Obtaining Vegetable Production Enriched with Minor Micronutrients Using Fullerene Derivatives

Round 1

Reviewer 1 Report

Comments for author File: Comments.pdf

Moderate editing of the English language is necessary.

Author Response

Response to Reviewer 1 Comments

Dear Reviewer! We are thankful for reviewing our manuscript and for the constructive comment.

Point 1: The submitted manuscript: "OBTAINING VEGETABLE PRODUCTION ENRICHED WITH MINOR MICRONUTRIENTS USING CARBON NANOSTRUCTURES BASED ON FULLERENE DERIVATIVES" represents a solid study for the biofortification of plants with microelements and could be of great importance in the agriculture practice for the improvement of the plant growth, productivity, and the quality. The following changes are recommended and some clarifications should be made:

 Modification in the section Abstract: The abstract section should be completely revised, since it is too generally written. The authors did not provide some key points used in this study, such as the plant species, treatments, parameters or methods, as well the main results and the conclusion.

 

Response 1: The abstract section was completely revised in accordance with the comments

 Point 2: Modification in the section Introduction –

Page 2, Line 60-64: The authors should provide appropriate citations for the following statements: “The above-mentioned diet nutrition includes the regular consumption of fruits and vegetables containing the necessary amounts of biologically active compounds with a complex of useful properties, including selenium and zinc. However, in order to obtain the recommended daily doses of useful micronutrients with these plants products, it is necessary to consume several kg or more per day, which is not possible for the human organism.”

 Response 2: Lines 81-88. The text of the statements is supplemented with numerical data for clarity and appropriate citations.

“…The above mentioned diet nutrition includes the regular consumption of fruits and vegetables containing the necessary amounts of biologically active compounds with a complex of useful properties, including selenium and zinc. However, in order to obtain the recommended daily doses (for example, selenium – ~0,030-0,055 mg/day; zinc – ~10-19 mg/day) [6, 9-11] of useful micronutrients with these plants products (for example, vegeta-ble crops contain, in mg per kg of the plants edible part: selenium - ~0.0001-0,022, zinc – ~0,8-9,1) [9, 12], it is necessary to consume several kg or more per day, which is not possi-ble for the human organism”.

 

Point 3: Modification in the section Introduction –

Page 2, Line 65-69: Please rephrase or split the following sentence: “One of the ways to solve this problem is to increase the production of plant raw materials and high-quality products containing minor micronutrients in quantities that are safe for the human organism, through the development and implementation in the agroindustrial complex of new environmentally friendly and highly effective biofortification’s methods, techniques, means, technologies and others.”

Response 3: Lines 89-93. Sentence was splited.

“…One of the ways to solve this problem is to increase the production of plant raw materials and high-quality products containing minor micronutrients in quantities that are safe for the human organism. The latter can be achieved through the development and implementation in the agroindustrial complex of new environmentally friendly and highly effective biofortification’s methods, techniques, means, technologies and others.”

 Point 4: Modification in the section Introduction –

Page 3, Line 126-131: The following sentence is not clear. Please, rephrase it: “Thus, in the works of various authors it was shown that polyhydroxylated fullerene at certain concentrations can damage onion cells [48], contribute to an increase in the density of the green algae culture Pseudokirchneriella subcapitata, accelerate the growth of the hypocotyl in Arabidopsis [47], and increase the biomass of bitter melon plants after seed treatment, increase its yield, the content of useful substances in fruits [49].”

Response 4: Lines 142-147: Sentence was rephrased.

“…Thus, in the works of various authors it was shown the effects of polyhydroxylated fullerene with different directions: from damaging onion cells [42] to an increasing in the densi-ty of the green algae culture Pseudokirchneriella subcapitata, as well as an accelerating the growth of the hypocotyl in Arabidopsis [41], increasing the biomass of bitter melon plants after seed treatment and its yield, the content of useful substances in fruits [43]”.

 Point 5: Modification in the section Introduction –

- Page 4, Line 155-162: Too long sentence, please rephrase it: “We have found that an increase in the net productivity of plants and their resistance to oxidative stress after the introduction of polyhydroxylated, carboxylated and amino acid derivatives of C60 into the root environment (nutrient solution for hydroponic plant cultivation) or after foliar treatment plants is associated with established changes in the structure and efficiency of the photosynthetic apparatus, as well as with the effect on the processes of plant metabolism, 2 exchange, their antioxidant defense systems, namely, the intensity of lipid peroxidation, the activity of superoxide dismutase, the generation of reactive oxygen species [53-55, 57, 62].”

Response 5: Lines 171-179: Sentence was rephrased.

“…We have found that an increase in the net productivity of plants and their resistance to ox-idative stress after the introduction of polyhydroxylated, carboxylated and amino acid de-rivatives of C₆₀ into the root environment (nutrient solution for hydroponic plant cultivation) or after foliar treatment plants is associated with established changes in the structure and efficiency of the photosynthetic apparatus, as well as with the effect on the processes of plant metabolism, exchange, their antioxidant defense systems. Indicators of the antioxidant defense systems state were the intensity of lipid peroxidation, the activity of superox-ide dismutase, the generation of reactive oxygen species [47-53].”

 

 Point 6: Modification in the section Introduction –

- Page 4, Line 163-167: The following statement is not clear, please rephrase it: “The revealed potential ability of fullerene derivatives to increase the content of macro- and microelements in the aerial parts of plants testified to the expediency of developing methods for introducing them into the soil (soil substitute) - plant system and using them as potential mediators that contribute to the enrichment of plants with physiologically active substances useful for humans.”

Response 6: Lines: 180-183: The statement was rephrased.

“...The revealed potential ability of fullerene derivatives to increase the content of macro- and microelements in the plants testified to the expediency of using them as potential me-diators that contribute to the enrichment of plants with physiologically active substances useful for humans”.

 Point 7: Modification in the section Introduction –

- Page 4, Line 168-174: The research hypothesis is not clearly presented.

Response 7: Lines: 184-190: The research hypothesis was corrected.

“…The research hypothesis: the compositions of water-soluble fullerenes and a micro-elements compounds having different charges are very effective in increasing the content of the applied microelement in the edible part of the used vegetable crops, and in significantly improving the physiological state plants, when their solutions are introduced into the soil (soil substitute) - plants system. The noted effects are expected to be observed at very low concentrations of compounds in solutions due to the above-mentioned unique properties of water-soluble fullerenes..”

 Point 8: Modification in the section Materials and Methods –

Figure 1: Please, revise the numeration of Figure 1 (а, б, в, г).

Response 8: The numeration of Figure 1 was corrected.

 Point 9: Modification in the section Materials and Methods –

Line 240-242: The control group should be mentioned here along with the treated group of plants (C60 fullerenol at concentrations of 1 mg/kg, 10 mg/kg and 100 mg/kg).

Response 9: Lines: 285-288: Information about control was added.

“…Variants of introduction into soil were the solutions of C₆₀ fullerenol at concentrations of 1 mg/kg, 10 mg/kg and 100 mg/kg of soil. In the control variant, distilled water was introduced into the soil in the same volume as the volume of C₆₀ fullerenol solutions introduced into the soil.”

Point 10: Modification in the section Materials and Methods –

Line 276-282: Please, check the way of citation in the following statements: “At the end of the experiment, pHKCl of the soil (A arable layer) was determined by the potentiometric method according to [77]” I suggest using this way of citation: “At the end of the experiment, pHKCl of the soil (A arable layer) was determined by the potentiometric method according to ISO 10390 [77].” or “At the end of the experiment, pHKCl of the soil (A arable layer) was determined by the potentiometric method [77].

Response 10: Lines: 299-302: It was corrected. A brief description of the essence of the method with references to literary sources is given in a separate subsection "Soil analyzes" on page 10.

 “…At the end of the experiment, pHKCl of the soil (A arable layer) was determined. In the soil, the content of ammonium nitrogen, nitrate nitrogen, mobile phosphorus and potas-sium, exchangeable calcium and magnesium, trace elements (mobile iron, mobile copper, mobile zinc, mobile manganese). “

 Point 11: Modification in the section Materials and Methods –

Line 317-321: Please, rephrase the following statement, since it is not clearly presented: “The effect for foliar treatment of cv. Natasha tomato plants with mono- and mixed solutions of C60-L-Gly or C60-L-Arg (0.0001%) and sodium selenate (0.0001% Na2SeO4) was evaluated in a vegetation experiment under favorable controlled conditions when growing plants under artificial light using the panoponics method on a thin-layer analog of the soils described above.”

Response 11: Lines: 328-330: Statement was rephrased.

“…The effect for foliar treatment of cv. Natasha tomato plants with mono- and mixed so-lutions of C60-L-Gly or C60-L-Arg (0.0001%) and sodium selenate (0.0001% Na2SeO4) was evaluated in a vegetation experiment under conditions shown in the Table 1.”

Point 12: Modification in the section Materials and Methods –

Line 318-324 and Line 344-348: Several statements in the subsection “Plants foliar treatment with solutions of the C60-L-Gly or C60-L-Arg and Na2SeO4“ and “Plants foliar treatment with solution of the fullerenol C60(OH)22-24 or C70(OH)12-14 and ZnSO4“ are completely the same and they should not be repeated – “The plants were grown using the panoponic method on a thin-layer analog of the soils described above. The latter is placed into the trays of plant growing light equipment. Seeds of plants are put into the layer (1-mm thick) of suspension based on the Cambrian clay, on the surface of material. The latter provides plant root systems with a nutrient solution circulating over the tray bottom through the flat slotted capillaries [14, 66, 83].”

Response 12: Statements were corrected.

Lines: 330-334

“…Seeds of plants are put into the layer (1-mm thick) of suspension based on the Cambrian clay, on the surface of mentioned above hydrophilic material placed in the plant growing equipment tray. The material provides plant root systems with a nutrient Knop's solution circulating over the tray bottom through the flat slotted capillaries [62, 63].”

Lines: 348-349

“…The conditions of the experiment are presented in Table 1. The method of growing plants - panoponics - is similar to that described above.”

 Point 13: Modification in the section Results -

Table 2: The authors should correct the Note in Table 2, since the comparisons are between control and treated groups, not between parameters. The term “Values in columns” should be replaced with “Values in rows”. The same should be implemented for Table 7 and Table 8.

 Response 13: Note in Table 2, Table 7 and Table 8 was corrected.

 

Point 14: Modification in the section Results –

Figure 3: Please indicate what is the meaning of different letters in the graphs.

Response 14: It was done. A sentence was added to the caption under the Figure 3

“…Bars with different letters are significantly different at p ≤ 0.05, as determined by Duncan’s multiple range test”.

 

Point 15: Modification in the section Results –

Why the authors noted “a significant decrease (up to 47%) in the activity of the oxidative enzyme peroxidase in leaves and roots”, since the POX activity in the root treated with 100 mg/kg C60 fullerenol is significantly increased compared to control? See Table 5.

Response 15: The Sentence was supplemented with information. Page 17

“…-        it mainly contributes in the form of a trend or a significant decrease (maximum change - by 47%) in the activity of the oxidative enzyme peroxidase in leaves and roots. The exception is the roots in the variant with the introduction of fullerenol in-to the soil at a concentration of 100 mg/kg, where the activity of peroxidase was higher than that in the control by 14%;”

 

Point 16: Modification in the section Results –

Any explanation for decreased LPO in treated plants with C60 fullerenol or any bibliographic data?

Response 16: Explanation for decreased LPO in treated plants with C60 fullerenol was added in Discussion. Page 27.

 

Point 17: Modification in the section Results –

The authors should check the value percentage in the following statement: “a significant increase (up to 647%) in the activity of the catalase enzyme (Table 5).”

Response 17: In the text on page 17, a clarifying phrases were added to the numerical data characterizing the maximum change in parameter values.

“…maximum change - by …%”.

 

Point 18: Modification in the section Results –

Figure 4: The note for Figure 4 should be deleted, since it should be mentioned in the section Materials and methods.

Response 18: The note for Figure 4 was deleted. It was added sentence – “…Bars with different letters are significantly different at p ≤ 0.05, as determined by Duncan’s multiple range test”.

 

Point 19: Modification in the section Results –

Figure 5: Please indicate what is the meaning of different letters in the graphs.

Response 19: It was done. It was added sentence -

“…Bars with different letters are significantly different at p ≤ 0.05, as determined by Duncan’s multiple range test.”

 

Point 20: Modification in the section Conclusion - The Conclusion section should be revised, since there are too general statements and the authors repeated the same sentences from the Abstract.

Response 20: The Conclusion section was revised.

 

Point 21: General comments: The manuscript needs significant improvements, especially in the section Results, because they are not clearly presented. Also, the authors should compare their results with other studies that are relevant to the current topic. I strongly suggest to the authors to improve the manuscript and to include Discussion section.

Response 21: We shortened the introduction a little, removing some links to Russian-language publications in journals that are difficult to find on the Internet. Added links to English-language publications. Edited a number of paragraphs in accordance with the comments of the reviewers, including the hypothesis, information about what is presented in the article and the purpose of the work.

In the "Materials and Methods" section, we tried to remove repetitions in the description of vegetation experiments and make the presentation of the material more clear.

Filled with relevant information Subsections were added to the “Materials and Methods” section:

- Experimental Design with subsubsection:  1. Evaluation of the influence of the C₆₀ fullerenol’s introduction into the soil on the cucumber plant and the content of the main macro- and microelements in the soil and plants;  2.    Study of the influence of introduction into the nutrient solution of the  C₆₀-L-Gly or C₆₀-L-Arg and Na₂SeO₄ compositions on the Chinese cabbage and the selenium content in the plants; 3.               Study of the influence of foliar treatment with the C₆₀-L-Gly or C₆₀-L-Arg and Na₂SeO₄ compositions on the tomato and the selenium content in its production; 4.                Study of the influence of foliar treatment with the compositions of the fullerenol C₆₀(OH)_22-24  or C₇₀(OH)_12-14 and ZnSO₄  on the cucumber and the zinc content in its production;

- Soil analyzes;

- Plant analyzes: Morphology measurements; photosynthetic pigment analysis; Activity of Antioxidant Systems; Quality and Safety Indicators of Plant Production;

- Statistical Analysis.

 

We added a Discussion section and edited the Results section accordingly.

The research is pioneering and there is nothing yet to compare the results of plant enrichment with fullerenes. It may be done only with our previous research results or with the results of studies on the effects of other non-fullerene nanoparticles on plantsю We trayed to do it in Discussiob section.

 

Completely revised Conclusion and Abstract.

 The bibliographical part of the manuscript was revised: some of the Russian-language publications were replaced by English-language ones and added some references.

 

Sincerely,

authors of the manuscript.

 

 

Author Response File: Author Response.pdf

Reviewer 2 Report

Dear authors

The following modifications are required:

Title

The title is not clear and should be changed

Abstract

ü  In general, this section is poorly written. It is written simply. This section should include the most important findings from this study. As a result, this section should be improved.

ü  Before describing the goal, the authors must define the issue in a single line and explain why they chose this approach to study this research.

ü  The aim is not correctly written

ü  No information about the method and results are available in this manuscript

ü  The authors should provide some quantitative scored data

ü  In the final line of the abstract, the authors should present a decisive conclusion derived from the research and provide a single line of future prospects.

Keywords

ü  The content of keywords did not reflect the content of this manuscript and the words used for forming the title should not be used as the keywords. So, the structure of keywords should be changed.

Introduction

ü  This section is generally poorly written. It is written in a straightforward manner. This section is too long and should be minimized

ü  The caption of Figure 1 is not representing the content of figure. The letter on the figure should be defined

ü  The authors should give some lines about the knowledge gap which their research has covered along with the hypothesis statement

ü  Also, the authors should provide a novelty statement at the end. What new things authors have done or correlated in this research compared to old ones?

ü  The general and specific aim should be inserted

Materials and Methods

ü  The plant materials are not correctly mentioned because the authors mentioned two types of plants: cabbage and cucumber, which one of them used by the authors?

ü  The preparation of fullerence is not clearly written

ü  The treatment of plants by fullerence is not clearly provided

ü  The type of experimental design is not available

ü  All procedures of measurements of the studied traits are not clearly presented

ü  The authors should be written the procedures in detail

ü  The authors should provide the measurements of traits (morphological and chemical parameters) in detail

ü  A section of statistical analysis with the experimental design used should be added in the end of the section MM

ü  All experiments conducted in the study should be detailed in clear manner.

Results and discussion

ü  In general, this section is too long and should be focused on the most important point of results

ü  L385-400: These sentences should be removed

ü  The ANOVA table should be inserted

ü  The authors should state the significant status of measured traits

ü  The results should be presented in detail

ü  The authors should provide some quantitative data

ü  The discussion is zero. The authors should interpret the results by adding the mechanism effects of factors used in this project on the studied traits.

Conclusion

ü  The structure of this section should be changed. The authors should focus on the conclusion of main results because this section is written as the results section

ü  The authors should include some information about future works in this field.

 

 

 

 

It needs extensive correction

 

Author Response

Response to Reviewer 2 Comments

 Dear Reviewer! We are thankful for reviewing our manuscript and for the constructive comment.

Point 1: Dear authors

The following modifications are required:

Title: The title is not clear and should be changed

Response 1: The title was changed

OBTAINING VEGETABLE PRODUCTION ENRICHED WITH MINOR MICRONUTRIENTS USING FULLERENE DERIVATIVES

 Point 2: Abstract -

ü  In general, this section is poorly written. It is written simply. This section should include the most important findings from this study. As a result, this section should be improved.

ü  Before describing the goal, the authors must define the issue in a single line and explain why they chose this approach to study this research.

ü  The aim is not correctly written

ü  No information about the method and results are available in this manuscript

ü  The authors should provide some quantitative scored data

ü  In the final line of the abstract, the authors should present a decisive conclusion derived from the research and provide a single line of future prospects.

Response 2: Lines: 13-58. The Abstract was revised in accordance of your comments.

“Abstract: Elaborating the methods and means of enriching nutrition’s, include the plants, with a number of microelements vital for humans is actually today due to the unresolved acute problem of micronutrient deficiency and imbalance in the predominant part of the population of various countries of the world. Promising for the implementation of biofortifica-tion in terms of safety, efficiency, nanosize, biocompatibility, transportability are water-soluble derivatives of C₆₀ or C₇₀ fullerene. By now the use of water-soluble fullerenes (С₆₀(ОН)_22-24 or C₇₀(OH)_12-14 fullerenols, C₆₀ fullerene with glycine or with arginine - C₆₀-L-Gly or C₆₀-L-Arg) with various functional groups for plant enrichment is pioneering. Ex-perimental research work was carried out at the agrobiopolygon of the Agrophysical Research Institute with controlled microclimate conditions. It was assessment the influence of С₆₀(ОН)_22-24 fullerenol introduction into the soil on content of macro- and microele-ments in the soil and in plants for example cucumber, as well as on the plants physiolog-ical state (photosynthetic pigments, the intensity of lipid peroxidation, the activity of pe-roxidase and catalase enzymes), growth and elements content. It was study possibility to enrich the plants production (Chinese cabbage, tomato, cucumber) with the compositions on basis of the fullerene derivatives (C₆₀-L-Gly or C₆₀-L-Arg, С₆₀(ОН)_22-24 or C₇₀(OH)_12-14 fullerenols) and selenium or zinc compounds by introducing them into a nutrient solu-tion or by foliar treatment of plants. It was revealed that the introduction of solutions of С60 fullerenol in various concentrations (1 mg/kg, 10 mg/kg, 100 mg/kg) into soddy-podzolic sandy loamy soil contributed to activates the processes of nitrogen transfor-mation in the soil, in particular, enhances the process of nitrification and to increase in the content of mobile forms of some macro- and microelements in the soil, as well as of the latter in plant organs, for example, cucumber plants, especially in their leaves. Along with this, the plants showed an increase in the content of photosynthetic pigments, mainly a decrease in the activity of the oxidative enzyme peroxidase, the intensity of lipid peroxidation, and an increase in the content of the reducing enzyme catalase. The im-provement in the physiological state of plants had a positive effect on the growth rates of cucumber plants. The compositions of solutions of amino acid fullerenes (C₆₀-L-Gly or C₆₀-L-Arg) and sodium selenate, as well as C₆₀ or C₇₀ fullerenols and zinc sulfate, selected on the basis of different charges of molecules or functional groups of fullerene deriva-tives, showed higher efficiency at low concentrations in enriching plant products in Chinese cabbage, tomato and cucumber with selenium and zinc, respectively, compared with mineral salts of the indicated elements and with control (edible part of Chinese cab-bage - by 31.0% -89.0% relative to that in the control and by 26.0%-81.0% relative to the treatment of plants with a sodium selenate; tomato fruits - by 33.7% - 42.2% relative to that in the control and by 10.2% - 17.2% relative to the treatment of plants with a sodium selenate; cucumber fruits - by 42.0% - 59.0% relative to that in the control and by 10.0% - 23.0% relative to the treatment of plants with a zinc sulfate).

At the same time, the quantitative characteristics of growth, productivity and / or quality of the obtained products increase and improve accordingly. The prospect of fur-ther research is an in-depth knowledge of the mechanisms of the compositions of fuller-ene derivatives and various compounds of trace elements influence on the plants, as well as the synthesis and study of the various exo- and endo derivatives of fullerenes proper-ties, including C60 complex compounds with transition metals, fullerenes containing in-side their carbon networks atoms of various chemical elements, such as lanthanum and others.”

 

Point 3: Keywords

ü  The content of keywords did not reflect the content of this manuscript and the words used for forming the title should not be used as the keywords. So, the structure of keywords should be changed.–

Response 3: Lines 60-62: structure of keywords was changed.

Keywords: biofortification, trace elements, selenium, zinc, Chinese cabbage, tomato, cucumber, polyhydroxylated fullerenes C60, C 70, amino acid derivatives C60, soil (soil sub-stitute) - plant system, plants growth, quality, productivity.

Point 4: Introduction

ü  This section is generally poorly written. It is written in a straightforward manner. This section is too long and should be minimized

ü  The caption of Figure 1 is not representing the content of figure. The letter on the figure should be defined

ü  The authors should give some lines about the knowledge gap which their research has covered along with the hypothesis statement

ü  Also, the authors should provide a novelty statement at the end. What new things authors have done or correlated in this research compared to old ones?

ü  The general and specific aim should be inserted

Response 4: We shortened the introduction a little, edited a number of paragraphs, gave some lines about the knowledge gap which their research has covered, revised the hypothesis, gave the  information about what is presented in the article and what is the novelty of research, edited the purpose of the work;

Figure 1 is in section “Materials and Methods”. We changed it on more informative ‘Scheme of obtaining C₆₀(OH)₂₄ and C₇₀(OH)₁₂ fullerenols (a) and C₆₀-L-Gly and C₆₀-L-Arg derivatives (b)”.

 

Point 5: Materials and Methods

ü  The plant materials are not correctly mentioned because the authors mentioned two types of plants: cabbage and cucumber, which one of them used by the authors?

ü  The preparation of fullerence is not clearly written

ü  The treatment of plants by fullerence is not clearly provided

ü  The type of experimental design is not available

ü  All procedures of measurements of the studied traits are not clearly presented

ü  The authors should be written the procedures in detail

ü  The authors should provide the measurements of traits (morphological and chemical parameters) in detail

ü  A section of statistical analysis with the experimental design used should be added in the end of the section MM

ü  All experiments conducted in the study should be detailed in clear manner.

 Response 5:  

The text about the objects of study has been corrected -

“…The objects of research were test vegetable crops on the example of different varieties or hybrids – Chinese cabbage Brassica rapasubsp. pekinensis (Lour.) Hanelt cv. Daqingkou, tomato Solanum lycopersicum L., cv. Natasha, cucumber Cucumis sativus L. (hybrid F1 Neva)”.

 Lines: 212-246: information about C₆₀ fullerene adducts synthesis was added to the their brief description.

 The text about the treatment of plants with mono- and mixed solutions of fullerene derivatives and sodium selenate, zinc sulfate was edited. According to the experimental design, all studies were divided into 4. The plants treatment forme, the names of plants and fullerene derivatives follow from the formulations of each from 4 direction.

“…Experimental Design

The following series of experiments were carried out:

1. Evaluation of the influence of the C₆₀ fullerenol’s introduction into the soil on the cucumber plant and the content of the main macro- and microelements in the soil and plants.
2. Study of the influence of introduction into the nutrient solution of the C₆₀-L-Gly or C₆₀-L-Arg and Na₂SeO₄ compositions on the Chinese cabbage and the selenium content in the plants.
3. Study of the influence of foliar treatment with the C₆₀-L-Gly or C₆₀-L-Arg and Na₂SeO₄ compositions on the tomato and the selenium content in its production.
4. Study of the influence of foliar treatment with the compositions of the fullerenol C60(OH)22-24 or C₇₀(OH)_12-14  and ZnSO₄ on the cucumber and the zinc content in its production.”

 Procedures of measurements of the studied traits were presented in detail in the subsection:

Soil analyzes; Plant analyzes: Morphology measurements; Photosynthetic pigments analysis; Activity of Antioxidant Systems; Quality and Safety Indicators of Plant Production

 A section of statistical analysis was added in the end of the section MM.

 

Point 6: Results and discussion

ü  In general, this section is too long and should be focused on the most important point of results

ü  L385-400: These sentences should be removed

ü  The ANOVA table should be inserted

ü  The authors should state the significant status of measured traits

ü  The results should be presented in detail

ü  The authors should provide some quantitative data

ü  The discussion is zero. The authors should interpret the results by adding the mechanism effects of factors used in this project on the studied traits.

Response 6: Section “Results and discussion” was divided on the section “Results” and section “Discussion”.

Lines: 385-402: Text was text moved to section “Discussion”.

The ANOVA tables were intoduced in file “Supplementary materials”.

All values in tables and in Figures were followed by letters (a–c). Values with different letters are significantly different at p ≤ 0.05, as determined by Duncan’s multiple range test.

The results have been more detailed in the section “Results”

The quantitative data were provide in the section “Results”.

In the Discussion section, the results were interpreted as far as possible, indicating the possible mechanisms of action of the assessed factors on the plants state.

  

Point 7: Conclusion

ü  The structure of this section should be changed. The authors should focus on the conclusion of main results because this section is written as the results section

ü  The authors should include some information about future works in this field.

Response 7: Section “Conclusion” was changed in accordance with your comments and comments of other reviewers.

The information about future works in this field was added.

 

Sincerely,

authors of the manuscript. 

 

Author Response File: Author Response.pdf

Reviewer 3 Report

The article tested the effect of water-soluble derivatives of C60 and C70 fullerenes on a number of vegetable crops and the content of trace elements in the products. The experiment design is good and the data is enough to prove its conclusions. 

 

Author Response

Response to Reviewer 3 Comments

 Point 1: Comments and Suggestions for Authors

The article tested the effect of water-soluble derivatives of C60 and C70 fullerenes on a number of vegetable crops and the content of trace elements in the products. The experiment design is good and the data is enough to prove its conclusions.

Response 1: Dear Reviewer! We are thankful for reviewing our manuscript and for the positive evaluation of our work.

 

Sincerely,

authors of the manuscript.

 

 

Author Response File: Author Response.pdf

Reviewer 4 Report

The authors did an exhaustive and interesting work.

In my opinion, the introduction is too much extensive. It would be better if you improve the clarity of the ideas and conciseness.

The section of materials and methods is adequately described. However, it lacks in a isolated sub-section about the methods used to quantify the indicators of soil and the fruit quality (like elemental composition, chemical composition).

The caption of Table 2 should be improved.

In Figures 2 and 4, add in the caption what a.d.m. means.

The English is fine. However, the authors could improve the grammar in some sentence construction.

Author Response

Response to Reviewer 4 Comments

 

Dear Reviewer! We are thankful for reviewing our manuscript and for the constructive comment.

 

Point 1: Comments and Suggestions for Authors

The authors did an exhaustive and interesting work.

In my opinion, the introduction is too much extensive. It would be better if you improve the clarity of the ideas and conciseness.

Response 1: We shortened the introduction a little, edited a number of paragraphs, revised the hypothesis, gave the information about what is presented in the article and what is the novelty of research, edited the purpose of the work.

 

Point 2: The section of materials and methods is adequately described. However, it lacks in a isolated sub-section about the methods used to quantify the indicators of soil and the fruit quality (like elemental composition, chemical composition).

Response 2: Filled with relevant information Subsections were added to the “Materials and Methods” section:

- Experimental Design with subsubsection:  1. Evaluation of the influence of the C₆₀ fullerenol’s introduction into the soil on the cucumber plant and the content of the main macro- and microelements in the soil and plants;  2.    Study of the influence of introduction into the nutrient solution of the  C₆₀-L-Gly or C₆₀-L-Arg and Na₂SeO₄ compositions on the Chinese cabbage and the selenium content in the plants; 3. Study of the influence of foliar treatment with the C₆₀-L-Gly or C₆₀-L-Arg and Na₂SeO₄ compositions on the tomato and the selenium content in its production; 4.  Study of the influence of foliar treatment with the compositions of the fullerenol C₆₀(OH)_22-24  or C₇₀(OH)_12-14 and ZnSO₄  on the cucumber and the zinc content in its production;

- Soil analyzes;

- Plant analyzes: Morphology measurements; photosynthetic pigment analysis; Activity of Antioxidant Systems; Quality and Safety Indicators of Plant Production;

- Statistical Analysis.

 

Point 3: The caption of Table 2 should be improved.

Response 3: Another table was added to the “Materials and Methods” section, so table 2 became table 3. The caption of Table 3 was changed.

“…Table 3. The influence of the C₆₀ fullerenol on some soil agrochemical indicators and elements content by introduction of its solutions in various concentrations into the soil”

 

Point 4: In Figures 2 and 4, add in the caption what a.d.m. means.

Response 4: It was added.

 

 Point 5: Comments on the Quality of English Language

The English is fine. However, the authors could improve the grammar in some sentence construction.

Response 5: The grammar in sentence construction was improve.

  

Sincerely,

authors of the manuscript.

 

 

 

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

The authors satisfactorily responded to all suggestions.  

Minor English language editing is necessary.

Reviewer 2 Report

Th authors have been addressed all comments

moderate correction is needed