The Alien Invader, Rhus typhina L., Outperforms Its Native Competitor in the Scenario of Nitrogen Deposition and Arbuscular Mycorrhizal Fungi (AMF) Inoculation
Round 1
Reviewer 1 Report
There are numerous articles referring to the invasive capacity of Rhus typhina L. and its competition with native species. The originality of this article lies in the incorporation of arbuscular mycorrhizal fungi and the addition of N in different amounts.
The introduction is written in an orderly manner and is based on appropriate bibliographical background.
The materials and methods present several methodological problems. The initial paragraph (lines 97 to 106) does not correspond to the methodology section, it could be incorporated into the introduction. Regarding the preparation of the material, the substrate used for sowing the seeds in the germination trays is not explained, nor are the environmental conditions in which the seedlings grow during those days.
After the transplant, the environmental conditions of the greenhouse were not explained either, which is what really matters.
The treatment of AMF inoculation just says (1) no AMF are inoculated and (2) AMF are inoculated. It is necessary to clarify what inoculum was used, whether it is a commercial inoculum or it was obtained from a natural environment, how it was obtained, what species of fungi were inoculated, how was the inoculation technique, with what amount of inoculum it was made. Too much information is missing regarding inoculation with AMF, which constitutes a fundamental part of the work.
The treatment of nitrogen deposition should also be clarified how it was carried out, at what time and with what product, it is also not understood why they use these units of measurement g N m-2·yr-1
When determining the AMF colonization, the immersion time in KOH must be precise, it is not the same if the roots are 20 minutes or 60 minutes in the treatment. Root staining is usually done with Tripan Blue because it is easier to visualize fungal structures, it is not used here. When quantifying AMF colonization, it is important to differentiate the different structures of the fungus (hyphae, arbuscules, vesicles), in this case it is not explained which structures of the fungus were counted.
Mycorrhizal colonization, as described in the methodology, was calculated as a %, so in the graph, in the results section, it should be expressed as a %. Nor is it indicated whether or not the addition of N modified mycorrhizal colonization, this is an important issue and has been discussed in numerous papers.
The results, in general, are written in an orderly and understandable manner, but the conclusions obtained lose credibility due to the number of methodological errors that are observed.
The bibliography should be reviewed
Author Response
There are numerous articles referring to the invasive capacity of Rhus typhina L. and its competition with native species. The originality of this article lies in the incorporation of arbuscular mycorrhizal fungi and the addition of N in different amounts.
The introduction is written in an orderly manner and is based on appropriate bibliographical background.
- The materials and methods present several methodological problems. The initial paragraph (lines 97 to 106) does not correspond to the methodology section, it could be incorporated into the introduction. Regarding the preparation of the material, the substrate used for sowing the seeds in the germination trays is not explained, nor are the environmental conditions in which the seedlings grow during those days.
Response：Thanks very much. We agree with you and have adjusted and modified it. The modification trace is shown on Line 127, 136-138, 145-147, respectively.
- After the transplant, the environmental conditions of the greenhouse were not explained either, which is what really matters.
Response：Thanks very much. Yes, we agree with you, and have added the temperature and humidity conditions in the artificial climate chamber and greenhouse. The modification trace is shown in line 145-147.
- The treatment of AMF inoculation just says (1) no AMF are inoculated and (2) AMF are inoculated. It is necessary to clarify what inoculum was used, whether it is a commercial inoculum or it was obtained from a natural environment, how it was obtained, what species of fungi were inoculated, how was the inoculation technique, with what amount of inoculum it was made. Too much information is missing regarding inoculation with AMF, which constitutes a fundamental part of the work.
Response：Thanks very much. We have added this information. The modification trace is shown on Line 139-142, 154-161.
- The treatment of nitrogen deposition should also be clarified how it was carried out, at what time and with what product, it is also not understood why they use these units of measurement g N m-2·yr-1
Response：Thanks very much. The nitrogen was added once every Monday at 5 PM, and 100 mL nitrogen solution was added in each pot each time. The nitrogen was added once a week and the prescription and concentrations of added solutions followed in Table 1 in the revised manuscript.
About the units of measurement g N m-2·yr-1, we refer to the literature of He, L., which uses the unit of measurement g N m-2·yr-1.
- When determining the AMF colonization, the immersion time in KOH must be precise, it is not the same if the roots are 20 minutes or 60 minutes in the treatment. Root staining is usually done with Tripan Blue because it is easier to visualize fungal structures, it is not used here. When quantifying AMF colonization, it is important to differentiate the different structures of the fungus (hyphae, arbuscules, vesicles), in this case it is not explained which structures of the fungus were counted.
Response：Thanks very much. We agree with you, but the time depends on the condition of the root. It takes longer for the root to be strong or hard, and less time for the root to be younger. Different samples take different amount of time. And we used the method of Liu and Chen, 2007, staining with acid fuchsin lactate glycerin staining solution, which could observe and distinguish different structures of fungus.
- Mycorrhizal colonization, as described in the methodology, was calculated as a %, so in the graph, in the results section, it should be expressed as a %.
Response：Thanks very much. Ok, we changed the representation of the result and replaced the figure. The modification trace is shown in figure 1.
- Nor is it indicated whether or not the addition of N modified mycorrhizal colonization, this is an important issue and has been discussed in numerous papers.
Response：Thanks very much. Sorry, I didn't consider this problem in my experiment, but I will consider it more in future experiments.
- The results, in general, are written in an orderly and understandable manner, but the conclusions obtained lose credibility due to the number of methodological errors that are observed.
Response：Thanks very much. We are sorry for that, but we used the method of Liu and Chen , 2007, which could observe and distinguish different structures of fungi.
- The bibliography should be reviewed
Response：Thanks very much. We reviewed the bibliography, and have added the doi of references. The modification trace is shown in references.
Reviewer 2 Report
Comments for author File: 
Comments.pdf
Author Response
Response to the Reviewer 2:
- Your research results are presented in a good way. Before publishing the paper, the authors may consider describing which species (one) or combination of several species of AMF they used to inoculate the plants. In my opinion, the manuscript is suitable (after minor corrections) for publication in journal Sustainability
Response：Thanks very much for your praise. And we have modified and added. We used graphs and tables and thus made their methods, experimental conditions, and results more understandable for the reader. The modification trace is shown on Line 140-143.
Reviewer 3 Report
This manuscript present interesting results related to Morphological traits determine the invasion success of Rhus typhina L.: However, it needs improvements before acceptance for publication (see comments below)
Title
The title is quite long and detailed, the title mentions several different components of your study, including the species of interest (Rhus typhina L.), the impact of nitrogen deposition, and the use of arbuscular mycorrhizal fungi. While these are all important factors, it may be helpful to highlight the most significant or novel aspect of your study in the title to draw readers' attention.
The term "invasion success" in the title may be unclear to readers who are not familiar with the concept. Consider using a more common term, such as "spread" or "establishment."
Abstract
The abstract could benefit from a clearer statement of the research question or hypothesis, as well as a more explicit statement of the main conclusions of the study.
Introduction
The introduction provides some information on the effects of nitrogen deposition and arbuscular mycorrhizal fungi (AMF) on plant competition, but it does not provide a comprehensive overview of the current state of knowledge on the topic. Providing more context on the existing research can help the reader better understand the significance of the study.
Material and Methods
No. of plant per pot?
Give more detail about the growth conditions such as light intensity, day/night temp, day length etc.
Discussion
Citing prior studies without integrating them effectively: While the author cites prior studies, they fail to integrate the findings from these studies effectively into their own discussion. Instead, they simply state the findings of these studies without explaining how they relate to their own results. The discussion lacks critical analysis of the results. The author simply describes the results without providing a deeper analysis of what they mean or their significance.
The author uses inconsistent language throughout the discussion, which makes it difficult to understand the points being made. For example, the author uses terms such as "competitive capacity" and "competitiveness for space and resource" interchangeably, without defining them or explaining their relationship.
Author Response
Response to the Reviewer 3:
This manuscript present interesting results related to Morphological traits determine the invasion success of Rhus typhina L.: However, it needs improvements before acceptance for publication (see comments below)
- Title
The title is quite long and detailed, the title mentions several different components of your study, including the species of interest (Rhus typhina L.), the impact of nitrogen deposition, and the use of arbuscular mycorrhizal fungi. While these are all important factors, it may be helpful to highlight the most significant or novel aspect of your study in the title to draw readers' attention.
Response: Thanks very much. We have accepted your suggestion and made modifications. We replaced the former title with “The alien invader, Rhus typhina L. outperforms its native competitor in the scenario of nitrogen deposition and arbuscular mycorrhizal fungi (AMF) inoculation”. And the new title more directly expresses the main finding of our study. The modification trace is shown on Line 1-4.
- The term "invasion success" in the title may be unclear to readers who are not familiar with the concept. Consider using a more common term, such as "spread" or "establishment."
Response: Thanks very much. We accept your comments and have revised the term.
- Abstract
The abstract could benefit from a clearer statement of the research question or hypothesis, as well as a more explicit statement of the main conclusions of the study.
Response: Thanks very much. We have improved the description of the research question, and gave a more explicit statement of the main conclusions of the study. The modification trace is shown on Line 19-22, 34-35.
- Introduction
The introduction provides some information on the effects of nitrogen deposition and arbuscular mycorrhizal fungi (AMF) on plant competition, but it does not provide a comprehensive overview of the current state of knowledge on the topic. Providing more context on the existing research can help the reader better understand the significance of the study.
Response: Thanks very much for this valuable comment. We have provided more context on the existing research in the introduction. The modification trace is shown on Line 40-55.
- Material and Methods
No. of plant per pot?
Response: Thanks very much. We added this information to the part of 2. Materials and methods. The modification trace is shown on Line 164-166.
- Give more detail about the growth conditions such as light intensity, day/night temp, day length etc.
Response: Thanks very much. We added the temperature and humidity conditions in the revised manuscript. The modification trace is shown on Line 146-148.
- Discussion
Citing prior studies without integrating them effectively: While the author cites prior studies, they fail to integrate the findings from these studies effectively into their own discussion. Instead, they simply state the findings of these studies without explaining how they relate to their own results. The discussion lacks critical analysis of the results. The author simply describes the results without providing a deeper analysis of what they mean or their significance.
The author uses inconsistent language throughout the discussion, which makes it difficult to understand the points being made. For example, the author uses terms such as "competitive capacity" and "competitiveness for space and resource" interchangeably, without defining them or explaining their relationship.
Response: Thanks very much for this valuable comment. We replaced “competitive capacity” with “competitiveness for space and resource” throughout the Discussion and made the term use more consistent. And we have revised and improved the discussion, and provided a deeper analysis. The modification trace is shown on Line 334-335, 346-349, 360-365, 377-382.
Round 2
Reviewer 1 Report
With the modifications made, the manuscript is ready to be published.
