PONE-D-25-15737Cadmium-induced impairment in growth, photosynthetic apparatus and redox regulation in green amaranth (Amaranthus viridis) plant attenuated by salicylic acid and methyl jasmonatePLOS ONE

Dear Dr. Haque,

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PLOS ONE

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Reviewers' comments:

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Reviewer #1: Yes

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: No

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1:  The work is interesting and may have practical implications. I would suggest authors improve the conclusions and add strong suggestions for other studies:

-The sentence starting with "Keywords;" should end with a period instead of a semicolon.

-There are abbreviations which need elaboration

-There is need to add hypothesis in the introduction.

-Please be consistent for general name of plants or scientific names throughout the article

- Scientific names of plant species must in italics. Check it throughout the manuscript.

-Sometime methods have not been fully explained and sometime proper references are missing in this section

-Ensure consistency in terminology and notation throughout the results section.

-Properties of soil especially initial soil selected properties can be added in one or two sentences in the article. Sometime sentences are too short in length. Check this thoroughly in the whole article.

-Treatment application method is not clear

-Make sure that each figure has a clear and concise caption or legend. This should briefly explain the key findings or context of the figure for readers who may not closely examine the results section.

-There are many subheadings in this section and most of the subheadings are lengthy

-Sentence structure is very poor in this section in results section

-In some instances, it would be beneficial to explain how the cited studies are relevant to the current study in discussion section.

Discussion should be rewritten to explain mechanistic explanation rather than superficially covering the topic

-Conclusion is too short.

-It's valuable to include a paragraph on the potential applications of the findings and suggest directions for future research. For example, how could this knowledge be applied in agriculture, and what specific experiments or investigations might be warranted?

-Figure looks properly prepared except missing legends in few figures

Kindly add following references to make MS more catchy

https://link.springer.com/article/10.1007/s12210-018-0702-y

https://doi.org/10.1007/978-3-030-27165-7_7

https://doi.org/10.1007/s00344-018-9854-3

https://doi.org/10.1007/s12298-019-00715-y

https://doi.org/10.3389/fpls.2022.895427

Reviewer #2:  Thanks to the authors for this manuscript. This is a well-written and informative piece of work. The introduction clearly establishes the context, problem, and purpose of the study. The methods sections is well-detailed and crafted, although some sentences are a bit long and could be slightly shortened for better readability (I recognize this is subjective). The discussion is well done and coherent with the results, although some phrases might be a bit speculative and I would either expand them with more information on the underlying mechanisms that support the considerations, or tone them down.

The manuscript could be published as it is, since it is of good quality, but I have some suggestions for minor improvements if the authors accept them:

"42 Increased population growth in Bangladesh

43 forces rapid industrialization and imprudent use of agrochemicals (heavy metal-containing

44 pesticides and fertilizers) "

I agree that the "imprudent use" aspect is important to emphasize, but it's not inherent to population growth. It might be more accurate to say that the increased demand for food production drives agrochemical use, which might be or not abused depending on various aspects (for example, local laws).

"45 Heavy metals incorporate into the food chain

46 through plant uptake via soil-root-crop or water-root-crop pathways triggering serious health

47 hazards for living organisms (4,5).

I would slightly rephrase this saying that heavy metals *can* incorporate (depending on the plant species, the metal, and soil conditions) and *might* trigger health hazards, also depending on amounts.

"A well-known

112 cultivated variety “Gheekanchan” from Lal Teer Seed Ltd. was chosen for this experiment."

Can you provide a brief justification for choosing this specific variety? e.g. is it known for its Cd accumulation capability, its popularity among local farmers, or some other relevant characteristic? Even if it is just because it was simple to get and manipulate.

Also, I suggest to invert the order of words as "For this experiment, a well-known etc." (stylistic choice)

"amaranth seeds were disinfected with Vitavax-200"

Can you provide a short description of this product? In particular, describing why you chose this one in particular, with which some scholars might be unfamiliar.

"116 (95%) solution was purchased from Sigma-Aldrich, Germany and 11.8 µL was used to prepare a 1

117 mM stock solution for 50 mL volume. The SA (Sigma Aldrich, Germany) stock solution (1 mML

118 -1) was made by dissolving 138.2 mg in 2 mL methanol and then adding distilled water as needed"

Could you verify thins? Is 11.8 µL of 95% MeJA in 50 mL truly 1 mM? Or should it be a different volume? Is 138.2 mg/L of SA correct, given the molecular weight (molar mass)?

"Fourteen seedlings were supported by a perforated

126 cork sheet in each tank and regarded as a single replicate. "

I suggest to expand this for researchers trying to replicate the study or use it as a base for their projects, so that they will be able to handle with flexibility the given info. Only fourteen seedlings in total might be considered a bit little for accurate statistics, while putting them in just one sheet might be a bit crowded (depending on the size of the sheet). Could you add some text specifying why the outcome should not be compromised and is well reliable?

"Cd stress was initiated in

131 hydroponic conditions (21 d old seedlings) by introducing 10 µM CdCl2·H2O in the nutrient

132 solution in all treatments except control and the duration of Cd exposure was three weeks."

I suggest to specify why you chose this specific cadmium concentration and why exactly 21 days seedlings, even if you just found it to be a practical and workable setup. You later say "The Cd concentration was chosen based on a prior trial.", is it published data that you could cite, or just internal screening? Could you give further info? This can be useful for other researchers wanting to replicate the experiment who might discuss whether to follow by the book the protocol or slightly alter it depending on the situation.

"The composition was kept in the dark to initiate the reaction and then the absorbance was recorded at 390 nm wavelength."

You mean, immediately put in the dark and then you measured absorbance?

"MeJA and SA 424 supplementation might counteract growth inhibition by facilitating cell division and enlargement 425 and avoiding cell damage."

Can you describe the known mechanisms of action for this and similar passages? This can be useful for some latter phrases which hypothesize possible consequences from it.

"Under Cd stress, MeJa may help in controlling other growth-promoting

426 hormones such as gibberellin and ethylene enabling plant growth and development (63)."

I suggest to replace "enabling" with "promoting".

"The supplemented MeJA and SA may have increased their

429 endogenous levels that would restore cell permeability modulating nutrient uptake, gene expression

430 and redox balance under Cd stress (65)"

This is a bit speculative, since you didn't measure that. I would rephrase in more cautious words, and give a possible mechanism supporting this idea.

"In addition, Cd stress may induce perturbation" [...] "The degraded photosynthetic pigments along with the impairment of the light-harvesting complexes

439 and both PSI and PSII under Cd stress may have resulted in a declined rate of photosynthesis (A)"

As above, these phrase could be a bit speculative. I recognize that they can be legitimate, so take my suggestion as a way to make your conclusions stronger and clearer for the reader.

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Reviewer #1: No

Reviewer #2: No

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Cadmium-induced impairment in growth, photosynthetic apparatus and redox regulation in green amaranth (Amaranthus viridis L.) plant attenuated by salicylic acid and methyl jasmonate

PONE-D-25-15737R1

Dear Dr. Haque,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Dr. Debasis Mitra

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer #1: Thanks for the nice discussion. The authors have satisfactorily replied all my queries. Once again thank you

Reviewer #2: I thank the authors for resubmitting the manuscript and addressing the proposed points. I'm particularly appreciating the verification of the calculations and the expansion of the discussion. While it is in their right to hypothesize potential underlying causes for the results, it is methodologically safer a more conservative approach of first standing on established mechanisms of action and staying within the scopes of the study, until more specifical physiological and molecular investigations are performed. This work is in general a good result and in my judgement could be published on the journal, if the editor agrees.

The last thing that might be done could be a final check of potential small grammar mistakes, mispellings, formatting issues etc. that went unnoticed, as in many manuscripts there are often some that escape even vigilant eyes double or triple checking. But I don't want to cause distress to the authors, to whom I give my best wishes in their future research.