Intercropping of Soybean and Common Millet—ARational Way of Forage Biomass Quality Enhancement

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This study explores the effects of different intercropping patterns and biofertilizers on the biomass and contents of medium and trace mineral elements of soybeans and common millet. 
From the experimental results, biofertilizers do not seem to have a significant impact on the yield and contents of medium and trace elements of the two crops. Therefore, it is suggested that biofertilizers should not be considered in this paper, and it would be clearer to evaluate only the yield and element contents under different intercropping measures.
In addition, the discussion section should not be divided by the two crops; it is recommended to discuss the two crops together. Specifically, the discussion should separately address: ① the effects of different intercropping patterns on yield, biomass, and land equivalent ratio (LER); ② the effects of different intercropping patterns on the absorption and utilization of medium and trace elements in the crops.
In the discussion section, the discussion on why intercropping patterns improve the accumulation of medium and trace elements is not in-depth enough, and the underlying reasons have not been profoundly analyzed.
An excessive number of references have been cited, and some references with low relevance can be removed.
There are still a few minor issues as follows:
1.The abbreviations for each treatment are not clear enough. It is suggested to change AS to S1M1, AS1 to S2M2, and AS2 to S2M4.
2.The abstract is not well-written. It only provides qualitative descriptions of the differences between various treatments, lacking quantitative descriptions and necessary data support. For example, it fails to specify by what percentage the element content has increased, and so on.

Author Response

Reviewer 1

Open Review

(x) I would not like to sign my review report
( ) I would like to sign my review report

Quality of English Language

(x) The English could be improved to more clearly express the research.
( ) The English is fine and does not require any improvement.

Comments and Suggestions for Authors

This study explores the effects of different intercropping patterns and biofertilizers on the biomass and contents of medium and trace mineral elements of soybeans and common millet. 
From the experimental results, biofertilizers do not seem to have a significant impact on the yield and contents of medium and trace elements of the two crops. Therefore, it is suggested that biofertilizers should not be considered in this paper, and it would be clearer to evaluate only the yield and element contents under different intercropping measures.

Reply: Thank you for the comment. Biofertilizer showed low effect in general, what is result of the research, too. Some significant effects of the biofertilizer were present regarding increase in Mg, P, and S concentration in soybean pods, as well as B, Mn, and Mo increase in millet panicle, and decrease in P, Cr, and Zn concentration in millet vegetative part, what could be of particular importance when animal feed is considered.

In addition, the discussion section should not be divided by the two crops; it is recommended to discuss the two crops together. Specifically, the discussion should separately address: ① the effects of different intercropping patterns on yield, biomass, and land equivalent ratio (LER); ② the effects of different intercropping patterns on the absorption and utilization of medium and trace elements in the crops.

Reply: The Discussion section was divided into two subsections, as it was suggested, with part 1 considering biomass yield parameters and LER and part 2, considering elemental composition of biomass.

In the discussion section, the discussion on why intercropping patterns improve the accumulation of medium and trace elements is not in-depth enough, and the underlying reasons have not been profoundly analyzed.

Reply: Thank you for the valuable comment. Discussion section was modified in suggested manner, focusing on mineral elements acquisition and their distribution across vegetative and reproductive parts of soybean and millet. Irrespective to the lack of literature data regarding medium and trace elements acquisition by intercrops, particularly millet, main mechanisms were covered.

An excessive number of references have been cited, and some references with low relevance can be removed.

Reply: Reference list was updated after manuscript modification.

There are still a few minor issues as follows:
1.The abbreviations for each treatment are not clear enough. It is suggested to change AS to S1M1, AS1 to S2M2, and AS2 to S2M4.

Reply: The abbreviations are changed, accordingly. Also, to be clearer, instead of S2 for sole millet, we changed abbreviation to M1.

2. 
   The abstract is not well-written. It only provides qualitative descriptions of the differences between various treatments, lacking quantitative descriptions and necessary data support. For example, it fails to specify by what percentage the element content has increased, and so on.

Reply: Thank you for the valuable comment. We included quantitative description of the results in the Abstract, as it was suggested.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript shows a well-designed study on the effect of intercropping of Glycine max (L.) and Panicum miliaceum (L.)  on  biomass accumulation and quality.  The methods applied in field and lab were suitable including precise techniques and equipment.  The results were well analyzed and broadly discussed and may ontribute  to the field of sustainable production of forages.  Also, the discussion is well supported by the results.

The only concern about this manuscript is that the title is quite similar to that of one article already published where the materials and methods were the same as in this manuscript:

Šenk, M.; Simić, M.; Milojković-Opsenica, D.; Brankov, M.; Tolimir, M.; Kodranov, I.; Dragičević, V. Common millet and soy-962 bean intercropping with bio-fertilizer as sustainable practice for managing grain yield and quality. Front. Nutr. 2023, 10, 963 1267928, doi:10.3389/fnut.2023.1267928.

Other than that, the manuscript may be published.

Author Response

Reviewer 2

Open Review

( ) I would not like to sign my review report
(x) I would like to sign my review report

Quality of English Language

( ) The English could be improved to more clearly express the research.
(x) The English is fine and does not require any improvement.

Comments and Suggestions for Authors

The manuscript shows a well-designed study on the effect of intercropping of Glycine max (L.) and Panicum miliaceum (L.)  on  biomass accumulation and quality.  The methods applied in field and lab were suitable including precise techniques and equipment.  The results were well analyzed and broadly discussed and may ontribute  to the field of sustainable production of forages.  Also, the discussion is well supported by the results.

The only concern about this manuscript is that the title is quite similar to that of one article already published where the materials and methods were the same as in this manuscript:

Šenk, M.; Simić, M.; Milojković-Opsenica, D.; Brankov, M.; Tolimir, M.; Kodranov, I.; Dragičević, V. Common millet and soy-962 bean intercropping with bio-fertilizer as sustainable practice for managing grain yield and quality. Front. Nutr. 2023, 10, 963 1267928, doi:10.3389/fnut.2023.1267928.

Other than that, the manuscript may be published.

Reply:

Thank you so much for the support, and understanding the importance of the present results. We changed the manuscript title accordingly: Intercropping of soybean and common millet - rational way of forage biomass quality enhancement

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

Review of the article entitled ‘Intercropping and biofertilizing as sustainable practices for managing the quality and quantity of soybean and common millet biomass (ID: agriculture 3869178)

The subject matter of the work is current and very important. It is part of the trend of sustainable agriculture. Increasing the biodiversity of an agroecosystem through intercropping affects its balance and productivity. The selection of plants for intercropping (soybean, millet) is. appropriate. There is little work on the joint cultivation of these two species. The effect of intercropping is enhanced by the use of biofertilizer. Particularly noteworthy is the extensive research programme and the parallel assessment of the content of elements in plants and in the soil. It is a pity, however, that no connection between them was made in the work. In general, it should be noted that the authors have done a lot of work in collecting, compiling and interpreting the research results. It was not an easy task.

My comments on the manuscript are as follows:

Abstract

It is well written. It presents the main content of the work in a synthetic way.

Introduction

I also have no major comments on the ‘Introduction’ chapter. It is well written. It justifies the research problem well undertaken, points out the shortcomings in this area and ends with a well-presented the aim of the work. However, I think that this chapter should be supplemented with content related to different methods of sowing plants in intercropping.

Lines 79 and 81. I don't understand what the R4 and R7 stages mean (I don't know them). Maybe it would be better to use the BBCH scale.

Material and methods

I have the most comments on this chapter. It lacks a lot of necessary information.

Line 115. Please describe the soil in detail  (based on the Word Reference Base for Soil – WRB).

Table 1 (header). I don't know which element kg ha^-1 ends  with and which element mg kg^-1 begins with. Please construct the headline in such a way that there are no doubts. This note also applies to other tables.

This chapter does not contain any information on when biofertilizer (BF) was applied. Was it spraying BF plants or was it applied to the soil. If to the soil, then to what depth. There is no information on the agrotechnology of crops grown in intercropping. To what depth was the soybean and millet sown. What fertilization was used. How weeds, diseases and pests were controlled.

Lines 144, 148. Please specify exactly when the plants were sown and when they were harvested (date).

Line 147. In what stage of development were soybean and millet (scale).

Subchapter ‘Statistical analysis’

Please clearly state which factors of the experiment have been given in the statistical analysis (intercropping, biofertilizer, years?). What test was used to determine significant differences between means? Why is there no statistical analysis in Tables 10, 11, 12, and there is a standard deviation? Why are the mean values from two years of research given in the Tables? After all, the importance of differences between years has been demonstrated.  In scientific papers, it is generally accepted that in the results of research in Tables and Figures, the largest value is marked with the letter 'a'. At work, it's the opposite.

Results

In subchapter 3.1.1 please write about the common millet yield. No statistically significant differences is also the result.  In the entire 'Results' chapter, please re-examine the description of the results of the research based on the statistical analysis. Example: lines 234, 235. 'The concentration of P and S in AS1 and S2 was a similar level (the same group ‘b’), so there are no differences between them (the differences are in the scope of statistical error). The same is true of the sentence in line 239, lines 270, 271, 274 and 275. These are just examples. There are many more such cases in the text.

Discussion

The Discussion is very extensive. I suggest shortening this chapter by at least one third. It also contains information that has not been the subject of research.  The authors interpreted the results of the study based on the average values from 2018 and 2020. Meanwhile, in a fragment of text placed between lines 456-466, they write about the effect of precipitation and temperatures on the soybean biomass. Meteorological conditions were not the subject of research. In addition, they were not analysed separately each year.  I suggest removing this part of the text. The same comment applies to the passage between lines 573-581 concerning millet.

Line 493. Please write what R5 (growing stage) means.

Line 622. Please explain what AMF stands for.

Conclusions

I propose to delete the sentence placed between lines 672-674. In my opinion, it is unnecessary and does not bring anything new and important.

Similarly, I propose to remove the fragment of the sentence between lines 677-680, concerning precipitation. Precipitation was not the subject of research.

Lines 687-689. I suggest removing this part of the text. The study was about intercropping, not the effect of weather on yield and element content in soybean and millet.

References

The references are well chosen and consistent with the content of the work. However, I suggest replacing items 26, 28, 29, 30 and 31 with newer literature.

I have included the remaining (minor) remarks in the manuscript.

Comments for author File: Comments.pdf

Author Response

Reviewer 3

Open Review

(x) I would not like to sign my review report
( ) I would like to sign my review report

Quality of English Language

( ) The English could be improved to more clearly express the research.
(x) The English is fine and does not require any improvement.

Comments and Suggestions for Authors

Review of the article entitled ‘Intercropping and biofertilizing as sustainable practices for managing the quality and quantity of soybean and common millet biomass (ID: agriculture 3869178)

The subject matter of the work is current and very important. It is part of the trend of sustainable agriculture. Increasing the biodiversity of an agroecosystem through intercropping affects its balance and productivity. The selection of plants for intercropping (soybean, millet) is. appropriate. There is little work on the joint cultivation of these two species. The effect of intercropping is enhanced by the use of biofertilizer. Particularly noteworthy is the extensive research programme and the parallel assessment of the content of elements in plants and in the soil. It is a pity, however, that no connection between them was made in the work. In general, it should be noted that the authors have done a lot of work in collecting, compiling and interpreting the research results. It was not an easy task.

Reply: Thank you for the support and valuable comments. Since mineral elements acquisition by the crop roots is complex process, depending on various factors, which were not covered by this study, our focus was accumulation of mineral elements in aboveground biomass of both intercrops, as an animal feed. Including more data could additionally burden the long text.

My comments on the manuscript are as follows:

Abstract

It is well written. It presents the main content of the work in a synthetic way.

Reply: Thank you for the valuable comment.

Introduction

I also have no major comments on the ‘Introduction’ chapter. It is well written. It justifies the research problem well undertaken, points out the shortcomings in this area and ends with a well-presented the aim of the work. However, I think that this chapter should be supplemented with content related to different methods of sowing plants in intercropping.

Reply: Thank you for the valuable comment We included content related to the different methods of sowing of intercropps: Depending on the purpose, different types of intercropping can be applied, such as row, mixed, strip, or relay intercropping. Additionally, based on row arrangement and crop proportion, intercropping systems can be classified as additive or replacement systems [5].

Lines 79 and 81. I don't understand what the R4 and R7 stages mean (I don't know them). Maybe it would be better to use the BBCH scale.

Reply: Thank you for the notice. We choose R (reproductive) scale, which was mostly used for soybean, as more precise; we also provided explanation in the text: However, maximum accumulation rate of elements tended to occur during R4 (full pod) growth stage (with exception of K and Fe) [15]. Thus, to provide high dry matter yield and forage quality, soybean harvesting should be completed before R7 (beginning maturity) stage.  

Material and methods

I have the most comments on this chapter. It lacks a lot of necessary information.

Line 115. Please describe the soil in detail (based on the Word Reference Base for Soil – WRB).

Reply: Thank you for the valuable comment. We commonly use USDA-NRCS base for the detailed description and classification of our soils, so we added description with the reference, if is it acceptable for you: The soil was slightly calcareous chernozem, Molcal silt loam (Coarse-loamy, mixed, superactive, mesic Vitrandic Calcixerolls) (USDA-NRCS, 2019).  

Table 1 (header). I don't know which element kg ha^-1 ends  with and which element mg kg^-1 begins with. Please construct the headline in such a way that there are no doubts. This note also applies to other tables.

Reply: We included the vertical line in this and all other following tables, to provide visibility and accuracy of the data.

This chapter does not contain any information on when biofertilizer (BF) was applied. Was it spraying BF plants or was it applied to the soil. If to the soil, then to what depth. There is no information on the agrotechnology of crops grown in intercropping. To what depth was the soybean and millet sown. What fertilization was used. How weeds, diseases and pests were controlled.

Reply: We applied the biofertilizer on seeds prior to sowing, so the sentence was modified to be clearer: Accordingly, 50 g of biofertilizer Coveron (Hello Nature International Srl, Italy), containing Glomus sp., Trichoderma atroviride and plant growth-promoting rhizobacteria, was dissolved in 800 mL of water and solution was applied per 100 kg of seeds, inoculating seed prior to sowing.

All information about agrotechnology of crops was included into text. The information about weed control was already present in the Chapter 2.2.

Lines 144, 148. Please specify exactly when the plants were sown and when they were harvested (date).

Reply: Sentence was modified accordingly: The sowing was performed on May 3, 2018, and April 22, 2020.

Line 147. In what stage of development were soybean and millet (scale).

Reply: Sentence was modified: Harvesting of green biomass was performed at the beginning of seed development for soybean (R5) and at the ripening stage for millet, i.e., on August 2, 2018, and on July 22, 2020.

We did not use scale for millet, but  described actual phenophases, because it is not commonly used in scientific literature. For soybean was used accepted and the most commonly used scale in literature to describe phenophases.

Subchapter ‘Statistical analysis’

Please clearly state which factors of the experiment have been given in the statistical analysis (intercropping, biofertilizer, years?).

Reply: Thank you for the comment, sentence was modified: The experimental data were processed using Multivariate analysis of variance (MANOVA) at the significance level of p < 0.05 (F test); three-way analysis including year, intercropping and biofertilizer, as well as their interactions.

What test was used to determine significant differences between means?

Reply: It was replied in the upper comment.

Why is there no statistical analysis in Tables 10, 11, 12, and there is a standard deviation?

Reply: We included letters describing the significant differences between values.

Why are the mean values from two years of research given in the Tables? After all, the importance of differences between years has been demonstrated.  In scientific papers, it is generally accepted that in the results of research in Tables and Figures, the largest value is marked with the letter 'a'. At work, it's the opposite.

Reply: Thank you for the valuable comment. Yes, you are right, there is significant impact of the years, but the focus of the research was on intercropping and biofertilizer, thus including year as a factor in tables will additionally broaden them. The impact of the year was shown in Tables 4-8, and discussed. While data in Tables 9-12 show integrated influence of intercropping and biofertilizer. Taking into account that in further, the shortening of the manuscript regarding impact of the year was suggested in Discussion, we kept the present form of the tables. The letters in the tables, ranging from the greatest to the lowest value were corrected.

Results

In subchapter 3.1.1 please write about the common millet yield. No statistically significant differences is also the result.

Reply: Accordingly to the results in Table 4, there was no significant differences in soybean yield regarding factors, thus the text was modified, accordingly: FBY and DBY of vegetative and reproductive parts of millet were significantly affected by intercropping, while sowing pattern did not have significant impact on soybean yields.

In the entire 'Results' chapter, please re-examine the description of the results of the research based on the statistical analysis. Example: lines 234, 235. 'The concentration of P and S in AS1 and S2 was a similar level (the same group ‘b’), so there are no differences between them (the differences are in the scope of statistical error). The same is true of the sentence in line 239, lines 270, 271, 274 and 275. These are just examples. There are many more such cases in the text.

Reply: Thank you for the comment. As there were no statistical differences between some values from the same group, we pointed in text only the highest values from tables.

Discussion

The Discussion is very extensive. I suggest shortening this chapter by at least one third. It also contains information that has not been the subject of research. 

Reply: Thank you for the comment; we shortened Discussion, excluding information which are not the subject of the research.

The authors interpreted the results of the study based on the average values from 2018 and 2020. Meanwhile, in a fragment of text placed between lines 456-466, they write about the effect of precipitation and temperatures on the soybean biomass. Meteorological conditions were not the subject of research. In addition, they were not analysed separately each year.  I suggest removing this part of the text. The same comment applies to the passage between lines 573-581 concerning millet.

Reply: Thank you for the valuable comment; we excluded the suggested parts of the text.

Line 493. Please write what R5 (growing stage) means.

Reply: It was added in Chapter 2.2.

Line 622. Please explain what AMF stands for.

Reply: The abbreviation was explained in Introduction: arbuscular mycorrhizal fungi.

Conclusions

I propose to delete the sentence placed between lines 672-674. In my opinion, it is unnecessary and does not bring anything new and important.

Reply: Thank you for the comment, we deleted this part of the text.

Similarly, I propose to remove the fragment of the sentence between lines 677-680, concerning precipitation. Precipitation was not the subject of research.

Reply: We agree, but regarding to the fact that there was no significant difference between years on LER value, it presents the important information on stability of examined intercropping system regarding yield across seasons.

Lines 687-689. I suggest removing this part of the text. The study was about intercropping, not the effect of weather on yield and element content in soybean and millet.

Reply: Thank you for the comment, this part was deleted.

References

The references are well chosen and consistent with the content of the work. However, I suggest replacing items 26, 28, 29, 30 and 31 with newer literature.

Reply: We agree that the references are old, but they were referred to the methods used in study. Ref 31 is the original reference regarding LER calculation.

I have included the remaining (minor) remarks in the manuscript.

Reply: Thank you for remarks, they were valuavble for manuscript improvement.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

Comments on the manuscript ‘Intercropping and biofertilizing as sustainable practices for managing the quality and quantity of soybean and common millet biomass’:

The manuscript addresses an interesting topic — intercropping —which is relevant to sustainable crop production. The results of this study can complement research on the integration of legume and cereal farming and be useful for relevant farmers.

The manuscript has several strengths, including collecting large amounts of data, especially on a wide range of nutrients, and using appropriate statistical methods.

However, to improve the quality of the paper, the following should be considered or corrected:

• The effect of BF is mentioned as small or minor in the abstract and conclusion sections, while its effects on the concentrations of Mg, P, and S (Table 6), P, Cr, and Zn (Table 7), and B, Al, Mn, and Mo (Table 8) are significant. Therefore, it is necessary to rewrite the abstract and conclusion in such a way that these important and meaningful effects are referred to in some way.
• The number of keywords is too high. Also, the "inductively coupled plasma (ICP)" is not necessary because it is only a laboratory method!

• Lines 102 and 103 (the last paragraph of the introduction section): It has been said that “Currently, there is a lack of information regarding the intercropping of soybean and common millet.”! While numerous articles have been published on this subject, including the reference articles in your manuscript, the sentence should therefore be corrected.

• Line 133: What bacteria did the PGPRs (plant growth-promoting rhizobacteria) in the biofertilizer formulation include? PGPRs cover a wide range of genera and species.

• Line 134: The details of the application of biofertilizer need further clarification and explanation. Was this a seed coating/inoculation treatment before sowing? Or was the solution applied to the field/soil at a certain rate? The exact method and application rate per plot or plant should be specified for reproducibility.

• Line 179 (statistical analysis): What statistical procedure was used to alphabetically group and compare treatment means?

• The result ‘LER>1’ in treatment AS1 is an important finding, and this should be clearly highlighted in the results section with values (1.10 for fresh, 1.12 for dry biomass) as presented in the abstract section.
• Although millet yield decreased in treatment AS1, land equivalence ratio (LER) and forage quality improved. It is recommended that the discussion section of the paper explain how the reduction in millet yield is related to the high LER and the improvement in element concentration in treatment AS1.

• Lines 550-552: The claim of “new insights” in this study into the uptake of elements under stress conditions requires further elaboration. What are the specific novel insights, and how do they challenge or extend existing knowledge?

Some additional comments regarding the structure of the manuscript:

• The introduction section is prepared in a structured manner and includes the importance of the topic, the statement of the problem and the necessities, and the objectives of the research.
• The order of presenting the results is appropriate.
• The conclusion section, after editing (according to comment 1), can be consistent with the presented results.
• The references are relevant, and most of them are recently published.

Comments on the Quality of English Language

It could be improved.

Author Response

Reviewer 4

Open Review

(x) I would not like to sign my review report
( ) I would like to sign my review report

Quality of English Language

(x) The English could be improved to more clearly express the research.
( ) The English is fine and does not require any improvement.

Comments and Suggestions for Authors

Comments on the manuscript ‘Intercropping and biofertilizing as sustainable practices for managing the quality and quantity of soybean and common millet biomass’:

The manuscript addresses an interesting topic — intercropping —which is relevant to sustainable crop production. The results of this study can complement research on the integration of legume and cereal farming and be useful for relevant farmers.

The manuscript has several strengths, including collecting large amounts of data, especially on a wide range of nutrients, and using appropriate statistical methods.

However, to improve the quality of the paper, the following should be considered or corrected: The effect of BF is mentioned as small or minor in the abstract and conclusion sections, while its effects on the concentrations of Mg, P, and S (Table 6), P, Cr, and Zn (Table 7), and B, Al, Mn, and Mo (Table 8) are significant. Therefore, it is necessary to rewrite the abstract and conclusion in such a way that these important and meaningful effects are referred to in some way.

Reply: Thank you for the notice, the sentence in Abstract regarding BF was modified accordingly: While BF had generally small impact on forage biomass yield and elements accumulation, it increased Mg, P and S concentration in soybean pods, as well as concentration of B, Mn and Mo in panicle, simultaneously decreasing P, Cr and Zn concentration in millet vegetative parts. Also sentence in Conclusion was modified: BF had a minor impact on elements accumulation (increasing only concentration of Mg, P and S in soybean pods, and concentration of B, Mn and Mo in panicle, while decreased P, Cr and Zn concentration in millet reproductive parts), indicating that nutrient acquisition is not entirely reliant on symbiosis with mycorrhiza.

The number of keywords is too high. Also, the "inductively coupled plasma (ICP)" is not necessary because it is only a laboratory method!

 Reply: Number of keywords was reduced and ICP was deleted.

Lines 102 and 103 (the last paragraph of the introduction section): It has been said that “Currently, there is a lack of information regarding the intercropping of soybean and common millet.”! While numerous articles have been published on this subject, including the reference articles in your manuscript, the sentence should therefore be corrected.

Reply: Thank you for the valuable insight, but most of the literature used in this manuscript and present in scientific literature are referred to the other millet types (which have similarities with common millet to the some extent), but not the common millet, what presents novelty of the research.

Line 133: What bacteria did the PGPRs (plant growth-promoting rhizobacteria) in the biofertilizer formulation include? PGPRs cover a wide range of genera and species.

Reply: Thank you for the notice, we corrected text acordingly: We applied the biofertilizer on seeds prior to sowing, so the sentence was modified to be clearer: Accordingly, 50 g of biofertilizer Coveron (Hello Nature International Srl, Italy), containing Glomus sp., Trichoderma atroviride and plant growth-promoting rhizobacteria, was dissolved in 800 mL of water and solution was applied per 100 kg of seeds, inoculating seed prior to sowing.

While the description of product included mycorrhizal fungi and PGPRs, we included all information from the product label (declaration). We supposed that some parts of the product were under protection, such as PGPR composition.

Line 134: The details of the application of biofertilizer need further clarification and explanation. Was this a seed coating/inoculation treatment before sowing? Or was the solution applied to the field/soil at a certain rate? The exact method and application rate per plot or plant should be specified for reproducibility.

Reply: We corrected the sentence (in upper comment). This means that there was no coating, but we mixed seeds with biofertilizer solution manually, prior to sowing.

Line 179 (statistical analysis): What statistical procedure was used to alphabetically group and compare treatment means?

 Reply: Thank you for the valuable comment, we modified sentence in 2.4 section: The experimental data were processed using Multivariate analysis of variance (MANOVA) at the significance level of p < 0.05 (F test); three-way analysis including year, intercropping and biofertilizer, as well as their interactions. LSD test (values) were used to determine significant differences between means.

The result ‘LER>1’ in treatment AS1 is an important finding, and this should be clearly highlighted in the results section with values (1.10 for fresh, 1.12 for dry biomass) as presented in the abstract section.

Reply: Thank you for the comment, the sentence in Chapter 3.1.1 was modified, accordingly: The highest, and the only ones > 1, were obtained in S2M2 combination, with value of 1.10 for fresh and 1.12 for dry biomass.

Although millet yield decreased in treatment AS1, land equivalence ratio (LER) and forage quality improved. It is recommended that the discussion section of the paper explain how the reduction in millet yield is related to the high LER and the improvement in element concentration in treatment AS1.

Reply: Thank you for the notice. Possible explanation is literature supported, thus the sentence in Discussion was modified: Jahanzad et al. [100] emphasized the advantage of an optimal soybean-pearl millet ratio on LER for higher exploitation when environmental resources are limited, what could present the explanation for LER>1 in S2M2 combination, irrespective to the lower millet yield.

Lines 550-552: The claim of “new insights” in this study into the uptake of elements under stress conditions requires further elaboration. What are the specific novel insights, and how do they challenge or extend existing knowledge?

Reply: We agree that the impact of stress has complex impact, making challenging any crop production, but sustainable strategies, such as intercropping could diminish such effect to some extent, particularly when absorption and partitioning of mineral elements is considered. Thus, the sentence was modified to be more accurate: Although the majority of research has focused on reduced mineral uptake under stress conditions, this research provides new insights in this field, regarding macro-and microelements acquisition and partitioning by different intercrop combinations and paves the way for future research endeavors. This could be of particular importance, taking into account climate change, and rational use of agroecosystem services toward increased forage and food quality.

Some additional comments regarding the structure of the manuscript:

The introduction section is prepared in a structured manner and includes the importance of the topic, the statement of the problem and the necessities, and the objectives of the research.

Reply: Thank you for the valuable comment.

The order of presenting the results is appropriate.

Reply: Thank you for the valuable comment.

The conclusion section, after editing (according to comment 1), can be consistent with the presented results.

Reply: Thank you for the valuable comment, we accepted all suggestions.

The references are relevant, and most of them are recently published.

Reply: Thank you for the valuable comment.

Comments on the Quality of English Language

It could be improved.

Reply: The manuscript was checked again, by language expert.

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

Dear Authors,

You have done a great and good  job. The article looks much better in its current form than in the previous version. All my comments included in the review have been taken into account.

I accept the article in its presented form.

Reviewer 4 Report

Comments and Suggestions for Authors

The corrections have been made.