Morpho-Physiological and Biochemical Responses in Prosopis laevigata Seedlings to Varied Nitrogen Sources

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The article “Morpho-physiological and biochemical responses in mesquite (Prosopis laevigata) seedlings to varied nitrogen supply” presents valuable research with significant implications for reforestation practices.

In my opinion, by addressing the areas mentioned below, the authors can improve the clarity, depth, and impact of their work, making it more compelling and useful for the scientific community and practitioners in the field.

In the title, please consider this suggestion that can help to better reflect the study's focus and findings: “Impact of different nitrogen sources on biomass allocation and nutrient use efficiency in Prosopis laevigata seedlings.”

In the Introduction, please expand the section related to information about Prosopis laevigata, as it is not a well-known species to the broader scientific audience. Therefore, I recommend dedicating more space to it for this reason.

Materials and Methods are ok in its current form.

In the Discussion section, please enhance the concept of the potential ecological and practical implications of reducing N use efficiency with high nitrogen inputs, especially in the context of reforestation in nutrient-poor soils.

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

I have  reviewed the manuscript entitled "Morpho-physiological and biochemical responses in mesquite (Prosopis laevigata) seedlings to varied nitrogen supply" and found the research to be of considerable interest. However, while the study contributes to the essential field of science, it lacks the degree of novelty that one might expect for publication in a high-impact journal. Despite these limitations, the research is sufficiently rigorous and well-articulated to recommend its publication, provided that the authors address certain issues during the revision process.

The study demonstrates satisfactory levels of conceptualization and presentation, particularly in its background and the methodological approaches employed. The authors have successfully presented a well-written introduction that effectively frames the current challenges related to nitrogen uptake, supplementation, and bioavailability, particularly within the context of forestry. However, there are several aspects of the manuscript that require improvement to enhance clarity and strengthen the overall narrative.

Please see below for details:

 

Introduction:

The authors have provided a comprehensive introduction that aptly contextualizes the study within the broader discourse on nitrogen dynamics, with specific relevance to forestry. The background information is articulated in a clear and concise manner, effectively setting the stage for the research. Nevertheless, the manuscript would benefit from a more explicit and precise delineation of the study’s aims, objectives, and research questions. The current presentation lacks a clear articulation of the specific research questions and hypotheses that the study seeks to address. This omission undermines the clarity of the study’s purpose and the framework within which the research was conducted. I strongly recommend that the authors revise this section to state the research questions and hypotheses tested, thereby providing a more robust foundation for the study.

 

Materials and Methods:

The Materials and Methods section is generally well-constructed, with the description of greenhouse conditions and plant harvesting protocols meeting high standards of scientific rigor. The meticulous attention to detail is evident, and the authors have clearly invested considerable effort in ensuring that the study is replicable. This is a commendable aspect of the manuscript, as it enhances the reliability and validity of the research findings.

However, there are a few areas that require clarification and revision. Specifically, in the subsection on statistical analysis [234-255], the authors mention the use of R software for data computation, yet only a limited number of R packages are referenced [248, 252, 255]. Given the complexity of the analyses conducted, it is evident that additional R packages were utilized. It is essential that all packages used in the analysis, including those for Kruskal-Wallis, Levene's, and Shapiro-Wilk tests, ANOVA, and any others, be properly cited. This not only ensures ethical citation practices but also allows reviewers to accurately assess the appropriateness of the methods employed. I strongly advise the authors to revise this section to include citations for all R packages used in their analyses.

 

Results:

The Results section is well-organized and adheres to the appropriate scientific conventions, ensuring that the findings are presented in a clear and coherent manner. The descriptive narrative effectively complements the data, making the results both engaging and easy to comprehend. No further revisions appear necessary in this section.

 

Discussion:

The Discussion section is cogently written, with a logical flow that effectively synthesizes the results with the study’s objectives and the broader literature. The authors have successfully linked their findings to existing research, thereby providing a meaningful contribution to the field. The section demonstrates a commendable level of scholarly rigor, and I do not find any significant issues that warrant correction.

 

Conclusions:

The conclusions drawn from the study are well-grounded in the data presented and reflect the implications of the findings. However, the section would benefit from a discussion of potential future research directions. Including suggestions for further studies would enhance the manuscript by providing a pathway for continued investigation in this area, thus offering valuable insights for future research endeavors.

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

Review:

Morpho-physiological and biochemical responses in mesquite (Prosopis laevigata) seedlings to varied nitrogen supply.

In the present manuscript the Authors assessed several morpho-physiological and biochemical responses of Prosopis laevigata seedlings to a high supply of N, provided as either inorganic (NH4NO3) or organic (amino acids).

 

General comment

The subject of the paper fits the aims of the Journal and results could be of interest for the scientific community. However, in my opinion the present manuscript does not meet the scientific standard of the Journal, for a number of main concerns.

 

For optimal plant growth, it is most important to maintain a balance between the content of nitrates and ammonium ions in the substrate. Moreover, according to Liebig's law, both too low and too high concentrations of any of these ions will have a negative effect on plants, due to insufficient nitrogen in the substrate or toxic effects of too high NO₃^- and NH₄⁺ concentrations. Unfortunately, the Authors in the manuscript omitted the content of macronucleus in the substrate, and as a consequence the results of the experiment cannot be correctly interpreted.

All plants studied so far, including cultivated species, have demonstrated the ability to take up free amino acids from the soil solution (Miller and Cramer, 2004). It is assumed that organic nitrogen compounds can be taken up and used by plants, but they do not constitute their primary source of this element (Näsholm et al., 2009). The Authors, however, assumed that the use of amino acids is a form of nitrogen fertilization. If so, please prove it. The Authors did not present what the research hypothesis exist. Therefore, it is difficult to unequivocally assess the value of the presented research results. For example optimal tomato growth, the ratio of nitrates to ammonium cations in the soil should be 3:1, and only at higher concentrations of NH4+ relative to NO3- do plants exhibit weakened growth (Crawford and Glass, 1998).

Detailed comment:

Lack of research methodology, lack of description.

Line 102: It is not clear whether the plants are grown from seed (up to 25 cells) or from seedlings.

Line 103: I understand that there were 25 plants in one tray and one tray (25 plants) was one repetition. So why was it calculated as 20 plants times three repetitions times three combinations for a total of 180 plants, shouldn't there be 225 plants?

Line 108: It is necessary to determine the content of general forms of macronutrients in the substrate and the C:N ratio, exchange sorption of the substrate.

Line 115: How the authors converted the additional dose of 90 mg nitrogen (in the form of NH4NH3) into nitrogen contained in amino acids (Naturamin®-118 WSP, DAYMSA, Spain)

Line 118: The amino acid composition (Naturamin®-118 WSP, DAYMSA, Spain) must be precisely specified.

Line 163: Please clarify what is meant by the term whole plant (leaves, stem and root)?

 

Results

Section 3.1. Since abbreviations are used in the table, abbreviations should also be used in the text, e.g. LMF, etc.

Line 266: Please explain what does rapid growth mean.

Line: 267: The explanations below the table do not compatible the data in the table 1

Line 274: Symbols p < 0.05, ** p < 0.01, ***  p < 0.001). are not consistent with the figure 1 and figure 2, likewise.

Figure 2. is unclear:

1. What is the difference between 2A and 2B, it should be clear from the drawing, it should be explained in the description of the axis.

2. It would be much easier to indicate the statistical difference using letter designations instead off square brackets.

Section 3.4: description in section 3.4 does not correspond to the data in Figure 4 and 5 abcd

 

Discussion

Line 337-339: This sentence should be for the introduction section

Line 341-345: I don't understand, the sentences are too long, ungrammatically formulated.

Line 354: The authors should decide whether root growths were present or not.

Line 401 -403: It is not clear.

Line 406: What factors do the Authors write about?

Line 401-412: Please explain: what organic forms of N are the Authors writing about? This is an overinterpretation if the authors do not take into account the C:N ratio in the substrate. It is necessary to present the content of available N before and after fertilization.

  A major overhaul is needed all the text.

Comments on the Quality of English Language

For example: Line 341-345: I don't understand, the sentences are too long, ungrammatically formulated.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

I have no substantive objections to the results of the experiment. The Authors responded and took into account all my suggestions.   I

 

Author Response

Thank you once again for your valuable feedback, which significantly enhanced our manuscript.