Enhancing Seed Potato Production from In Vitro Plantlets and Microtubers through Biofertilizer Application: Investigating Effects on Plant Growth, Tuber Yield, Size, and Quality

Round 1

Reviewer 1 Report

Dear authors,

It is a well-written paper. However the statistical analysis and presentation as well as interpretation of the results still need to be revised appropriately. The experiment cannot be analyzed as a two-factor experiment, but it has to be analyzed as a three-factor experiment. The second factor called Cultivars actually consists of two independent variables, i.e. cultivars (consisting of two cultivars, i.e. Spunta & Russet) and types of seedlings (consisting of two types, i.e. microtuber and plantlet). 

If these two independent variables are combined into only one factor consisting of 4 levels, i.e. Spunta plantlets, Spunta microtubers, Russet plantlets and Russet microtubers, then it means that the authors are trying to do multiple comparison among Spunta plantlets, Spunta microtubers, Russet plantlets and Russet microtubers. Statistically, this comparison cannot be done because Spunta microtubers and Russet plantlets or Spunta plantlets and Russet microtubers are not comparable, so they cannot be scientifically compared, because they are different in both cultivars and types of seedlings. If they are significant, what could be the reason? Was it because of the difference in cultivars or the difference in types of seedlings? Nobody can answer it.

Therefore, the experiment and the data have to be analyzed using a three-way ANOVA, which enables the authors to find out the effects of biofertilizers, the effects of cultivars, and the effects of different types of seedlings, as well as the interaction effects among these three experimental factors.

Please review the reviewer's comments on the manuscript, which I think can help the authors revise this manuscript to be relevant for publication in Agronomy.

Comments for author File: Comments.PDF

Author Response

Dear Reviewer,

Thank you for your three-way ANOVA statistical analysis suggestion. Indeed, the article has been revised, and this has become much more valuable. The results were analyzed as a three-factor experiment as below. Statistical analysis and presentation; tables (Tables 3, 4, 5, 6, 7) and graphs (Figures 3 and 4), and interpretation of the results, are revised appropriately. Newly added revisions to the manuscript are written in red color.

Three-Way-ANOVA has been done as follows:

Cultivar (2)

Seed Material (2)

Biofertilizer (5)

Cultivar x Seed Material (2*2=4)

Cultivar x Biofetilizer (2*5=10)

Seed Material x Biofertilizer (2*5=10)

Cultivar x Seed Material x Biofertilizer (2*2*5= 20)

Also, thank you very much for your good comment on the English evaluation of our article.

THE ANSWERS TO THE QUESTIONS ON THE PDF MANUSCRIPT ARE BELOW:

Comment [WW3] Five plants of uniform growth were randomly selected for each treatment. How many plants were grown per treatment?

Amended, Line 205: A total of 30 plants per treatment were grown. Five plants of uniform growth were randomly selected for each treatment.

Comment [WW5]: Why per liter? Wasn’t the sample ovendried leaf?

Amended : The revised manuscript used the tuber mineral element unit as mg kg^‒1.

All comments regarding statistical analysis on the PDF manuscript were answered by performing a three-way-ANOVA statistical analysis with new tables (Tables 3, 4, 5,5 6, 7) , graphs (Figures 3 and 4) and full text (See revised manuscript).

Author Response File: Author Response.pdf

Reviewer 2 Report

1.The paper is too long, it should be shortened by reducing some data and description.

2.Ihe important of Seed Potato industry in local should be shown in the introduction.

3.The differences between control and T2, T3, T4 are small, discussion and conclusion should be careful and find out the reasons. I think maybe more nutrients in the substrate worked. 

4.To some extent, vermicompost is one of the best organic fertilizer, compare to the mycorrhiza, beneficial bacteria, its component is more like a fertilizer, not microbial agent, so when test them together, the difference is great. 

The quality of paper is not so high, the experiment is too simple and less enlightenment for readers. 

Author Response

Dear Reviewer,

Thank you for your evaluation and suggestions for our Manuscript. Our point-by-point answers to your evaluations and comments are below. Newly added revisions to the manuscript are written in red color.

1.The paper is too long. It should be shortened by reducing some data and description.

Reply: Upon your suggestion, leaf mineral element analysis results (N, P, K, Ca, Mg, Fe, Mn, Zn, and Cu) and seed mini tuber microelements (Fe, Mn, Zn and Cu) have been removed from the manuscript.

2.The important of the Seed Potato industry in local should be shown in the introduction.

Reply: Lines 49-52 The local importance of potatoes for Tunisia is now mentioned.

3.The differences between control and T2, T3, T4 are small, discussion and conclusion should be careful and find out the reasons. I think maybe more nutrients in the substrate worked. 

Reply: Lines 485-488 Amended: Three-way-ANOVA statistical analysis was re-done on the suggestion of the first reviewer, and necessary comparisons were made about biofertilizers (T1, T2, T3, T4, T5) in the rewritten "Results" section. In addition, in the “Discussion section”, the nutritional content and microorganism presence of vermicompost, which is different from other biofertilizers, is given.

4.To some extent, vermicompost is one of the best organic fertilizer, compare to the mycorrhiza, beneficial bacteria, its component is more like a fertilizer, not microbial agent, so when test them together, the difference is great. 

Reply: We agree entirely with your comment about vermicompost. However, it is known that there are billions of beneficial bacteria, mycorrhizae and other microorganisms in vermicompost.

Author Response File: Author Response.pdf

Reviewer 3 Report

Question 1: Line 1, The title should be in a more organized way.

Question 2: Line 214, Describe the analysis of starch content, should be clearly written form.

Question 3: Need to upgrade grammatical errors.

Question 4: Introduction and discussion part should be upgraded?

Need to upgrade grammatical errors.

Author Response

Dear Reviewer,

Thank you for your evaluation and suggestions for our Manuscript. Our point-by-point answers to your evaluations and comments are below. Newly added revisions to the manuscript are written in red color.

Question 1: Line 1, The title should be in a more organized way.

Amended new title: Enhancing Seed Potato Production from in vitro Plantlets and Microtubers through Biofertilizer Application: Investigating Effects on Plant Growth, Tuber Yield, Size, and Quality

Question 2: Line 214, Describe the analysis of starch content, should be clearly written form.

Reply: Lines 243-253: Starch analysis is written in detail.

Question 3: Need to upgrade grammatical errors.

Reply: Grammatical errors are corrected now throughout the article.

Question 4: Introduction and discussion part should be upgraded?

Reply: The introduction and discussion parts were upgraded and as highlighted in the revised manuscript with red text.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Dear Authors, you have done a significant revision to significantly improve this paper. However, please read again carefully and do some more correction and necessary revisions. I also think as what I usually do, to show interaction effect very clearly, the easiest way to show and to make correct interpretation is using bar chart like in Figure 3, but with SE error bars. This is especially true when showing and interpreting a three-way interaction effect, at least for the parameters that are the focus of this study. In addition, based on the statistics convention, which is highly scientifically logical, when the three-way interaction effects are significant, the the conclusion must be drawn from the three-way interaction because other effects (main effects and the two-way interaction) become ungeneralizable.  

Comments for author File: Comments.PDF

Please check carefully, some statements need to be revised in order to clearly state the correct conditions or treatments. 

Author Response

Dear Reviewer,

Thank you very much for your valuable suggestions. SE error bars have been added to Figures 3 and 4 in the manuscript. Titles of the tables were corrected according to your suggestions. All of your recommendations on MS have been implemented, and below the point-by-point responses are shown. As your suggestion “Conclusion” and “Abstract” sections have been drawn from the three-way interactions. Newly added revisions to the manuscript are written in blue color on MS. 27.09.2023

Thanks again to your valuable contributions, the manuscript has greatly improved in quality.

Best regards

Prof. Dr. Hayriye Yildiz DASGAN

Corresponding author

RESPONSES TO COMMENTS ON MANUSCRIPT

Commented [MC1]: Please rewrite the contents of the Abstract based on the revised conclusion.

Reply: According to the revised "Conclusion," the Abstract was also revised and rewritten. Please see blue color highligts in Abstract.

Commented [MC2]: 60% from what? Please explain how this % was calculated?

Reply: 60% was written unnecessarily; I deleted it in Table 2

Commented [MC3]: Phosphor oxide is not K2O (this is Potasium)…

Reply: It was corrected as “potassium” in Table 2.

Commented [MC4]: Peaks or a peak?

Reply: It was corrected as “peak” in Line 238.

Commented [MC5]: Please do not confuse the readers, please fix the title of Table 4, because Table 4 is only showing the main effect of each factor and the two-way interaction effects of all factors.

Reply: The title of Table 4 has been corrected as “Main effect of cultivar, in vitro material, biofertilizer and the two-way interaction effects of all factors on plant growth parameters”.

Commented [MC6]: Please do not confuse the readers, please fix the title of Table 5, because Table 5 is only showing the three-way interaction effect.

Reply: The title of Table 5 has been corrected as “Three-way-interactions effects of cultivar, in vitro material, biofertilizer on plant growth parameters.” 

Commented [MC7]: Throughout the Results (and Discussion), when referring to interaction of the factors (two-way or thgree-way interaction), it is OK to use “x”, BUT in referring to combination of treatments, such as ‘Spunta’ grown from microtubers, please use “+”, such as ‘Spunta+microtuber’ combination. This needs to be done in order this paper not to confuse the readers between the interaction (the factors) and the combination (the treatment

applied).

Reply: All the corrections have been made throughout the manuscript and are highlighted in blue.

Commented [MC8]: Please place the 40% appropriately and please fix this kind of statement because this statement says that control used 40% mineral fertilizer, which is not correct because control used 100%, while those using 40% was the biofertilizer treatments (as stated in Table 2).

Reply: Amended Line 360 as 100%.

Commented [MC9]: In order not to confused with interaction, this should be written

Spunta+microtuber+vermicompost combination, or explain further: “Spunta variety grown from microtuber fertilized with vermicompost”

Reply: Amended Line 367-368-371-72 and the others….

Commented [MC10]: Please do not confuse the readers, please fix the title of Table 6, because Table 6 is only showing the main effect of each factor and the two-way

interaction effects of all factors.

Reply: The title of Table 6 has been corrected as “Main effect of cultivar, in vitro material, biofertilizer and the two-way interactions on tuber’s sugar, starch and nutrients.”

Commented [MC11]: Please do not confuse the readers, please fix the title of Table 7, because Table 7 is only showing the three-way interaction effect.

Reply: Reply: The title of Table 7 has been corrected as “Three-way-interactions effects of cultivar, in vitro material, biofertilizer on tuber’s sugar, starch and nutrients.”.

Commented [MC12]: Since the three-way interaction effects were significant on tuber yield, most yield components, and mineral contents, then the most valid for the conclusion is the result of the three-way interaction effect. Main effects and the two-way interaction effects then cannot be generalized. Please make sure this is done when the three-way interaction effects are significant. Please use the revised conclusion to rewrite the contents of the Abstract.

Reply: The conclusion was restructured according to the referee's suggestion, Please see blue color highligts in Conclusion.

Commented [MC13]: Please state clearly “when the variety used was Spunta”. When using Russet variety, the second highest was in mycorrhiza biofertilizer.

Reply: Amenden; the statement was not clear so the relevant sentence was deleted from the 'Conclusion' section.

Author Response File: Author Response.pdf

Reviewer 2 Report

Authors have revised the manuscript carefully and strictly, the quality improved, I suggest accepting to publish.

Author Response

Dear Reviewer,

We would like to express our sincere gratitude for your appreciation of the revisions we made to our manuscript, and for your recommendation to accept the article. Thanks to your valuable contributions, the manuscript has greatly improved in quality. We are truly thankful for your support.

Sincerely

Best regards

Prof. Dr. Hayriye Yildiz DASGAN

Corresponding author

Reviewer-2 commenst:

Comments and Suggestions for Authors: Authors have revised the manuscript carefully and strictly, the quality improved, I suggest accepting to publish.

Author Response File: Author Response.pdf