Human-Mediated Dispersal of Plantago asiatica Mucilaginous Seeds in Urban Environments

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript (ID: seeds-3580874) addresses the dispersion of Plantago asiatica seeds by human walking because the seeds attach to the shoes by mucilage production. The article demonstrates that this type of dispersion occurs up to 1000 m on asphalt or grass paths and with different footwear types. The manuscript is well-structured, the experimental design is well-described, and the English writing is satisfactory. However, the authors must address and revise specific issues to enhance the manuscript. Moreover, the Results and Discussion sections require further revision and improvement.

1. a) The Introduction section seems complete about seed dispersion mechanisms, dispersion by humans and animals and the role of mucilage in dispersion. However, more context is needed about the plant species, their weedy behavior in urban areas, and the issues related to potential or factual invasions of this species. A brief description of the plant species is found in lines 66 and 67 of the Materials and Methods section. This description should be moved to the Introduction section and expanded. Furthermore, I would like to read the descriptions of the plant stands chosen to be stomped in the “Study Species” subsection within the Materials and Methods section.
2. b) The results obtained warrant further description in the Results section. The authors concentrated on the fraction of seeds attached to shoes after 1000 m in both experiments. However, there are interesting results that are not commented on. The dynamics of seed loosening during walking differed significantly between the grass and asphalt paths, and this difference is more evident than what was recorded at 1000 m. Half of the seeds (50%) or more (I do not have the raw data) detached in the grassy path within only 10 m, while nearly 50 m is required to reach the same fraction in the asphalt road (Figure 4). This difference has implications for the seed dispersion of this species, depending on the nature of the path, and deserves to be highlighted. A similar observation applies to the results of the shoe-comparison experiment. There is no description of the differences in seed-loosening dynamics. The differences found at 1000 m among shoe types show changes throughout the walking experiment (Figure 6). These results also merit further commentary and discussion.
3. c) The Discussion and Conclusions section is unsatisfactory. First, fragments from the Materials and Methods and Results sections would be more appropriate in the Discussion section (lines 93-97, 137-140 and 155-156). Second, only the first paragraph (lines 162-166) discusses the obtained results, where the emphasis is only on the 1 km dispersion potential of the P. asiatica seeds in various road and shoe types. I would like to read comparisons among the obtained results and some of the cited references that address the seed dispersion caused by humans (approximately 19 references), the distances recorded, and the efficiency of mucilage as a dispersion mechanism compared to other mechanisms or species with mucilage (which has about 30 references on this last topic). By doing this, you will strengthen your results and the entire work. Third, the second paragraph of this section (lines 167-183) serves as a disclaimer, but it can not be the primary focus of this section. Additionally, a text fragment between lines 167 and 174 resembles a passage from Abe et al. 2022 (reference number 15). I recommend significantly reducing this paragraph and concentrating on discussing your results. Lastly, I do not find a clear conclusion in this section.

There are a few minor observations:

Figures 1, 2 and 3 are illustrative, but the reference letters (a, b, c) are below the photos. In some cases, such as Figure 3, I found some difficulty in separating a) from b). I suggest putting the reference letters on top and left of each photo or group of photos, as usual in scientific publications and distinguishing the group of photos more clearly. Moreover, Figures 4 and 6 display the averaged results for each treatment; however, no error bars or references are provided, although 20 repetitions for each treatment have been done. Could you include error bars in the results? This addition would help the reader better appreciate the reliability of your work.

There are fragments of the text which are sustained by a high number of references. This citation method may be suitable in some ways because the reader has multiple options to continue their bibliographic search. However, the drawback of doing this is the lack of hierarchization of the references. The readers do not know which of the tens of references is the most appropriate for a topic, and if they are interested, they would spend so much time searching, which would make the experience inconvenient. I suggest selecting the most appropriate references for each sentence or text fragment and drastically reducing the number of references.

Author Response

Please find our responses in the attached file.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Comments for Author

The study seems “OK” for seed ecologists after major revision. I wish to see that all my comments are correctly answered. In my eyes, it needs major revision before publication.

1. The title should indicate not only “seeds”, but “mucilaginous seeds” and indicate the season, “simulated rain”

Please rephrase the title and add the above terminologies to improve your title.

2. Reduce the number of keywords. line 21 -22, remove human-vectored dispersal, anthropochory; animal dispersal;  plant-animal interactions; However, you need to add the location (the study area) and the species name under the keyword section.
3. Replace “human-vectored dispersal” with “human-mediated dispersal” throughout the manuscript.
4. Plant-animal interaction is a very broad story; therefore, you need to replace it with other terminology that directly fits your study topic.
5. Please note that the following terminologies throughout your manuscript: seeds mucilage = myxospermy, and fruit or pericarp( mucilage) is myxocarpy. This must be differentiated.
6. The introduction section should tell us some of the polysaccharides i.e., cellulose, hemi-cellulose, and pectin mucilage. Note that some mucilage types are important to anchor to the ground, others to adhere to animals… others to retain much water. Therefore, this should be introduced in a few sentences. Kindly see Mohammed and Mummenhoff (2024). Functional divergence exists in mucilage-mediated seed dispersal, but not in germination of myxospermic Lepidium campestre and Lepidium draba (Brassicaceae)
7. I suggest that you introduce short and long-distance dispersal. How is long-distance dispersal defined in the context of seed ecology? Just a sentence or two.
8. I am wondering if possible to include the walking speed for each interval and to the final destination (1000 m)??? Also, the age or the weight of the people who walk through it???
9. Most of the experiments were carried out on the asphalt road. Do you think that it is possible to compare the asphalt road and grass road with different sample sizes? I think this study did not use the sample size for the two road experiments.
10. What is the practical implication of using different sole shoes? This should be mentioned. What does it tell us?
11. Figure 2a and b should be the same-sized photos. Also, Figure 3 (the shoe photos) should have the same size.
12. Line 138-139 remove “From these results, we concluded that seeds attached to the shoes were dispersed after walking on different types of roads in realistic urban environments.” This is a conclusion, not a result.
13. Line 155-156, remove “From these results, we concluded that all types of shoes investigated here were effective vectors of asiatica seeds.” This is a conclusion, not a result.
14. Assume that your readers do not know your study species (Plantago asiatica,). Therefore, it must be introduced in the introduction section or create a topic in the method section “Description of the study species,” just a few sentences about the species. If it has some economic or medicinal importance, mention this. Then your discussion and conclusion sections should be written in the context of this.
15. Remove “we” and “our” throughout the manuscript instead say “the current study” or “the present study”….
16. Is the study species is indehiscent or dehiscent fruit-producing species? If the species produces dehiscence fruit, this makes sense because the fruit will fall to the ground upon maturity, and human-mediated dispersal could be possible upon wetting (rainy season). If the study species produces indehiscent fruit, this does not make sense because the whole fruit is a dispersal unit in this context. If the former is true, then indicate that the seeds are true dispersal units of the study species (probably when you describe the species)
17. The discussion and conclusion section mainly talks about the limitations of the study; however, it is important to tell the readers about the practical implications of your study. This is important for us. Discuss your findings in the context of what tells your result. Discuss your result instead of discussing the limitation. What is the take-home message from the study? This should be discussed and concluded in a way that the readers learn from the study. For example, if the species is a medicinal plant, consider this when you discuss and conclude. This means the discussion and conclusion must be written in the context of the species. This section needs major revision.

Author Response

Please find our responses in the attached file.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

This is an interesting work concerning epizoochorous dispersal of mucilaginous diaspores. 

I have only a few comments/questions to this manuscript.

 - In the introduction are missing some ecological information concerning the studied species e.g. habitat, life form, mode of dispersal (endozoochory?), invasiveness – if any. In the abstract the authors are speaking about the vegetation in urban and protected areas. Is this species in Japan an invasive plant?

- The conditions of this two types of roads (grass, asphalt) were always comparable? It means – the experiments were performed always on dry roads or/and wet  (e.g. after the rain)? 

Comments on the Quality of English Language

English is not my mother language so I do not feel competent to evaluate it. 

Author Response

Please find our responses in the attached file.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

The study addresses an ecologically relevant topic by investigating human-mediated seed dispersal of Plantago asiatica via shoe attachment under urban conditions. The experiments are well-structured, and the findings contribute to understanding anthropogenic impacts on plant dispersal. However, the manuscript requires revisions to improve clarity, methodological transparency, and contextualization of results.

Specific Comments

1. Title and Abstract

The phrase "effectively dispersed by humans in diverse urban environments" is vague. Specify the quantitative results (e.g., retention rates, distances).

2. Introduction

The rationale for focusing solely on Plantago asiatica is unclear. Justify why this species was chosen over others with similar mucilage traits. Hypotheses are implicit but not explicitly stated. Clearly outline hypotheses for both experiments.

 

3 Methods

Sample Size: Justify why 20 trials per condition were chosen. A power analysis would strengthen this.

Shoe Types: While all shoes had synthetic rubber soles, sole patterns (e.g., tread depth) likely influence seed retention. This variable is not quantified or controlled.

Hydration Method: Artificial hydration (tap water) may not mimic natural rainfall. Discuss potential biases.

4. Statistical Analysis:

Clarify why a Poisson GLMM was used for initial seed counts (count data) but a binomial GLMM for retention fractions. Ensure model assumptions (e.g., overdispersion) are validated. Report effect sizes (e.g., odds ratios) alongside p-values for meaningful interpretation.

5. Results

Figure 4: The y-axis label ("Fraction of seeds remained") is grammatically incorrect. Revise to "Fraction of seeds remaining."

Figure 5: Median values are mentioned in the text but not visualized in the boxplot. Add medians or convert to violin/beeswarm plots.

Statistical Reporting: For non-significant comparisons (e.g., leather vs. kakkusu shoes: p = 0.07), discuss potential biological relevance despite statistical thresholds.

6 Discussion

Overgeneralization: The conclusion that "all types of shoes... were effective vectors" conflicts with the significantly lower retention in leather/rubber boots (3–4% vs. 12–15%). Emphasize variability among shoe types.

Limitations:

The study ignores seed attachment to socks/trousers, which may enhance dispersal in natural settings (as noted by Mount & Pickering, 2009). Discuss how this affects extrapolation.

The focus on a single species limits generalizability. Acknowledge this and suggest future multi-species comparisons.

Contextualization: Compare retention rates with prior studies (e.g., Abe et al., 2022) more critically. Why did asphalt/grass lanes show similar retention despite differing textures?

Author Response

Please find our responses in the attached file.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors did a very good job for overcome the observations. I am satisfied with the new version of the manuscript.

Reviewer 4 Report

Comments and Suggestions for Authors

I am glad to review the revised manuscript again. I found that the authors have made a substantial effort to address all my previous comments, and I have no further comments at this stage. Light is green from me.