PONE-D-21-33766Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat and turnipweed in wheatPLOS ONE

Dear Dr. Rastgoo,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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We look forward to receiving your revised manuscript.

Kind regards,

Ahmet Uludag, Ph.D.

Academic Editor

PLOS ONE

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Additional Editor Comments:

Both reviewers have given very good hints to make this paper more valuable. I hope you will follow their suggestions. I think a month is enough to make all these. However, I can give you longer duration if you need. I am not going to rewrite reviewers words here.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

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?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

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: Unpredicted variability in temperature is associated with frequent extreme low-temperature events. And it is threatening global food security. Due to the lack of information on the efficacy of herbicides under freezing stress, this research was conducted to evaluate the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat and turnipweed under freezing stress before and after herbicide application. The research results have guiding significance for production, but the article still needs to be revised and improved.

（1）The introduction part should increase the impact of global climate change on the occurrence of low temperature adversity events. And its impact on wheat production.

（2）Experimental site and plant material part：Explain the amount of fertilizer used and the ratio of N, P, K.

（3）The source of the meteorological data in Figure 1 should be clearly explained.

（4）Results part: The description of the results needs to be more in-depth. For example, how much does FBS significantly increase or decrease compared to NF?

（5）The discussion part is not deep enough, some physiological discussions should be added.

（6）The description in the conclusion part needs to be refined to increase readability.

（7）There are grammatical and editorial issues somewhere in this paper. Many of them could cause difficulties to the readers.

（8）The article does not have data on wheat yield. It would be very good if we could increase the data on the effects of herbicides before and after low temperature on wheat yield.

Reviewer #2: Dear Authors,

Reviewer comments PONE-D-21-33766

The manuscript entitled „Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat and turnipweed in wheat“ represents a useful study focused on the impact of freezing stress on the efficiency of two herbicides treatments, clodinafop-propargyl and 2,4-D plus MCPA treatments, resulting in the statement that freezing stress attenuates the efficiency of herbicides in weed elimination as indicated by a decreased selectivity index. This result also indicates that freezing increases potential harmfulness of these herbicides for wheat as a crop.

I have only a few minor comments on the present manuscript:

In Materials and methods, Statistical analysis, as well as in Results, Table 1, a kind of a post-hoc test following ANOVA analysis used for determination of significant differences, has to be specified both in Materials and methods, Statistical analysis section and in results, Table 1 legend. Moreover, in Table 1 legend, it has to be clearly stated that the presented data represent mean±SE, and the number of replicates has to be specified.

Further formal comments on the manuscript text:

Abstract, lines 37-38: Add a comma both preceding and following the words „for 2,4-D plus MCPA,…“

Introduction, line 53: Use the term „winter cereals“ instead of „autumn cereals“ for autumn-sown cereals which have vernalization requirement thus they require winter temperatures prior to flowering.

Introduction, line 59: Use the term „optimum climatic conditions“ instead of „normal climatic conditions“.

Materials and methods, line 109: Replace the verb „obtain“ with „induce“ in the statement „To induce plant cold acclimation, the plants were kept outdoors,…“

Materials and methods, line 125: The authors wrote that „Freezing treatment was performed using a thermogradient freezer.“ However, basic information on the freezer has to be given including the manufacturer (company). Was the freezing treatment always just to -4 °C?

Line 175: Remove „the“ preceding the term „relative potency“.

Line 182: Mdify the statement as follows: „Relative potency < 1 indicates the decreased effectiveness of the herbicides under freezing stress with respect to optimum conditions.“

Results, Figure 4 legend, line 209; Figure 5 legend, line 246: Write rather „64 g ha-1“, not „64 g ai ha-1“.

Results, line 252: Use a plural form of the verb „were“ instead of „was“ in the statement „The GR50 of NF, FBS, and FAS were 176, 299, and 208 g ha-1, respectively…“

Line 263: Remove „the“ preceding the term „selectivity index“ and modify the statement as follows: „As shown in Table 2, selectivity index values for clodinafop-propargyl in NF, FBS, and FAS treatments were 2.4, 0.91, and 0.78, respectively, and, for 2,4-D plus MCPA, they were 2.6, 0.12, and 0.88, respectively.“

Discussion, line 299: Remove „the“ Preceding the chemical name „clodinafop propargyl“.

Discussion, line 332: Replace the word „less“ with „lower“ in the statement „The relative potency values of clodinafop-propargyl and 2,4-D plus MCPA were lower than one under both FBS and FAS treatments in the present experiment.“

Discussion, line 343: Modify the statement into a plural form as follows: „The LD10 values of 2,4-D plus MCPA in NF were 89% and 48% higher compared to FBS and FAS, respectively.“

Final recommendation: Accept after a minor revision.

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Reviewer #1: No

Reviewer #2: No

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PONE-D-21-33766R1Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat and turnipweed in wheatPLOS ONE

Dear Dr. Rastgoo,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Your reasoning why you have chosen mix of two auxinic ingredients and discussion needs to be improved although it has been successful already. You need to use latin names of weeds in title and abstract as well. You need further information to be given about herbicides such as safener in clodinafop if there is, acid equvalent rates of auxicinic herbicides, separated amounts of 2,4-D and MCPA. I am not sure if you need both ED50 and GR50. By the way, no GR50 calculation in your materials and methods.  Please see comments in this e mail and attched sanitized manuscripts.  

Please submit your revised manuscript by 19 May 2022. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Ahmet Uludag, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments (if provided):

The topic and manuscript are good in general. However, there are some more issues needs to be changed in your manuscript and issues needs to be explained further. I was not able to find out the number of weeds (density) in experimental area. This can help you explain what you thought 93% control is not acceptable if you have very high numbers of weeds. Especially suggestions of reviewer three on discussion session needs special care.

Please do not skip my personal suggestions either.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Partly

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

Reviewer #2: Yes

Reviewer #3: Yes

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

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #3: Yes

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6. 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 #2: Reviewer comments PONE-D-21-33766R1

The revised manuscript entitled „Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat and turnipweed in wheat“ was significantly improved by the authors according to my previous comments.

I think that the revised manuscript is suitable for publication in PLoS One.

I have only a few formal comments on the revised manuscript which are provided below:

Abstract, line 23: The abbreviations „2,4-D“ and „MCPA“ have to be explained when used for the first time.

Abstract, line 29 and the further text including the figures: The „ai“ used in the „g ai ha-1“ has to be explained. What is the difference between „g ai ha-1“ and „g ha-1“??

Line 134: Add a comma following the words „Each morning at 5:30 A.M., …“

Line 337: Use the term „optimal non-freezing conditions“ instead of „normal conditions.“

Final recommendation: Accept after a formal revision.

Reviewer #3: In general, the paper is interesting and the topic nice. However, several serious issues ought to be addressed. First of all, I absolutely disagree with statements of the authors (in the abstract and the text) that the occurrence of freezing stress around herbicides application is one of the most important factors influencing their performance. Many more factors (rate, growth stage, adjuvant, weed species, water volume, pressure etc) play a significant role on that. Moreover, survival 7% means efficacy of 93%, farmers do not need to double the rate to achieve a complete control (usually farmers are satisfied with an efficacy of 90-95%). In wild oat authors say that under no freezing (NF) and freezing after spray (FAS) conditions, winter wild oat was completely controlled with the recommended dose of clodinafop-propargyl, while at FBS control was 93%. Why the ranking of LD50 and GR50 was different (meaning the FBS between the two others)? There is an important confusion. Furthermore, I would encourage authors a) to justify their choice to evaluate an ACCase inhibitor alone and a mixture of auxinic herbicides (and not an individual)? and b) substantially revise and improve the discussion section in order to talk more about the potential mechanism behind that reduced efficacy and the potential generalization of their results (since in many cases farmers chose to spray under low temperatures since the notice an increased susceptibility of several weeds to glyphosate and other herbicides). Consequently, I recommend a major revision of the paper.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes: Klára Kosová

Reviewer #3: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

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Submitted filename: Reviewer comments PONE-D-21-33766R1.docx

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PONE-D-21-33766R2Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat (Avena ludoviciana Durieu) and turnipweed [Rapistrum rugosum (L.) All.] in wheatPLOS ONE

Dear Dr. Rastgoo,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

You will see my comments  below part of this e mail. I would like to say repeating issue is  still acceptable bu Plosone. Could you please check all suggestions and recheck any missing point from the earliest reviews. 

Please submit your revised manuscript by 30.09.2022. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Ahmet Uludag, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (if provided):

In each cycle, a reviewer ended up with major revision. It think the reason is writing style, statistical problems, not strong interpretations. It does not mean totally unacceptable but, makes your paper problematic. I think you need some editorial improvement in your manuscript as reviewer's mentioned although this is not a problem of text throughout. Could you please check again your statistical analyses with a help a biometrician to make sure your interpretations. In addition, reviewer 4 has pointed out a few misleading statements in the manuscript, which need to be revised according to my suggestions. Furthermore, one more time visit earlier suggestions on earlier version's of the manuscript.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #4: (No Response)

Reviewer #5: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #4: Partly

Reviewer #5: Yes

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

Reviewer #4: No

Reviewer #5: Yes

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

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #4: Yes

Reviewer #5: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #4: No

Reviewer #5: Yes

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6. 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 #4: The manuscript investigated the response of two problematic weed species to herbicides under freezing conditions. I have realized that some other colleagues had reviewed the manuscript. I also realized that one of the reviewers raised a concern regarding the language of the manuscript, but the authors rebutted this comment and claimed that their manuscript had been edited by “a Native English-speaker colleague”. Please note that being a native English speaker is not synonymous with being a good writer. After reading the manuscript, I also feel the language is not comprehensive in many places and does not meet the minimum standard required by international journals. Hence, I recommend editing help from a language service.

Specific comments

Lines 24 and 68- Please spell out these abbreviated herbicide names. The idea is that the manuscript should be comprehensive to any readers, not only to those who share the same area of expertise as the authors.

Line 100 “seeds of winter wild oat and turnipweed” How many plants did you sample? Did the samples size sufficiently represent the population?

Line 105 “The seeds were placed in” I assume this refers to weed seed germination. How about wheat?

Line 109 “same time on October 21 (first run) and 24 (second run) in 2019”. I would not call this “a repeat in time”. We usually repeat experiments in time to cover sufficient spatial and temporal variation. Repeating an experiment 3 days after the other does not meet this criterion.

Line 111 Nutrient deficiency can affect herbicide efficiency. The authors claimed they had not added any fertilizer to the potting media. This needs to be justified in the manuscript; otherwise, it is a major flaw.

Line 112 Inconsistency. In Line 106-108, the authors claimed that they had pregerminated the seed of each weed species, and subsequently, they transplanted the seedlings. But here, they claimed they planted the seed directly. Please justify.

Line 113 Only eight plants per replicate? Would this number of plants sufficiently represent the population sample? Since the authors estimated the LD50, I suspect the low number of plants used in this experiment would sufficiently represent the variability among individuals of sampled populations? How many pregerminated seeds did you place in each pot for turnipweed?

Line 115 “irrigated according to daily requirements” How?

Line 114 How did you grow wheat?

Lines 132-133 “The pots were kept at -4℃ for 1 h and then removed from the freezer (Fig. 2).” I can`t follow this part. The plants were supposed to be kept in a freezer from 7pm to 5 am. But the authors remove the plants after 1 h exposure to -4 °C. Does this mean that the temperature reached to 4 °C at about 4 am?

Line 157 “herbicides” Please give more details than “herbicides”. What was the formulation? Did you use any wetting agents? What was the trade name of each herbicide? What was the portion of each herbicide in the mixture of 2,4-D and MCPA?

Line 174 “the curve around e, and e is the effective dose”, The letter “e” is used in two different places in the equation. Which one represents the “effective dose”?

Line 195- 196 “Afterward, differences between treatments …” This is not really a solid approach to compare treatments statistically.

Line 197-198 Pooling data from two experiments is a poor practice and results in missing many effects that are needed. The most appropriate statistical approach is to use a “mixed model analysis”. Please see the manuscript below for more information.

Piepho, H. P., A. Bu¨chse, and K. Emrich, 2003: A hitchhiker’s guide to the mixed model analysis of

randomized experiments. J. Agron. Crop Sci. 189, 310—322.

Line 207, As one of the reviewers noted, 7% is really negligible in terms of herbicide efficacy. But As noted by the authors (Table 2), it appears that freezing treatments reduced the tolerance of wheat to the ACCase inhibiting herbicide, suggesting using 2-fold the recommended rate would definitely damage the crop.

Line 228 In Table 1, the authors indicated the significance levels using asterisks; however, it is not clear what the asterisks represent. I assume they represent the significance of the parameters in the model rather than the differences between treatments. The authors need to use an appropriate statistical analysis to compare the estimated parameters between treatments, and the significant differences between treatments can be demonstrated using letters “e.g. a, b, etc.).

Line 272 “However, LD10 of wheat in” The authors must include the dose-response results for wheat treated with both herbicides under all three conditions.

Line 286 “nights before and after spray reduces the efficacy of this herbicide compared to NF.” This statement is misleading. First of all, the differences in LD50 values appear to be negligible. Secondly, there is natural variation among the individuals of a population resulting in varied responses to herbicides. So such a low difference can be due to variability among individuals.

Lines 289-289 “Freezing treatment for three nights before and after spraying of clethodim herbicide (the mode of action similar to clodinafop-propargyl) led to an increase in LD50 “. In the manuscript by Saini et al. 2017, the authors claimed that “Addition of frost treatments did not markedly affect survival of the S population” and eventually noted no significant differences. I feel in the current manuscript, the authors have misinterpreted their results.

Line 300- 301 “Herbicide translocation in the phloem in freezing conditions could decline due to damage to the phloem cells” The freezing treatment only lasted for 3 days; thus, I suspect impaired translocation could explain the outcome of this research. I guess in your case, freezing temperature reduced herbicide absorption.

Lines 314-322, 333-337 and 360-361 The physiological mechanisms underlying the response of plants to frost are different from those of low (cold) temperatures. So the response of plants to herbicides would be different under freezing versus low temperatures. For instance, for turnipweed, the freezing treatment before herbicide application reduced the efficacy of 2,4-d and MCPA treatment later. As there were no herbicide molecules in the plants at the time of freezing treatment, reduced translocation of the herbicide can not solely explain reduced herbicide efficacy in FBS plants.

Line 370 “their effectiveness in controlling narrow leaf…” you only studied one grass species. So this may not be true for other “narrow-leaved” species.

Line 374-375 “Therefore, the use of these herbicides in the mentioned conditions (FBS for clodinafop-propargyl and FAS for 2,4-D plus MCPA) is recommended.” This is misleading, as it appears the desirable crop (wheat) loses its tolerance to both herbicides under both freezing conditions. So, it is not a good idea to use these herbicides under freezing conditions to control weed species in wheat. Please revise accordingly.

Reviewer #5: The authors incorporated the comments of both reviewers in true sense. The revised draft shaped well and can be accepted for publication.

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Reviewer #4: No

Reviewer #5: No

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PONE-D-21-33766R3Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat (Avena ludoviciana Durieu) and turnipweed [Rapistrum rugosum (L.) All.] in wheatPLOS ONE

Dear Dr. Rastgoo,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

I have read your manuscript carefully. I have seen some points confusing. I hope you can explain them or make necessary changes. Your manuscript is almost ready for acceptance.

Please submit your revised manuscript by 20 September 2022. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

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We look forward to receiving your revised manuscript.

Kind regards,

Ahmet Uludag, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments:

I have shown my questions and concerns on the manuscript named editor which is attached. You can avoid concerns of reviewer 5 about tables and figures that are checked by I am. By the way you have criticized me on different reviewers. Unfortunately the same reviewers did not reviewed again. This is the biggest problem of academic editors which colleagues like publishing but not reviewing. An academic editor does not gain anything from reviewing except happiness from contributing colleagues. If I can get responds to my concerns on the text clearly and you can make necessary changes I will accept your paper immediately.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #6: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #6: Partly

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

Reviewer #2: Yes

Reviewer #6: Yes

**********

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

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #6: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #6: No

**********

6. 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 #2: Reviewer comments PONE-D-21-33766R3

The revised manuscript entitled „Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat (Avena ludoviciana Durieu) and turnipweed (Rapistrum rugosum (L.) All.) in wheat“ was improved by the authors in accordnace with my and other reviewers´ comments.

I have no further major comment on the revised manuscript.

I have only a few formal comments on the revised manuscript which are given below:

Introduction, line 64: Add the word „fact“ in the statement „Based on this fact,…“

Materials and methods, line 116: Add the verb „were“ in the statement „…were transferred to pots…“

Discussion, line 310: Add the word „respectively“ at the end of the statemnt „the efficacy of 2,4-D plus MCPA under FBS and FAS was 1.6 and 1.1 times lower compared to NF, respectively.“

Final recommendation: Accept.

Reviewer #6: Line 160-161: This sentence is very confusing. How did you measure the survivability % by placing the samples in an oven?

Table 1: This table is very confusing. The table mentioned “the dose of clodinafop-propargyl and 2,4-D plus MCPA required for 50% mortality (LD50) and the dose that caused 50% inhibition of growth noted by biomass (GR50) of winter wild oat and turnipweed under different freezing treatments. But where is the data for the two weeds in the table?

Table 2: Confusing. The table mentioned “sensitivity of plant species of wheat, winter wild oat, and turnipweed to clodinafop-propargyl and 2,4-D plus MCPA under different freezing treatments”. But the table presents wheat and weed data for the freezing treatment.

Figure 4: Survival (A) and dry weight (B) of winter wild oat treated with various doses of clodinafop-propargyl under different freezing treatments but why these data were not presented for the turnipweed?

Figure 5: Survival (A) and dry weight (B) of turnipweed treated with various doses of 2,4-D plus MCPA under different freezing treatments but why these data were not presented for the weed wild oat?

It is very difficult to understand the message from the current conclusion. Please reduce the length of the conclusion and focus on the most important finding of the research and take-home message.

**********

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Reviewer #2: Yes: Klára Kosová, Ph.D.

Reviewer #6: No

**********

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Attachments

Attachment

Submitted filename: Reviewer comments PONE-D-21-33766R3.docx

Attachment

Submitted filename: PONE-D-21-33766_R3 editor.pdf

PONE-D-21-33766R4Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat (Avena ludoviciana Durieu) and turnipweed [Rapistrum rugosum (L.) All.] in wheat (Triticum aestivum L.)PLOS ONE

Dear Dr. Rastgoo,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

You will see my concerns on the attached document. SOme of them can be skipped but the others should be changed. You can differentiate. The most important point should be added is the role of safener on efficacy of clodinafop. I recommend you check old papers related to former safener of fenoxaprop.

Please submit your revised manuscript by 30 October 2022. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Ahmet Uludag, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments:

Dear colleague, I have checked your manuscript myself because most of the issues about former version were mine. I read again your manuscript and showed my suggestions. My aim is to make this invaluable data more attractive. However, the role of safener for clodinafop efficacy and effect was skipped in the manuscript. It is definitely needed. I am not suggesting an experiment on this stage but you should discuss this and add all relevant places of the manuscript. I also wonder if there is a role of formulation especially for hormon-like herbicide. I have no idea about it, it will be nice to add such discussion points to paper if possible.

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Reviewers' comments:

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachments

Attachment

Submitted filename: PONE-D-21-33766_R4 Academic editor.pdf

Freezing stress affects the efficacy of clodinafop-propargyl and 2,4-D plus MCPA on wild oat (Avena ludoviciana Durieu) and turnipweed [Rapistrum rugosum (L.) All.] in wheat (Triticum aestivum L.)

PONE-D-21-33766R5

Dear Dr. Rastgoo,

Thank you for submitting your revised manuscript to PLOS ONE; I sincerely apologise for the unusually delayed review timeframe. 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.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Emily Chenette

Editor in Chief

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

**********

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

Reviewer #2: Yes

**********

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

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

**********

6. 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 #2: I have no further comments on the revised manuscript. I can now recommend its prompt publication in PLoS One.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

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Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes: Klára Kosová, Ph.D.

**********