Expression of Secondary Sexual Dimorphism in the Diurnal Course of Leaf Gas Exchanges Is Modified by the Rhythmic Growth of Ilex paraguariensis Under Monoculture and Agroforestry

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Dear authors should pay attention to the revisions, addressing each and every comment:

1. Add a paragraph importance of sexual dimorphism leaf gas exchange of lex paraguariensis with global CO₂ enhancement.

2. Figures quality must be improved for better understanding.

3. The conclusion is currently unclear and needs revision. The authors should enhance scientific accuracy, clearly articulate the novelty of their perspective, and address the existing gaps in the research.

4. In respect to references, Scientific names ethics formatting, proper spelling, line spacing and paragraphing can improve overall manuscript quality. I suggest to authors, re-check the reference formatting carefully. 

Comments on the Quality of English Language

The English could be improved to more clearly express the research.

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

( ) The quality of English does not limit my understanding of the research.
(x) The English could be improved to more clearly express the research.

 

	Yes	Can be improved	Must be improved	Not applicable
Does the introduction provide sufficient background and include all relevant references?	( )	(x)	( )	( )
Is the research design appropriate?	(x)	( )	( )	( )
Are the methods adequately described?	(x)	( )	( )	( )
Are the results clearly presented?	( )	(x)	( )	( )
Are the conclusions supported by the results?	( )	( )	(x)	( )

 

Comments and Suggestions for Authors

 

Dear authors should pay attention to the revisions, addressing each and every comment:

 

Authors: All changes recommended by the Reviewer #1 are highlighted in red, except for language modifications and slight restructuring.

 

1. Add a paragraph importance of sexual dimorphism leaf gas exchange of lex paraguariensis with global CO₂

 

Authors: None experiment with Ilex paraguariensis under elevated CO₂ conditions had never been performed in the world. However, we added some words about Salix glauca, Populus cathayana and yerba mate in the scope of climate changes, giving special attention to leaf gas exchange and SSD (Lines 527-539):

 

“Salix glauca females have lower drought tolerance than males under years of extreme aridity, while males grew at a consistent rate regardless of habitat aridity, which was followed during ten year period [70]. Such responses indicate that one annual cycle probably would not be enough for a full generalization about yerba mate. The spatial gender segregation could shift under global warming and female plants of various dioecious species would lose their dominance in high resource habitats, and males would increase their dominance in relatively low resource habitats [70]. Accordingly, males of P. cathayana exhibit higher A_net, biomass accumulation and carbon balance mechanism under elevated CO₂ than females [71]. No experiment about physiology of yerba mate was performed under elevated CO₂, but some simulations projected that current favorable areas for yerba mate cultivation will reduce significantly in Brazil [3, 16]. How gender dominance in yerba mate carbon acquisition and WUE would change under elevated CO₂ is a topic for further research.”

 

2. Figures quality must be improved for better understanding.

 

Authors: We improved the structure and quality of all figures in the manuscript. Additionally, we sent the high quality (high resolution figures) together with the manuscript to the MDPI.

 

3. The conclusion is currently unclear and needs revision. The authors should enhance scientific accuracy, clearly articulate the novelty of their perspective, and address the existing gaps in the research.

 

Authors: We worked on conclusions to underline the novelty of our paper and improve this section (Lines 561-564):

 

“As novelty, we found a modification of SSD expression in leaf gas exchange traits over the rhythmic growth period, which was unknown in yerba mate. Then, our initial hypothesis that females would have higher photosynthetic performance than males was strongly modified by growth rhythmicity. In fact, A_net, A_net/C_i and WUE were higher in females than males during the summer rest (R1) and spring growth unit (GU2) periods. Also, g_s and E were higher in females than males during the summer R1. Oppositely, higher WUE in males than in females was observed during the fall growth unit (GU1) and winter rest (R2), together with A_net/C_i ratio during the winter R2, and E during the spring GU2 periods. This means that the gender dominance changed over the growth periods and phenology of yerba mate. All those leaf gas exchange traits were modified by light environment, with higher values found generally under MO as compared with AFS. Despite the strong effect of cultivation system on LAI and leaf gas exchange traits over the diurnal course, SSD expression was rarely modified by the cultivation system, being expressed for E and WUE only in MO during spring GU2 and winter R2, respectively. Higher WUE of females during early vegetative (R1) and early reproductive (GU2) growth periods would confer an ecological advantage during warmer periods. On the other hand, higher WUE in males during fall GU1 and winter R2 would benefit plants during colder and drier annual periods. The superior A_net in females during early vegetative (R1) and early reproductive (GU2) stages can be underlined as a generalized SSD expression in yerba mate and could be considered as a fitness strategy of female plants in their additional reproduction efforts. While long-term experiments would reveal if the ontogeny changes the gender dominance in leaf gas exchanges of yerba mate, our study highlights the importance of the period of rhythmic growth when the ecophysiological traits were evaluated.”

 

4. In respect to references, Scientific names ethics formatting, proper spelling, line spacing and paragraphing can improve overall manuscript quality. I suggest to authors, re-check the reference formatting carefully.

 

Authors: All references were carefully checked and corrected. By five previous experiences with MDPI, the publishing team always adapt slightly the form to the proper journal/paper. Two recent publications (references) were updated with volume and e-number.

 

Comments on the Quality of English Language

 

The English could be improved to more clearly express the research.

 

Authors: Language was carefully checked and corrected when needed.

 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Review Comments – Manuscript Forests-3386839

Type of manuscript: Article

Title: Expression of Secondary Sexual Dimorphism in Diurnal Course of Leaf Gas Exchanges is Modified by the Rhythmic Growth of Ilex paraguariensis Under Monoculture and Agroforestry

Authors: Miroslava Rakočević, et al.

 

          The manuscript provides valuable insights into the ecophysiological responses of Ilex paraguariensis under different cultivation systems and the implications of secondary sexual dimorphism. The results are well-supported by comprehensive data and analyses, which contribute significantly to understanding the interaction of cultivation systems, seasonal rhythms, and gender-based physiological differences.

The manuscript is generally well-written and structured, though there are areas where improvements can be made for clarity and completeness. The results presented in the paper are of general interest of Forests journal.

  

The text

1. I propose considering the inclusion of information on hormonal mechanisms and biochemical stress markers. This could strengthen the interpretation of physiological responses to environmental stress in different cultivation systems. Current sections (e.g., lines 15–35, 280–304) discuss stress primarily in terms of physiological traits, but there is no reference to biochemical markers​

​2. What do the authors think about expanding the discussion on the influence of genotypic variability on SSD? Adding such data could help explain the physiological differences between genders.

3. I propose considering adding comparisons of yerba mate results with other dioecious species in the context of environmental stress. Do the observed SSD differences in MO and AFS align with findings for other dioecious plants? Currently, no such comparisons are present; the context is limited to SSD in yerba mate (lines 450–479)​

​4. What do the authors think about discussing seasonal changes in traits in more detail, particularly in relation to potential ecological advantages of each gender? This could enhance the interpretation of the findings.

​5. I suggest clarifying the application of mixed-effects models and discussing significant interactions between factors (e.g., gender × season), particularly in the context of presenting results in tables and figures.

 6. What is the way forward to this study?

 

Figures

1. I suggest in Figure 1 increasing the font on the axis descriptions to make them more readable.

 

Equations

1. Why is equation number 1 in bold?

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

(x) The quality of English does not limit my understanding of the research.
( ) The English could be improved to more clearly express the research.

 

	Yes	Can be improved	Must be improved	Not applicable
Does the introduction provide sufficient background and include all relevant references?	( )	(x)	( )	( )
Is the research design appropriate?	(x)	( )	( )	( )
Are the methods adequately described?	(x)	( )	( )	( )
Are the results clearly presented?	( )	(x)	( )	( )
Are the conclusions supported by the results?	( )	( )	(x)	( )

 

Comments and Suggestions for Authors

 

Review Comments – Manuscript Forests-3386839

Type of manuscript: Article

Title: Expression of Secondary Sexual Dimorphism in Diurnal Course of Leaf Gas Exchanges is Modified by the Rhythmic Growth of Ilex paraguariensis Under Monoculture and Agroforestry

Authors: Miroslava Rakočević, et al.

 

The manuscript provides valuable insights into the ecophysiological responses of Ilex paraguariensis under different cultivation systems and the implications of secondary sexual dimorphism. The results are well-supported by comprehensive data and analyses, which contribute significantly to understanding the interaction of cultivation systems, seasonal rhythms, and gender-based physiological differences. The manuscript is generally well-written and structured, though there are areas where improvements can be made for clarity and completeness. The results presented in the paper are of general interest of Forests journal.

 

Authors: Thank you. We revised the entire text to improve its structure, fluency and clarity. Such improvements, together with those related to the language, had not been highlighted. However, all other modifications requested by the Reviewer #2 were marked in blue along with the new version.

 

1. I propose considering the inclusion of information on hormonal mechanisms and biochemical stress markers. This could strengthen the interpretation of physiological responses to environmental stress in different cultivation systems. Current sections (e.g., lines 15–35, 280–304) discuss stress primarily in terms of physiological traits, but there is no reference to biochemical markers​.

 

Authors: In fact, our paper is about the sexual dimorphism (SSD) expression and the environmental regulation is a secondary topic herein. For sure, an experiment on stress physiology and SSD would be quite interesting as well as the role of genes and growth. We added some words about this. Thank you. See lines 96-99:

 

“The complex sexual phenotypic differentiation in angiosperms can be understood through the genetic regulation and the spatial and temporal role of plant growth regulators [33]. The single-gene sex determining genetic systems could be common in dioecious species that evolved via monoecy [34].”

 

2. What do the authors think about expanding the discussion on the influence of genotypic variability on SSD? Adding such data could help explain the physiological differences between genders.

 

Authors: Thank you. We added (Lines 458-462):

 

“Despite that genetic variation could have a prominent role in sex expression in dioecious species [21, 35], the findings from three independent experiments with varying genetic material indicate a generalized SSD expression in yerba mate and this could be associated with as a fitness strategy of female plants in their additional reproduction efforts.”

 

3. I propose considering adding comparisons of yerba mate results with other dioecious species in the context of environmental stress.

 

Authors: As commented previously, our paper is not about stress physiology and for this we do not have enough data for a strong discussion and comparison. Anyway, about 51% of our discussion addresses species other than yerba mate, as shown in lines 398-400; 424-426; 434-442; 458-462; 469-474; 484-490; 499-518; 519-520; 521-522; 523-524; 525-536.

 

4. Do the observed SSD differences in MO and AFS align with findings for other dioecious plants? Currently, no such comparisons are present; the context is limited to SSD in yerba mate (lines 450–479).

 

Authors: There is no similar study done with other species under field conditions about leaf gas exchanges under MO and AFS, or pure stand and AFS with focus on SSD. Trying to follow the reviewer’s advice, we mentioned papers about Silene latifolia and Amaranhus palmeri (lines 434-442):

 

Gas exchange of two genders of Silene latifolia do not respond differently to low resource availability (light, water, nitrogen, phosphorus, and potassium), and higher female reproductive effort relative to males does not differentially affect their ability to assimilate carbon, despite that male A_net and g_s were slightly, but consistently, higher than those of females [55]. Varying gender acclimation to low light can be observed in some dioecious species, as in Amaranthus palmeri females [56]. They respond to shading by stem elongation, whereas the male plants respond by increasing specific leaf area. A. palmeri genders showed a differential response to stressful conditions because of differences in their ontogeny and physiology, and possibly due to the cost of reproduction.

 

Lines 469-472 (about Amaranthus palmeri):

 

“Similarly, the greater photosynthetic capacity of female plants compared with male ones was noticed under high light conditions (compared here to MO) in Amaranthus palmeri, and such differential photosynthetic performance diminished in both genders with progressed phenology [60]..”

 

… and about Populus cathayana (lines 512-515):

 

“Female roots of P. cathayana release diverse phenolic allelochemicals into the soil environment, resulting in growth inhibition of the same sex neighbors when grown in sex monoculture, but their growth with males is consistently enhanced, especially the root growth [67].”

 

5. What do the authors think about discussing seasonal changes in traits in more detail, particularly in relation to potential ecological advantages of each gender? This could enhance the interpretation of the findings.

 

Authors: Our sampling along seasons was done to capture changes in SSD expression instead of studying plant responses to seasonal modifications in light, temperature and water. However, we improved the text for clarifying the ecological advantages due to variations in A_net, E and WUE. Please, see below.

 

Lines 32-36:

 

Authors: “High WUE in males during the winter would benefit plants during cold and dry periods, improving the balance between carbon acquisition and water loss through transpiration. On the other hand, high A_net during the summer and spring could be considered as a general fitness strategy of female plants to improve photoassimilate supplying and support their additional reproduction costs”.

 

Lines  490-595:

Authors: “In our experiment, WUE was higher in females than in males during the early vegetative (summer R1) and early reproductive (spring GU2) stages periods (Figure 6), conferring an ecological advantage of female yerba mate plants. On the other hand, higher WUE was observed in males in fall (GU1) and winter (R2), conferring the ecological advantage of male plants due to a higher competition for water uptake during colder and drier periods (Figure 7).”

 

Lines  555-561:

 

Authors: “Higher WUE of females during early vegetative (R1) and early reproductive (GU2) growth periods would confer an ecological advantage during warmer periods. On the other hand, higher WUE in males during fall GU1 and winter R2 would benefit plants during colder and drier annual periods. The superior A_net in females during early vegetative (R1) and early reproductive (GU2) stages can be underlined as a generalized SSD expression in yerba mate and could be considered as a fitness strategy of female plants in their additional reproduction efforts.”

 

6. I suggest clarifying the application of mixed-effects models and discussing significant interactions between factors (e.g., gender × season), particularly in the context of presenting results in tables and figures.

 

Authors: We studied interactions between genders and cultivation systems over the diurnal cycle, while the seasonal effects were analyzed as a simple factor. Significant interaction between cultivation system and gender was observed in only one case. For resuming our findings, we prepared Figure 7.

 

7. What is the way forward to this study?

 

Authors: Great, please see the text below (lines 538-539):

 

“How gender dominance in yerba mate carbon acquisition and WUE would change under elevated CO₂ is a topic for further research.”

 

Figures

I suggest in Figure 1 increasing the font on the axis descriptions to make them more readable.

 

Authors: Thank you. The axis descriptors were increased. And many other details were improved in Figure 1 and all figures.

 

Equations

Why is equation number 1 in bold?

 

Authors: Thank you for your observation. It was an error of typing. The equation was rewritten without bold letters.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The work "Expression of Secondary Sexual Dimorphism in Diurnal Course of Leaf Gas Exchanges is Modified by the Rhythmic Growth of Ilex paraguariensis Under Monoculture and Agroforestry" is very interesting from a scientific point of view.

It brings new knowledge on leaf gas exchange in a dioecious woody plant such as Yerbe mate. The work is a complex, multifactorial field experiment. Due to this, the methodological assumptions, and especially the presentation and interpretation of results, are difficult.

Therefore, the graphs are very extensive, complex, contain a lot of information and are therefore difficult to interpret. Despite this, I believe that they are appropriate for the collected empirical data. With this type of empirical data, the use of advanced multifactorial analysis models would be an even greater challenge.

The work is scientifically valuable and therefore I recommend it for publication after correction in accordance with the comments that are posted as "comments" directly in the work (file forests-3386839-review-v1_29 12 2024). I posted the comments in the following lines of the work: 20, 40, 44, 126, 131, 137, 141, 158, 220, 267, 301, 330, 494 and 509.

Comments for author File: Comments.pdf

Author Response

REVIEWER #3

 

The work "Expression of Secondary Sexual Dimorphism in Diurnal Course of Leaf Gas Exchanges is Modified by the Rhythmic Growth of Ilex paraguariensis Under Monoculture and Agroforestry" is very interesting from a scientific point of view.

It brings new knowledge on leaf gas exchange in a dioecious woody plant such as Yerbe mate. The work is a complex, multifactorial field experiment. Due to this, the methodological assumptions, and especially the presentation and interpretation of results, are difficult.

Authors: Thank you. The assumptions and experimental work were complex. We made the update of all figures and the whole text, and manuscript is much easier to follow. All responses made to Reviewer #3 were marked in the text in green color.

Therefore, the graphs are very extensive, complex, contain a lot of information and are therefore difficult to interpret. Despite this, I believe that they are appropriate for the collected empirical data. With this type of empirical data, the use of advanced multifactorial analysis models would be an even greater challenge.

Authors: Thank you. We applied multifactorial models because of complex questions that were posed.

The work is scientifically valuable and therefore I recommend it for publication after correction in accordance with the comments that are posted as "comments" directly in the work (file forests-3386839-review-v1_29 12 2024). I posted the comments in the following lines of the work: 20, 40, 44, 126, 131, 137, 141, 158, 220, 267, 301, 330, 494 and 509.

Authors: Thank you. We followed your suggestions and added the adequate text over the manuscript.

Line 20: information was added in the line 22 of the actual text.:

“…intercellular CO₂ concentration (C_i)…”

Line 40: information about the natural habitat of Ilex paraguariensis can be found (in detail) in lines 47-51 and 52-63.

Line 44: Authors - we added:

 “and row material and is used intensively for tea, medicines and pharmaceuticals”.

Line 126: The share with other species was not the aim of this study and had not been studied. But we explained the arrangement (Lines 147-148). The forest where the yerba mate was grown was complex and the composition was explained in lines 137-141.

Line 131: Authors: We added the explanations:

“In both cultivation systems, young clonal plants issued from vegetative reproduction were planted in June 2018. They were of ~10 cm of height, and each clone originated from one plant of the same sex. The planting arrangement was about 3 m distance between rows and 1.5 m between plants in rows.”

Line 137:

Authors: “… with the regrowth where the branches of the 1^st order tend to grow perpendicular to the supporting structure [10].”

Line 141:

Authors: We gave very detailed explanations about the measurements in the text that followed the first paragraph of the session 2.2. The results that are presented are from 18-72 means (of such number of days) of 10 seconds registers integrated on hourly values - lines 165-170 :

“PPFD values were recorded with a datalogger (H21-USB Data Logger USB, HOBO, Bourne MA, USA), every 10 seconds. Here, we used hourly mean values. As we had only one datalogger, the sensors were moved every two-three weeks from one to the other system, AFS and MO, along the experimental period. Simultaneously with PPFD measurements, air temperature was registered in both systems and during the four seasons, with the U23-003 HOBO 2x external temperature data logger (HOBO, Bourne MA, USA, Figure S1).”

…” Mean ± SE (n = 18-72)” (Line 245)”…

Line 158:

Authors –  LAI is a proxy of plant size and canopy when measurements were taken. In fact, LAI is determined by the genetic of populations/clones and environmental conditions. We did not cut, pruned or collected any leaves. LAI is the measure of the whole plant area of leaves projected on the soil surface.

Line 220:

Authors –  thank you, we added from 10:45h to 14:45h (actual line 232).

Line 267:

Authors –  thank you, we added “in the summer” in the actual line 288.

                Line 301:

Authors –  thank you for your observation. But only in AFS (which is written), where the interaction exists between the Hour and the cultivation system, and we added:

… “ interaction Sys x Hour” in the line 316-317.

                Line 330:

Authors: Thank you, we added “statistically” (actual line 353)

Line 494: We added the complete description of the growth periods in yerba mate in ‘Conclusions’.

Line 509: Other three reviewers also had the opinions about the Conclusions and in this revised version we finished with projections of physiological studies end SSD. In the ‘Conclusions’ we conclude what we did, and we did not evaluate economical values.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

The paper addresses an interesting topic: in how the SSD approach has been used to demonstrate leaf gas exchange as modified by rhythmic growth under climatic conditions. However, certain aspects need to be revised and improved to make a significant impact on the research.

 

In the introduction, the research's theoretical aspects should be considered to challenge existing views or provide new insights into the topic. For example, existing literature on sexual dimorphism in dioecious plants, particularly its relation to gas exchange and rhythmic growth, should be addressed and adjusted in lines 50 to 88.

 

Missing citations for some sentences or definitions need to be checked by the authors.

 

In the methods section, the authors should provide detailed descriptions of the experimental area. Suggestion: the author may consider making a flowchart that clearly shows the research step.

 

The article provides a solid overview of the results with an analysis of the dataset. Is it necessary to explain or map the LAI to understand the changes? This may relate to the scalability across different climatic conditions and geographic regions. Also, how to validate the resulting dataset in terms of the specific parameters derived from the climate measurement.

 

Finally, the discussion section should be strengthened by addressing the implications of these findings for existing literature.

Author Response

REVIEWER #4

 

Open Review

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

 

Quality of English Language

(x) The quality of English does not limit my understanding of the research.
( ) The English could be improved to more clearly express the research.

 

 

	Yes	Can be improved	Must be improved	Not applicable
Does the introduction provide sufficient background and include all relevant references?	( )	(x)	( )	( )
Is the research design appropriate?	( )	(x)	( )	( )
Are the methods adequately described?	( )	(x)	( )	( )
Are the results clearly presented?	( )	(x)	( )	( )
Are the conclusions supported by the results?	( )	(x)	( )	( )

 

Comments and Suggestions for Authors

The paper addresses an interesting topic: in how the SSD approach has been used to demonstrate leaf gas exchange as modified by rhythmic growth under climatic conditions. However, certain aspects need to be revised and improved to make a significant impact on the research.

 

Authors: Thank you for your positive comments. All changes made along with the paper were highlighted in violet.

 

In the introduction, the research's theoretical aspects should be considered to challenge existing views or provide new insights into the topic. For example, existing literature on sexual dimorphism in dioecious plants, particularly its relation to gas exchange and rhythmic growth, should be addressed and adjusted in lines 50 to 88.

 

Authors: We added some existing literature about SSD, leaf gas exchange and seasonality (lines 85-94 and 104-107):

 

“The sexual reproduction in Sagittaria latifolia imposes asymmetry between the genders, with greater biomass costs for females and greater nitrogen costs for males [24]. Curiously, this difference in nitrogen costs between the genders was not associated with differences in photosynthetic rates [24]. On the other hand, in Silene latifolia where males produce abundancy of small flowers, genetic correlations between floral display and leaf gas exchanges, especially expressed in higher stomatal conductance (g_s) and leaf transpiration (E) in males than in females, leading to a higher cost of reproduction for males [25]. In some dioic species and environments, biomass allocation to reproduction is three-times higher in female than in male plants [22], while in other species, as in Fraxinus mandshurica, no biomass difference between the genders is observed during early ontogeny [26].”

 

“Increasing net photosynthetic rate (A_net) in dioecious Populus spp during the early summer is related to an increase in the chlorophyll pool during leaf development [38]. However, differences between genders in terms of photosynthetic gas exchanges are not always observed [38].”

 

Missing citations for some sentences or definitions need to be checked by the authors.

 

Authors: Thank you. References were added to some phrases where they were missing.

 

In the methods section, the authors should provide detailed descriptions of the experimental area. Suggestion: the author may consider making a flowchart that clearly shows the research step.

 

Authors: One flowchart was added to the Supplementary Material, as Figure S1, showing the research steps.

 

Figure S1. Flowchart showing measurements of light (photosynthetically active photon flux density, PPFD) and A_net/PPFD curves for further modeling the instantaneous leaf gas exchanges along the diurnal period in monoculture (MO) and agroforestry (AFS).

 

The article provides a solid overview of the results with an analysis of the dataset.

 

Authors: Thank you.

 

Is it necessary to explain or map the LAI to understand the changes? This may relate to the scalability across different climatic conditions and geographic regions. Also, how to validate the resulting dataset in terms of the specific parameters derived from the climate measurement.

 

Authors: We are not sure about the point raised by the reviewer. Here, LAI is a proxy of plant size and canopy when measurements were taken. In fact, LAI is determined by the genetic of populations/clones and environmental conditions. We added some words about this (lines 421-423):

 

“Such difference was less pronounced herein (Figure 2), which is likely due to the methods and equipment used for evaluating plants, to the genetic background of populations/clones and environmental conditions.”

 

 

As in other modeling studies, we tried to represent a given process based on assumptions and then reveal plant responses to the surrounding environment.  Regarding the data validation, this is a topic for further research as there is no similar study done with this species under field conditions.

 

Finally, the discussion section should be strengthened by addressing the implications of these findings for existing literature.

 

Authors: Various papers and their contributions were added to discussion, as suggested by you and the reviewers, #1 (red),  #2 (blue), and #3 (green).

 

 

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

I share my consent as 'Accept'

Comments on the Quality of English Language

OKAY

Author Response

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 quality of English does not limit my understanding of the research.
(x) The English could be improved to more clearly express the research.

 

	Yes	Can be improved	Must be improved	Not applicable
Does the introduction provide sufficient background and include all relevant references?	(x)	( )	( )	( )
Is the research design appropriate?	( )	(x)	( )	( )
Are the methods adequately described?	(x)	( )	( )	( )
Are the results clearly presented?	(x)	( )	( )	( )
Are the conclusions supported by the results?	(x)	( )	( )	( )

Comments and Suggestions for Authors

I share my consent as 'Accept'

Authors – Thank you very much

Comments on the Quality of English Language: OKAY

Authors – Thank you very much

Author Response File: Author Response.docx

Reviewer 4 Report

Comments and Suggestions for Authors

The revised manuscript is now clearer and more detailed in several aspects.

Author Response

Open Review

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

Quality of English Language

(x) The quality of English does not limit my understanding of the research.
( ) The English could be improved to more clearly express the research.

 

	Yes	Can be improved			Must be improved		Not applicable
Does the introduction provide sufficient background and include all relevant references?	(x)		( )	( )		( )
Is the research design appropriate?	(x)		( )	( )		( )
Are the methods adequately described?	( )		(x)	( )		( )
Are the results clearly presented?	(x)		( )	( )		( )
Are the conclusions supported by the results?	(x)		( )	( )		( )

Comments and Suggestions for Authors

The revised manuscript is now clearer and more detailed in several aspects.

Authors – Thank you very much

Author Response File: Author Response.docx