New Approaches on Micropropagation of Arracacia xanthorrhiza (“Arracacha”): In Vitro Establishment, Senescence Reduction and Plant Growth Regulators Balance

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Suggestions/Comments to the Author:

Dear Authors, 

The manuscript (Horticulturae-3848191) presents a comprehensive study on the use of antibiotics and silver nitrate to improve in vitro culture establishment and reduce premature senescence in Arracacia xanthorrhiza. The experimental design is sound, and the results are clearly presented, providing valuable insights into optimizing culture asepsis and shoot regeneration. The discussion effectively integrates previous literature, highlighting both the benefits and potential limitations of the applied treatments. Overall, the study contributes meaningfully to the field of plant tissue culture and offers practical recommendations for improving propagation protocols.

However, some aspects of the manuscript require minor improvements. Certain references contain typographical errors, inconsistencies in formatting, or minor inaccuracies in spelling and spacing that should be corrected to meet journal standards. Additionally, clarifying the rationale for specific antibiotic and silver nitrate concentrations, as well as ensuring that all cited literature directly supports the conclusions drawn, will enhance the clarity and impact of the manuscript.

Suggestions for Improvement:

• Page 1, Lines 29-32: Please include a concluding remarks at the end of your abstract.
• Page 1: Restructure the abstract so that the three aspects in the title—in vitro establishment, senescence reduction, and plant growth regulator balance—are explicitly highlighted as separate outcomes.
• Page 2, Line 56-63; Line 64-72: Sections on cultural, economic, and industrial uses are somewhat fragmented. Group these under broader themes like economic importance and industrial potential for smoother reading.
• Page 2, Lines 66-72: Production numbers (hectares, tonnage, revenue) can be condensed, giving more space to explain how these figures relate to the need for improved propagation methods.
• Page 2, Lines 73-82: This section should be revised for better grammar and clarity. The content should be written in normal text and avoid using formulaic expressions or reference-style formatting, which will help improve readability and flow.
• Page 3-4, Line 136-145, 170-171, 178-181: Standardize units, concentrations, and terminology (lines 136–145, 170–171, 178–181) – Ensure consistent formatting of units (µM, g L⁻¹, %, etc.) and chemical names (e.g., 6-Benzylaminopurin →BAP) throughout. Avoid mixing text with parentheses excessively.
• Page 7-8, Line 276-305: Use consistent units and formatting (lines 275–305) – Some concentrations are written as “mg L⁻¹” (AC) and others as “µM” (AgNO₃) with inconsistent spacing. Ensure uniform formatting for clarity.
• Pages 6-12: Please see the figure legends specially their formatting style for example ‘Figure 4. Morphological aspects of A. xanthorrhizain vitro plantlets cultivated under dif-399 ferent dosages 6-benzylamylopurine (BAP), kinetin (KIN) and meta-topolin (mT): A –ex-400 perimental control (no cytokinins supplementation); B –emerging sprout feasible of iso-401 lation to form a new in vitro propagule; C –limb malformation induce by 27 µM of BAP; 402 D –BAP at 1 µM; E –KIN at 1 µM; F –mT at 1 µM; G –BAP at 3 µM; H –KIN at 3 µM; I –403 mT at 3 µM; J –BAP at 9 µM; K –KIN at 9 µM; L –mT at 9 µM.’ while ‘Figure 1. xanthorrhiza in vitro cultures under antibiotic treatment: A – newly introduced 232 steam apex along with paper impregnated with ampicillin (100 mg L-1); B –aseptic plantlet 233 regenerated after 21 days of culture.’ See the differences in formatting.
• Page 5; 7; 8, Line 215-221; 279-280; 303-304: Highlight key quantitative results clearly (lines 215–221, 279–280, 303–304)– Use concise tables or bullet points to show survival, contamination, shoot, and sprout numbers. Currently, numeric data are embedded in long sentences, making it harder for readers to grasp trends quickly.
• Page 5; Page 7, Line 207-271: The explanation of ampicillin and tetracycline modes of action, historical references, and literature examples is very detailed. Consider summarizing these points and focusing mainly on your own experimental findings and key comparisons.
• Page 7, Line 273-297: Clarify interpretation of silver nitrate vs. activated charcoal– The discussion about AC adsorption properties is informative but could be shortened. Emphasize your own results and the comparative effectiveness of AgNO₃ vs AC, keeping mechanistic explanation concise.
• Most references are fine, but watch for typos, spacing errors, and consistency for online sources and database citations. Some may need verification to ensure they actually support the text (Reference 52 (lines 614–617): “tow pepper varieties” → should be “two pepper varieties.”; Reference 30 (line 561–562): Check “The development of third-generation tetracycline antibiotics and new perspectives.” You cite it for tetracycline mode of action in plants; verify if the reference actually contains plant-relevant info; Reference 56 (lines 625–627): “Elec J Biotech” → typically “Electronic Journal of Biotechnology” or “ J. Biotechnol” depending on journal style).

 
The manuscript provides a strong research foundation, and addressing the suggestions above will improve its clarity and depth. I look forward to seeing your revised version.

Comments on the Quality of English Language

The language is generally accurate and well-structured. Minor edits, if any, are sufficient, but no major revision is necessary.

Author Response

Dear reviewer, we appreciate your contributions. All suggestions were incorporated into the manuscript.

Best regards

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

To authors:

In this manuscript “New Approaches on Micropropagation of Arracacia Xanthorrhiza (“Arracacha”): In Vitro Establishment, Senescence Reduction and Plant Growth Regulators Balance” by Marques et al., This paper investigates the issues of severe bacterial contamination, rapid aging, and non-traditional cytokinins during the in vitro cultivation of Peruvian carrots. The effect of growth inhibitors was evaluated by in vitro introduction experiments of ampicillin and tetracycline embedded in paper sheets, and by adding activated carbon (1.75 g L-1) and silver nitrate to the culture medium. At the same time, five different concentrations (0, 1, 3, 9, and 27 µ M) of three aromatic cytokinins (6-benzylaminopurine, BAP) were evaluated; The effects of kinins, kinins, and meta topolin (mT) on plant morphogenesis. The research results provide important data support for in vitro culture or microproduction studies of Peruvian carrots.

Moreover, I have some other concerns, listed as following:

1. The introduction section about Peruvian carrots can be simplified appropriately to get straight to the topic and clarify the research background and significance.
2. Line 170-172, “…….the BMS was supplemented with 0, 5, 15, or 45 µM of filter sterilized AgNO3 solutions”, how is the concentration of silver nitrate defined, why are these concentrations chosen, and based on what. The same as the cytokinins concentrations (0, 1, 3, 9, and 27 µM).
3. Suggest writing the results and discussion sections separately, so that the results of this study can be clearly seen. Mixing them together as they are currently, can easily lead to misunderstandings and make it difficult to distinguish between the results of this study and those of previous studies. For example, in Table 3, it is difficult to distinguish what the author is trying to express and what the meaning behind it is.
4. The author should carefully verify whether all the variance analyses in the table are correct, especially the letters indicating significant differences. In addition, letters with significant differences should also be added to the graph.
5. The conclusion section can be appropriately simplified to directly clarify the main results and significance of this study.
6. As a research paper, the references should be selected from classic and closely related papers for application, without citing too many and removing some irrelevant ones.
7. Authors should refer to the format requirements of the journal, correctly and properly cite references.

Author Response

Dear reviewer, we appreciate your contributions. All suggestions were incorporated into the manuscript.

Best regards

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The author has made corresponding modifications according to the comments. It is worth noting that the author should check again whether the analysis of variance in Table 1 is correct and whether the lowercase letters representing the significance of the difference are wrong.

Author Response

Please verify the letter to the reviewer.

Thanks with regards

Author Response File: Author Response.pdf