Review Reports - Vigour Index on Time Basis Calculation on Agastache mexicana Subsp. mexicana Throughout Induced Hydric Stress: SiO2 and Artificial Shade Application Effects

Round 1

Reviewer 1 Report (New Reviewer)

Comments and Suggestions for Authors

Please see the attachment.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

In my opinion the English needs only minor revision. I have indicated necessary correction.

Author Response

Dear reviewer, we appreciate your recommendations for improving our manuscript. You will find the changes made and the responses in this letter in the manuscript in blue.

Comments and Suggestions for Authors

TO THE AUTHORS

The paper describes an experiment examining the survival and overall condition of seedlings of Agastache mexicana subsp. mexicana (Amm) subjected to water deficit at different shade levels and silicon dioxide doses. It seems that the aim of the experiment was to determine the resistance of the seedlings in the face of drought stress at different germination conditions.

However, the Authors express the aim in different ways: (1) in Abstract, “to assess the quality of a seed lot of Amm” (lines 26-27), and (2) in the Introduction, “to calculate an optimal Vigor Index (VI) for Amm seeds under adverse conditions” (lines 87-88).

We did the changes, please see the Abstract and Introduction.

The study is interesting. The experiment was well designed and the material and methods are mostly well described. The results in the manuscript are presented in numerous figures and tables, and analyzed using appropriate statistical methods. However, the mode of description of the research problem is week. The manuscript would be appropriate to be published in 'Stresses' after a major revision.

The main problems are as follows:

The Authors have developed a new Vigor Index (VI) in order to assess the overall condition of seedlings of Amm subjected to water deficit at different shade levels and silicon dioxide doses. If it was the main aim of the study, the Authors should have compared the results of their VI calculation to the results calculated using other Vigor Indices (other mathematical formulas) indicated in the references shown in line The alternative way is tho remove the lines 80-86 from Introduction chapter and move them to the Discussion chapter. Then, in the Discussion chapter the Authors can clearly explain why they have developed a new calculation of VI instead of using the previously used calculations. This novelty should be also emphasized in the Conclusions.
We did the changes. Please, see the new discussion.
The Introduction should be rewritten. Here the influence of drought, shading and SiO2 addition on seedlings growth and survival should be consistently The aim of the study should be clearly introduced in the last paragraph.
In the new introduction we talk about these issues.
The Discussion chapter needs to be Please comment on (1) ASL effect, then (2) SDL effect based on Tables 1-3 and then (3) discussing the irrigation effect would be appropriate. Here the discussion on different VI calculations should be included.
Done, we write base don Table 1-3 and Figures

Minor remarks are as follows:

Abstract: please indicate that a new calculation of Vigor Index was introduces in this
Done. Please, see the new redaction
Line 86, Introduction: unnecessary
Thank you, done
Lines 87, 296, 508, ...: please be consistent and used italics for “Amm”.
Done
Lines 178-193, Results: the description of Figures should be located in Figure Please comment here on the results shown in the Figures.
Done, please see the new redaction
Lines 209-210, Results: please move this comment to the
Done, please see the new discussion
Lines 243-244. This phrase is not
Thank you Deleted
Lines 267-270: it seems that ASL and SDL are not mentioned
Thank you, done
Lines 290-292: please move this phrase to the Figure 7 R. Done.
Figure 7: the graph in not enough clear, it would be better to differentiate the dash lines and/or points for treatments 1-4, 5-8, and 9-12.
Las líneas dash son distintas para cada grupo de tratamientos
Line 320, Discussion: please correct LDS and MDS according to description in line
Thank you, done.
Lines 347-351: these statements are not related to the Authors' findings, in such a case they should be moved to Introduction.
Moved to introduction, please see the new introduction
Lines 352-353: these statement is not related to the Authors' findings, in such a case it should be moved to Introduction.
Moved to introduction, please see the new introduction
Lines 357-369: this paragraph is not essential for this If the Authors aim to show the background of their research please do it in Introduction.
We have deleted these paragraphs
Lines 380 397, M&M: the number of a Table is not
Thank you, it was changed to Table 4
Line 454: “and” instead of “y” should be
Done, thank you
Line 508, Conclusions: in 38% ASL it was ascertained only to some extent, see significance of differences in Table 1, line 129.
True, thanks, see new conclusions
Lines 523-525: according to Table 2 and 3 there was no effect of SiO2 addition, these findings were not confirmed statistically.
We rewrite the paragraph
Reference No 30: please provide the full information on this
May be, we don´t understand, the reference is complete or may be is another reference
Reference No 31: please add the English

Done

I recommend to enlarge Figures if posible.
We will request it to the Stresses Journal
Supplementary materials:

Section 2: please indicate which levels of artificial shading are shown in Tables 1-12.

Done, please see the new supplementary material

Comments on the Quality of English Language

In my opinion the English needs only minor revision. I have indicated necessary correction.

We have done correction at all manuscript

Author Response File: Author Response.pdf

Reviewer 2 Report (New Reviewer)

Comments and Suggestions for Authors

The manuscript entitled “Vigor Index calculation as a function of the time on Agastache mexicana subsp. mexicana throughout induced hydric stress: SiO2 and Artificial Shade application effects” is well written. However, few major questions need to be addressed before publishing in reputed journal.

The manuscript is well organized; however, it does not fully align with the primary objectives of the journal. The content leans more toward agronomical aspects, with limited emphasis on stress-related parameters and insufficient discussion of stress responses. As a result, the manuscript slightly misses the intended focus of the journal. In my view, it should be reoriented and revised to better address the journal’s objectives.

In line no. 382–392, the authors mentioned, using unsterilized Canadian peat/peat moss for growing plant seeds under greenhouse conditions. Could the authors clarify why the growth medium was not sterilized? If the peat/peat moss was contaminated, it could have influenced plant growth, making it difficult to distinguish whether the observed effects were due to contamination, irrigation, SiO₂, or artificial shade.

In Figure 2, the authors present data on the leaves number in relation to irrigation levels. However, the manuscript does not provide information on the moisture conditions of the Canadian peat/peat moss under control treatment, nor on its key physicochemical parameters, such as organic carbon content. Could the authors clarify whether the organic carbon content of the Canadian peat/peat moss and soil was comparable? Since humic acids in soil contain a high percentage of carbon, which directly influences plant growth, this information is critical for proper interpretation of the results.

Comments for author File: Comments.pdf

Author Response

Dear reviewer, we appreciate your recommendations for improving our manuscript. You will find the changes made and the responses in this letter in the manuscript in blue. In the discussion section, you will find the justifications of some bibliographic sources for the manuscript's findings.

Comments and Suggestions for Authors

We have included general responses to the stresses of waterlogging and drought in the introduction. Measurements of the vigor index and its behavior in the experimental stages constitute the response to stresses, and we include a comparison with VI measurements that do not consider time and other factors. We reorganize the discussion by proposing the deduction of the effect of water stress, the effect of SiO2 and shade on plants under water stress, these effects in red. These effects include leaf number, plant growth, survival, and mortality because of these characteristics are the source of the VI calculation.

The Canadian peat moss data sheet states that it is free of pests, weeds, and diseases. However, the method used to achieve this condition is not specified, which is important, as it would allow the probability of contamination to be estimated.

Although the experiment was conducted in a closed biospace, it had exterior ventilation. Therefore, there is a possibility that insects, parasites, fungi, or bacteria could have entered the environment through outside dust. However, none of these contaminants were detected during the experiment.

Recommended Microbiological Tests

Microbiological tests are suggested both at the beginning and end of the experiment to clarify any questions related to contamination. Recommended tests include:

Fungal and yeast count by plate spread, using potato dextrose or Sabouraud agar.
Aerobic mesophilic count.
Total coliform count, a simple test to perform, similar to the previous ones.

Identification of bacteria using biochemical or PCR testing should only be considered if microbiological contamination is confirmed, as these tests require a considerable investment of time.

The soil is exclusively composed of Peat Moss, whose physicochemical parameters are found in the Table 27 into Supplementary material. We present it also here.

Table 27. Physicochemical parameters of the soil

		Símbolo	Peat moss	Units
1.	Hydrogen Potential	pH	5.5-6.5	Adi.
2.	Electrical conductivity	σ	0.6-0.8	(mΩ⸱cm)^-1 S/m, Ω⁻¹/m
3.	Moisture holding capacity (dry weight)		8x a 10x	%m/m
4.	Initial Moisture content (105°C / 24 h)	H	45 a 50	%m/m
5.	Initial content of organic materia		68 a 82	%m/m
6.	Initial ashes content		18 a 32	%m/m
7.	Initial specifical density	σ	130	kg/m²
8.	Moisture holding capacity on a dry basis		8x a 10x	%m/m
9.	Fiber ratio Mesh +9		19 a 24 21.5	%m/m
10.	Coarse fiber ratio Mesh +20		22 a 39 30.5	%m/m
11.	Medium fiber content +100 mesh		38 a 54 41.5	%m/m
12.	Fine fiber ratio 9+20+100+>100 mesh		100	%m/m

Notes:

It refers to the activity of protomes and not to the concentration
“S” it refers to Siemens, “Ω” electrical resistance

“8x” means 7g of water in a total of 8g and substrate mix, after this amount of moisture the water starts to drain at room temperature
“%m/m” refers to 45g of water per 100g of water plus dry matter

Author Response File: Author Response.pdf

Reviewer 3 Report (New Reviewer)

Comments and Suggestions for Authors

The article “Vigor Index calculation as a function of the time on Agastache mexicana subsp. mexicana throughout induced hydric stress: SiO2 and Artificial Shade application effects provide understanding about the hydric stress on the medicinal plants. However, there are some changes needed to be done:

Major comments:

First the work is worthy, but I did not got the experiment design, such as no clear method for Waterlogging stress.
I think the did something else and wrote something else, I cant understand the writing style for this manuscript, need completely revised by native English speaker. The current version even hard for me to read and get the idea what the author want to convey the messge.
The way the author wrote the results is quite different, such as its strange to start from Figure 1, 2, 3, 4 and shows....the author need to read some high quality article and follow the writing style for results section.
The results evaluation and comparing and justification has not been discussed in discussion section. The discussion section needed to be improve.

Minor comments:

Line 44, Only waterlogging stress is ok.
Line 44, Only drought stress is fine
Please remove ‘in plant’ from key words
Line 122, whats mean by statistical results. Write heading that’s represent the whole significant results for this section.
There are so many rough and unscientific words and sentences which has been used. A serious revision for the whole manuscript is needed.

Comments on the Quality of English Language

Need 100% revision. the current version is not clear at all.

Author Response

Dear reviewer, we appreciate your recommendations for improving our manuscript. You will find the changes made and the responses in this letter in the manuscript in blue.

Comments and Suggestions for Authors

Major comments:

First the work is worthy, but I did not got the experiment design, such as no clear method for Waterlogging stress.

The water regime design is completely random. In the discussion, we deduced the different states of hydric stress, considering the VI and SI values, as well as fluctuations in leaf number and plant height. We modified Table 5 showing the water regimes, although we insist on calling them stages because the application times are also different.

I think the did something else and wrote something else, I cant understand the writing style for this manuscript, need completely revised by native English speaker. The current version even hard for me to read and get the idea what the author want to convey the messge.

The manuscript was completely revised and rewritten. We sent the proofreading copy to the editor. You may request it.

The way the author wrote the results is quite different, such as its strange to start from Figure 1, 2, 3, 4 and shows...the author need to read some high quality article and follow the writing style for results section.

We greatly appreciate this contribution. We have modified the structure and wording, and we have learned a lot about writing better.

The results evaluation and comparing and justification has not been discussed in discussion section. The discussion section needed to be improve.

We've reorganized the wording, included a comparison of VI calculation formulas from previous research, and emphasized the VI measurement over time. In addition, we separated the effects of SiO2 and shade; we hope the new structure will be satisfactory.

Minor comments:

Line 44, Only waterlogging stress is ok. R. Done, please see highlights
Line 44, Only drought stress is fine. R. Done, please see highlights
Please remove ‘in plant’ from key words. R. Done, please see highlights
Line 122, whats mean by statistical results. Write heading that’s represent the whole significant results for this section. R. Done, please see the new heading
There are so many rough and unscientific words and sentences which has been used. A serious revision for the whole manuscript is needed.

R.Thank you for your clarification, which you subsequently sent to the editor. We read and learned from high-level articles to improve the structure of the manuscript.

Comments on the Quality of English Language

Need 100% revision. the current version is not clear at all.

Done, thanks

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report (New Reviewer)

Comments and Suggestions for Authors

Please find my remarks in the attachement.

Comments for author File: Comments.pdf

Author Response

To the Reviewer

Dear reviewer, we deeply appreciate your work. It opened the door for us to publish our manuscript. We also believe it has been an invaluable guide for writing and clarity. We also learned a lot! We don't have the honor of meeting you personally, and although we would like to, we know it's not possible, but we send you our warmest regards. Thank you very much…

Turning to the scientific questions, we've written our responses in blue.

I appreciate the Authors' efforts to improve their manuscript and I have found some valuable amendments. However, in my opinion numerous paragraphs were changed inappropriately. Moreover, Introduction and Discussion are poor and still do not justify properly the experiment and analyses done by the Authors, which I find very valuable. Many paragraphs are not placed properly,

i.e. some part of Discussion should be moved to Results etc. I wish the Authors can publish their results, however, still major revision is necessary.

The first problem is Introduction: it became completely messy.

The response to SiO2 is placed just after the description of the species although SiO2 addition was not the main problem of the paper (it a growth of seedlings under waterlogging and then drought),
We have included bibliographical to explain our research findings. Please read lines 68-92 in introduction, the new results and discussion with seedlings under waterlogging and drought stresses.
The description of response to SiO2 is doubled (lines 54-61 and 65-74) while the summarizing phrase is inserted just between, it makes no sense,
We corrected the duplication and done a new paragraph shorter. Please read lines 106-115.
Other issues showing the context of this research are mixed and introduced in an unclear manner,
We corrected with your recommendations, please read.
Lines 88-92 should be moved to M&M.
We have moved it to the lines 629-631

I suggest rewrite again the Introduction in this order:

I recommend to save the first part of the Introduction as it was done in the FIRST VERSION of the manuscript (which I have reviewed on 8 September), i.e. I recommend to leave lines 48-79 or shorten if recommended by another Reviewer.
We leave the shorten lines, please read the new introduction
Then write “connecting” phrases in a new paragraph like these: “Several environmental factors affect initial growth of seedlings and may evoke a lethal These are drought, waterlogging, insufficient or excess light.”
Done, please read lines 68-69
Then explain the effects of drought on juvenile plants, next describe effects of waterlogging, next explain the role of light but also indicate that some species need shading when at germination/juvenile growth stage (here the lines 83-87 from the SECONDARY VERSION of the manuscript are appropriate).
Done, please read lines 70-92
Next, introduce the reason why silicon was added, it may sound like this: “To assess properly the optimum conditions for seedling growth several stress factors at different levels should be applied. Moreover, then producing healthy seedlings that can be successfully transplanted from seedbeds to open fields some additional procedures may be applied like

silicon fertilization.” and here the lines 54-61 and 65-74 from the SECOND VERSION (combined paragraphs describing silicon actions in plants) should be placed.

Done, please read lines 99-116, besides, we include a new paragraph to highlight the joint effect of shade and water stress, see lines 117-122.
Next, explain that “To summarize survival and growth parameters different Vigor Indices were developed [Ref 11-16 from the FIRST VERSION]. Next, continue lines 107-109 from the SECOND Next, insert a phrase like this: “Likewise, mathematical modeling can help interpret data.” and then continue with lines 110-116 from the SECOND VERSION.
Done, please see lines 123-135
Finally, indicate the aims of the study listed as follows: (1) to track the Amm seedlings growth under changing waterlogging/drought conditions, (2) to evaluate the effect of different levels of artificial shading and SiO2 fertilization on enhancing healthy seedlings production, (3) to evaluate Vigor Index on time basis, VI (or maybe VIT) developed by the Authors, which couples together survival rate, basic growth parameters and time
Done, see lines 136-141

Second, I enhance the Authors to name their vigor index as “vigor index on time basis”, VIT. My suggestion is to help readers and followers in the future to distinguish this VIT from other VIs developed by other researchers.

Thank you very much, we already include it

Other main problems are as follows:

Line 175: please insert again the commentary to the Figure 1, i.e. lines 166-168 from the FIRST In my opinion, phrases like “Figure 1 shows” make no sense because any Figure caption should explain what is shown in the Figure. (However, do not remove them from this manuscript). The description and interpretation of the plots in the Figures is of crucial meaning in Results chapter.
Thak you very much, done.
Line 179: please insert again the lines 173-174 from the FIRST
Thak you very much, done. Lines 201-202
Line 206: please explain the abbreviation VIg (or VIg), the format term “general values” in

the FIRST VERSION was clearer.

We discard VIg, we have corrected and again we explain how VI “general values”

Line 295: Please justify more clearly why the Authors have developed a new For

example, insert the comment: “The previously developed vigor indices do not take into account the role of time in seedlings survival.” Please explain here why you find necessary

to include time in VI calculation. It is important to justify why the Authors have developed a new parameter. Moreover, I suggest to abbreviate this Vitality Index as VIT just to differentiate it from other formerly developed VIs.

To justify this recommendation, we have removed lines 499-502 from the first version and we have added new explanations in a clearer manner. Please read lines 418-465.
Lines 306-342: please move these Sections (3.2 and 3.3) to Results. There is no discussion (no reference to other papers) in these paragraphs. The Discussion chapter should always contain the comparison of the Authors' results to findings from other Or explanations of unexplained issues in the Authors' experiment based on the findings from other authors. So, please show here your results of ASL and SiO2 in a very condensed manner and REFER them to other papers.
We've added bibliographic references for the discussion and moved sections 3.2 and 3.3 to results. We actually rewritten the new results and discussion sections for clarity. They're not exactly the same as before, so please read the new sections.
Line 477: please insert here the lines 438-441 from the FIRST VERSION, it is necessary to explain here PH, Ln, Spn and St.
Done,
Lines 522-531: I strongly recommend to move this Section to Discussion and explain clearly why previous indices are not so suitable to be used in this experiment. See lines 499-502 in the FIRST VERSION of the manuscript.
Done, see the entire section 3.1 lines 418-465.

Minor remarks are as follows:

Line 110: References 38-39 refer to silicon application not to VI evaluation.
When rewriting the new introduction, these references were removed and new ones were included.
Lines 284-285: please move this description to the Figure 7
Done, we added a brief description of the results
Lines 537: please remove “38%” according to Results shown in the Table
Done
Line 599: I propose to promote VIT as a novel individual index so that it might be written as

follows: “[...] a novel method for calculating VI on time basis as VIT.”.

Thanks for your suggestion, we incorporated it to the manuscript.
Reference 13 is doubled as No. 16.
Corrected
No 41: please provide the full information on this publication: Publisher or
Done, now the reference number is 47

Reviewer 2 Report (New Reviewer)

Comments and Suggestions for Authors

Authors have made extensive corrections, according to the suggestions. I recommend this manuscript be published in its current version.

Author Response

Dear reviewer.

Thank you very much, we have improved the version with the corrections and suggestions of other reviewers. We expect that this new version being of your pleasantness Comments and Suggestions for Authors

Best regards

Reviewer 3 Report (New Reviewer)

Comments and Suggestions for Authors

I had previously recommended a major revision for the manuscript ID stresses-3867893 entitled “Vigor Index calculation as a function of the time on Agastache mexicana subsp. mexicana throughout induced hydric stress: SiO2 and Artificial Shade application effects.” However, after reviewing the revised version, I find that the changes made by the authors have not substantially improved the scientific quality, clarity, or readability of the manuscript. Therefore, I strongly recommend that this article should not proceed further in the review process.

While the authors have introduced some modifications, these revisions do not adequately address the concerns raised in the previous round. In many instances, the revisions appear superficial rather than effective in enhancing the manuscript’s quality. For example, the title has been changed to “Vigor Index calculation as a function of the time on Agastache mexicana subsp. mexicana throughout induced hydric stress: effects of SiO2 and artificial shade application.” However, the revised title remains unclear and difficult to interpret, and similar issues are evident in the abstract, introduction, and other sections of the manuscript.

In summary, the revisions have not resulted in meaningful improvements, and the manuscript still lacks the necessary scientific rigor, fluency, and clarity required for publication.

Comments on the Quality of English Language

english language is not fine.

Author Response

Dear reviewer, We are grateful for your comments, which helped us improve a lot in the first instance. You will find our response to your comments in blue.

In summary, the revisions have not resulted in meaningful improvements, and the manuscript still lacks the necessary scientific rigor, fluency, and clarity required for publication.

Comments on the Quality of English Language

english language is not fine.

The editor asked us to take into account the comments of other reviewers so that we could improved the manuscript. We did whatever they asked us. We strongly believe that the improvement is enough in the scientific rigor, fluency, and clarity. If you want, can read the new version. Regarding the English version, we sent the editor proof of the previous review done by a native speaker. You can request it upon request.

Round 3

Reviewer 1 Report (New Reviewer)

Comments and Suggestions for Authors

Please see the attachement.

Comments for author File: Comments.pdf

Author Response

TO THE REVIEWER

Dear Reviewer, once again we deeply appreciate your corrections and suggestions, which we have implemented in full.

The Authors' have made a great work to improve their manuscript and I really appreciate their efforts. Now, the manuscript is written quite well. My congratulations! It is also a satisfaction for me to contribute to a good paper. However, I still find some amendments, technical rather, to be done. I will mark them as “minor revision” but I hope the Authors will pay attention to them.

I accept the change of the title (not necessary in my opinion) but please check the English
We changed the previous title again, but redefined the VIT. It really was better.
Lines 99-102 and 103-105: it would be better to put two paragraphs into
Done
Line 106: I am not sure if plants require It would be better to write something like this: “Although silicon is not essential for plants, beneficial effects of SiO2 application in stressed plants were previously found.”
Thank you, done
Lines 123-125 and 126-127: please connect them into one
Done
I encourage to extend a paragraph in lines 123-127 and to indicate that previously developed Vigor Indices do not refer to changes in seedlings’ traits during the seedlings’ This may lead to overestimation of seedlings’ growth potential. I guess that such inaccurate assessment becomes often a reason for dissatisfaction. Maybe References No. 7-10 are adequate here, please check.
Done, we added a new reference
Lines 423-425: please note that in the formulas (1) and (2) a dash symbol is used instead of a short symbol of hyphen. This is strongly confusing because it suggests a substraction: VI “minus” I suggest to use brackets, for example VI(DW), or a subscript, like VIDW, both in the main text and in the formulas. Or ultimately a hyphen, but consequently both in the main text and in the formulas.
In the cited reference, they are written that way; we want to keep the original form; however, the suggestion seems extremely appropriate to us, and we have changed it.
Line 547: this sentence seems to be
Done, we rewritten it
Lines 579-582: Please correct: SDL (not ASL!) had not a significant The level 0.0% refers to SDL levels.
Thank you, corrected
Line 712: be consistent when using VI
R. We were several changes, see VI_T in blue across over manuscript

Author Response File: Author Response.pdf

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The paper should be improved before considering for publicatuion. The paper is not well organized. Section such as literature review and data and methods were not described.

1. The aim in introduction and abstract should be the same. The study focuses on the adaptation changes of Amm plants, encompassing mortality events, fluctuations in growth rate, leaf loss and recovery, and most importantly, the ability to survive over time. And aim from abstract: This study specifically examines hydric stress, and its effects on the emergence, seedlings and plants growth of Agastache mexicana subsp. mexicana (Amm) in a controlled environment.

2. The abstract should be divided into following sections: background, aim, merhods, results and conclusions.

3. The introduction should be focused on research gap. What is new in your research compared to existing knowledge in this field? What is research hypothesis? How your hypothesis was veryfied? At the end of introduction please write how your paper is organized?

4. There is no literatute review section. If you increse literature review section you will have to use more scientific literature and your paper will be more recognizeable.

5. There is no section data and methods. Some parts from introduction can be mooved to such section.

6. The best elaborated chapter is results and it does not need improvements.

7. Conclusions needs to be improved and the information about hypothesis veryfication should be placed.

8. More scientific literature should be used.

Author Response

R1-Open Review
Dear reviewer, we deeply appreciate your feedback on improving our manuscript. You'll
find the responses and changes in blue.
The paper should be improved before considering for publication. The paper is not well
organized. Section such as literature review and data and methods were not described.
1. The aim in introduction and abstract should be the same. The study focuses on the
adaptation changes of Amm plants, encompassing mortality events, fluctuations in growth
rate, leaf loss and recovery, and most importantly, the ability to survive over time. And aim
from abstract: This study specifically examines hydric stress, and its effects on the
emergence, seedlings and plants growth of Agastache mexicana subsp. mexicana (Amm)
in a controlled environment.
R. Done. Please, see the new Abstract and Introduction
2. The abstract should be divided into following sections: background, aim, methods,
results and conclusions.
R. Done. Please, see the new Abstract

Abstract: Understanding the impact of hydric stress on medicinal plants in
the context of climate change is gaining increasing importance. This study
aimed to assess the quality of a seed lot of Agastache mexicana subsp.
mexicana (Amm) by calculating the Vigor Index (VI) across six different
irrigation levels applied to seedlings and plants over time. We examined the
relationships among plant height, leaf count, and survival rates to achieve
this.
To maximize the survival of seedlings and plants, we implemented Artificial
Shade Levels (ASL) of 38, 87, and 94 %, as well as Silicon Dioxide Levels
(SDL) of 0.0, 0.2, 0.4, and 0.8 %. These treatments were selected for their
potential to enhance water retention and photosynthetic capacity, among
other benefits.
The findings reveal that plants adapt to hydric stress by shedding leaves and
exhibiting slower height growth, followed by a phase of regrowth until reaching
senescence and, ultimately, death. Our observations indicated that the
proposed irrigation levels resulted in waterlogging stress, which was evident
from the lowest Survival Index (SI) and the slow growth of height and leaves
during stages 1 and 2. Additionally, we noted drought stress, characterized by
a sustained decline in VI values from stages four to six.
The VI measurements were graphed using spline interpolation, a method that
provides a novel approach to effectively visualize and quantify the adverse
conditions impacting Amm's development over time. The VI results
demonstrated a statistically significant effect of ASL at certain stages, with the
highest values observed at ASL = 94%. However, the SDL did not show
statistical significance; nonetheless, it was evident that plants without SiO2
exhibited more consistent growth.

3. The introduction should be focused on research gap. What is new in your research
compared to existing knowledge in this field? What is research hypothesis? How your

2
hypothesis was verified? At the end of introduction please write how your paper is
organized?
R. Done, please see the new introduction section
4. There is no literature review section. If you increase literature review section you will
have to use more scientific literature and your paper will be more recognizable.
The section mentioned is content after section 5 Conclusions. We have aggregated 11
new references as literature to explain in a better way our findings, discussion and results.
5. There is no section data and methods. Some parts from introduction can be moved to
such section.
This section is the number 4 into manuscript after Results and Discussion sections,
besides, we have aggregate Supplementary material with data. We have removed one
Table to this section. New numeration in tables can be seen in the new version of our
manuscript.
6. The best elaborated chapter is results and it does not need improvements.
R. Thank you
7. Conclusions needs to be improved and the information about hypothesis verification
should be placed.
R. Done, please see the new Conclusions
8. More scientific literature should be used.
R. We have aggregate more literature in the Introduction and discussion sections
principally
Submission Date
04 July 2025
Date of this review
13 Jul 2025 10:55:59

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The present work try to correlate the effect of the abiotic stress on vigor index of the Agastache mexicana (medicinal plant). The stresses applied were artificial shade level and addition of silicon dioxide level. Which why those stresses were selected are not explained,

then is confused when describe artificial shade, related with stop watering and mixed that this caused waterlogging. does no explain the relation of shade-stop watering-with waterlogging

Title:

Should be rewrriten:

Effect of induced hydric stress on Agas…

In tiltle first indicates SiO2

In abstract first describes artificial shade then SiO2

Abstract:

Why SiO2 function is support survival, never explain this

Irrigatin level was reduced, but this produced waterlogging and rought stresses.. not clear if water was reduced why caused waterlogging. In all case, then, the work studied artificial shade that is related with reduction of light for photosynthesis, drought, and waterlogging, as well as addition of SiO2.

I see that the work was many variables,

Conclusion: plants adapt to grow under shading, but.. is also on waerlogging?

Keywords. Describe drought stress and abstract is waterlogging

On the other hand. Why or what is the use of SiO2 is not explained

So the title should be artificial shade, waterlogging and drought stresses?

Lines 89-90

Describe that in separate research study (is double the word, research or study= the same)

In local environment..but is not described wich conditions are local environment.

Lines 113 and 114. Authors describe that ASL and SDL can have an effect to support the plants survival.. but never explain the actions on plants

Line 122-129

It needs a rewrite the paragraph

Explain more clear,

Title

Table 1. Effect of vigor Index values by artificial shade level

In the table footnote,

-data are the mean of the measures of x replicates. Same letters shows no stastistical differences among means. ASL= artificial shade level; W=week

ASL

(%)

Line 131.. W9=W9…?

Table 2. Effect of silicon dioxide level on vigor index

SDL

(%)

0.0

0.2

0.4 if letter is the same, it is no necessary to add it

0.8

Table footnote. Should be as:

SDL=silicon dioxide level; W=weak. Data shows mean values of n=??. Multiple comparison by Tukey´s test showed no significant differences among values

Line 142-143.. the p values of the ..

This information should be part of M&M section

Figure 3a is the same as figure 1

Lines 328 and 329, seems is missing a reference.

Line 305-306. …Amm plants adapted and thrive in the face of fluctuations .. of what.. in plant height…

Is confuse this sentence authors should rewrite,

Line 354 and 355..

The cultivation of medicinal plants… dependent on the quality of the produce..in general this last paragrah, has little in relation to the work.

the quality of medicinal plants are the phytochemicals they produce, and yes growth conditions could affect the quality of the plants, but not only vigor of seeds, but the compunds they produce. Work lack of measured the important part, the phytochemiclas under the conditions.

Comments on the Quality of English Language

One of the problems to follow the manuscript is that ideas are confuse or the english is not very good.

Author Response

R2-Open Review
Dear reviewer, we deeply appreciate your feedback on improving our manuscript. You'll find the
responses and changes in blue.
Comments and Suggestions for Authors
The present work try to correlate the effect of the abiotic stress on vigor index of the Agastache
mexicana (medicinal plant).
The stresses applied were artificial shade level and addition of silicon dioxide level. Which why
those stresses were selected are not explained, then is confused when describe artificial shade,
related with stop watering and mixed that this caused waterlogging. does no explain the relation
of shade-stop watering-with waterlogging
R. The stress was hydric stress only.
R. We have done changes in the Introduction to explain in a better way:

This study aimed to calculate the Vigor Index (VI) of Amm seeds
under challenging conditions. These conditions included the use of an
unsterilized substrate with limited nutrients and the evaluation of
induced abiotic stress effects on germination, seedlings, and plants
through varying irrigation levels.
We focused on two key input factors to enhance plant survival
and growth: Silicon Dioxide Levels (SDL) at 0.0, 0.2, 0.4, and 0.8 %,
and Artificial Shade Levels (ASL) at 38, 87, and 94 %. By exploring
the interplay between irrigation levels and these input factors, we
aimed to calculate an optimal VI for the seedlings and plants. To
enhance seed survival, various studies investigated the impact of
different light levels or shade, along with silicon application rates, on
plant growth, especially under drought stress [11,12, 17-20,]. The
studies included improvement in root and shoot length, root and shoot
dry weight, leafs and shoot fresh weight, and total leaf area. Besides,
researchers have examined the relationship between plant height and
the number of leaves in the plant species Amm to predict successful
cultivation [21,22,23].
The combination of SDL and ASL was chosen for their ability to
enhance water retention, photosynthetic capacity, drought stress
resistance, and protection of plants from high temperatures
respectively(ranging from 24 to 35°C) [24-27].
The study investigates the adaptive changes in Amm seedlings
and plants, examining factors such as mortality events, variations in
growth rates, leaf loss and recovery, and most importantly, their
capacity for long-term survival. These elements highlight a gap in
several existing studies that have not explored the time scales of
these phenomena. To achieve our objective, we employed the
equations presented in section 4.5 of this manuscript. Ultimately, the
calculations of VI provide support for our hypothesis.
On the other hand, interpolation is a technique used to insert new
data points within a range of known data points. It can fill in missing
data, smooth existing data, and make predictions. The spline function

involves fitting a series of cubic polynomials. This method has been
used to track corn growth over time, including dry weight and total leaf
area per plant [28]. Additionally, it has been applied to humidity control
to enhance plant quality, [29] and to estimate water consumption in
the water cycle, specifically the reference evapotranspiration, which
enables more accurate prediction and planning of water resources
[30]. This research proposes the use of interpolation to connect
discrete points into a continuous curve function, which provides
valuable insights for developing techniques in the germination and
emergence processes. This method aims to produce healthy
seedlings for crop propagation. Unlike other methods that only take
into account germination percentage, emergence percentage, and
seedling size, this approach offers a more comprehensive analysis.

Title:
Should be rewrriten:
Effect of induced hydric stress on Agas…
In tiltle first indicates SiO2
In abstract first describes artificial shade then SiO2
R. Done. Please, see the new title
Abstract:
Why SiO2 function is support survival, never explain this
Irrigation level was reduced, but this produced waterlogging and drought stresses.. not clear if
water was reduced why caused waterlogging. In all case, then, the work studied artificial shade
that is related with reduction of light for photosynthesis, drought, and waterlogging, as well as
addition of SiO2.
I see that the work was many variables,
R. Done, please see the new Abstract and Introduction sections.
Conclusion: plants adapt to grow under shading, but.. is also on waterlogging?
R. Done, please see new redaction
Keywords. Describe drought stress and abstract is waterlogging
R. Done, the keywords are now: seed vigor in plants; waterlogging stress in plants; drought stress
in plants; artificial shade in plants; vigor index in plants; silicon as fertilizer.
On the other hand. Why or what is the use of SiO2 is not explained
So the title should be artificial shade, waterlogging and drought stresses?
R. Done, we have renamed the manuscript with a new title.
Lines 89-90. Describe that in separate research study (is double the word, research or study= the
same) in local environment..but is not described which conditions are local environment.
A. We removed the paragraph because it did not add value to the manuscript, and it did not have
the specific reference.
Lines 113 and 114. Authors describe that ASL and SDL can have an effect to support the plants
survival.. but never explain the actions on plants
R. Done

We focused on two key input factors to enhance plant survival
and growth: Silicon Dioxide Levels (SDL) at 0.0, 0.2, 0.4, and 0.8 %,
and Artificial Shade Levels (ASL) at 38, 87, and 94 %. By exploring
the interplay between irrigation levels and these input factors, we
aimed to calculate an optimal VI for the seedlings and plants. To
enhance seed survival, various studies investigated the impact of
different light levels or shade, along with silicon application rates, on
plant growth, especially under drought stress [11,12, 17-20,]. The
studies included improvement in root and shoot length, root and shoot
dry weight, leafs and shoot fresh weight, and total leaf area. Besides,
researchers have examined the relationship between plant height and
the number of leaves in the plant species Amm to predict successful
cultivation [21,22,23].
The combination of SDL and ASL was chosen for their ability to
enhance water retention, photosynthetic capacity, drought stress
resistance, and protection of plants from high temperatures
respectively(ranging from 24 to 35°C) [24-27].

Line 122-129. It needs a rewrite the paragraph
Explain more clear,
R. Done, please see lines 133-138

Title Table 1. Effect of vigor Index values by artificial shade level. In the table footnote, -data are
the mean of the measures of x replicates. Same letters shows no statistical differences among
means. ASL= artificial shade level; W=week
ASL
(%)
38
87
94
Line 131.. W9=W9…?
R. Done. Thank you for your help
Table 2. Effect of silicon dioxide level on vigor index
SDL
(%)
0.0
0.2
0.4 if letter is the same, it is no necessary to add it
0.8
Table footnote. Should be as: SDL=silicon dioxide level; W=weak. Data shows mean values of n=??.
Multiple comparison by Tukey´s test showed no significant differences among values
R. Done. Thank you for your help
Line 142-143.. the p values of the ..
This information should be part of M&M section

R. Done, we have made the changed. Thank you for your help
Figure 3a is the same as figure 1
R. Yes, however we have used the same graphics to explain different things because we
considered it necessary.

Lines 328 and 329, seems is missing a reference.
R. The references are correct, the paragraph is a little along only. The references in the new
version of the manuscript are 33 and 34
Line 305-306. …Amm plants adapted and thrive in the face of fluctuations .. of what.. in plant
height…Is confuse this sentence authors should rewrite,
R. Done, please see lines 319 and 320
Line 354 and 355..
The cultivation of medicinal plants… dependent on the quality of the produce..in general this last
paragraph, has little in relation to the work.
the quality of medicinal plants are the phytochemicals they produce, and yes growth conditions
could affect the quality of the plants, but not only vigor of seeds, but the compounds they
produce. Work lack of measured the important part, the phytochemicals under the conditions.
R. We have eliminated the paragraph. Thank you for your help.
Comments on the Quality of English Language
One of the problems to follow the manuscript is that ideas are confuse or the english is not very
good.
R. Done, we have thoroughly reviewed the manuscript and made the relevant changes in the
English language.
Submission Date
04 July 2025
Date of this review
16 Jul 2025 22:57:29

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

I think the autora sufficiently imprpoved the paper. It can be published.

Author Response

R1-Open Review

Dear reviewer, we deeply appreciate your feedback on improving our manuscript. You'll find the responses and changes in blue.

The paper should be improved before considering for publication. The paper is not well organized. Section such as literature review and data and methods were not described.

R. Done. Please, see the new Abstract and Introduction

2. The abstract should be divided into following sections: background, aim, methods, results and conclusions.

R. Done. Please, see the new Abstract

Abstract: Understanding the impact of hydric stress on medicinal plants in the context of climate change is gaining increasing importance. This study aimed to assess the quality of a seed lot of Agastache mexicana subsp. mexicana (Amm) by calculating the Vigor Index (VI) across six different irrigation levels applied to seedlings and plants over time. We examined the relationships among plant height, leaf count, and survival rates to achieve this.

To maximize the survival of seedlings and plants, we implemented Artificial Shade Levels (ASL) of 38, 87, and 94 %, as well as Silicon Dioxide Levels (SDL) of 0.0, 0.2, 0.4, and 0.8 %. These treatments were selected for their potential to enhance water retention and photosynthetic capacity, among other benefits.

The findings reveal that plants adapt to hydric stress by shedding leaves and exhibiting slower height growth, followed by a phase of regrowth until reaching senescence and, ultimately, death. Our observations indicated that the proposed irrigation levels resulted in waterlogging stress, which was evident from the lowest Survival Index (SI) and the slow growth of height and leaves during stages 1 and 2. Additionally, we noted drought stress, characterized by a sustained decline in VI values from stages four to six.

The VI measurements were graphed using spline interpolation, a method that provides a novel approach to effectively visualize and quantify the adverse conditions impacting Amm's development over time. The VI results demonstrated a statistically significant effect of ASL at certain stages, with the highest values observed at ASL = 94%. However, the SDL did not show statistical significance; nonetheless, it was evident that plants without SiO2 exhibited more consistent growth.

3. The introduction should be focused on research gap. What is new in your research compared to existing knowledge in this field? What is research hypothesis? How your hypothesis was verified? At the end of introduction please write how your paper is organized?

R. Done, please see the new introduction section

4. There is no literature review section. If you increase literature review section you will have to use more scientific literature and your paper will be more recognizable.

R. The section mentioned is content after section 5 Conclusions. We have aggregated 11 new references as literature to explain in a better way our findings, discussion and results.

5. There is no section data and methods. Some parts from introduction can be moved to such section.

R. This section is the number 4 into manuscript after Results and Discussion sections, besides, we have aggregate Supplementary material with data. We have removed one Table to this section. New numeration in tables can be seen in the new version of our manuscript.

6. The best elaborated chapter is results and it does not need improvements.

R. Thank you

7. Conclusions needs to be improved and the information about hypothesis verification should be placed.

R. Done, please see the new Conclusions

8. More scientific literature should be used.

R. We have aggregate more literature in the Introduction and discussion sections principally

Submission Date

04 July 2025

Date of this review

13 Jul 2025 10:55:59

Reviewer 2 Report

Comments and Suggestions for Authors

In general, the work is extensive using lot of formulas

some figures seems duplicated,

no more comments, decision will be taken by editor

Comments on the Quality of English Language

no further comments

Author Response

R2-Open Review

Dear reviewer, we deeply appreciate your feedback on improving our manuscript. You'll find the responses and changes in blue.

Comments and Suggestions for Authors

The present work try to correlate the effect of the abiotic stress on vigor index of the Agastache mexicana (medicinal plant).

The stresses applied were artificial shade level and addition of silicon dioxide level. Which why those stresses were selected are not explained, then is confused when describe artificial shade, related with stop watering and mixed that this caused waterlogging. does no explain the relation of shade-stop watering-with waterlogging

1. The stress was hydric stress only.

R. We have done changes in the Introduction to explain in a better way

Title:

Should be rewrriten:

Effect of induced hydric stress on Agas…

In tiltle first indicates SiO2

In abstract first describes artificial shade then SiO2

R. Done. Please, see the new title

Abstract:

Why SiO2 function is support survival, never explain this

Irrigation level was reduced, but this produced waterlogging and drought stresses.. not clear if water was reduced why caused waterlogging. In all case, then, the work studied artificial shade that is related with reduction of light for photosynthesis, drought, and waterlogging, as well as addition of SiO2.

I see that the work was many variables,

R. Done, please see the new Abstract and Introduction sections.

Conclusion: plants adapt to grow under shading, but.. is also on waterlogging?

R. Done, please see new redaction

Keywords. Describe drought stress and abstract is waterlogging

R. Done, the keywords are now: seed vigor in plants; waterlogging stress in plants; drought stress in plants; artificial shade in plants; vigor index in plants; silicon as fertilizer.

On the other hand. Why or what is the use of SiO2 is not explained

So the title should be artificial shade, waterlogging and drought stresses?

R. Done, we have renamed the manuscript with a new title.

Lines 89-90. Describe that in separate research study (is double the word, research or study= the same) in local environment..but is not described which conditions are local environment.

R. We removed the paragraph because it did not add value to the manuscript, and it did not have the specific reference.

Lines 113 and 114. Authors describe that ASL and SDL can have an effect to support the plants survival.. but never explain the actions on plants

R. Done. We have re-written the manuscript, you can read that the new version contains this explanation along all manuscript.

Line 122-129. It needs a rewrite the paragraph

Explain more clear,

R. Done, please see lines 133-138

Title Table 1. Effect of vigor Index values by artificial shade level. In the table footnote, -data are the mean of the measures of x replicates. Same letters shows no statistical differences among means. ASL= artificial shade level; W=week

ASL

(%)

Line 131.. W9=W9…?

R. Done. Thank you for your help

Table 2. Effect of silicon dioxide level on vigor index

SDL

(%)

0.0

0.2

0.4 if letter is the same, it is no necessary to add it

0.8

Table footnote. Should be as: SDL=silicon dioxide level; W=weak. Data shows mean values of n=??. Multiple comparison by Tukey´s test showed no significant differences among values

R. Done. Thank you for your help

Line 142-143.. the p values of the ..

This information should be part of M&M section

R. Done, we have made the changed. Thank you for your help

Figure 3a is the same as figure 1

R. Yes, however we have used the same graphics to explain different things because we considered it necessary.

Lines 328 and 329, seems is missing a reference.

R. The references are correct, the paragraph is a little along only. The references in the new version of the manuscript are 33 and 34

Line 305-306. …Amm plants adapted and thrive in the face of fluctuations .. of what.. in plant height…Is confuse this sentence authors should rewrite,

R. Done, please see lines 319 and 320

Line 354 and 355..

The cultivation of medicinal plants… dependent on the quality of the produce..in general this last paragraph, has little in relation to the work.

the quality of medicinal plants are the phytochemicals they produce, and yes growth conditions could affect the quality of the plants, but not only vigor of seeds, but the compounds they produce. Work lack of measured the important part, the phytochemicals under the conditions.

R. We have eliminated the paragraph. Thank you for your help.

Comments on the Quality of English Language

One of the problems to follow the manuscript is that ideas are confuse or the english is not very good.

R. Done, we have thoroughly reviewed the manuscript and made the relevant changes in the English language.

Submission Date

04 July 2025

Date of this review

16 Jul 2025 22:57:29