Foliar Application of Salicylic Acid Improved Growth, Yield, Quality and Photosynthesis of Pea (Pisum sativum L.) by Improving Antioxidant Defense Mechanism under Saline Conditions

Round 1

Reviewer 1 Report

Comments for author File: Comments.pdf

Author Response

Dear Editor

We are pleased to re-submit our revised manuscript, “Foliar application of salicylic acid improved growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions” Ms. Ref. No.: Sustainability-1910631) to be considered for publication in “Sustainability”.

Thank you very much for making a critical assessment of the original version of our manuscript. We are glad to learn that the editor as well as the reviewer(s) considered our findings meaningful. However, the reviewer(s) provided some valuable comments and suggestions for the potential improvement of the manuscript. Based on the comments, you encouraged us to re-submit a revised version, which takes into account all of the points raised by the reviewer(s). We have now completed a thorough revision as per the recommendations of the reviewer(s). Revised portions are marked in red in the paper. Our response to reviewers’ comments has been listed below point-by-point.

Reviewer 1

Comments and Suggestions for Authors

The manuscript entitles “Foliar application of salicylic acid improves growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions” has been written well and have some comments below:

In the abstract after sentence line 31, please add a sentence to explain the importance of using salicylic acid.

Response back: Line 32-33; Importance of salicylic acid is added as per reviewer suggestion.

Rewrite the sentence line 39-41 to avoid repetition

Response back: Line 39-43; corrections have been to avoid repetition within the manuscript.

In introduction part:

Line 60-62: Please add more details to explain how to manage salt stress using for example ; macro and micronutrients, proteomic, many proteins…

Response back: Line 61-66; Description has been added and mentioned in red color in the manuscript.

Line 66: I think that salt toxicity in plants is due to the accumulation of Na and Cl ions and especially the Na ion which has a unidirectional transport from the roots and accumulates in the leaves. Please verify this information.

Response back: Line 69-71; Corrections have been made as per suggestions and mentioned in red color in the manuscript.

Line 96: write (greater than 1mM instead of than 1.

Response back: line 100; Suggested line has been incorporated.

The section of Material and Methods should be edited in detail to explain better the experiments developed;

The technique of sowing (section 2.2.) is not clear. it is a question of sowing for two years and making three pickings but the results express only one picking. Please, explain this or rewrite the method.

Response back: section 2.2. has been improved. Line 123-124; Three pickings were in each season (year) and average was computed. However, corrections have been inserted in the text.

It is the same for the experimental setup (section 2.4). The concentrations applied for salt treatment are control, 50, 100, 150 and 200 mM but in the results there is only three concentrations. The same for salicylic acid application the concentrations levels are different than in the results.

Response back: Line 133; There are three levels i.e. control, 25, 50, and 100 mM.

More details are needed to explain how to apply salinity treatment. Do you use the salt solution at each irrigation? What is the volume you use to spray the salicylic acid solution.

Response back: Line 135; All the irrigations were made with saline soils except control (0 mM NaCl). Volume of salicylic acid solution was 10 mL and add these informations are added in the manuscript.

Line 131 write in past the verb start.

Response back: Line 132; corrections have been made.

 For carotenoids measurement, the samples are the grains or the leaves. It is not clear, please do you explain ?

Response back: Line 145; Samples were the leaves and information added in the manuscript.

In line 131, what does [0.5M] indicate?

Response back: Line 145; corrections have been made as 0.5 g of leaf samples

You should indicate the number of repetitions for each experiment.

Response back: corrected as there were three replications the each treatment.

In the results part

In table 1, can you explain how the difference between the Ec of the soil for the different salinity concentration are low ( 1.76 -1.84) and the differences of Na and Cl concentrations are high.

Response back: Line 196-197; Description is added within the manuscript because EC indicate the level of salinity in soil. However, soils can be categorized on the basis of EC.

Please correct the shift on Y axis of all the figures.

Response back: Axis has been changed

Line 276, 278, 312, 314, and 316, rewrite the sentences to avoid repetition of mM.

Response back: Line 283, 285, 319, 321, 323, Repetition of mM has been removed from the manuscript

In the discussion part Ref 31 is missing.

Response back: This was typo, thank you for correction.

Please review this part (line 400-408) of the discussion as it is not clear and contradictory.

Response back: Line 405-410; This part has been rephrased to avoid contradiction.

 

Reviewer 2 Report

See attaced file

Comments for author File: Comments.pdf

Author Response

Dear Editor

We are pleased to re-submit our revised manuscript, “Foliar application of salicylic acid improved growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions” Ms. Ref. No.: Sustainability-1910631) to be considered for publication in “Sustainability”.

Thank you very much for making a critical assessment of the original version of our manuscript. We are glad to learn that the editor as well as the reviewer(s) considered our findings meaningful. However, the reviewer(s) provided some valuable comments and suggestions for the potential improvement of the manuscript. Based on the comments, you encouraged us to re-submit a revised version, which takes into account all of the points raised by the reviewer(s). We have now completed a thorough revision as per the recommendations of the reviewer(s). Revised portions are marked in red in the paper. Our response to reviewers’ comments has been listed below point-by-point.

 

Reviewer 2

Review Report Sustainability

Manuscript No.  1910631

Title: Foliar application of salicylic acid improves growth, yield, quality, and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions

 

The study evaluated the application of salicylic acid on growth, yield, quality, photosynthetic pigments, and physiological traits in peas under saline conditions.  Four levels of each factor (exogenous application of salicylic acid – control, 1, 2, 3 μM,  and NaCl in irrigation water – control, 50, 100, 150 mM) were studied. The study was carried out during two seasons under greenhouse conditions in a completely randomized design in pots filled with 8 kg of soil with three replicates.

 

 The growth and yield were significantly reduced at 100 mM NaCl compared to control and 50 mM salinity level while the application of salicylic acid at 3 μM concentration mitigated the harmful effects. The chlorophyll a, chlorophyll b, total chlorophyll, photosynthesis, stomatal conductance, the content of carotenoids, ascorbic acid, and phenolic were decreased at 100 mM NaCl, but were significantly improved under salicylic acid application of 3 μM. Similarly, an increase was also observed in SOD, CAT, POD, and APX under 100 mM NaCl and 3 μM salicylic acid treatment.

 

Though the article presents good results but has some limitations:

The study was carried out in small pots with a limited amount (8 kg) and volume of soil, hence its validity under field conditions is necessary before application of these results;

Both the factors studied are quantitative and hence in case of significant effect of individual factors, it is desirable to discuss results by regression analysis instead of LSD qualitative test and in case of a significant interaction effect between factors, interpretation of results by response surface is commonly recommended (e.g. results presented in Table 2 show a significant interactive effect of NaCl concentration between irrigation water x application of salicylic acid) on all yield traits;

While for yield traits information about the isolated and interactive effects is provided at the end of Table 2 but such information (summary of ANOVA) is missing for the other traits – growth, quality, photosynthetic pigments, enzymatic assays, etc. In the Figures presented comparison is made only by LSD.

Besides this the paper needs a thorough revision, particulaily regarding the  sentences in the following lines:

Line 59/60 ........irrigated water or irrigation water;

Response back: Line 60-61; This is irrigation water however corrected in the manuscript.

Line 74 .............. numerous others effective for.......;

Response back:Line 78; Corrcted as per directions of reviewer

Line 91 .......... stomatal conduce....;

Response back: Line 95; Corrected as suggested.

Line 129 ............ remained continues;

Response back:Line 134; Corrected as per directions

Line 131 ...... salicylic acid starts after;

Response back:Line 136; corection has been made.

Line 157 ............. pigments, a described method by;

Response back: Line 163;Corrcted as per directions of reviewer

Line 189 ...... due to irrigation salinity levels;

Response back: Line 195; Corrrected as per suggestions of reviewer

Figures 2, 3, 4, and 5(wrongly identified as Figure 2 Line 372)  in axis Y avoid overwriting;

Response back: Numbering of figures correcteed

Line 407 ....... by little ...

Response back: Line 413;Corrected

Line 432 ....... increased by the increased

Response back: Line 438; Sentense has been rephrased

Line 437 ... conductance, decreased and transpiration rate increased in pea plants treated with 100 mM  (Needs to be reevaluation and reconsideration).

Response back:line 443-444; Line has been repharsed

 In view of the aforesaid, the paper can not be accepted in the current form and needs a thorough revision and resubmission for evaluation.  

 

Reviewer 3 Report

Dear Authors,

The work entitled: "Foliar application of salicylic acid improves growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defence mechanism under saline conditions" is interesting and addresses not only an important research problem, but also a practical one. However, it requires some important corrections before being accepted for printing. Here are the detailed notes:

1)     The title is not grammatically correct.

2)     Abstract should be structured: background, purpose of work, shortly methodology, results, and general conclusion. There is no clear purpose of the work in the abstract

3)     The introduction does not contain a clear purpose of the work as an alternative hypothesis to the null hypothesis and verification of this hypothesis.

4)     Chapter "Material and Methods". The propagating material should have a health certificate from the state seed inspection or state seed laboratory. The mere purchase of seeds from a seed company does not guarantee anything.

5)     Subchapter 2.2. The soil should be better described, its physico-chemical properties and enzymatic activity should be provided.

6)     Subchapter 2.3 The care of plants in pots is enigmatically characterized, it should be supplemented.

7)     Describe in detail the modification to Ruck's method in Section 2.7.

8)     Subsection 2.11. Statistical calculations should be better described. Statistical software is only a calculation tool, but it should be described how the sources of variation were tested, and what confidence intervals were used for the comparisons of means. What does it mean "Intermediate treatment was separated with a 5% probability of the LSD test" - explain

9)     Chapter "Results". The test results in Table 1 have not been properly discussed and interpreted. These test results should be used properly.

10)   The discussion of the results in Figure 2 is sketchy, and the figure does not provide all the information as to the significance of the influence of the studied factors. I am asking for a solid discussion of the results in this figure

11)   The titles of subsections 3.1, 3.2, 3.3, 3.4, 3.5 are not appropriate. They should not suggest conclusions, because not all levels of the application factor of salicylic acid improved the examined traits, some caused their deterioration. Therefore, the titles and subtitles of the chapters should be formulated carefully, preferably using neutral phrases, e.g., "Effect of salicylic acid on the quality of Pisum sativum traits"

12)   The test results presented in Figures 2 and 3 are chaotically described and poorly interpreted.

13)   The commentary on subchapter 3.6 is insufficient. Please discuss the research results thoroughly.

14)   The discussion of the results is cursory and generalizing. It requires deepening and using the latest literature.

15) More specialized vocabulary should be used in the 'Conclusions' section. Not the "machinery" of photosynthesis, but the "machinery" of photosynthesis. Corollary 2 is too generalized. Authors should add a conclusion for agricultural practice.

Comments for author File: Comments.pdf

Author Response

 

Dear Editor

We are pleased to re-submit our revised manuscript, “Foliar application of salicylic acid improved growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions” (Ms. Ref. No.: sustainability-1910631) to be considered for publication in “sustainability”.

Thank you very much for making a critical assessment of the original version of our manuscript. We are glad to learn that the editor as well as the reviewer(s) considered our findings meaningful. However, the reviewer(s) provided some valuable comments and suggestions for the potential improvement of the manuscript. Based on the comments, you encouraged us to re-submit a revised version, which takes into account all of the points raised by the reviewer(s). We have now completed a thorough revision as per the recommendations of the reviewer(s). Revised portions are marked in red in the paper. Our response to reviewers’ comments has been listed below point-by-point.

Reviewer 3

The work entitled: "Foliar application of salicylic acid improved growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defence mechanism under saline conditions" is interesting and addresses not only an important research problem, but also a practical one. However, it requires some important corrections before being accepted for printing. Here are the detailed notes:

1)     The title is not grammatically correct.

Response back: Title correction has been made

2)     Abstract should be structured: background, purpose of work, shortly methodology, results, and general conclusion. There is no clear purpose of the work in the abstract

Response back: Corrections have been made in the abstract as per directions of the reviewers.

3)     The introduction does not contain a clear purpose of the work as an alternative hypothesis to the null hypothesis and verification of this hypothesis.

Response back: Line 107-111; corrections have been made as per suggestions of reviewer.

4)     Chapter "Material and Methods". The propagating material should have a health certificate from the state seed inspection or state seed laboratory. The mere purchase of seeds from a seed company does not guarantee anything.

Response back: Company may get seed health certificate from state seed laboratory. However, authors get seed from the seed company.

5)     Subchapter 2.2. The soil should be better described, its physico-chemical properties and enzymatic activity should be provided.

Response back: Physico-chemical properties of soils were described in the Table 1.

6)     Subchapter 2.3 The care of plants in pots is enigmatically characterized, it should be supplemented.

Response back: Corrected as per suggestions of reviewer.

7)     Describe in detail the modification to Ruck's method in Section 2.7.

Response back: Line 153-161; Detailed modifications were added in the manuscript.

8)     Subsection 2.11. Statistical calculations should be better described. Statistical software is only a calculation tool, but it should be described how the sources of variation were tested, and what confidence intervals were used for the comparisons of means. What does it mean "Intermediate treatment was separated with a 5% probability of the LSD test" – explain

Response back: Line 187- 190; Comparison made under least significance difference (LSD) test and some modifications are also added.

9)     Chapter "Results". The test results in Table 1 have not been properly discussed and interpreted. These test results should be used properly.

Response back: Table 1 was developed and all the analysis were made from water and soil testing laboratory.

10)   The discussion of the results in Figure 2 is sketchy, and the figure does not provide all the information as to the significance of the influence of the studied factors. I am asking for a solid discussion of the results in this figure

Response back: Line 401-404; The discussion in Figure 2 has been improved

11)   The titles of subsections 3.1, 3.2, 3.3, 3.4, 3.5 are not appropriate. They should not suggest conclusions, because not all levels of the application factor of salicylic acid improved the examined traits, some caused their deterioration. Therefore, the titles and subtitles of the chapters should be formulated carefully, preferably using neutral phrases, e.g., "Effect of salicylic acid on the quality of Pisum sativum traits"

Response back: Subsections have been corrected

12)   The test results presented in Figures 2 and 3 are chaotically described and poorly interpreted.

Response back: some more information is added in the manuscript.

13)   The commentary on subchapter 3.6 is insufficient. Please discuss the research results thoroughly.

Response back: Results are thoroughly described in the results section

14)   The discussion of the results is cursory and generalizing. It requires deepening and using the latest literature.

Response back: Latest references are added in the manuscript.

15) More specialized vocabulary should be used in the 'Conclusions' section. Not the "machinery" of photosynthesis, but the "machinery" of photosynthesis. Corollary 2 is too generalized. Authors should add a conclusion for agricultural practice.

Response back: Line 455-468; Some modifications have been made in the manuscript conclusion as suggested by reviewer.

 

Round 2

Reviewer 2 Report

Although the authors have made changes in the manuscript, but the basic question remains unsolved. As mentioned in my earlier review report, the evaluated factors (irrigation water salinity - EC and salicylic acid concentrations applied are quantitative, so results should have been analyzed by regression analysis and not by comparison of means and in case of significant effect of the interaction by means of the response surface. Further, information about ANOVA is missing for most of the evaluated traits.

Author Response

Dear Editor

We are pleased to re-submit our revised manuscript, “Foliar application of salicylic acid improved growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions” Ms. Ref. No.: Sustainability-1910631R2) to be considered for publication in “Sustainability”.

Thank you very much for making a critical assessment of the original version of our manuscript. We are glad to learn that the editor as well as the reviewer(s) considered our findings meaningful. However, the reviewer(s) provided some valuable comments and suggestions for the potential improvement of the manuscript. Based on the comments, you encouraged us to re-submit a revised version, which takes into account all of the points raised by the reviewer(s). We have now completed a thorough revision as per the recommendations of the reviewer(s). Revised portions are marked in red in the paper. Our response to reviewers’ comments has been listed below point-by-point.

 

Reviewer 2

Although the authors have made changes in the manuscript, but the basic question remains unsolved. As mentioned in my earlier review report, the evaluated factors (irrigation water salinity - EC and salicylic acid concentrations applied are quantitative, so results should have been analyzed by regression analysis and not by comparison of means and in case of significant effect of the interaction by means of the response surface. Further, information about ANOVA is missing for most of the evaluated traits.

Response back: ANOVA of the applied treatments is listed below the LSD tables and mentioned in red color.

Reviewer 3 Report

Dear Authors,

The work has been greatly improved. However, not all answers are complete.

For example, the answer to question 4 is incorrect. It was necessary to provide the degree of seed qualification, not to write evasively and to blame the company trading in propagating material.

The answer to question 5 is evasive. Although the physico-chemical characteristics of the soil are listed in Table 1, they had to be interpreted reliably.

The answer to question 8 is unclear. Do the authors not know the rules of experimentation?

The authors did not answer question 9, but indicate who performed the analysis. However, the Authors are responsible for the results of the work, and they should interpret them.

Answer 15 is evasive. I am asking for a full answer also in the text of the work.

Author Response

 

Dear Editor

We are pleased to re-submit our revised manuscript, “Foliar application of salicylic acid improved growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions” (Ms. Ref. No.: sustainability-1910631R2) to be considered for publication in “sustainability”.

Thank you very much for making a critical assessment of the original version of our manuscript. We are glad to learn that the editor as well as the reviewer(s) considered our findings meaningful. However, the reviewer(s) provided some valuable comments and suggestions for the potential improvement of the manuscript. Based on the comments, you encouraged us to re-submit a revised version, which takes into account all of the points raised by the reviewer(s). We have now completed a thorough revision as per the recommendations of the reviewer(s). Revised portions are marked in red in the paper. Our response to reviewers’ comments has been listed below point-by-point.

Reviewer 3

he work has been greatly improved. However, not all answers are complete.

For example, the answer to question 4 is incorrect. It was necessary to provide the degree of seed qualification, not to write evasively and to blame the company trading in propagating material.

Response back: All the commercial companies had their seed qualification certificate of their propagated material. Moreover, auhors were also assurred the quality of seeds of propagating material 

The answer to question 5 is evasive. Although the physico-chemical characteristics of the soil are listed in Table 1, they had to be interpreted reliably.

Response back: Descripton has been added in the mauscript and mentioned in red color.

The answer to question 8 is unclear. Do the authors not know the rules of experimentation?

Response back: Some modifications have been made. 

The authors did not answer question 9, but indicate who performed the analysis. However, the Authors are responsible for the results of the work, and they should interpret them.

Response back: Detailed interpretation has been addded in the manuscriot and highlighted with red color.

Answer 15 is evasive. I am asking for a full answer also in the text of the work.

Response back: Interreation has been added in the the sggested section and mentioned in blue color.

 

Round 3

Reviewer 2 Report

As mentioned in my earlier review reports, the authors studied two quantitative factors (salicylic acid concentration and irrigation water salinity, hence the results should be presented and discussed by means of regression analysis (if the effect of only one factor is significant on a variable (trait) analyzed or by response surface in case the interactive effect of factors studied is significant. Looks like the problem lies in the experimental design chosen (a completely randomized design instead of split-plot). The advantage of analyzing results by regression analysis or by response surface it is possible to determine precisely to what level of salicylic acid concentration or electrical conductivity the effect is positive or negative in quantitative terms. 

Besides this problem, the article still needs a thorough and critical revision, for example, the footnote below Table 1 mentions means sharing the same letters ....., while no letters follow the means (line 200);  ...... using a procedure of Singleton and Rossi [24] procedure by ....  ( line 153); some places the authors use ..... leaf greenness .... (line 142/217), to define the level of significance, the authors use at some places p at other places P, it should be standardized as per norms.  Many more similar examples can be cited. 

Under such circumstances, I recommend to reconsider the contribution only after major revision. 

Author Response

 

Dear Editor

We are pleased to re-submit our revised manuscript, “Foliar application of salicylic acid improved growth, yield, quality and photosynthesis of pea (Pisum sativum L) by improving antioxidant defense mechanism under saline conditions” (Ms. Ref. No.: sustainability-1910631R3) to be considered for publication in “sustainability”.

Thank you very much for making a critical assessment of the original version of our manuscript. We are glad to learn that the editor as well as the reviewer(s) considered our findings meaningful. However, the reviewer(s) provided some valuable comments and suggestions for the potential improvement of the manuscript. Based on the comments, you encouraged us to re-submit a revised version, which takes into account all of the points raised by the reviewer(s). We have now completed a thorough revision as per the recommendations of the reviewer(s). Revised portions are marked in red in the paper. Our response to reviewers’ comments has been listed below point-by-point.

Reviewer 3:

As mentioned in my earlier review reports, the authors studied two quantitative factors (salicylic acid concentration and irrigation water salinity, hence the results should be presented and discussed by means of regression analysis (if the effect of only one factor is significant on a variable (trait) analyzed or by response surface in case the interactive effect of factors studied is significant. Looks like the problem lies in the experimental design chosen (a completely randomized design instead of split-plot). The advantage of analyzing results by regression analysis or by response surface it is possible to determine precisely to what level of salicylic acid concentration or electrical conductivity the effect is positive or negative in quantitative terms. 

Response back: Authors strongly agreed to the comments sent by reviewer but at this time authors can apologies to work on split plot and regression analysis. Both analysis are informative. Here I am trying to explain the concerns.  The objective of the study was to determine salicylic effects on respective crop. Salicylic acid improves the crop growth, yield and defense related parameters as described in the present results. One more thing, by application of split plot there will a major concern related to statistical analysis. Whole manuscript will be changes which get too much time and not easy for authors to re-write in the light of split plot design. Moreover, split plot is mostly applied on field crop, while current study was conducted in the pots. CRD design is more appropriate which is already applied and results are suitably well written according to this design. A lot of research trails were conducted on these type of statistical analysis to determine the increase in growth and yield under salinity conditions. So, hoping good authors explanation will be accepted.

Khalifa, G.S., Abdelrassoul, M., Hegazi, A.M. and Elsherif, M.H., 2016. Attenuation of negative effects of saline stress in two lettuce cultivars by salicylic acid and glycine betaine. Gesunde Pflanzen, 68(4), pp.177-189.

Yildirim, E., Ekinci, M., Turan, M., Dursun, A., Kul, R. and Parlakova, F., 2015. Roles of glycine betaine in mitigating deleterious effect of salt stress on lettuce (Lactuca sativa L.). Archives of Agronomy and Soil Science, 61(12), pp.1673-1689.

Dehghan, S., Sadeghi, M., Pöppel, A., Fischer, R., Lakes-Harlan, R., Kavousi, H.R., Vilcinskas, A. and Rahnamaeian, M., 2014. Differential inductions of phenylalanine ammonia-lyase and chalcone synthase during wounding, salicylic acid treatment, and salinity stress in safflower, Carthamus tinctorius. Bioscience reports, 34(3).

Ardebili, N.O., Saadatmand, S., Niknam, V. and Khavari-Nejad, R.A., 2014. The alleviating effects of selenium and salicylic acid in salinity exposed soybean. Acta Physiologiae Plantarum, 36(12), pp.3199-3205.

Naz, S., Mushtaq, A., Ali, S., Muhammad, H.M.D., Saddiq, B., Ahmad, R., Zulfiqar, F., Hayat, F., Tiwari, R.K., Lal, M.K. and Altaf, M.A., 2022. Foliar application of ascorbic acid enhances growth and yield of lettuce (Lactuca sativa) under saline conditions by improving antioxidant defence mechanism. Functional Plant Biology.

Besides this problem, the article still needs a thorough and critical revision, for example,

the footnote below Table 1 mentions means sharing the same letters ....., while no letters follow the means (line 200);  ...... using a procedure of Singleton and Rossi [24] procedure by ....  ( line 153); some places the authors use ..... leaf greenness .... (line 142/217), to define the level of significance, the authors use at some places p at other places P, it should be standardized as per norms.  Many more similar examples can be cited. 

Response back: the footnote below Table 1 mentions means sharing the same letters “corrections has been made and mention in red color. Line 200 removed whcih was irrlevant. All procedures have been correct as per suggstion. P has been replaced with p in uniform style.

Under such circumstances, I recommend to reconsider the contribution only after major revision. 

 

Round 4

Reviewer 2 Report

Comments for author File: Comments.pdf

Author Response

I revised all comments..

Author Response File: Author Response.pdf