Can Moringa Leaf Spray Treatment Increase the Nutraceutical Properties of Radish Baby Leaf?
Round 1
Reviewer 1 Report
Undoubtedly, an important aspect of the research is the use of plant extracts as possible growth stimulants to improve the chemical properties of plants.
The Authors undertook to evaluate the effects of MLE on young radish leaves, which is interesting due to their use as a food product (using potential plant waste) and by the stimulant itself. The article is written correctly; however, I have some major and minor comments:
Enter affiliation.
Is there a waiting period after spraying before consuming the leaves? Has it been checked?
Continuing with the question above, is this extract safe to consume at the dosages used?
How was the MLE dose determined?
Why were these spraying times chosen?
Have all measurements been made after the entire spraying series? Why was it used three times with the preparation, finally research was carried out? As it makes sense, one dose could have had the same effect. With the possible use of such sprayers for cultivation practice, it is unjustified from an economic point of view.
I am asking for a better description of the substrate used for the research (content of micro and macro elements, pH, etc.)
Please complete the number of measurements (n =?), This applies to the entire manuscript.
Please correct the language of the pictures to English.
Letters indicating differences in relevant statistics are always ranked from the highest value, please correct.
Please provide numerical values up to three significant places (applies to the entire work).
Please correct the conclusions. Indicate in detail in which cases the dose of 1:30 was indicated, and in which 1:40. It is impossible to say which one is better.
Author Response
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 and highlighted in yellow. We hereby provide a point-by-point answer.
The authors
Enter affiliation.
A.A.: Sorry for the mistake. The information were added.
Is there a waiting period after spraying before consuming the leaves? Has it been checked?
A.A.: The plants were sprayed three times with moringa leaf extract (MLE), at 15, 30 and 2 days before harvest (45 days after planting). We have added this information to the text.
Continuing with the question above, is this extract safe to consume at the dosages used?
A.A.: We do not think that the fresh leaf extract used is harmful to health. The leaves of this plant, in fact, are a rich source of nutrients such as protein, carbohydrates, fibre, vitamin C, β carotene and minerals such as calcium, potassium, iron and phosphorus. The leaves contain antioxidant compounds of different kinds, such as ascorbic acid, flavonoids, carotenoids and phenolic compounds, and function as a natural antioxidant (Joshi P, Mehta D. Effect of dehydration on the nutritive value of drumstick leaves. Journal of Metabolomics and Systems Biology 2010, 1(1):5-9.)
How was the MLE dose determined?
Thanks for the question. The doses of moringa leaves were determined according to the information available in literature (Zulfiqar, F., Casadesús, A., Brockman, H., & Munné-Bosch, S. (2020). An overview of plant-based natural biostimulants for sustainable horticulture with a particular focus on moringa leaf extracts. Plant Science, 295, 110194), but we believe that this aspect deserves to be further investigated. At the moment, we are carrying out a trial on baby leaf, where different doses (from 1:10 to 1:50) of moringa leaves are analysed. In the conclusion of the paper this aspect (dose and modality of application of moringa) was underlined as object of future investigation.
Why were these spraying times chosen?
A.A.: The better performance of foliar applications has been attributed to the immediate interaction with the plant tissues because foliar absorption happens almost immediately. The optimal application times for these extracts were determined to be around every 10–14 days for provoking the best plant responses (Alì et al. 2021. Biostimulant Properties of Seaweed Extracts in Plants: Implications towards Sustainable Crop Production)
Have all measurements been made after the entire spraying series? Why was it used three times with the preparation, finally research was carried out? As it makes sense, one dose could have had the same effect. With the possible use of such sprayers for cultivation practice, it is unjustified from an economic point of view.
In literature normally the treatments of Moringa leaf extract were reiterated many times (see Zulfiqar et al., 2020 already mentioned); the reason is linked to characteristics of natural stimulants. We did not analyse the economic aspect of the treatment. Your observation will be useful for further trials, aimed at analysing the influence, in addition to the dose, of the number of treatments.
I am asking for a better description of the substrate used for the research (content of micro and macro elements, pH, etc.)
The substrate that was used was a commercial substrate from Vigorplant Italia SRL. The information on the packaging are: pH 5,5 — 6,5; EC 0,15 - 0,25 dS/m. These information were reported in the text. We do not have the composition of the micro and macro elements, and we had not investigated because the utilized substrate was the same for all treatments. We will consider this suggestion for future trials.
Please complete the number of measurements (n =?), This applies to the entire manuscript.
A.A.: Done. The information were added.
Please correct the language of the pictures to English.
A.A.: Sorry for the mistake. The word was corrected.
Letters indicating differences in relevant statistics are always ranked from the highest value, please correct.
A.A.: Sorry for the mistake; I was wrong in calculating the standard error in Table1. Now I have corrected the data in the whole table.
Please provide numerical values up to three significant places (applies to the entire work).
A.A.: The three significant places were added to all tables.
Please correct the conclusions. Indicate in detail in which cases the dose of 1:30 was indicated, and in which 1:40. It is impossible to say which one is better.
A.A.: More information in the conclusions were added.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The manuscript titled "Can moringa leaf spray treatment increase the nutraceutical characteristics in radish baby leaf?" deals with rather interesting topic in both agriculture and horticulture that is application of various plant extracts as potential biostimulants to other plant species, here, in particular, aqueous extract of Moringa oleifera. The manuscript is well structured, the problem and the basic hypothesis are properly presented in Introduction, the M&M section describes assays very well although some clarifications are still needed, statistical analysis seems to be correctly done, and Results and Discussion contain enough information to derive the answer posed in the main title. However, the manuscript suffers from mild language usage problems, which are highlighted yellow and here-and-there interposed with my comments in the attached file. One of the main concerns might be statistical significance in Table 1 for the values of Unit leaf area and Wy. These values are labeled with different significance letters, while standard errors seem to be large enough to reach other values. My advice it to recheck these values and to conduct another testing if necessary.
Comments for author File: Comments.pdf
Author Response
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 and highlighted in yellow. We hereby provide a point-by-point answer.
The authors
One of the main concerns might be statistical significance in Table 1 for the values of Unit leaf area and Wy. These values are labeled with different significance letters, while standard errors seem to be large enough to reach other values. My advice it to recheck these values and to conduct another testing if necessary.
A.A.: Sorry for the mistake; I was wrong in calculating the standard error. Now I have corrected the data in the whole table
Line 4: 1
A.A.: Done
Line 9: Erroneous article (the, a, an) usage throughout the whole manuscript.
A.A.: The manuscript has been accordingly revised also in English language certified by MDPI.
Line 9: belonging to the
A.A.: Done
Line 23: compressions are not used in scientific literature.
A.A.: Done
Line 64: All these compounds along with flavonoids make zeatin? This is not true and the sentence is too vague to be understood. Please rewrite to clarify.
A.A.: The sentence was rewrite
Line 68: what are "biochemical metabolites"? Did the authors mean "specialized metabolites"?
A.A.: The sentence was modified.
Line 69: Please note that there are hundreds of scientific papers dealing with this topic (https://scholar.google.com/scholar?hl=sr&as_sdt=0%2C43&q=moringa+leaf+extract+biostimulant&btnG=)
A.A.: The sentence was rewrite
Line 74: Too subjective claim for scientific literature.
A.A.: A reference was added that underline the ornamental value of moringa
Line 86: Please specify "ordinary techniques".
A.A.: Done
Line 92: or
A.A.: Done
Line 116: This cannot be a subtitle for the M&M section. Maybe "Quantification of chlorophyll and carotenoids"?
A.A.: Thanks for their comment. The sentence was modified in “Quantification of chlorophyll and carotenoids”.
Line 126: Please do not start a sentence with a digit.
A.A.: Done
Line 129: Please rephrase to input more information.
A.A.: More information were added.
Line 132: Please note that content cannot be extracted but total phenolics. Please rephrase.
A.A.: The sentence was rewritten
Line 133: Should there be information on duration in hours?
A.A.: Sorry for the mistake. The sentence was modified.
Line 140: Please merge these two paragraphs.
A.A.: Done
Line 153: What is "Asc"?
A.A.: Sorry for the mistake. The sentence was rewritten.
Line 160: ... by the method developed (or reported) by Cataldo [26].
A.A.: Done
Line 163: Term repetition.
A.A.: The sentence was rewrite.
Line 192: shift the marked text after the bracket.
A.A.: Done
Line 209: Superscript
A.A.: Done
Line 219: of what?
A.A.: The sentence was rewrite.
Line 222.: capitalization?
A.A.: Yes. Done
Line 243: merge first three paragraphs
A.A.: Done
Line 248: please provide more references here since this scientific topic is highly explored in the modern agriculture, e.g.,
A.A.: More references were added.
Line 255: Too incomprehensible. Please rephrase.
A.A.: The sentence was rewrite
Line 260: What is LME? Shouldn't it be MLE?
AA: Sorry for the mistake. The correction was found.
Line 267: Please avoid this expression in scientific literature. It usually mean you wanted to say more but you didn't.
A.A.: The sentence was rewrite.
Line 269: Please define "LAI"
A.A.: The definition of LAI was added
Line 304: What is ONE?
A.A.: The definition of ONE was added
Line 317: in full "aqueous extracts of moringa leaf extract". "Extract" repeated. Please rephrase.
A.A.: The sentence was rewrite.
Line 324: All baby leaves?
A.A.: The entire paragraph of conclusions was modified.
Author Response File: Author Response.pdf
Reviewer 3 Report
Due to the need for restriction of agrochemicals use in agricultural production is looking for various kind of preparations that can replace their action are sought. Frequently there are extracts with interesting chemical composition, obtaining from the different species and plant organs. Belongs to them also Moringa oleifera Lam. Therefore the scientific problem which was undertaken by Authors is interesting and justified. However, so that the effects of these preparations action were authoritative and convincing the studies have to be properly performed and obtained results are not in doubt.
My suggestions to improve the paper:
1. In the paper is not give the dose of spray (for instance per 1 plant or per 10 plants), therefore is hard to refer the obtained results to cultivation of radish in the field conditions.
2. Why to the studies are selected only two extract concentrations 1:30 and 1:40 and not more?
3. It should be shorten and generalized the title of all Tables. For instance in the title of Table 1 it is necessary to write that it contains data for mass and other parameters of radish leaves.
4. When the measurements of leaves and harvests of radish leaves were performed (i.e. haw many days from the spraying),
5. Why only aboveground part of radish was analyzed without analyses of roots? Because in effect of Moringa oleifera Lam. extract the parameters of leaves changed so, it is possible to suppose that the root’s parameters were also changed.
6. It should be more precisely define what means a statement “The seedlings were irrigated based on ordinary techniques”.
7. What means the word “Controllo” on the Fig. 1-3?
8. A statement in the Discussion „Now-a-days, farmers and researches are frequently using biostimulant plant extracts to improve crops in normal and stress conditions” should be supported with literature.
9. There is lack of chemical content of mentioned extract which would make the results more believable and widen knowledge on the mechanism of extract from Moringa oleifera Lam. leaves. If the analysis of this extract were not perform it should be give the proper literature with its content. It is basic information and very important in aspect the evaluation which substance has this kind of effect and finally - practical recommendations/ application.
10. It should be considered whether the data presented on the Figures 1-3 should be presented in the tables because at present the differences are very little visible.
11. In the Results should be post information which could not be shown in these research.
Author Response
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 and highlighted in yellow. We hereby provide a point-by-point answer.
The authors
My suggestions to improve the paper:
- In the paper is not give the dose of spray (for instance per 1 plant or per 10 plants), therefore is hard to refer the obtained results to cultivation of radish in the field conditions.
A.A.: The plants have been sprayed until dripping. The information was added to the text.
- Why to the studies are selected only two extract concentrations 1:30 and 1:40 and not more?
A.A.: According to information present in the literature, they are the concentrations more used. We are currently investigating the question of dose and method of distribution
- It should be shorten and generalized the title of all Tables. For instance in the title of Table 1 it is necessary to write that it contains data for mass and other parameters of radish leaves.
A.A.: Thanks for the suggestion. More information were added to the tables.
- When the measurements of leaves and harvests of radish leaves were performed (i.e. how many days from the spraying),
A.A.: The plants were sprayed three times with moringa leaf extract (MLE), at 15, 30 and 2 days before harvest (45 days after planting). We have added this information to the text.
- Why only aboveground part of radish was analyzed without analyses of roots? Because in effect of Moringa oleifera Lam. extract the parameters of leaves changed so, it is possible to suppose that the root’s parameters were also changed.
A.A.: In the present trial, we wanted to analyze only the aerial part of the plant, because we mainly wanted to analyze the effects of moringa on the nutraceutical components of the edible portion of the product, therefore considering only the leaves. We consider your suggestion for future trials.
- It should be more precisely define what means a statement “The seedlings were irrigated based on ordinary techniques”.
A.A.: The sentence was rewrite.
- What means the word “Controllo” on the Fig. 1-3?
A.A.: Sorry for the mistake. The word was corrected
- A statement in the Discussion “Nowadays, farmers and researches are frequently using biostimulant plant extracts to improve crops in normal and stress conditions” should be supported with literature.
A.A.: The references were added.
- There is lack of chemical content of mentioned extract which would make the results more believable and widen knowledge on the mechanism of extract from Moringa oleifera Lam. leaves. If the analysis of this extract were not perform it should be give the proper literature with its content. It is basic information and very important in aspect the evaluation which substance has this kind of effect and finally - practical recommendations/ application.
A.A.: The moringa extract of this trial is the same that was used in the paper by Toscano et al., 2020. Its chemical content has been analyzed and the values are reported in this Table. In the text, the reference to the paper has been added.
Chemical composition of principal component of Moringa oleifera leaves.
|
Component |
Value |
|
Phosphorus (P) |
4.9 g kg−1 DW |
|
Potassium (K) |
16.0 g kg−1 DW |
|
Calcium (Ca) |
16.2 g kg−1 DW |
|
Iron (Fe) |
0.2 g kg−1 DW |
|
Magnesium (Mg) |
3.0 g kg−1 DW |
|
DPPH 1 |
130 mg TE g−1 DW |
|
Total polyphenols |
22.6 mg GAE g−1 DW |
|
Chlorophyll a |
1.2 µg mg−1 FW |
|
Chlorophyll b |
1.4 µg mg−1 FW |
|
Carotenoids |
0.10 µg mg−1 FW |
|
Nitrate concentration |
515.5 mg kg−1 FW |
1 Diphenyl-1-Picrylhydrazyl Radical-Scavenging Activity (DPPH)
- It should be considered whether the data presented on the Figures 1-3 should be presented in the tables because at present the differences are very little visible.
A.A.: AA: Thanks for their comment. Rather than reporting the values in the table, we have modified the axes of the figures, thus making the values clearer. We believe they can now be clear.
- In the Results should be post information which could not be shown in these research.
AA: At the end of the discussion we reported the future investigation that we will carry out. In particular:
“We plan to conduct future research to analyze whether the response is species specific, to identify the optimal dose using a wider range of MLE treatments, and to identify the limit dose. The effect of repeated treatments over time should be the focus of future research to understand the influence of the additive effect on the nutraceutical properties of plants.”
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
The Authors significantly improved the quality of the manuscript and answered my questions comprehensively. Recommends the manuscript for publication in its current form.
Reviewer 3 Report
I would like to inform, that Authors sufficiently took due account of my comments contained in the review.
Therefore this paper in the present form can be published in the Horticulturae journal.
