Use of Plant Regulators for Activation of Antioxidant Enzymes in Basil Plants under Water Deficit Conditions

Round 1

Reviewer 1 Report

The work essentially lacks novelty because it is a repeat of studies already done with other plants.  And only a small fraction of what could be analyzed has been examined.  The enzymes assayed represent only one aspect of adaption to water deficits in plants.  How is this modified in the water stress tolerant basil to start with?   Many comments as sticky notes.

Also you argue why basil is an important crop. Yet the key factor  its essential oil content is not examined   maybe the plants do better but that would not matter if there are consequences to the oil content. 

no information on the basic parameters of growth/ water content etc  either. 

Comments for author File: Comments.pdf

Author Response

Point 1: The work essentially lacks novelty because it is a repeat of studies already done with other plants.  And only a small fraction of what could be analyzed has been examined.  The enzymes assayed represent only one aspect of adaption to water deficits in plants.  How is this modified in the water stress tolerant basil to start with?   Many comments as sticky notes.

Response point 1: Despite several studies, the objective of this work was to analyze the use of a plant regulator that has not been studied in aromatic or medicinal plants, such as basil. Other analyses, such as photosynthesis and fluorescence, were carried out, but as it had already been published, it was not possible to present these results in the present work. Some points of sticky notes in the manuscript are in the word file attached here

Point 2: Also you argue why basil is an important crop. Yet the key factor  its essential oil content is not examined   maybe the plants do better but that would not matter if there are consequences to the oil content. 

Response point 2: The essential oil was extracted during the experiment but the technician responsible for the extraction passed on the information that the results obtained were not enough to carry out the analysis as they would be too fragile data to present in the work.

Point 3: No information on the basic parameters of growth/ water content etc  either. 

Response point 3: information on basic growth parameters has been added in the manuscript. The evaluated parameters were leaf number, leaf area, diameter, dry mass and plant height. But the last two parameters (dry mass and plant height) did not show significance and therefore were not added to the work. Information on water content has not been evaluated.

 

Note: Attached are some responses and questions about the comments added in the sticky notes of the manuscript.

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript presents the physiological effects of the application of plant regulators in basil plants suffering from water deficit. Although the results properly analyzed, the main problem is that it is completely a descriptive report with no insight into the mechanism of the action of antioxidative enzymes. There are many similar articles already published in literature.

Some shortcomings were found as well.

The language needs improvement.

The title should be changed. The results did not clearly confirm the information contained in the title. Moreover, there is no information on plant development in the manuscript.

Introduction: The background is not clearly and sufficiently defined in the context of the topic. There is no clearly defined goal as well.

Results and discussion are very simple and descriptive.  The issue of plant regulators was completely omitted (kinetin (Kt, cytokinin),  indolylbutyric acid (IBA, auxin), and gibberellic acid (GA3, gibberellin).

Materials and Methods:

The conditions for plant growth in the greenhouse have not been specified (temperature and air humidity are factors that affect the intensity of drought, they can intensify it). It is difficult to compare your own results with others, if you do not know the conditions of plant growth.

How was the development of plants determined? (reference to the title).

What was the final water deficit determined by stomatal conductivity measurement?

There is no information on the dates of application of the plant regulator. How many times has plant regulator been applied? There are different dates (DAT) in the description of the tables and figures.

Conclusion section should be the implication of this research, justified by results.

Author Response

Point 1: The manuscript presents the physiological effects of the application of plant regulators in basil plants suffering from water deficit. Although the results properly analyzed, the main problem is that it is completely a descriptive report with no insight into the mechanism of the action of antioxidative enzymes. There are many similar articles already published in literature.

Response point 1: The intention of the research was to bring studies of the application of plant regulators in aromatic plants because they only had work on the effect of water stress but not solutions to alleviate them. Some other analyzes were performed, such as photosynthesis and fluorescence, but due to a conflict of interest, they need to be left out of this article.

Point 2: The language needs improvement.

Response 2: Enclosed the certificate of proficiency in the English language.

Point 3: The title should be changed. The results did not clearly confirm the information contained in the title. Moreover, there is no information on plant development in the manuscript.

Response 3: The title has been changed and is found in the manuscript underlined in yellow with the other changes (Plant regulators in the development of basil under conditions of water deficiency).

Point 4: Introduction: The background is not clearly and sufficiently defined in the context of the topic. There is no clearly defined goal as well.

Response 4: I brought more information in the introduction, mainly about the usefulness of the plant hormones that make up Stimulate. Change is in the manuscript.

Point 5: Results and discussion are very simple and descriptive.  The issue of plant regulators was completely omitted (kinetin (Kt, cytokinin),  indolylbutyric acid (IBA, auxin), and gibberellic acid (GA3, gibberellin).

Response point 5: The use of each of the plant hormones has been added to the introduction.

Point 6: Materials and Methods: The conditions for plant growth in the greenhouse have not been specified (temperature and air humidity are factors that affect the intensity of drought, they can intensify it). It is difficult to compare your own results with others, if you do not know the conditions of plant growth.

Response point 6: Temperature and humidity were added to the material and methods, in which the temperature and humidity variation within the greenhouse was averaged.

Point 7: How was the development of plants determined? (reference to the title).

Response 7: Culture development was performed using biochemical analysis, growth analysis (which were added to the manuscript) and photosynthesis and fluorescence analysis, the latter data not being added because they were already used in another article.

Point 8: What was the final water deficit determined by stomatal conductivity measurement?

Response point 8: Added information in the manuscript.

Point 9: There is no information on the dates of application of the plant regulator. How many times has plant regulator been applied? There are different dates (DAT) in the description of the tables and figures.

Response 9: The plant regulator was always applied one day before the analyses. Application dates were added to the manuscript and DAT differences between figures and tables were corrected.

Point 10: Conclusion section should be the implication of this research, justified by results.

Response 10: The conclusion of the work was redone, in order to present the results of the research.

 

Note: Attached is the certificate of proficiency in English, which was made based on the initial title of the work.

Author Response File: Author Response.pdf

Reviewer 3 Report

Dear authors

 

This manuscript regarding “Plant regulators activate antioxidant system and increase the development of basil under water deficit”. This manuscript is well performed with clear objectives and discussion; the manuscript might be a good contribution to researchers working on this research related to water deficit stress.

However, the authors have performed only an enzymatic Antioxidants essay and they must be performed a few Non-enzymatic biochemical (antioxidants) essays such as phenolic and ascorbate content. Further, the author may include Other Biochemical Parameters such as Malondialdehyde Content. Thus, I would like to recommend this manuscript for publication after major revision.

A few other comments are below for the advancement of the manuscript:

The reference should be arranged according to the journal format style. Kindly check the author's guidelines and format the entire reference accordingly.

The scientific name should be in italic in the entire manuscript including references such as Spondias tuberosa in line no 569.

Author Response

Point 1: This manuscript regarding “Plant regulators activate antioxidant system and increase the development of basil under water deficit”. This manuscript is well performed with clear objectives and discussion; the manuscript might be a good contribution to researchers working on this research related to water deficit stress.

Response point 1: We appreciate the feedback on the research carried out and also the correction suggestions made.

Point 2: However, the authors have performed only an enzymatic Antioxidants essay and they must be performed a few Non-enzymatic biochemical (antioxidants) essays such as phenolic and ascorbate content. Further, the author may include Other Biochemical Parameters such as Malondialdehyde Content. Thus, I would like to recommend this manuscript for publication after major revision.

Response point 2: The work was carried out in 2020 and therefore it will not be possible to carry out the analyzes that were indicated because at the time, only the analyzes present in the article were carried out. Regarding the content of malondialdehyde, this analysis is nothing more than what is presented in the work as lipid peroxidation, in which we performed it through the contents of malondialdehyde (MDA).

Point 3: The reference should be arranged according to the journal format style. Kindly check the author's guidelines and format the entire reference accordingly.

Response point 3: References have been checked and corrected according to guidelines and are in the manuscript.

Point 4: The scientific name should be in italic in the entire manuscript including references such as Spondias tuberosa in line no 569.

Response point 4: The scientific names have been corrected and placed in italics, as stated in the guidelines. The corrections are in the manuscript and underlined in yellow for later correction by the reviewers.

 

Note: Attached are some responses and questions about the comments added in the sticky notes of the manuscript.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The authors have made changes but again the grammar is poor which makes reading of the paper difficult.

the methods of showing data are not optimal   

the crucial factor of oil is not measured.

In the introduction a reference indicates similar work already published

 

 

Comments for author File: Comments.pdf

Author Response

Ponto 1: Os autores fizeram alterações, mas novamente a gramática é ruim, o que dificulta a leitura do artigo.

Ponto de resposta 1: Gostaríamos de nos desculpar pela inconveniência em relação ao idioma inglês. Pedimos uma revisão detalhada para o responsável pela tradução. Mas gostaríamos de reforçar que a pessoa possui fluência no idioma e também experiência em tradução de artigos. Caso ainda haja dificuldades de entendimento, entre em contato conosco para que possamos buscar mais alternativas de revisão.

Ponto 2: os métodos de exibição de dados não são ideais   

Response point 2: We appreciate the indication of improvement in statistics. But we would like to understand a little more about the point to be improved. Does it indicate that we added the lines, pointers? Because a reviewer in the area of ​​statistics pointed out to us that quantitative data should be presented in regression. We are waiting for your feedback to improve the presentation of the data.

Point 3: the crucial factor of oil is not measured.

Response point 3: The data obtained from essential oil were not enough to be analyzed and discussed.

Point 4: In the introduction a reference indicates similar work already published

Ponto de resposta 4: O ponto principal do trabalho foi trazer estudos não apenas relacionados à atividade de enzimas antioxidantes, mas principalmente sobre marcadores de estresse. Trouxemos alguns pontos para o manuscrito para melhorar o entendimento da obra.

Em anexo estão as respostas sobre os pontos levantados no manuscrito

Author Response File: Author Response.docx

Reviewer 2 Report

Title: Plant regulators in defense mechanism of basil under water deficit conditions

The title is too general and does not correspond to the content of the work.

In the paper too little emphasis is placed on the influence of growth regulators on defense mechanisms.

The authors supplemented the introduction and introduced important references.

The hypothesis was clearly stated.

However, the work still contains many shortcomings.

The number of features compared in the work is small. I do not understand why the authors do not want to show other results they  obtained (e.g. regarding morphometric features). The description of the plant growth results is very short. Moreover, the date of measurement is not specified in Figures 5 and 6.

The discussion still lacks reference to plant regulators that have been applied to plants in the form of a specific commercial product (cytokinin, auxin and gibberellin).

There is also no correlation between the studied features. Perhaps the relationships between the features would explain the ambiguous results regarding enzyme activity (CAT, POD).

The methodology includes chapter "4.3. Total soluble proteins". There is no reference to this point in other parts of the work.

Why stomatal conductance value = 50% was considered water stress for basil? In plant physiological studies on drought stress, soil moisture is often used to determine water conditions. What was the relationship between stomatal conductance = 50% and soil moisture?

The obtained results do not clearly confirm the first sentence of the conclusion.

Author Response

Point 1: The title is too general and does not correspond to the content of the work.

Response point 1: Dear reviewer, the title has been changed as per guidelines.

Point 2: In the paper too little emphasis is placed on the influence of growth regulators on defense mechanisms.

Response point 2: As a suggestion, I brought points to the article that emphasize more the action of regulators in the plant's defense mechanism.

Point 3: The authors supplemented the introduction and introduced important references. The hypothesis was clearly stated. However, the work still contains many shortcomings.

Response point 3: We appreciate the suggestions regarding the work and we try to bring more discussions to improve the work

Point 4: The number of features compared in the work is small. I do not understand why the authors do not want to show other results they  obtained (e.g. regarding morphometric features). The description of the plant growth results is very short. Moreover, the date of measurement is not specified in Figures 5 and 6.

Response point 4: All results obtained in the work were presented. We understand the absence of some data but these were not collected throughout the experiment and therefore were not presented as requested. We apologize for the absence of these data and hope to be able to adjust this for the next experiments carried out. The other questions have already been changed in the manuscript.

Point 5: The discussion still lacks reference to plant regulators that have been applied to plants in the form of a specific commercial product (cytokinin, auxin and gibberellin).

Response point 5: We added to the discussion some points of isolated regulators and how they act in culture.

Point 6: There is also no correlation between the studied features. Perhaps the relationships between the features would explain the ambiguous results regarding enzyme activity (CAT, POD).

Response point 6: The enzymes were analyzed separately, despite the relationship between them. And that's why we weren't able to bring this point of correlation that was presented to us as an improvement.

Point 6: The methodology includes chapter "4.3. Total soluble proteins". There is no reference to this point in other parts of the work.

Response point 6: The topic of total soluble proteins is used to calculate the other enzymes. It is presented in the text only to show how it was done and does not enter as an isolated analysis.

Point 7: Why stomatal conductance value = 50% was considered water stress for basil? In plant physiological studies on drought stress, soil moisture is often used to determine water conditions. What was the relationship between stomatal conductance = 50% and soil moisture?

Response point 7: We adopted a conductance of 50% for the basil crop because a review of work already carried out in similar cultures and work carried out in the group was carried out, and we agreed to use this stomatal conductance. We understand the importance of soil sampling but unfortunately it was not performed during the conduct of the experiment.

Point 8: The obtained results do not clearly confirm the first sentence of the conclusion.

Response point 8: Completion has been changed as indicated.

 

Attached are the answers regarding the points raised in the manuscript

Author Response File: Author Response.docx

Reviewer 3 Report

Dear Authors,

Revised manuscripts have been much improved. Although, I am still not satisfied with the response to one of my comments. After all, I would like to recommend it for acceptance in its current form with agreeing of the other reviewers only.

Author Response

Point 1: Dear Authors,

Revised manuscripts have been much improved. Although, I am still not satisfied with the response to one of my comments. After all, I would like to recommend it for acceptance in its current form with agreeing of the other reviewers only.

Response 1: Dear reviewer,

Thank you for all your contributions to our work. We would like to ask you to send us again which comment you were not satisfied with. We are willing to revise to improve the published article.

 

Attached are the answers regarding the points raised in the manuscript

Author Response File: Author Response.docx

Round 3

Reviewer 1 Report

The paper is not written well  and this detracts from any science understanding

Comments for author File: Comments.pdf

Author Response

Dear reviewer,

We appreciate all the corrections and suggestions you are sending us so that the article gets even better for the future publication. We are working hard to get all fixes right.
As for the English translation, we understand that you are having difficulty understanding and we sincerely apologize. The person responsible for the translation has references and experience with translating articles, and has already made other articles for us to publish in MDPI. In addition, at all stages, he was available to revise whatever was needed.

We would like to find a solution together to adapt in the best way. We believe that for you, the best indication would be to do the translation through the website platform. We did the simulation to analyze the value and would like to apologize because unfortunately we do not have the financial conditions to cover this expense. We are from a public university in Brazil, a country in which it is developing and unfortunately investment in education and research is very scarce. We were unable to obtain grants or resources to pay for publications or anything else related to the research. All our efforts to bring more and more innovation to research have been paid with our own money, the same money we use to pay our bills and food. We are very sorry and hope you understand the situation in which Brazilian students and researchers find themselves.
As we have already informed, we are always willing to improve the article to be published and we are open to possible solutions so that both sides can leave satisfied and with the article published in a prestigious journal, such as yours.

Furthermore, we thank you for the opportunity and are available to answer any questions you may have.

Reviewer 2 Report

Line 298-292

Title: Use of plant regulators for activation of antioxidant enzymes in basil plants under water deficit conditions

The title has been corrected. The authors supplemented the introduction and introduced important references. However, there are still shortcomings in the manuscript that should be corrected.

There are no conclusions in the summary.

Line: 288 - 293. There is no reference to the table or figure presenting the results of research regarding the height and dry weight of plants. If the authors used the results in another publication, they should refer to this publication. In the presented form (no data, no statistical evaluation), the description is not very credible. The Results section should contain only descriptions that directly characterize the published results.

Description of chapter "4.5. Growth measurements" is unclear and needs to be clarified. Were biometric measurements (the number of leaves, plant height, and stem diameter) determined only after the plants were dried?

Author Response

Point 1: There are no conclusions in the summary.

Response point 1: The conclusion has been added to the abstract.

Point 2: Line: 288 - 293. There is no reference to the table or figure presenting the results of research regarding the height and dry weight of plants. If the authors used the results in another publication, they should refer to this publication. In the presented form (no data, no statistical evaluation), the description is not very credible. The Results section should contain only descriptions that directly characterize the published results.

Response point 2: Height and dry weight of plants did not show significant results in the statistical analysis and therefore are not present in the work, because there would be no way to discuss. In order to avoid misinterpretation by the reader in this regard, we removed the part where we cited these analyses, just to introduce that they did not show significant results. We also took from the material and methods to avoid problems in this regard.

Point 3: Description of chapter "4.5. Growth measurements" is unclear and needs to be clarified. Were biometric measurements (the number of leaves, plant height, and stem diameter) determined only after the plants were dried?

Response point 3: This topic has been corrected in the manuscript. We appreciate the guidance to improve our work.