Susceptibility of Selected Crops to Simulated Imazethapyr Carryover: A Morpho-Anatomical Analysis
Round 1
Reviewer 1 Report
The reported study compares sensitivity of different crop species to herbicide residue in soil. Herbicide carryover is an important issue that must be considered when planning crop rotation. The reported results are interesting from the point of view of plant biology and have application potential. The results can be used as guidelines for conducting bioassays of potentially contaminated soils. Overall, the text is clear and the visual material is very qualitative. I would suggest, as an improvement, to discuss more the possible differences of response in different varieties of the sensitive and less sensitive crops.
Some comments on the text:
Ln122 "crop varieties" - probably, crop species? Different varieties of the same crop may also have different sensitivity to herbicide residues
Ln131 “glass pot” is not necessary, in my opinion it is sufficient to write that the sand was in a glass Petri dish
Ln141 "cross-sections" were these not longitudinal sections?
Table1: it is a little misleading that the order is ED50 – ED10 – ED20, an increasing or decreasing order would be more logical
Ln215 and 219 “cultivar" is used in the first case and “variety” in the other case, as far as I know, there are different varieties of oilseed rape.
In the paragraph the ED10 and ED20 refer to the ED values for root length, this can be stated in Table 2 (although this is mentioned in the method description, still it will facilitate reading).
Ln246 “very significant” - does this mean that the difference was statistically significant (in which case “very” is unnecessary) or that the difference was both statistically significant and large? The p values can be reported in the text.
(same comment to Ln 282, 284, 310 etc)
I detected no significant problems
Author Response
Dear Reviewer,
Thank you for your positive feedback and helpful comments.
Below are our responses to your comments.
Comment 1. I would suggest, as an improvement, to discuss more the possible differences of response in different varieties of the sensitive and less sensitive crops.
Answer: We added these sentences in the discussion: It is well known that different plant varieties respond differently to herbicides in terms of selectivity or phytotoxicity [57,86]. However, it is not clear whether this phenomenon is due to micromorphological characteristics, as in some weeds [90], subtle differences in physiology, gene expression, or other factors, making this topic an interesting opportunity for further research. (lines 237-241)
Comment 2. Ln122 "crop varieties" - probably, crop species? Different varieties of the same crop may also have different sensitivity to herbicide residues
Answer: We agree, corrected as suggested (line 124).
Comment 3. Ln131 “glass pot” is not necessary, in my opinion it is sufficient to write that the sand was in a glass Petri dish
Answer: We agree, corrected as suggested (line 133).
Comment 4. Ln141 "cross-sections" were these not longitudinal sections?
Answer: Sorry for omission, corrected as suggested (line 147).
Comment 5. Table1: it is a little misleading that the order is ED50 – ED10 – ED20, an increasing or decreasing order would be more logical
Answer: We agree, corrected as suggested
Comment 6. Ln215 and 219 “cultivar" is used in the first case and “variety” in the other case, as far as I know, there are different varieties of oilseed rape.
Answer: We agree, corrected as suggested (line 223).
Comment 7. In the paragraph the ED10 and ED20 refer to the ED values for root length, this can be stated in Table 2 (although this is mentioned in the method description, still it will facilitate reading).
Answer: Corrected as suggested (lines 295-296).
Comment 8. Ln246 “very significant” - does this mean that the difference was statistically significant (in which case “very” is unnecessary) or that the difference was both statistically significant and large? The p values can be reported in the text.
Answer: The differences were statistically significant. We have corrected this throughout the text. The p-values are now included in the text.
same comment to Ln 282, 284, 310 etc
Answer: Same as above for Ln246
We thank you for the time and effort you took to review our manuscript. We hope that we were able to improve our manuscript based on your suggestions.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments to assist with final editing:
Line 29 -- spelling of "acetolactate"
Line 246 -- Use "highly" instead of "very"
Line 248 -- Use "highly" instead of "very"
Line 262 -- Use "root growth not inhibited" instead of "roots not worn down"
Line 263/264 -- Meaning not clear: "...fewer cells were worn...."
Line 353: Spelling of "imazethapyr"
Line 356: Spelling "roots" instead of "root"
In general, I commend your good work. Anatomical investigations are rarely conducted -- yours is appropriate and good quality work.
Generally of good quality -- the few minor aspects requiring attention/correction have been pointed out to the authors.
Author Response
Dear Reviewer, Thank you for your positive feedback, commends and helpful comments. We have made corrections based on your suggestions and hopefully improved our manuscript.
Below are our responses to your comments.
Comment 1. Line 29 -- spelling of "acetolactate"
Answer: Corrected as suggested
Comment 2. Line 246 -- Use "highly" instead of "very"
Answer: The differences were statistically significant. We have corrected this throughout the text.
Comment 3. Line 248 -- Use "highly" instead of "very"
Answer: The differences were statistically significant. We have corrected this throughout the text.
Comment 4. Line 262 -- Use "root growth not inhibited" instead of "roots not worn down"
Answer: Corrected and clarified as suggested
Comment 5. Line 263/264 -- Meaning not clear: "...fewer cells were worn...."
Answer: Corrected and clarified as suggested
Comment 6. Line 353: Spelling of "imazethapyr"
Answer: Corrected as suggested
Comment 7. Line 356: Spelling "roots" instead of "root"
Answer: Corrected as suggested
Author Response File: Author Response.pdf
Reviewer 3 Report
The manuscript presented for a review contains the results of one series of laboratory experiments verifying the ED doses of imazethapyr residues that do not cause phytotoxic effects on crops. It also analyses anatomical changes in root tissues of crops that have contact with this herbicide to find a safe dose of herbicide, not phytotoxic to the succeeding crops.
Comments:
1. Introduction. Add WSSA/HRAC group for the mode of action of this herbicide when you characterize it.
2. Ln. 80 - geographic coordinates of the soil collection place?
Ln. 81 - what was the source for the classification of soil? Why was this soil type chosen? I think better would be to use two contrasting soils - heavy and light- for a better comparison, i.e., in the light of different sorption capacities and pH.
Ln. 98 How was the field capacity measured? How much was it?
Ln. 100, were the crops sown together (different species) in pots or separately?
Fog. 1. Explain ppbw in footnote.
Tables 1 and 2 - what is n=?
Ln. 206 - slight differences. Were they significant?
Ln. 246 and 784 - what do "very significant differences" mean?
Are the Figures representative of each treatment?
Table 2 does not present any internal differentiation (SD or ED), which is why it is difficult to conclude the value of the results.
Tables - for decimals, please use dots instead of commas.
In conclusion, I would add specific crops that the measurements that were carried out. Not just generally - more and less susceptible to imazethapyr.
Author Response
Dear Reviewer,
Thank you for your positive feedback and helpful comments. We hope that our answers/corrections will meet your expectations and improve our manuscript.
Below are our responses to your comments.
Comment 1. 1. Introduction. Add WSSA/HRAC group for the mode of action of this herbicide when you characterize it.
Answer: Corrected as suggested (line 26).
Comment 2. 2. Ln. 80 - geographic coordinates of the soil collection place?
Answer: We have included these data in the text, as suggested (lines 81 - 82).
Comment 3. Ln. 81 - what was the source for the classification of soil? Why was this soil type chosen? I think better would be to use two contrasting soils - heavy and light- for a better comparison, i.e., in the light of different sorption capacities and pH.
Answer: The source for the soil classification was the USDA Texture Calculator (https://www.nrcs.usda.gov/resources/education-and-teaching-materials/soil-texture-calculator). All soil analyzes were performed at the Department of Pedology and Geology, Faculty of Agriculture, University of Belgrade.
This soil type was used because it is very common in areas with intensive agricultural production in our country. The research topic of this study focused primarily on investigating the specific effects of bioavailable herbicide residues on selected crops. Thank you for your suggestion of using two contrasting soils. The soil properties you mentioned are very important to the fate and behavior of herbicides in the soil, and some of our future studies may incorporate this objective.
Comment 4. Ln. 98 - How was the field capacity measured? How much was it?
Answer: The field capacity was measured by the core method with water retention at -33 kPa (Carter, M. Soil sampling and methods of analysis. Lewis Publishers; Boca Raton, Florida, 1993; pp. 569-579). The field capacity of the soil was 24%vol. We have included these data in the Material and Method section of the manuscript (lines 101-102).
Comment 5. Ln. 100 - were the crops sown together (different species) in pots or separately?
Answer: The crops were sown separately. I have rephrased this sentence so that hopefully there is no longer any doubt. The corrected version is now in lines 98-101.
Comment 6. Fig. 1. Explain ppbw in footnote.
Answer: Corrected as suggested. The corrected version is now in lines 202-203.
Comment 7. Tables 1 and 2 - what is n=?
Answer: Corrected as suggested. Table 1 now contains the n values.
In Table 2, the number of plants per plant species and per treatment is given in Table caption. Because both permanent and temporary microscopic preparations were made, n is not identical for each anatomical parameter. For permanent microscopic preparations, 20 roots per plant species and treatment (5 plants/roots per plant species and treatment, with 4 replications) were used, and for temporary microscopic preparations, 40 roots per plant species and treatment (10 plants/roots per plant species and treatment, with 4 replications) were used. The latter information can be found in the Materials and Methods section (lines 141-145).
Comment 8. Ln. 206 - slight differences. Were they significant?
Answer: This has now been clarified. The corrected version is now in lines 214-217.
Comment 9. Ln. 246 and 784 - what do "very significant differences" mean?
Answer: The differences were statistically significant. Where necessary, corrections were made throughout the text.
Comment 10. Are the Figures representative of each treatment?
Answer: Yes they are.
Comment 11. Table 2 does not present any internal differentiation (SD or ED), which is why it is difficult to conclude the value of the results.
Answer: Corrected as suggested. The SD values are now included (mean±SD) as well as the SE.
Comment 12. Tables - for decimals, please use dots instead of commas.
Answer: Corrected as suggested
Comment 13. In conclusion, I would add specific crops that the measurements that were carried out. Not just generally - more and less susceptible to imazethapyr.
Answer: Corrected as suggested
We thank you for the time and effort you took to review our manuscript.
Author Response File: Author Response.pdf
