Effect of Pb Stress on Ionome Variations and Biomass in Rhus chinensis Mill
Round 1
Reviewer 1 Report
Comments
The manuscript entitled “Effect of Pb stress on ionome variations and biomass in Rhus chinensis Mill is interesting but it only mentions the ionomic changes in Rhus chinensis in response to Pb stress. At present, there are too many research reports in this field, but the innovation is not high. Furthermore, the content of the discussion is too simple. In the present form, the work is not acceptable as it requires changes and careful edition.
Some comments and corrections are given below as examples, without being exhaustive.
Line 32: Give full spelling of As (Arsenic) as it is represented first time and later you can use the symbol throughout the manuscript
Line 73: Italicize the word Anacardaceae
Line 92: Please provide quantity detail of plant replicates used for statistical analysis. Please provide photos of experimental setup.
Line 93: Delete the word “was from”
Line 96: Why plants were grown in 500 & 1000 mg/kg Pb? Have you noted this limit of Pb-contamination?
Line 98-99: Delete the repeated sentence “Pb(NO3)2 solution was prepared and sprayed on the soil, and then the soil was equilibrated for one month”
Line 113: The Pb accumulation and translocation factor (TF) should also give more details about Quality Assurance and Quality Control. Authors should indicate the obtained accuracy values. Were used plant reference materials? If so, these should be listed. It would be also interesting to provide the reader with limits of detection/determination of analyzed element
Line 160-161: Please justify why TF is high (0.411) in control compared to treated samples
Line 180-181: Discuss this sentence “This might be because Pb stress reduced biomass but did not prevent the roots from absorbing and transferring nutrient elements, which caused the accumulation of nutrient elements” in the discussion part.
Line 287: Mention the name of plants
In the introduction part, could you please describe the Pb-detoxification mechanism and how it affects the growth and development of the plant?
In conclusion, you should comment on one possible limitation of the study, and make suggestions in the future study.
Please improve the overall language of the manuscript.
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
The authors investigated the effects of Pb contamination on the ionome and stoichiometry of CNP and the biomass of different organs of Chinese sumac. The study was carried out in the context of remediation and conservation efforts to remove heavy metals from soils. Nevertheless, a very strong and difficult to overestimate aspect of this study is the sheer demonstration of the variability in the elemental composition and proportions of the elements that make up the different plant organs. The conservation context is an additional asset of the study. In my opinion, the study will be of interest to a wide audience, from general ecologists working on ecological stoichiometry and ionomics in the context of ecosystem functioning and food webs, to specialists focusing on conservation. The weaknesses of the manuscript are:
(1) failure to introduce the reader to the topic of ionomics and stoichiometric ecology, and thus to discuss the results in a general context, which is important to realise the full potential of this study and increase its impact.
(2) methodological ambiguities, mainly related to the methods of elemental analysis, which require further explanation.
I believe that this study has a lot of potential and in my comments below I have focused on improving aspects related to the weaknesses. The manuscript should be ready for publication if the authors fully address the weaknesses of the manuscript by taking all my comments fully into account when rewriting the manuscript text.
Introduction
This section lacks a first short paragraph introducing the reader to the frameworks of ionomics and ecological stoichiometry. Such an introduction would be necessary for the reader to understand why the authors are studying the concentrations and ratios of elements in different organs, what biological significance these concentrations and ratios have in the context of the functioning of the whole organism, and, finally, what general research question the authors are asking and why this is worthwhile. At the end of this section, there should be a statement of the general research question that the authors address in their study.
In order to streamline and shorten the peer review process, I am providing information below on three resources where the authors can find the information they need:
Filipiak, M. & Filipiak, Z.M. (2022). Application of ionomics and ecological stoichiometry in conservation biology: Nutrient demand and supply in a changing environment. Biol. Conserv., 272, 109622. https://doi.org/10.1016/j.biocon.2022.109622
Fernández‐Martínez, M. (2022). From atoms to ecosystems: elementome diversity meets ecosystem functioning. New Phytol., 234, 35–42. https://doi.org/10.1111/nph.17864
Kaspari, M. (2021). The Invisible Hand of the Periodic Table: How Micronutrients Shape Ecology. Annu. Rev. Ecol. Evol. Syst., 52, 199–219. https://doi.org/10.1146/annurev-ecolsys-012021-090118
Materials and methods
Lines 112 – 125 – Analysis of elemental concentrations
I do not understand the authors' reasons for the choice of the Kjeldalh method for the analysis of nitrogen with the CHNS analyser at their disposal. The CHNS analyser, while measuring the concentration of C in the samples, is also supposed to give results for N and S. This method is more reliable than the one used by the authors because it does not require digesting the samples. The authors used a mixture of sulphuric acid and hydrogen peroxide to digest the samples. Such a mixture is not strong enough for complete digestion of plant tissues. Perchloric acid is needed. I therefore ask the authors to provide a detailed justification for their approach in the text of the manuscript.
Line 113. It is not sufficient to state that the pretreatment of plant samples has been described previously. In the text, please describe the method used in such a way that the whole study can be easily reproduced.
Lines 94-99. There is something wrong with this text - the sentence is repeated. Please correct it.
Discussion
What is missing from the text is a discussion of the general differences in the concentrations and ratios of the elements in the different plant organs. Such a discussion should apply only to the data from the T0 treatment - without the addition of Pb. In the discussion, the authors should refer to the available literature on ionomics and ecological stoichiometry. They should compare the results obtained with the literature data. In this context, they should point out a rather important aspect of this study - that elemental concentrations and elemental ratios vary from one plant to another and from one organ to another within a plant. This has important implications for ecologists working in ionomics and ecological stoichiometry: elemental contents and ratios in a plant cannot be compared when measured in different organs.
Author Response
Please see the attachment.
Author Response File: Author Response.docx
Round 2
Reviewer 2 Report
The authors have considered the comments made by the reviewers and have responded to them accordingly. The authors' response to the reviewers' comments is an adequate explanation of the inaccuracies and there have been sufficient revisions to the manuscript in response to the reviewers' comments. I am satisfied with the current version of the manuscript.
Author Response
Thank you for your review of this manuscript and for your recognition!
