Spermidine Modify Antioxidant Activity in Cucumber Exposed to Salinity Stress

Round 1

Reviewer 1 Report

The authors investigated the effects of short-term 48h-long NaCl-stress and spermidine level modification on polyamines level and antioxidant status in the cucumber leaves. Polyamine synthesis inhibitor slightly decreased spermidine and spermine level during salinity and reversed the antioxidant activity mobilization. This work was found to be potentially interesting and worthy of publication in Agronomy. However, some of the details of the manuscript need to be explained further. The authors should use more microscopic characterizations to prove the conclusions in the experiments. Specific comments are listed as follows.

1. Line 11, Page 1: Do not use first person (we, our, us) in text.

2. The authors use a lot of abbreviations, please define carefully in the manuscript.

3. Line 36, Page 2: Please delete “authors of” in the manuscript.

4. Lines 69-72, Page 4: The purpose of this study is not described in detail. The authors should list the aims point by point.

5. Lines 75, Page 4: Please check this sentence carefully.

6. Lines 132-137, Page 6: It struck me as odd that the analysis of 0 h, 24 h and 48 h seem to be irregular.

7. The figures in this manuscript should be drawn by Origin or SigmaPlot software.

8. The sentence “The spermidine treatment additionally increased the activity of this enzyme, … in the control plants after 24 and 48 h.” More discussion is needed to clarify the details.

9. The authors should reduce the description of references in discussion section.

10. Lines 210-217, Pages 12-13: This is stated as if it is a clear conclusion, but what data supports this interpretation/conclusion?

11. The conclusion part should be separated from the discussion part.

Author Response

Dear Reviewer 1/Editor,

I would like to submit revised MS

Manuscript ID agronomy-1755668

Type Article

Title Spermidine modify antioxidant activity in cucumber exposed to salinity stress

 

Authors Agata Korbas , Jan Kubiś * , Magdalena Rybus-Zając , Tamara Chadzinikolau

Department of Plant Physiology, Poznan University of Life Sciences, ul. Wołyńska 35,

60-637 Poznań, Poland,

:

The authors should use more microscopic characterizations to prove the conclusions in the experiments. Specific comments are listed as follows.

Answer to Reviewer`s 1 suggestions

1. Line 11, Page 1: Do not use first person (we, our, us) in text. It has been changed into passive form and 3 person.
2. The authors use a lot of abbreviations, please define carefully in the manuscript.

These abbreviation have been carefully defined in MS, especially PA/PAs were consequently used.

3. Line 36, Page 2: Please delete “authors of” in the manuscript.

This word was removed.

4. Lines 69-72, Page 4: The purpose of this study is not described in detail. The authors should list the aims point by point.

Cucumber cultivar Dar was chosen, because it still exist on the list of cultivars allowed for cultivation in Poland, it is open ground cultivar.

5. Lines 75, Page 4: Please check this sentence carefully.

The additional spaces was removed. The English language will be corrected.

6. Lines 132-137, Page 6: It struck me as odd that the analysis of 0 h, 24 h and 48 h seem to be irregular.

Every treatment caused some disturbances in loss of water (RWC) especially under salt stress conditions - 1mM Spd and MGBG. It was almost not visible in well-watered seedlings.

7. The figures in this manuscript should be drawn by Origin or SigmaPlot software.

If there will be necessary by editing it would be changed.

8. The sentence “The spermidine treatment additionally increased the activity of this enzyme, … in the control plants after 24 and 48 h.” More discussion is needed to clarify the details.

I guessed, SOD activity…

Lines 177-180, Page 9. I was additionally explained – correlation coefficient was given. Line 233, 234 Page 13 some explanation and correlation as well.

9. The authors should reduce the description of references in discussion section.

The all data are not clear, and I tried to explained it in details.

10. Lines 210-217, Pages 12-13: This is stated as if it is a clear conclusion, but what data supports this interpretation/conclusion?

At the beginning of DISCUSSION I tried to place some general statement, PAs accumulation [19, 22,37] and PAs action [38-41] and the data citation as well. Details of precise mode of action, by ROS accumulation and enzyme activities description discussion and available literature citation, some position were added.

11. The conclusion part should be separated from the discussion part.

There were no intention to separate some final conclusion.

Reviewer 2 Report

Dear editor

Thank you for the opportunity to review this manuscript. In my opinion, the novelty of current study is a big problem, and the current data provided very limited information on this topic. The manuscript can be improved, and I recommend authors to revise the manuscript following the suggestions below.

1. I recommend authors to illustrate the background and significance of the study in abstract.

2. The accumulation of spermidine and spermine was observed after 24 h of the stress. After 48 h the PA levels were lower, similarly to the control plants. Please explain it.

3. I do not see the data of spermidine accumulation-denpendent antioxidant enzymes activities and ROS 198 accumulation. Where is the data of Table1？

4. I think the relationship between the salinity stress with antioxidant activity is a common sense. What are the scientific problems of the current study?

 

Author Response

Dear Reviewer 1/Editor,

I would like to submit revised MS

Manuscript ID agronomy-1755668

Type Article

Title Spermidine modify antioxidant activity in cucumber exposed to salinity stress

 

Authors Agata Korbas , Jan Kubiś * , Magdalena Rybus-Zając , Tamara Chadzinikolau

Department of Plant Physiology, Poznan University of Life Sciences, ul. Wołyńska 35,

60-637 Poznań, Poland,

Reviewer`s suggestions:

1. According to Reviewer suggestion I placed the background and significance of this study in abstract – the last sentences.
2. The accumulations of spermidine and spermine was observed after 24h of stress, but after 48h PA levels were lower… This effect was observed, significant accumulation in early hours of stress(12h, 24), than Spd level decreased in the leaves of F1 cucumber cultivars Meteor, Huzar, Avatar, Poznański. Korbas A. Doctoral thesis, Poznań 2020. Similar effects were observed in rice. Lefevre et al. 2001. Plant Science 161(5) 943-952. The dynamics of accumulation can be species or even cultivars or organ specific and was not the main task of this study.
3. The data of the Table 1 were placed at &RESULTS. Positive correlation coefficient between Spd accumulation and antioxidant enzymes activities (positive but ns for APX) at section enzymes antioxidative activity . Negative correlation Spd versus ROS generation at ROS section.
4. The main problem of our study… relationship between salinity stress, tolerance to salinity by ROS detoxifying possibilities, it was to ABSTRACT added.

-the novelty of current study, the citation for the papers published in the recent 3 years… was increased –  19, 23, 37, 38, 53, 55 and according cucumber 25, 26 – 19.05.22. And now, review 25.05.22, another 3 articles have been added – 7, 9, 46 dated 2021, 2019, 2018.

 

 

Author Response File: Author Response.pdf

Reviewer 3 Report

Abstract

"salinized with 50 mM NaCl" - treated 50 mM NaCl

Introduction   What was the basis to select 50 mM NaCl concentration to induce salinity stress? In the introduction, the authors have provided several examples of studies with different concentrations of NaCl. For instance - Zhao et al. examined the influence of a wide range of NaCl concentrations (0-300 mM) on PA levels in barley seedlings.   "By contrast, putrescine accumulated in wheat cultivars, whereas the spermidine content decreased; the change was less pronounced in salt-sensitive varieties" - Not clear   "Although the accumulation of polyamines is considered to be a general response to abiotic stress factors, the 56 close relationship between the PA accumulation and protection still remains unclear [20]. Researchers 57 observed different polyamine levels," - need to use PA abbreviation uniformly throughout the MS.   The Authors need to improve the flow of information to connect the paragraph "Cucumber is an important and widespread vegetable species.." with the preceding paragraphs.   "3. Results  131 RWC. The salt stress caused a water deficit" - here I guess "RWC" is the subheading. Authors need to provide well-elaborated subheadings    The discussion section is inadequate and has too much redundancy with the results. 

Author Response

Dear Reviewer 3/Editor,

I would like to submit revised MS

Manuscript ID agronomy-1755668

Type Article

Title Spermidine modify antioxidant activity in cucumber exposed to salinity stress

Authors Agata Korbas , Jan Kubiś * , Magdalena Rybus-Zając , Tamara Chadzinikolau

Department of Plant Physiology, Poznan University of Life Sciences, ul. Wołyńska 35,

60-637 Poznań, Poland,

Reviewer`s suggestions:

1. salinized was changed into treated
2. What was the basis to select 50mM NaCl – M&M, Plant material and treatment, The salt concentration were chosen on the base of initial experiments. Start from bio test method with the wide range (0-200mM) NaCl concentrations and experiments carried on five F1 cucumber cultivars treated 50, 75, 100, 150mM. Korbas A. Doctoral thesis, Poznań 2020.
3. The authors have provided several examples with different concentration of NaCl. It is the fact observed in several researches, salt concentration and duration of experiments and obtained results are often opposite. But general response – accumulation of PAs, less-pronounced changes in salt-sensitive cultivars, more-pronounced in salt tolerant cultivars and species as well and specific role of Spd. But all these data are not absolutely clear. This summarizing sentence is present, especially in reviews article.
4. PA/PAs abbreviation was used uniformly at MS
5. I tried to improve the flow of information to connect cucumber information with paragraph, The experiment was carried out on cucumber… was added.
6. Results RWC, salt stress caused water deficit… measured on the base of water content – RWC decrease, I mean.
7. The discussion section has too much redundancy. In my opinion discussion is the specific paragraph which not only comments and comperes obtained in this study results. It is the place for some speculation as well, the function of polyamines still remained unclear and a lot of researchers were engaged in this filed, and obtained results are sometimes not clear or even opposite.

Author Response File: Author Response.pdf

Reviewer 4 Report

The paper is interesting: my only suggestions are the following: please, the authors must better explain the novelty, the implications in the conclusion and consider a more recente and innovative international literature.

Author Response

Dear Reviewer 4/Editor,

I would like to submit revised MS

Manuscript ID agronomy-1755668

Type Article

Title Spermidine modify antioxidant activity in cucumber exposed to salinity stress

 

Authors Agata Korbas , Jan Kubiś * , Magdalena Rybus-Zając , Tamara Chadzinikolau

Department of Plant Physiology, Poznan University of Life Sciences, ul. Wołyńska 35,

60-637 Poznań, Poland,

Reviewer`s suggestions:

The paper is interesting: my only suggestions are the following: please, the authors must better explain the novelty, the implications in the conclusion and consider a more recente and innovative international literature.

The novelty, the implications in the conclusion and consider a more recent and innovative international literature. … was added  –  [19, 23, 37, 38, 53, 55] and according cucumber [25, 26] – 19.05.22. And now, review 23.05.22, another 3 articles have been added – [7, 9, 46] dated 2021, 2019, 2018.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Congratulation

Author Response

Thank you for congratulation