Review Reports - Deciphering Seed Pre-Treatment and Soil Amendment Effects on the Germination and Early Growth of Radhachura/Peacock Flower (<i>Caesalpinia pulcherrima</i> L.)

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Title: Radhachura appears like a local name for the plant; Is peacock flower a better word in the title?

Abstract: ok

Keywords: adequate

Section 2.1 In the paragraph nothing was described about the plant material.

Section 2.2 line 123. How many days of drying specifically?

Section 2.4 The notation should be Ni instead of ni in equation (iii)

Section 2.5 what does SPAD stands for?

Can the authors cite references for calculating specific leaf area (SLA)?

Results: I couldn't find figures 3 and 4. Where are they?

Discussion: The discussion section must be condensed. It is way too much.

Conclusions: follows results

References:

Way too many; I am not sure if all of them are needed.

Names of journals are written in full for some and abbreviated for few others; the authors need to follow clearly the format required for the journal

Author Response

Response 1: [Thank you for this valuable suggestion on the title. We agree that "Peacock Flower" improves accessibility for international readers and have revised the title accordingly to include both the local name "Radhachura" and "Peacock Flower" for clarity.]

Comments 2: Abstract: ok

Response 2: [Thank you for your positive assessment of the Abstract.]

Comments 3: Keywords: adequate

Response 3: [Thank you for confirming the Keywords are adequate.]

Comments 4: Section 2.1 In the paragraph nothing was described about the plant material.

Response 4: [We are very grateful to the reviewer for taking the time to review our manuscript and for providing thoughtful and constructive feedback. We have rewritten the sentence and added it to Section 2.1.]

Comments 5: Section 2.2 line 123. How many days of drying specifically?

Response 5: [Thank you for this helpful comment. We have now specified the exact drying duration in Section 2.2, line 119-121. The sentence has been revised to: “After collection, the pods were sun‑dried for a week to facilitate seed separation, and the extracted seeds were stored in airtight zip‑lock bags prior to pre‑treatment and sowing in pots.”]

Comments 6: Section 2.4. The notation should be Ni instead of ni in equation (iii)

Response 6: [Thank you, that’s our mistake. “ni” has been replaced by “Ni” in the manuscript (Section 2.5).]

Comments 7: Section 2.5 what does SPAD stands for?

Response 7: [Thank you for this observation. SPAD stands for Soil Plant Analysis Development, a standard chlorophyll meter used for non-destructive leaf chlorophyll estimation. We have added this definition as an acronym expansion in the manuscript (Section 2.6) for clarity.]

Comments 8: Can the authors cite references for calculating specific leaf area (SLA)?

Response 8: [Thank you for this helpful suggestion. We have added citations in the manuscript for the specific leaf area (SLA) calculation method, including the reference by Garnier et al. (2001) https://doi.org/10.1046/j.0269-8463.2001.00563.x, which provides the standard protocol we followed.]

Comments 9: Results: I couldn't find figures 3 and 4. Where are they?

Response 9: [Thank you for noting this issue. Figures 3 and 4 were included in the original submission but appear to have been mislabeled or overlooked during formatting. We have corrected the figure names, numbering, and placement in the revised manuscript to ensure they are clearly visible and properly referenced.]

Comments 10: Discussion: The discussion section must be condensed. It is way too much.

Response 10: [Thank you for this suggestion. We have condensed the Discussion section by removing redundant points and focusing on the most relevant interpretations, reducing its length while retaining key scientific insights.]

Comments 11: Conclusions: follows results

Response 11: [Thank you for pointing this out. We have revised the Conclusions section to more directly reflect and follow from the results presented, ensuring alignment between findings and interpretations.]

Comments 12: References: Way too many; I am not sure if all of them are needed.

Response 12: [We thank the reviewer for this comment. We retained references that are essential to support the background, methodology, and interpretation of the present study, as well as to acknowledge the most relevant prior work. We believe the revised reference list now better balances completeness and conciseness.]

Comments 13: Names of journals are written in full for some and abbreviated for few others; the authors need to follow clearly the format required for the journal.

Response 13: [Thank you for this helpful comment. We have revised the references and followed the journal’s required format consistently for all journal names.]

Reviewer 2 Report

Comments and Suggestions for Authors

Please see the attached review report.

Comments for author File: Comments.pdf

Author Response

Manuscript ID: nitrogen-4245717

Title: Deciphering Seed Pre-Treatment and Soil Amendment Effect on the Germination and

Early Growth of Radhachura (Caesalpinia pulcherrima L.)

General Comment

The paper addresses a relevant and potentially useful applied question in nursery propagation: how seed pre-treatments and soil amendments affect germination and early seedling growth of Caesalpinia pulcherrima. The factorial design is a strength, and the inclusion of correlation and PCA suggests an effort to move beyond simple trait-by-trait reporting. However, the current version has several substantive problems in internal consistency, figure labeling, statistical description, and interpretation that must be resolved before the work can be considered.

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted in the re-submitted files.

Major comments

Comments 1: The abstract states that germination indices did not vary significantly in response to seed pre-treatment, whereas the Results section reports a significant pre-treatment effect on germination percentage, with T1 and T2 highest and T3 lowest. Authors should present one coherent interpretation across the abstract, results, discussion, and conclusion.

Response 1: [We thank the reviewer for carefully noting this inconsistency. In our ANOVA, germination percentage (GP) was significantly affected by seed pre-treatment, whereas the other germination indices were not. To ensure a coherent interpretation throughout the manuscript, we have revised the Abstract, Results, Discussion, and Conclusions to state explicitly that seed pre-treatment significantly influenced GP only, while soil amendments primarily affected the other germination indices. The previous wording suggesting no significant pre-treatment effect on germination indices in general has been corrected to reflect this more precise interpretation. Plz find the correction in Line 225-228]

Comments 2: In Section 3.2, plant-height results are cited as Figure 1A and Figure 1B, even though Figure 1 is the germination figure. In addition, the caption for Figure 2 says panels C F are leaf number, leaf area index, and specific leaf area, while the adjacent text immediately below discusses root length, plant length, plant height:root length, and chlorophyll content. This creates major uncertainty about which figure panels correspond to which variables.

Response 2: [We thank the reviewer for carefully identifying these errors in our figure citations and captions. We acknowledge that plant height was incorrectly cited as Figure 1A and 1B in Section 3.2, although Figure 1 presents germination data, and (ii) the original caption for Figure 2 did not match the variables shown in the panels. In the revised manuscript, we have corrected all in‑text references so that plant height results are now cited as Figure 2A (pre‑treatments) and Figure 2B (soil amendments). Replaced the previous Figure 2 caption with the following accurate description that matches the actual panels (A–I) in the figure:

“Figure 2. Effects of soil amendments and seed pre-treatments on growth related traits of radhachura. (A) Effects of seed pre-treatments on plant height, (B) Effects of soil amendments on plant height, (C) Effects of seed pre-treatments and soil amendments on root length at 90 DAS, (D) Effects of seed pre-treatments and soil amendments on Plant Length at 90 DAS, (E) Effects of seed pre-treatments and soil amendments on ratio of plant height to root length at 90 DAS, and (F) Effects of seed pre-treatments and soil amendments on chlorophyll content at 90 DAS, (G) Effects of seed pre-treatments and soil amendments on shoot fresh weight at 90 DAS, (H) Effects of seed pre-treatments and soil amendments on root fresh weight at 90 DAS, and (I) Effects of seed pre-treatments and soil amendments on ratio of shoot dry weight (SDW) to root dry weight (RDW) at 90 DAS. (C-I) The left graph represents different pre-treatment while the right graph has different soil amendments. Where T₁ is unscarified (control), T₂ is scarification, T₃ is hot water with scarification, T₄ is sulfuric acid with scarification, F₁ is soil with cowdung, F₂ is soil with compost, F₃ is soil with vermicompost, F₄ is soil with biochar and F₅ is soil with NPK. Error bars represent standard errors of the mean. Similar letter(s) found in a figure means they do not differ significantly. On the contrary, having different letter(s) in a figure (as per Tukey HSD test) signifies differences at a 5% level of probability.” These corrections remove the ambiguity and the figure is now fully consistent with the text in Section 3.2.]

Comments 3: Authors describe the study as a 4 × 5 factorial CRD with three replications and provide treatment descriptions, pot size, and measured traits. However, it does not clearly state the number of seeds sown per pot, the number of pots per treatment combination, the experimental unit used in the analysis, or enough nursery-management detail to allow replication. These details should be added.

Response 3: [We thank the reviewer for highlighting the need for greater clarity on the experimental layout and nursery management. In the revised Materials and Methods, we now specify that 4 seeds were sown per pot, with three pots per treatment combination in the 4 × 5 factorial CRD, and that the pot was considered the experimental unit for all statistical analyses. We have also added nursery management details, including a sowing depth of 2 cm, the watering as needed, the use of a transparent polythene roof without side walls to provide shade, and the use of amended soil without additional fertilization, so that the experiment can be replicated accurately.]

Comments 4: Authors state that only ANOVA and Tukey HSD were used in Minitab, R Studio, and Excel. It should clarify the factorial model, whether interaction terms were formally tested throughout, whether assumptions were checked, and how repeated measurements at 15-day intervals were handled analytically.

Response 4: [We thank the reviewer for this helpful comment and the opportunity to clarify our analytical procedures. Germination indices were calculated in RStudio using publicly available code from GitHub (GerminaR and Germinationmatrix library), while one‑way and factorial ANOVA followed by Tukey’s HSD tests were performed in Minitab 18, and graphs were prepared in Microsoft Excel. In all cases, we used a two-factor factorial model with seed pre-treatment and soil amendment as fixed effects and their interaction term included; interaction terms were formally tested for each response variable and reported when significant. For parameters recorded at 15‑day intervals (plant height, leaf number), measurements at each time point were analyzed separately. We have added a detailed description of these procedures to the Materials and Methods section 2.7 in the revised manuscript]

Comments 5: The correlation/PCA section refers to “genotypes” and even to “panicle length” as an indicator of vigor yet Figure 5 lists the analyzed variables and panicle length is not one of them. This wording does not match the treatment-based design of the present study and should be corrected throughout.

Response 5: [Thank you for pointing this out. We sincerely apologize for the inconsistency in wording. We have corrected the correlation/PCA section throughout the manuscript and we have replaced the incorrect wording “genotypes” and “panicle length.”]

Comments 6: Authors often emphasize vermicompost and biochar as broadly superior, but the reported results are more mixed: vermicompost is strong for several germination responses, compost is highest for some root and plant-length traits, and NPK records the highest peak value. The conclusion also extends toward sustainable soil-health management and longer-term traits such as flowering, canopy development, and carbon sequestration, even though the experiment lasted three months and focused on early nursery performance. The discussion should distinguish clearly between measured findings and broader hypotheses.

Response 6: [We thank the reviewer for this thoughtful and important comment. We agree that our original wording occasionally overstated vermicompost and biochar as universally superior, whereas the results show a more nuanced pattern in which vermicompost performs particularly well for several germination indices, compost is strongest for certain root and plant-length traits, and NPK yields the highest peak value. In the revised Discussion and Conclusion, we now describe each amendment’s strengths and limitations more cautiously, emphasizing treatment‑specific effects rather than broad superiority claims (Section 4, lines 469–479).

We also concur that our previous conclusion extended beyond the temporal and experimental scope of this three‑month nursery study. We have therefore revised the Discussion and Conclusion to clearly distinguish between (i) findings directly supported by our data (early germination behavior, seedling growth, and biomass under different pre-treatments and amendments) and (ii) broader implications for sustainable soil health, flowering, canopy development, and carbon sequestration, which we now explicitly present as hypotheses and priorities for future long‑term field research rather than as demonstrated outcomes (Conclusion lines 489-498). We hope these revisions clarify the boundaries of the current evidence while still indicating the potential applications of our results.]

Minor comments

Comments 1: Because the manuscript examines two classes of treatments, “Effects” would read better than “Effect” in the title.

Response 1: [Thank you for this helpful suggestion regarding the title. We agree that "Effects" better reflects the examination of two classes of treatments and have revised the title accordingly to read: "Deciphering Seed Pre-Treatment and Soil Amendment Effects on the Germination and Early Growth of Radhachura or Peacock Flower (Caesalpinia pulcherrima L.)."]

Comments 2: Check section numbering.

Response 2: [Thank you for noting this issue. We have carefully reviewed and corrected the section numbering throughout the manuscript to ensure consistent and sequential formatting.]

Comments 3: A full English edit would improve readability and professionalism.

Response 3: [Thank you for this valuable suggestion. We have conducted a thorough English language edit throughout the manuscript to enhance readability, clarity, and professionalism.]

Comments 4: Because the study was conducted over about three months under controlled nursery conditions, the paper should explicitly frame the findings as relevant to germination and early seedling performance rather than to field establishment or long-term ecosystem outcomes.

Response 4: [Thank you for this insightful suggestion. We fully agree with the reviewer’s assessment regarding the study timeframe and conditions, and we have revised the title, Abstract, Introduction, Discussion, and Conclusions to explicitly frame all findings as relevant to germination and early seedling performance under controlled nursery conditions, rather than field establishment or long-term ecosystem outcomes.]

Reviewer 3 Report

Comments and Suggestions for Authors

This manuscript describes germination/establishment radhachura. The study is limited to one seed source at one time and the results are from a single (replicated) trial. The single trial is quite well documented and for that I give the authors credit. Because of the specific nature of the trial I question whether it merits publication in Nitrogen.

I consider this study as an excellent pilot study but a greater range of times and seed sources should be included in a larger study.

Comments for author File: Comments.pdf

Author Response

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted in the re-submitted files.

Comments 1: This manuscript describes germination/establishment radhachura. The study is limited to one seed source at one time and the results are from a single (replicated) trial. The single trial is quite well documented and for that I give the authors credit. Because of the specific nature of the trial I question whether it merits publication in Nitrogen.

I consider this study as an excellent pilot study but a greater range of times and seed sources should be included in a larger study.

Response 1: [We sincerely thank the reviewer for their thoughtful assessment and for recognizing that our single trial is "quite well documented" and worthy of credit, as well as for describing this work as an "excellent pilot study." We fully acknowledge the limitation of using one seed source and time point, which allowed us to establish a controlled baseline for Radhachura germination and early growth under specific pre-treatment and soil amendment conditions. Moreover, Bangladesh having subtropical climate and the species mostly planted in urban areas for beautification might yield almost similar source effect.

These preliminary findings are important for publication in Nitrogen because they provide novel insights into how nitrogen-related soil amendments enhance seed establishment in this understudied flowering shrub, offering practical guidance for farmers and researchers in nitrogen-limited environments. While we agree that broader replication across seed sources and times would strengthen generalizability, this foundational pilot identifies key trends and methodologies that directly address pre-treatments and soil amendment dynamics in plant establishment aligning with the journal's scope and sets the stage for our ongoing larger-scale studies.]

Comments 2: Line 14: some perennial...[some species do not need it]

Response 2: [We appreciate the reviewer’s comment and agree that not all perennial species require this process. We have therefore revised the sentence in line 14]

Comments 3: Line 25: which had a similare effect to....]

Response 3: [We thank the reviewer for pointing this out. The phrase has been corrected in line 24 to “which had a similar effect to …” to improve the grammar and clarity.]

Comments 4: Line 30: and root development

Response 4: [We thank the reviewer for this suggestion. The phrase around line 29–30 has been revised to clearly include “and root development.]

Comments 5: 50: ...used for treating swellings.....

Response 5: [We thank the reviewer for pointing this out. The phrase around line 49 has been revised to correctly state that the plant is also “used for treating swellings,” improving the grammatical accuracy of the sentence]

Comments 6: 51: nd it is also used for the treatment of...

Response 6: [We thank the reviewer for this correction. The sentence around line 50 has been revised to “and it is also used for the treatment of…”]

Comments 7: Line: 64-65: This is a bit redundant as in line 56

Response 7: [We thank the reviewer for noting this redundancy. The text around lines 62–63 has been revised to avoid repeating the information already stated in line 56, so that this point is now mentioned only once in a more concise form.]

Comments 8: Line: 68: The sentencs as it stands is not correct - perhpas start as 'The potential amendments includeboth organic and inorganic chemiical as well as biologically active additions.

Response 8: [We appreciate the reviewer’s helpful suggestion. The sentence in lines 66–68 has been rewritten to start with “The potential amendments include both organic and inorganic chemical, as well as biologically active, additions …,” which corrects the grammar and more clearly describes the different types of amendments considered.]

Comments :9 Line: 82-83: estblishment of some species. Radachura produces seeds....

Response 9: [We thank the reviewer for pointing out this unclear sentence. The text in lines 80–81 has been revised.]

Comments 10: Line: 86: overcome

Response 10: [We thank the reviewer for this language suggestion. The informal phrase “get around” in line 84 has been replaced with “overcome” to use a more appropriate academic wording.]

Comments 11: Line: 93: benefits from such treatments

Response 11: [We appreciate the reviewer’s suggestion. The phrase in line 91 has been revised to “benefits from such treatments,” following the reviewer’s wording to improve clarity and precision.]

Comments 12: Line: 93: species-dependent pre-treatment...

Response 12: [We thank the reviewer for this helpful clarification. The phrase in lines 91–92 has been revised from “pre-treatment” to “species-dependent pre-treatment”.]

Comments 13: Line: 127: Do you mean cylindrical pots 18 cm tall and 15 cm diameter? Please give clearer details.

Response 13: [We thank the reviewer for requesting this clarification. Yes, we used cylindrical pots with dimensions of 18 cm in height and 15 cm in diameter, and this information has been specified in the revised text line 126.

Comments 14: Line: 128: ratios of additions to soil weight..??

Response 14: [We thank the reviewer for this question. Here, ratios mean soil mixtures. For instance, soil: cowdung/compost/vermicompost was applied in a ratio of 3:1. hese mixture ratios are now clearly described in Section 2.3 (lines 139–142)]

Comments 15: Line: 132: Why was that desgn chosen rather than a randomised complete block design?

Response 15: [Response: We thank the reviewer for this important question regarding the experimental design. Because the trial was conducted in a small, net‑house like structure with minimal environmental heterogeneity, and involved a single species with relatively small experimental units, we considered the experimental units sufficiently homogeneous to use a completely randomized design (CRD) rather than a randomized complete block design (RCBD).]

Comments 16: Line: 146: delete

Response 16: [We agree with the reviewer’s suggestion. The sentence “Data were collected at 15-day intervals after placing the radhachura seeds in pots” has been deleted from line 146 in the revised manuscript.]

Comments 17: Line: 147: Following planting, observations were made...

Response 17: [We thank the reviewer for this useful wording suggestion. The sentence in line 151 has been revised to “Following planting, observations were made …” to clearly indicate that observations started after planting.]

Comments 18: Line: 165: I think Nmax is the total number of germinated seeds

Response 18: [ We thank the reviewer for this comment. In the context of the germinationmetrics package used in our analysis, N_max is defined as the maximum number of seeds germinated per interval, rather than the total number of germinated seeds over the experiment. This follows the original definition in the germinationmetrics documentation and associated vignette]

Comments 19: Line: 167: Simplere to have GSi = ni/Nmax perhaps divided by 15 to put on a per day basis

Response 19: [We thank the reviewer for this suggestion. We agree that expressing GS_i as n_i/N_max would be a simple alternative, but in this study, we followed the germination indices as implemented in the R package “germinationmetrics,” which directly adopts formulae from the underlying germination literature. To maintain consistency with these published definitions and with the software used for our analyses, we have retained the original formulae in the manuscript, while clarifying in the Methods that all germination metrics were computed using “germinationmetrics” in R]

Comments 20: Line: 171: the upper and lower case symbols are confusing

Response 20: [We thank the reviewer for this helpful observation. To avoid confusion between upper- and lower-case symbols, we have revised the equation and its notation in line 174 so that all symbols are now defined clearly and used consistently throughout the manuscript.]

Comments 21: Line: 185: Ti = 15 i

Response 21: [We thank the reviewer for this comment. As defined earlier (line 167), Ti denotes the time from the start of the experiment to the ith interval, following the definition used in the R package “germinationmetrics.” In our experiment, germination counts were made every 15 days, so Ti takes the values 15, 30, 45, … days]

Comments 22: Line: 186: KK to K

Response 22: [We thank the reviewer for pointing out this typographical error. The incorrect symbol “KK” in line 186 has been corrected to “K” in the revised manuscript]

Comments 23: Line: 190: Days after sowing

Response 23: [We thank the reviewer for this clarification request, DAS means Days after sowing which has been elaborated previously before using the abbreviated form.]

Comments 24: Line: 194: Is shoot length the same as PH?

Response 24: [We thank the reviewer for this question. In our study, “shoot length” refers to the same measurement as plant height (PH), and we have revised the text around line 198 to use PH consistently and to clarify that it represents shoot length]

Comments 25: Line: 195: Is mean height average across three replicates?

Response 25: [We thank the reviewer for raising this point. Yes, “mean height” refers to the average plant height calculated across the three replicates, and we have revised the text around line 199.]

Comments 26: Line: 208: An impressive array of statisitical packages were tried. But there some other considerations that need to be considered.

Germination % will have binomial errors so a generalised linear regression should have been used. Similarly, for leaf count the errors would have been Poisson. Were tests of normality and constant varience considered? Graphical plots would have been useful.

Response 26: [We thank the reviewer for these valuable suggestions regarding the statistical treatment of the germination and count data. We agree that germination percentage arises from binomial data and that generalized linear models (GLMs) with an appropriate link function provide a rigorous alternative to ANOVA for such responses, and likewise that Poisson‑type models are often suitable for count variables such as leaf number. In the present analysis, we followed a more traditional approach used in many seed‑germination and nursery studies and analyzed germination percentage and other continuous traits using factorial ANOVA after verifying model assumptions. Specifically, we examined residual plots to assess normality and homogeneity of variance, and applied transformations when necessary to improve adherence to ANOVA assumptions. Leaf number showed approximately symmetric residuals and no strong heteroscedasticity in these diagnostics, so we retained the ANOVA framework for consistency with the other traits. We fully recognize that binomial and Poisson GLMs would offer a more distribution-appropriate framework for germination percentage and count data, as highlighted by recent methodological work. Given the scope and timeframe of the current revision, we have not re‑implemented the entire analysis using GLMs, but we have now clarified our use of ANOVA, and the assumption checks performed.]

Comments 27: Line: 211: Which mean values do you mean, e.g mean acroos replicates ( average of 3) or mean across soil amendments or mean across scarification treatments

Response 27: [We thank the reviewer for raising this point. Mean across treatments i.e., mean across soil amendments or mean across pre- treatments.]

Comments 28: Line: 217: Noting that none of the interactions were significant, so main effects are important. These are shown in Figure 1.

Response 28: [We thank the reviewer for raising this point. Yes, the interactions were not significant. As the study was a factorial one, to have better clarity of the finding’s interaction tables were presented in supplementary materials and cited in the main text.]

Comments 29: Line: 276-279: That would be a direct result of the treatments - basically mass balance!

Response 29: [We thank the reviewer for this question. Yes, it’s basically the result of the treatments.]

Comments 30: Line: 315: if this is the common name, no capital. If the binomial (Latin) name it needs to be in italics

Response 30: [We thank the reviewer for this helpful remark. In this context we are using the common name, so we have revised the text to “radhachura” (lower case, non‑italicized) in line 331 to follow the standard convention for common plant names]

Comments 31: Line: 326: The first two components have a reasonable interpretation, but together expalined less than 50%

Response 31: [We thank the reviewer for this helpful comment. It is correct that PC1 and PC2 together explain less than 50% of the total variance (29.6% and 14.6%, respectively; 44.3% cumulative). In our view, this level of variance capture is typical for multivariate morphological datasets and still provides useful insight into how growth and biomass traits co‑vary among treatments, even though it does not represent the full phenotypic variation.]

Comments 32: Line: 344: (C. pulcherrima L.)

Response 32: [We thank the reviewer for this suggestion. “(C. pulcherrima L.)” has been removed from line 344 in the revised manuscript.]

Comments 33: Line: 349: Pretreatment did not have a large effect (Table S1).

Response 33: [We thank the reviewer for raising this point. Yes, pre-treatment does not have a large effect, that’s why we have written that their impacts were substantially different. We have revised the discussion part as well.]

Comments 34: Line: 356: Was this statitically significant, seeing the distribution would not have been normal.

Response 34: [We thank the reviewer for this important point. Seed germination percentage of radhachura in the scarified hot water treatment differed significantly from that of other scarified and non-scarified treatments.]

Comments 35: Line: 372: This seems very precise when observations were every 15 days (see line 146)

Response 35: [Many thanks for bringing this to our attention. Actually, in line 146, we mentioned data collection on some of the growth parameters (e.g., plant height, number of leaves) which were taken at 15-day intervals. On the other hand, PGT, a germination index, indicates that soil amendment with biochar delays germination (7 days) or has no stimulant effect.]

Comments 36: Line: 389: But the differences were not statistically significant.

Response 36: [Many thanks for the suggestion, we have rewritten the sentence for clarity based on findings. We have re-worded as- “Overall, these results indicate that seed pre-treatments had identical effects on radhachura seed germination, but soil amendments………………”]

Comments 37: Line: 435: water holding capacity

Response 37: [We thank the reviewer for this helpful clarification. In this context, “water content” was intended to mean “water holding capacity”, and we have revised the wording accordingly in line 413 to use the correct term]

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Is T2 in figure 1 sandpaper scarification? If so, write that explicitly. Also in line 274.

Captions of figures 2 E and i, if there are ratios write them clearly.

Author Response

Comment 1: Is T₂ in figure 1 sandpaper scarification? If so, write that explicitly. Also in line 274.

Response 1: [We thank the reviewer for this helpful suggestion. T₂ indeed corresponds to the sandpaper scarification treatment, and we have now stated this explicitly in the legends of Figures 1 and 2, as well as in the text around lines 245–246 and 274–275, so that the treatment labels are clear without needing to refer back to the Methods section.]

Comments 2: Captions of figures 2 E and i, if there are ratios write them clearly.

Response 2: [We thank the reviewer for this helpful suggestion. In the revised Figure 2, we now state the ratios explicitly as follows: panel (E), line 267, “Effects of seed pre-treatments and soil amendments on the ratio of plant height/root length at 90 DAS” and panel (I), line 271, “Effects of seed pre-treatments and soil amendments on the ratio of shoot dry weight/root dry weight at 90 DAS”]

Reviewer 2 Report

Comments and Suggestions for Authors

No further comment

Author Response

Comment 1: No further comment.

Response 1: [We thank the reviewer for carefully evaluating our manuscript and are pleased that they have no further comments at this stage.]

Reviewer 3 Report

Comments and Suggestions for Authors

The paper has been tidied up and apart from some minor details is ready for publication.

The only matter that needs to be changed is the interpretation of the scree plot - even if there is no structure in the data some eigen values would be greater than one - the scree plot shows there is little gain after 3 components. I would like to see that matter rectified.

It is a matter for the editorial board if they consider results from a single glasshouse trial warrants publication.

Two small points in the abstract - it should be 'Timson's', also I don't think it is correct to refer to initial germination - once a seed is considered germinated that is it!

Author Response

Comment 1: The paper has been tidied up and apart from some minor details is ready for publication.

Response 1: [We thank the reviewer for this positive evaluation and are pleased that the manuscript is considered essentially ready for publication. We have carefully addressed all minor details raised in the subsequent comments and revised the manuscript accordingly, which we believe has further improved the clarity and presentation of the work.]

Comments 2: The only matter that needs to be changed is the interpretation of the scree plot - even if there is no structure in the data some eigen values would be greater than one - the scree plot shows there is little gain after 3 components. I would like to see that matter rectified.

Response 2: [We thank the reviewer for this important clarification regarding the interpretation of the scree plot. We have revised the description of the scree plot in Figure 4A. Because eigenvalues of 1.0 or greater are often considered meaningful, we retain this information in the manuscript. At the same time, the scree plot shows a clear elbow after the third principal component, and PC1–PC3 together explain 54.1% of the total variance, indicating that most of the meaningful structure in the data is captured by these three components. Accordingly, and in line with the reviewer’s suggestion, our interpretation in the Results section was focused on PC1–PC3.]

Comments 3: It is a matter for the editorial board if they consider results from a single glasshouse trial warrants publication.

Response 3: [We appreciate the reviewer’s remark regarding the scope of inference from a single glasshouse experiment. Our intention in this study is to provide an initial, controlled evaluation of seed pre‑treatments and soil amendments on radhachura germination and early growth, recognizing high environmental control but limited environmental generality.]

Comments 4: Two small points in the abstract - it should be 'Timson's', also I don't think it is correct to refer to initial germination - once a seed is considered germinated that is it!

Response 4: [We thank the reviewer for carefully checking these details in the abstract. First, we have corrected the term to “Timson’s index,” consistent with the original germination literature and the “germinationmetrics” R package documentation. Second, we agree that the phrase “initial germination” is imprecise, since germination is a one‑time event for each seed. We have therefore replaced “initial germination” with “germination,” in the abstract.]