Different Oligosaccharides Induce Coordination and Promotion of Root Growth and Leaf Senescence during Strawberry and Cucumber Growth

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Comments on the manuscript with the title “Different Oligosaccharides Induce Coordination and Promotion of Root Growth and Leaf Senescence during Strawberry (Fragaria × ananassa Duch) and Cucumber (Cucumis sativus L.) Growth”.

 

Abstract is too long, which makes the main message to be lost. I suggest to keep in the abstract only the most relevant or novel findings from the study that can be attractive for readers. Please consider that the abstract is the first which the readers see. I do not find relevant to mention the years of the experiment in the abstract (2021-2022).

 

Introduction

– the authors after species do not have to be italics (Duch and L.), also if authors indeed want to be accurate, the name has to be given according to updated nomenclature after reputable sources. E.g. I just checked in IPNI and I found it as Fragaria × ananassa (Weston) Duchesne ex Rozier.

 

Page 3 – what does “creature compatibility” refers to?

 

Material and Method

Authors mention they applied two-way analyses of variance ANOVA, followed by Duncan. Two way suggest that experiment was bifactorial – two factors each with its own gradients. Yet, Table 1 shows variants as treatments, evident by Figures from Results sections that indicate Duncan was applied separately for strawberry and separately for cucumber.

I strongly suggest at experimental design to clarify the number of experimental variants. 

Two-factors for a Two-way ANOVA would be something like: Factor A) the crop with two levels a1 – cucumber a2-srawbery; Factor B) the treatment with 4 levels b1- …, b2- …. From the combination of the two factors A x B resulted 8 experimental variants.

Perhaps two sets of a nonfactorial experiments with four variants each was the design – hence one-way ANOVA should have been applied?

This needs to be clarified and to be in line with the results. Please name the experimental variants and factors considered.

 

Results

Figure captions have to mention what columns and whiskers represent: means/SE/CI…

 

Table 4 – please name under the table what Type of correlations were used, Pearson?

 

 

Conclusion – I suggest authors to renounce to the use of acronyms in the conclusions section. Also, I suggest them to create a structure that mirrors the aim and objectives in the same order. In this way it will be very clear for any reader what authors intended and how they solved each objective (these were given at the end of the introduction). The conclusions should mirror those.  

 

References list 37 sources relevant for the topic. Both old and recent sources were used.

 

Best regards.

Comments on the Quality of English Language

minor English syntax and grammar improvements are recommended.

Author Response

Manuscript Title：Different oligosaccharides induce coordination and promotion of root growth

and leaf senescence during strawberry and cucumber growth

 

Response to Reviewer 1 Comments

 

Dear Reviewer,

Thank you for your helpful comments and suggestions on our manuscript. We have modified the manuscript accordingly, and detailed corrections are listed below, point by point.

 

Comments 1: Abstract is too long, which makes the main message to be lost. I suggest to keep in the abstract only the most relevant or novel findings from the study that can be attractive for readers. Please consider that the abstract is the first which the readers see. I do not find relevant to mention the years of the experiment in the abstract (2021-2022).

Response 1: Thanks for the comments. We have simplified the abstract in the revised manuscript according to expert suggestions, which mainly showed the effects of the mixed oligosaccharides on crop physiology. Please check the details in revised draft, page 2 with line 34-36.

Comments 2: Introduction– the authors after species do not have to be italics (Duch and L.), also if authors indeed want to be accurate, the name has to be given according to updated nomenclature after reputable sources. E.g. I just checked in IPNI and I found it as Fragaria × ananassa (Weston) Duchesne ex Rozier.

Response 2: Thanks for the comments. We have deleted the accurate specie names for simplification. Please check the details in revised draft, page 1 with line 1-2, page 2 with line 43.

Comments 3: Page 3 – what does “creature compatibility” refers to?

Response 3: Thanks for the comments. The “creature compatibility” refers to that the oligosaccharide derivatives extracted from chitosan as natural substance, are more easily absorbed by plants.

Comments 4: Material and Method Authors mention they applied two-way analyses of variance ANOVA, followed by Duncan. Two way suggest that experiment was bifactorial – two factors each with its own gradients. Yet, Table 1 shows variants as treatments, evident by Figures from Results sections that indicate Duncan was applied separately for strawberry and separately for cucumber. I strongly suggest at experimental design to clarify the number of experimental variants. Two-factors for a Two-way ANOVA would be something like: Factor A) the crop with two levels a1 – cucumber a2-srawbery; Factor B) the treatment with 4 levels b1- …, b2- …. From the combination of the two factors A x B resulted 8 experimental variants. Perhaps two sets of a nonfactorial experiments with four variants each was the design – hence one-way ANOVA should have been applied? This needs to be clarified and to be in line with the results. Please name the experimental variants and factors considered.

Response 4: Thanks for the comments. For statistical analysis, Two-way analyses of variance (ANOVAs) were performed in the research, one factor referred to different treatments; the other referred to different biological replicates: For the strawberry experiment, four plots were conducted, each plot contained a treatment (nine beds of strawberry plants) with three biological replicates, each biological replicate included three beds of strawberry plants and separated by a bed. For the cucumber experiment, sixteen plots were conducted, each treatment had four biological replicates, each biological replicate was a plot with four beds of cucumber plants.

Comments 5: Results-Figure captions have to mention what columns and whiskers represent: means/SE/CI…

Response 5: Thanks for the comments. The columns presented different treatments, and the error bars in figures presented standard deviation among replicates. Please check the details in revised draft, page 9 with line 222, page 11 with line 261, page 13 with line 304, page 14 with line 312, page 15 with line 326, page 1 with line 1-2.

Comments 6: Table 4 – please name under the table what Type of correlations were used, Pearson?

Response 6: Thanks for the comments. We have added the type of correlations under the table, which indicated the Pearson correlation analysis. Please check the details in revised draft, page 15 with line 331.

Comments 7: Conclusion – I suggest authors to renounce to the use of acronyms in the conclusions section. Also, I suggest them to create a structure that mirrors the aim and objectives in the same order. In this way it will be very clear for any reader what authors intended and how they solved each objective (these were given at the end of the introduction). The conclusions should mirror those.

Response 7: Thanks for the comments. We have renounced the use of acronyms in the conclusions section. Please check the details in revised draft, page 20 with line 469. The clear structure in the conclusion section could be seen. Firstly, the regulatory functions of foliar spraying with chitosan oligosaccharide and mixed oligosaccharides were showed through special indicators, such as chlorophyll content, integrated chloroplast structure, maximum photochemical efficiency and photachemical quenching coefficiency, etc. Secondly, the regulation effects of two oligosaccharide regulators on crop yield were compared. Thirdly, the responses of strawberry and cucumber to chemical regulators were compared. In the end, a hypothesis was made for the regulation, and pointing the deeply research direction in the future.

 

 

We look forward to your positive response.

Yours sincerely,

Corresponding Author

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

thank you for the opportunity to meet the manuscript entitled: "Different Oligosaccharides Induce Coordination and Promotion of Root Growth and Leaf Senescence during Strawberry (Fragaria × ananassa Duch) and Cucumber (Cucumis sativus L.) Growth".

In their research, the authors focused on the influence of oligosaccharides on physiological and morphometric parameters, as well as the yield of strawberries and cucumbers.

One of the main goals of the experiment was the analysis of the internal relationship of the root–shoot growth coordination promotion. However, the evaluation of this analysis is absent in the results of the study, and some of the results are presented only in the discussion.

In addition, the authors missed the opportunity to evaluate the effect of foliar treatment on fruit quality. I consider this a lack of research in relation to the crops studied.

Comments:

I recommend modifying the title of the manuscript, as oligosaccharides are a huge group and it would be more appropriate to define in the title which one from the group of oligosaccharides was investigated.

The authors state in chapter MM that the experiments were established during 2021-2022. In figure 1, the temperature and humidity during the experiments are presented, but in the years 2022 and 2023. This needs to be adjusted.

In the MM chapter, I ask to describe the CM treatment in detail. Without basic information, it is not possible to scientifically verify and compare the effect of different treatments.

In the case of chitosan oligosaccharides, was a commercial preparation used?

MixOS treatment needs to be characterized in detail in relation to reproduction and comparisons of future research. This means what components were included, in what proportion, etc.

L130 The authors state a dose of 1100 kg.ha-1. However, the application was foliar. So what was the dose of water, or the concentration?

L168 The authors report that a Two-way Anova was used. However, only one factor was investigated in the experiment.

L278 – wrongly marked table.

In the conclusions, the authors assume that the application of oligosaccharides contributes to the antibacterial properties. However, this has not been studied, so I recommend removing this wording.

Author Response

Manuscript Title：Different oligosaccharides induce coordination and promotion of root growth

and leaf senescence during strawberry and cucumber growth

 

Response to Reviewer 2 Comments

 

Dear Reviewer,

Thank you for your helpful comments and suggestions on our manuscript. We have modified the manuscript accordingly, and detailed corrections are listed below, point by point.

Comments 1: One of the main goals of the experiment was the analysis of the internal relationship of the root–shoot growth coordination promotion. However, the evaluation of this analysis is absent in the results of the study, and some of the results are presented only in the discussion.

Response 1: Thanks for the comments. We have added the analysis of the internal relationship of the root–shoot growth coordination promotion in the conclusion section. Please check the details in revised draft, page 20 with line 461-462.

Comments 2: In addition, the authors missed the opportunity to evaluate the effect of foliar treatment on fruit quality. I consider this a lack of research in relation to the crops studied.

Response 2: Thanks for the comments. The study mainly focused on regulatory effects of chemical regulators on plant physiologies of strawberry and cucumber during growth period, the fruit quality analysis was also conducted, but will be deeply clarified in the next paper.

Comments 3: I recommend modifying the title of the manuscript, as oligosaccharides are a huge group and it would be more appropriate to define in the title which one from the group of oligosaccharides was investigated.

Response 3: Thanks for the comments. The oligosaccharides indicate the type of chemical regulators used in our research, including chitosan oligosaccharide (CSOS) and mixed oligosaccharides (MixOS).

Comments 4: The authors state in chapter MM that the experiments were established during 2021-2022. In figure 1, the temperature and humidity during the experiments are presented, but in the years 2022 and 2023. This needs to be adjusted.

Response 4: Thanks for the comments. We have revised the year information in figure 1. Please check the details in revised draft, page 5 with line 120-121

Comments 5: In the MM chapter, I ask to describe the CM treatment in detail. Without basic information, it is not possible to scientifically verify and compare the effect of different treatments.

Response 5: Thanks for the comments. The CM treatments used the conventional management of farmers, and the quantities of irrigation, fertilization, pesticides, and fungicides were all completely consistent with those used by local farmers. The fungicide used in the experiment is difenoconazole, and foliar spray based on recommended concentration with 30-125 g∙ha⁻¹ before flowering stage.

Comments 6: In the case of chitosan oligosaccharides, was a commercial preparation used?

Response 6: Thanks for the comments. Chitosan oligosaccharide (CSOS) regulator tested in the study is a commercial product with the single source of chitosan oligosaccharide.

Comments 7: MixOS treatment needs to be characterized in detail in relation to reproduction and comparisons of future research. This means what components were included, in what proportion, etc.

Response 7: Thanks for the comments. Mixed oligosaccharides chemical regulator (MixOS) tested in the study was jointly developed by the Institute of Environment and Sustainable Development in Agriculture of the Chinese Academy of Agricultural Sciences and Resonac Corporation (Showa Denko K.K.), which was supported by the National Key Research and Development Program of China, as well as funded by International Cooperation Key Project. It is widely used in facility agriculture in Japan with the trade name KROPICO. Mixed oligosaccharides regulator (MixOS) contains various single oligosaccharides from multiple sources, including cello-oligosaccharide, chitosan oligosaccharide and xylo-oligosaccharide, which is a multiple functional product. For the detailed information about MixOS mentioned in the reviewer comments, we have clearly analyzed the monosaccharide compositions, molecular weight, functional group valence bond and glucoside bond of the MixOS through ion chromatography, infrared spectroscopy and nuclear magnetic resonance and other techniques. However, the above information is inconvenient to publish because of the involvement of the confidential content of both cooperation parties, and still in the Chinese patent application stage.

Comments 8: L130 The authors state a dose of 1100 kg.ha-1. However, the application was foliar. So what was the dose of water, or the concentration?

Response 8: Thanks for the comments. The mixed oligosaccharides regulator (MixOS) tested in the study is a commercial product named KROPICO, was foliar sprayed in 1000-fold diluted KROPICO based on the recommended concentration and the results of a large number of our previous laboratory concentration screening tests. The chitosan oligosaccharide was foliar sprayed in 500-fold diluted single chitosan oligosaccharide regulator based on the recommended concentration. The doses of the two regulators could be calculated based on application area, total application amount and recommended concentration.

Comments 9: L168 The authors report that a Two-way Anova was used. However, only one factor was investigated in the experiment.

Response 9: Thanks for the comments. For statistical analysis, Two-way analyses of variance (ANOVAs) were performed in the research, one factor referred to different treatments; the other referred to different biological replicates: For the strawberry experiment, four plots were conducted, each plot contained a treatment (nine beds of strawberry plants) with three biological replicates, each biological replicate included three beds of strawberry plants and separated by a bed. For the cucumber experiment, sixteen plots were conducted, each treatment had four biological replicates, each biological replicate was a plot with four beds of cucumber plants.

Comments 10: In the conclusions, the authors assume that the application of oligosaccharides contributes to the antibacterial properties. However, this has not been studied, so I recommend removing this wording.

Response 10: Thanks for the comments. We have removed the information of antibacterial properties in the conclusions. Please check the details in revised draft, page 20 with line 455-468.

 

We look forward to your positive response.

Yours sincerely,

Corresponding Author

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Dear authors,

 

Thank you for your responses to the comments. I appreciate the effort in responding the best to each of them, and I read each of them.

 

One major issue I insist on is the statistical analyses. The response I received from you is that “Two-way analyses of variance (ANOVAs) were performed in the research, one factor referred to different treatments, the other referred to different biological replicates”. I don’t think I ever heard of a instance when replicates are taken as factors, this is why I believed that two separate one way ANOVA tests were performed for strawberry and cucumber separately. Hence, I strongly advise you to ask the opinion of a biostatistics expert of colleague to ensure the statistical tests are accurately applied.

 

Best regards.

Comments on the Quality of English Language

minor syntax and grammar revision recommended

Author Response

Manuscript Title：Different oligosaccharides induce coordination and promotion of root growth

and leaf senescence during strawberry and cucumber growth

 

Response to Reviewer 1 Comments

 

Dear Reviewer,

Thank you for your helpful comments and suggestions on our manuscript. We have modified the manuscript accordingly, and detailed corrections are listed below, point by point.

 

Comments 1: One major issue I insist on is the statistical analyses. The response I received from you is that “Two-way analyses of variance (ANOVAs) were performed in the research, one factor referred to different treatments, the other referred to different biological replicates”. I don’t think I ever heard of an instance when replicates are taken as factors, this is why I believed that two separate one way ANOVA tests were performed for strawberry and cucumber separately. Hence, I strongly advise you to ask the opinion of a biostatistics expert of colleague to ensure the statistical tests are accurately applied.

Response 1: Thanks for the comments. We agree with this comment. We have revised the statistical analysis method that “One-way analyses of variance (ANOVAs)”. Please check the details in revised draft, page 7 with line 189.

 

 

We look forward to your positive response.

Yours sincerely,

Corresponding Author

Author Response File: Author Response.pdf

Round 3

Reviewer 1 Report

Comments and Suggestions for Authors

Dear authors,

I see the issue was fixed.

Best regards.

Comments on the Quality of English Language

minor English style improvements are welcome