Enhancing the Quality of Two Species of Baby Leaves Sprayed with Moringa Leaf Extract as Biostimulant
Round 1
Reviewer 1 Report
The article "Enhancing the quality of two species of baby leaves sprayed with moringa leaf extract as biostimulant" with authors Stefania Toscano, Antonio Ferrante and Daniela Romano, presents new information regarding the possibilities for application of leaf extract from Moringa oleifera Lam. The article is well structured and the experiments are logically designed. However there are some weak points which need to be addressed:
Abstract - The abstract is informative enough, however the concluding sentence (L# 24-26) is not convincingly supported by the text.
The use of abbreviation MLE >< LME needs to be unified through the entire article.
Introduction - The introductory information is well presented and the problem is comprehensively described.
Lines# 40-42: Please rewrite this sentence.
Materials and Methods - The methods are thoroughly described. However, reading this part some questions pop-up and the answers to some of them is good to be mentioned in the article:
The authors state that the initial MLE extract was 25% (why 25% but not more?), then some dilution was made. So, I wonder what was the final concentration used? Also, is there a recommended dosing for application of MLE, and how the authors have chosen the treatment dose for their experiments? And another one, what is the rationale to shade dry the leaves and then wet them back? A final one, is there a particular reason the authors to choose these two varieties of these plant species, so to compare the effects of MLE on them?
Another remark, it is not clear how many days after the last treatment the samples for analyses were taken.
Statistics - The authors indicate that the data were analyzed by two-way ANOVA. In part of the Figures one-way ANOVA is mentioned. Please justify which-way ANOVA was used.
Results - This section in my opinion needs an extended revision. My concerns are related mainly to the way the data are presented, especially in the Tables. The data described in the tables bring some difficulties to understand from where the data originate: For example, in Table 2 the first two rows represent the genotypes BN and CL which (logically for me) have to be non-treated. Then what the next two rows stand for? There are Control and Treatment. They should be associated to a Genotype, but to which one? If they are not related to a Genotype, then please explain what they are. This is valid for all Tables excluding Table 1.
Please rewrite this part and try not to present same data as tables and figures.
Lines# 216-218. There is not much logic the CL-Control to be increased in relation to BN-Treatment, because: a). It is a Control. b). It is not a BN Control. Please switch the point of view and rewrite this part.
Regarding Figures - the data presented in figures are much more understandable than the tables. My recommendation would be the authors to use a more distinct color scheme (or add patterns) of the greens, which both I like very much but some readers may find it difficult to distinguish the treatments.
Line #242 Table 3 --> Table 4.
Line # 244 Figure 2 --> Figure 3. Please renumber all following Figures in figure captures and in the text.
Discussion - The Discussion section covers the problem and the data presented. My recommendation would be the discussion to be adapted to the new version of the Results section (if the authors decide to rewrite this).
Conclusions - Lines #437-440 need revision.
The manuscript is written in a good English language which in some parts needs a little polishing.
My overall impression of the article is positive. The problem discussed is important as biostimulants of natural origin are a perspective area of interest which definitely needs to be investigated.
Author Response
The article "Enhancing the quality of two species of baby leaves sprayed with moringa leaf extract as biostimulant" with authors Stefania Toscano, Antonio Ferrante and Daniela Romano, presents new information regarding the possibilities for application of leaf extract from Moringa oleifera Lam. The article is well structured and the experiments are logically designed. However there are some weak points which need to be addressed:
Dear reviewer,
The authors would like to thank the reviewers for their comments. The manuscript has been accordingly revised. Corrections and suggestions have been implemented in the current version of the manuscript. We hereby provide a point-by-point answer.
The authors
R: Abstract - The abstract is informative enough, however the concluding sentence (L# 24-26) is not convincingly supported by the text.
AA: The concluding sentence was modified according your suggestion.
R: The use of abbreviation MLE >< LME needs to be unified through the entire article.
AA: Sorry for he mistakes. The corrections were found in all manuscript.
R: Introduction - The introductory information is well presented and the problem is comprehensively described.
AA: Thanks for their comment.
R: Lines# 40-42: Please rewrite this sentence.
AA: The sentence was rewrite.
R: Materials and Methods - The methods are thoroughly described. However, reading this part some questions pop-up and the answers to some of them is good to be mentioned in the article:
The authors state that the initial MLE extract was 25% (why 25% but not more?), then some dilution was made. So, I wonder what was the final concentration used? Also, is there a recommended dosing for application of MLE, and how the authors have chosen the treatment dose for their experiments?
AA: The extract obtained was indicated as 25% extract which was further diluted to the ratio 1:30 (v/v). Tween 20 (0.05%) was added to spray solutions as a wetting agent. The concentration of moringa leaf extract (MLE) was of 200 mg L-1. The choice was done on the basis of the literature. In particular, the same concentration used in the work of Sakr et al., (2021) was used on brassica plants.
R: And another one, what is the rationale to shade dry the leaves and then wet them back?
AA: The procedure for preparing the moringa extract followed what was reported by Pervez et al., (2017). In particular, the collected leaves were dried in the shade and finely ground with an electric grinder. 50 g of the plant material was immersed in 200 ml of distilled water. The mixture was kept at 25 ° C for 48 hours and filtered using Whatman No. 1 filter paper. The dilution required to obtain a concentration of 200 mg L was then carried out (Sakr et al., 2021).
R: A final one, is there a particular reason the authors to choose these two varieties of these plant species, so to compare the effects of MLE on them?
AA: The choice of the two genotypes was made on the basis of previous paper (Di Bella et al 2019). In particular, these are two genotypes that have different leaf characteristics for which we wanted to investigate a possible different response to the treatment.
R: Another remark, it is not clear how many days after the last treatment the samples for analyses were taken.
AA: The plants were sprayed three times with moringa leaf extract (MLE), at 15, 30 and 1 day before harvest (43 days after planting). This information relative to treatment has been added.
R: Statistics - The authors indicate that the data were analyzed by two-way ANOVA. In part of the Figures one-way ANOVA is mentioned. Please justify which-way ANOVA was used.
AA: Sorry for the mistake, a typo has been reported. A two-way ANOVA was carried out, and in the figures the interaction was calculated.
R: My concerns are related mainly to the way the data are presented, especially in the Tables. The data described in the tables bring some difficulties to understand from where the data originate: For example, in Table 2 the first two rows represent the genotypes BN and CL which (logically for me) have to be non-treated. Then what the next two rows stand for? There are Control and Treatment. They should be associated to a Genotype, but to which one? If they are not related to a Genotype, then please explain what they are. This is valid for all Tables excluding Table 1.
AA: thank you for the comments. We have reported the mean of the data for each genotype including the control and MLE treatment for both BN and CL. However, as requested by the reviewer, data has been included as suggested in addition to the overall means.
R: Please rewrite this part and try not to present same data as tables and figures.
AA: The text has been revised after the table modification.
R: Lines# 216-218. There is not much logic the CL-Control to be increased in relation to BN-Treatment, because: a). It is a Control. b). It is not a BN Control. Please switch the point of view and rewrite this part.
AA: The sentence was rewrite.
R: Regarding Figures - the data presented in figures are much more understandable than the tables. My recommendation would be the authors to use a more distinct color scheme (or add patterns) of the greens, which both I like very much but some readers may find it difficult to distinguish the treatments.
AA: The figure colors have been changed to make them more understandable.
R: Line #242 Table 3 --> Table 4.
AA: Sorry for the mistake. The correction was found.
R: Line # 244 Figure 2 --> Figure 3. Please renumber all following Figures in figure captures and in the text.
AA: Sorry for the mistakes. The corrections were found.
R: Discussion - The Discussion section covers the problem and the data presented. My recommendation would be the discussion to be adapted to the new version of the Results section (if the authors decide to rewrite this).
AA: The discussion has been revised after the result modification.
R: Conclusions - Lines #437-440 need revision.
AA: The conclusion was rewrite according to your comment.
The manuscript is written in a good English language which in some parts needs a little polishing.
My overall impression of the article is positive. The problem discussed is important as biostimulants of natural origin are a perspective area of interest which definitely needs to be investigated.
Author Response File: 
Author Response.docx
Reviewer 2 Report
The obtained results concerning the effect of leaf extract of moringa on baby leaf characteristics of two genotypes of Brassica: kale and broccoli are well interpreted and compared with the literature data. The authors showed a different response of the assessed genotypes to the experimental factor. The very beneficial effect of leaf extract of moringa on the health properties of kale indicates the purposefulness of the research undertaken. The authors rightly pointed out that the biostimulant can have a variable effect from species to species, which limits the possibility of generalizations and at the same time indicates the need for further research. The authors' conclusion is correct that the work should be continued in order to determine the dose of the applied biostimulator. The work does not require major changes, and editorial notes are marked in the text. In table 7., I propose to keep the order as in tables 2, 3, 4, 5, 6, where BN is placed before CL. A sentence from lines 226-227: an increase in CL compared to BN genotype was found (P <0.05); I suggest writing like this: significantly more leaves were registered on CL than BN genotype (P <0.05);
Comments for author File: 
Comments.pdf
Author Response
The obtained results concerning the effect of leaf extract of moringa on baby leaf characteristics of two genotypes of Brassica: kale and broccoli are well interpreted and compared with the literature data. The authors showed a different response of the assessed genotypes to the experimental factor. The very beneficial effect of leaf extract of moringa on the health properties of kale indicates the purposefulness of the research undertaken. The authors rightly pointed out that the biostimulant can have a variable effect from species to species, which limits the possibility of generalizations and at the same time indicates the need for further research. The authors' conclusion is correct that the work should be continued in order to determine the dose of the applied biostimulator. The work does not require major changes, and editorial notes are marked in the text.
Reviewer #2:
Dear reviewer,
The authors would like to thank the reviewers for their comments. The manuscript has been accordingly revised. Corrections and suggestions have been implemented in the current version of the manuscript. We hereby provide a point-by-point answer.
R: In table 7., I propose to keep the order as in tables 2, 3, 4, 5, 6, where BN is placed before CL.
AA: The revision was found.
R: A sentence from lines 226-227: an increase in CL compared to BN genotype was found (P <0.05); I suggest writing like this: significantly more leaves were registered on CL than BN genotype (P <0.05);
AA: The sentence was modified.
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
Dear colleagues,
I have no more remarks on your paper.
Thanks for answering my questions.
Best regards,
Reviewer XY
