manuscript: "To investigate the regulatory mechanism of MtTGA1, a 2000 bp sequence was obtained at the 5' upstream end of MtTGA1 located on genome chromosome 3."and the relevant modifications can be displayed in revision mode.

2. Summarize the legends of the y-charts in Figure 4.

We have summarized the legends of the y-chart in Figure 4,Thank you for your reminder.

3. As part of the real-time PCR analysis, which gene is the housekeeper's gene?

The AtUBQ10 gene is the housekeeper's gene. We have added this information to Materials and Methods, thank you for your suggestions.

4. I think that the abstract should be written more precisely, for example, the difference in gene expression should be included in the abstract.

We've revised the summary to make it more precise, thank you for your reminder.

5. The discussion was not deep enough. Reorganize and rewrite.

We think this is an excellent suggestion.We have re-written this part .

We tried our best to improve the manuscript and made some changes marked in revised paper which will not influence the content and framework of the paper. We appreciate for Editors/Reviewers’ warm work earnestly, and hope the correction will meet with approval. Once again, thank you very much for your comments and suggestions.

 

Best regards,

Shuwen Li

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript entitled “MtTGA1 Transcription Factor Enhances Salt Tolerance through Hormonal Regulation and Antioxidant Enzyme Activity in Medicago truncatul” is well written. However, a few major necessary questions need to be addressed.

 

 In the result section, the authors have included antioxidant enzymes such as APX, POD and SOD, but why not stress markers like TBARS and ROS? Which factors induce activities of enzymes without stress markers is very difficult to understand.

 

In Figure 5., the authors incorporated two times POD in Figure? Also, replace “leave” with “leaves”

 

Figure 10. The picture quality is poor.

 

Why did the authors have only selected salt stress? There are so many other factors, like drought and temperatures affect different signalling pathways in plants.

 

 

 

 

 

Minor changes like spelling mistakes are required to correct.

Author Response

Dear reviewer,

Thank you very much for your hard work and valuable comments. These opinions help to improve academic rigor of our paper. Based on your suggestion and request,we have made corrected modifications on the revised manuscript. We hope that our work can be improved again. Point-by-point replies were listed as follows.

1. In the result section, the authors have included antioxidant enzymes such as APX, POD and SOD, but why not stress markers like TBARS and ROS? Which factors induce activities of enzymes without stress markers is very difficult to understand.

We think this is an excellent suggestion. So, we added expression levels of key enzymes for detoxification of ROS in the results section, including: Ascorbic Acid(AA) and Monodehydroascorbate Reductase(MDHAR). Relevant content can be identified in the revised manuscript.

2. In Figure 5., the authors incorporated two times POD in Figure? Also, replace “leave” with “leaves”

We were really sorry for our careless mistakes.We have modified Figure 5 and replaced “leave” with “leaves”. All the modifications could be identified in the manuscript in track-changed mode. Thank you for your reminder.

3. Figure 10. The picture quality is poor.

We sincerely thank the reviewer for careful reading. As suggested by the reviewer, we have enhanced Figure 10.

4. Why did the authors have only selected salt stress? There are so many other factors, like drought and temperatures affect different signalling pathways in plants.

We performed pre-experiments on salt stress and drought treatment on the transgenic plants, and found that the phenotype of salt stress treatment was obvious, but after drought stress treatment, no obvious phenotype was found, so we focused on the effect of salt stress treatment on MtTGA1. Other experiments related to abiotic stress will be explored step by step. 

We tried our best to improve the manuscript and made some changes marked in revised paper which will not influence the content and framework of the paper. We appreciate for Editors/Reviewers’ warm work earnestly, and hope the correction will meet with approval. Once again, thank you very much for your comments and suggestions.

 

Best regards,

Shuwen Li

Author Response File: Author Response.docx

Reviewer 3 Report

The authors need to see the comments on the attached manuscript PDF. They need to see comments on yellow-highlighted text and comment boxes. The authors need to include all suggestions before resubmitting the revised version of the manuscript.

Comments for author File: Comments.pdf

Minor editing of the English language is required.

Author Response

Dear reviewer,

Thank you for your letter and for the reviewers’ comments. Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made correction which we hope meet with approval. Revised portion are marked the manuscript. The main corrections in the manuscript and the responds to the reviewer’s comments are as flowing:

1. Correct title: "MtTGA1 Transcription Factor Enhances Salt Tolerance through Hormonal Regulation and Antioxidant Enzyme Activity in Barrel Medical (Medicago truncatula Gaertn.)"

Thanks for the above suggestion. "Medicago truncatula" is the Latin name and in most cases "Medicago truncatula" is used to represent the plant. For example,

[1] Ariel, F., Diet, A., Verdenaud, M., Gruber, V., Frugier, F., Chan, R., and Crespi, M. (2010). Environmental regulation of lateral root emergence in Medicago truncatula requires the HD-Zip I transcription factor HB1. Plant Cell 22: 2171–2183.

[2] Cosson, V., Durand, P., d'Erfurth, I., Kondorosi, A., and Ratet, P.(2006). Medicago truncatula transformation using leaf explants. Methods Mol. Biol. 343: 115–127.

[3] Cheng, X., Peng, J., Ma, J., Tang, Y., Chen, R., Mysore, K.S., and Wen, J. (2012). NO APICAL MERISTEM (MtNAM ) regulates floral organ identity and lateral organ separation in Medicago truncatula.New Phytol. 195: 71–84.

[4] Rakocevic, A., Mondy, S., Tirichine, L., Cosson, V., Brocard, L., Iantcheva, A., Cayrel, A., Devier, B., Abu El-Heba, G.A., and Ratet, P.(2009). MERE1, a low-copy-number copia-type retroelement in Medicago truncatula active during tissue culture. Plant Physiol. 151: 1250–1263.

So, “MtTGA1 Transcription Factor Enhances Salt Tolerance through Hormonal Regulation and Antioxidant Enzyme Activity in Medicago truncatula” may be more suitable as the title of the manuscript.

2. Abstract: When and where was this study carried out?How did you carry out this study, and what is the objective of this study?

In accordance with your suggestion, we have revised the abstract and the relevant modifications can be displayed in revision mode.

3. Add' Medicago truncatula' as one of the keywords.

As suggested by the reviewer, we have add' Medicago truncatula' as one of the keywords.Thank you for your reminder.

4. Write clearly when was this experiment carried out?Write below subheading and its detail in last portion of Materials and Methods; Statistical analysis: Which statistical test you perfomed and which parameters you analyzed ?, Which statistical software or which verson of Excel you used?

We have supplemented “Statistical Analysis” in our manuscript. Thank you for your kind suggestion, which is valuable for improving the accuracy of the manuscript.

5. Figure 4: Provide the name of the X-axis; the text on the Y-axis overlaps with numericals, so make these figures again.What does the error bar indicate for what information? Are they the standard error of the mean?

We have modified Figure 4 and labeled the name of X-axis, with error bars representing the standard error of the mean.Thank you for your reminder.

6. Figure 5,Figure 6 and Figure 10 :What does the error bar indicate for what information? Are they the standard error of the mean?It would be better to make color diagram instead of black and white.

Error bars representing the standard error of the mean.We think this is an excellent suggestion.We have replaced the black and white chart with a color chart.

7. Figure 11: Do not keep any figures in the discussion section.

We have moved Figure 11 to the results section.

8. References: All references should follow the journal guidelines. Add DIO for journal references.

Thank you for your comment for the references, all the revised have been included in the revised manuscript.

We sincerely hope that this revised manuscript has addressed all your comments and suggestions.We appreciated for reviewers’ warm work earnestly,and hope that the correction will meet with approval. Once again, thank you very much for your comments and suggestions.

 

Best regards,

Shuwen Li

Author Response File: Author Response.docx