Parental effects of Bt toxin and vitamin A on Helicoverpa armigera

Carmen López; Daniela Zanga; Alejandro Juárez-Escario; Pilar Muñoz; Matilde Eizaguirre

doi:10.1371/journal.pone.0269585

Peer Review History

Original SubmissionFebruary 14, 2022
6 Apr 2022 Decision Letter - Patrizia Falabella, Editor PONE-D-22-04499Parental effects of Bt toxin and vitamin A on Helicoverpa armigeraPLOS ONE Dear Dr. Malak 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 manuscript has now been reviewed, and the reviewers' comments are appended below. You will see that, while they find your work of interest, they have raised some points that need to be addressed before we can make a decision on publication. Please submit your revised manuscript by May 21, 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. 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The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf. 2. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section. 3. Thank you for stating the following in the Acknowledgments Section of your manuscript: [We are grateful to Ministerio de Ciencia e Innovación (Spanish government) for financial support. The authors thank Teresa Estela her administrative support and Joan Safont for his invaluable help in the general rearing of insects. Thanks to Professor Ramon Albajes for his unconditional support.] We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: [The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.] Please include your amended statements within your cover letter; we will change the online submission form on your behalf. 4. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well. [Note: HTML markup is below. Please do not edit.] 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 ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 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 ****** 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: No Reviewer #2: Yes ******** 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: Comments: This paper by López et al. “Parental effects of Bt toxin and vitamin A on Helicoverpa armigera” has researched the effect of vitamin A (β-carotene) and Bt protein on H. armigera larvae and their offspring. The work could be helpful to better understand the transgenerational effect of vitamin and Bt protein, and the potential resistance of H. armigera to Bt crops. In general, the experimental design of the paper is good and the data provided consequent, the information amassed is interesting. However, the manuscript is not acceptable before some experiment should be added. Meanwhile, the language quality of the MS should be improved. I hope my comments could help this interesting work Introduction The paragraphs of the Introduction is much too long. And there are too many long sentences, many of them are hard to understand. Materials and methods Insect rearing “H. armigera larvae were collected in maize or alfalfa crops in Lleida”. Host adaptability of insect has been found to be dependent on many factors, the authors need to be clear that the larvae were collected from maize? or alfalfa crops? Diets “……, a 25% of maize flour was replaced by lyophilized Bt or non-Bt maize leaves, adding or not 0.6 % of β-Carotene”. The author needs to explain that why diets and 25% maize folur, 0.6% β-Carotene. Is there any literature on this? What was the dosage of Cry1Ab protein in 25% of Bt maize flour? Experiments Experiments with the F1 larvae and adults “Larval mortality, duration of larval development and pupal weight were recorded”. Were the insect observed and recorded the mortality daily? What is the standard of death? Why were there 6 females and 5 males in the mating cage? Please clearly state the number of insect for each experiment. Statistical analysis “using a generalized linear model with a binomial distribution” corrected to “using a generalized linear model (GLM) with a binomial distribution”. I recommend that you use this symbol instead of the common abbreviations throughout the manuscript. Results F1 mating and fertility “……but neither β-carotene nor the interaction Bt-β-carotene had any effect”, the result of this sentence was not show in Fig. 4. F2 Development and Mortality Can the authors clarify why they did not research on the β-carotene to F2 development and mortality? Figure note should be added in Fig. 7. Discussion The paragraphs of the Discussion is much too long “However, the life-history-traits affected by phytophagous feeding……”, please cite literature references. “……in previous work with the same species [8] and with other nearby species such as H. zea [22]”, the authors should be discussed the differences between references and this research. “Maternal o paternal effects due to……” corrected to “Maternal or paternal effects due to……” Reviewer #2: Reviewer Comments for PONE-D-22-04499 The manuscript entitled “Parental effects of Bt toxin and vitamin A on Helicoverpa armigera” by Carmen López, Daniela Zanga, Alejandro Juárez-Escario, Pilar Muñoz, Matilde Eizaguirre provides advances in the existing knowledge on Bt toxins for crop protection. Introduction developed instead of development Add more reference against each already mentioned “H. armigera is a serious cosmopolitan insect pest whose larvae are polyphagous [9] and have a high capacity to develop resistance to insecticides [10] and to the Bt toxin [11]. Rephrase “that feed on maize mainly on the silks (styles) among many other crops such as cotton or alfalfa.” (to the Bt toxin) and although part of the larvae feed on transgenic maize diet, another considerable part survives and pupates. were collected from maize or alfalfa crops in Lleida 16:8 light: dark (h) photoperiod relative humidity (>60%) the pupae were placed in the adult-emerging conditions at 25ºC Rephrase this “(cylindrical methacrylate tubes of 25 cm Ø and 40 cm high, closed at the base with an expanded polystyrene lid and at the top with a white gauze) with a drinker with honey solution and a cotton ball for egg laying.” 80⁰C until use. diet made with non-Bt leaves (non-Bt), rewrite in scientific way (5.5 cm ø x 3cm high) and use X from symbols not from alphabets correct the space between degree and Celsius throughout the text of manuscript (e.g., 25 º C) Better to use symbolic multiplication instead of others……for (6 cages* 6 females/cage) and 30 males (6 cages* 5 males/cage) for each condition. & ANOVA was used (the factors were, diet * sex) Maintain space with * for two-way (parentsdiet) ANOVA on development duration Maintain space (Bt β-carotene). Space between fig and number (e.g., Fig1 = Fig 1) Results Percent mortality of the Helicoverpa Discussion Please update with latest references Overall, please carefully crosscheck the reference mentioned in the text and list. ******** 6. 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 #1: No Reviewer #2: Yes:** Abid Ali [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. https://doi.org/10.1371/journal.pone.0269585.r001
Revision 1
19 May 2022 Author Response PONE-D-22-04499 Parental effects of Bt toxin and vitamin A on Helicoverpa armigera PLOS ONE Patrizia Falabella Academic Editor and Reviewer's Questions PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf. 2. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section. Now Corrected 3. Thank you for stating the following in the Acknowledgments Section of your manuscript: [We are grateful to Ministerio de Ciencia e Innovación (Spanish government) for financial support. The authors thank Teresa Estela her administrative support and Joan Safont for his invaluable help in the general rearing of insects. Thanks to Professor Ramon Albajes for his unconditional support.] We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: [The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.] Please include your amended statements within your cover letter; we will change the online submission form on your behalf. Done L 394 4. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well. Done, Line 108 [Note: HTML markup is below. Please do not edit.] 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 ________________________________________ 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ________________________________________ 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 ________________________________________ 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: No The document has now been revised by an official English Editing services. We have the certificate showing it. Reviewer #2: Yes ________________________________________ 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: Comments: This paper by López et al. “Parental effects of Bt toxin and vitamin A on Helicoverpa armigera” has researched the effect of vitamin A (β-carotene) and Bt protein on H. armigera larvae and their offspring. The work could be helpful to better understand the transgenerational effect of vitamin and Bt protein, and the potential resistance of H. armigera to Bt crops. In general, the experimental design of the paper is good and the data provided consequent, the information amassed is interesting. However, the manuscript is not acceptable before some experiment should be added. Meanwhile, the language quality of the MS should be improved. I hope my comments could help this interesting work Introduction The paragraphs of the Introduction is much too long. And there are too many long sentences, many of them are hard to understand. Done. The document has now been revised by an official English Editing services Materials and methods Insect rearing “H. armigera larvae were collected in maize or alfalfa crops in Lleida”. Host adaptability of insect has been found to be dependent on many factors, the authors need to be clear that the larvae were collected from maize? or alfalfa crops? The larvae of all experiments were collected only in alfalfa fields. Now this point has been corrected in M&M Line 108 Diets “……, a 25% of maize flour was replaced by lyophilized Bt or non-Bt maize leaves, adding or not 0.6 % of β-Carotene”. The author needs to explain that why diets and 25% maize folur, 0.6% β-Carotene. Is there any literature on this? What was the dosage of Cry1Ab protein in 25% of Bt maize flour? Done. We have rewritten lines 129-136 adding the data and necessary references Experiments Experiments with the F1 larvae and adults “Larval mortality, duration of larval development and pupal weight were recorded”. Were the insect observed and recorded the mortality daily? Yes, daily, line 149. What is the standard of death? We consider the standard mortality, the mortality on the non-Bt diet Why were there 6 females and 5 males in the mating cage? In the M&M, Insect General Rearing we have added the explanation with the reference of why we used 6 females and 5 males per cage, lines 114-115 Please clearly state the number of insect for each experiment. Each experiment indicates the number on insects tested. Lines 145, 171 Statistical analysis “using a generalized linear model with a binomial distribution” corrected to “using a generalized linear model (GLM) with a binomial distribution”. I recommend that you use this symbol instead of the common abbreviations throughout the manuscript. Done Results F1 mating and fertility “……but neither β-carotene nor the interaction Bt-β-carotene had any effect”, the result of this sentence was not show in Fig. 4. We apologized for the unclear presentation of these results. Now, we have added the statistics of the effect of β-carotene in lines 244-246, we have also modified the X-axis the Figure 4 and the Figure 4 caption (lines 251-253) to explain in a better way. F2 Development and Mortality Can the authors clarify why they did not research on the β-carotene to F2 development and mortality? Figure note should be added in Fig. 7. Again, we apologized for the unclear presentation of these results. Now, we have added the statistics of the effect of β-carotene in lines 298-299, we have also modified the X-axis in the Figure 7, and the Figure 7 caption (lines 304-306) to explain in a better way. Discussion The paragraphs of the Discussion is much too long Done. The document has been revised. “However, the life-history-traits affected by phytophagous feeding……”, please cite literature references. The literature references appear in line 314 “……in previous work with the same species [8] and with other nearby species such as H. zea [22]”, the authors should be discussed the differences between references and this research. Done, lines 329-332 “Maternal o paternal effects due to……” corrected to “Maternal or paternal effects due to……” Changed in the new version, line 360 Reviewer #2: Reviewer Comments for PONE-D-22-04499 The manuscript entitled “Parental effects of Bt toxin and vitamin A on Helicoverpa armigera” by Carmen López, Daniela Zanga, Alejandro Juárez-Escario, Pilar Muñoz, Matilde Eizaguirre provides advances in the existing knowledge on Bt toxins for crop protection. Introduction developed instead of development Changed in the new version Add more reference against each already mentioned “H. armigera is a serious cosmopolitan insect pest whose larvae are polyphagous [9] and have a high capacity to develop resistance to insecticides [10] and to the Bt toxin [11]. Done, added more references. L74-77 Rephrase “that feed on maize mainly on the silks (styles) among many other crops such as cotton or alfalfa.” Eliminated in the new version (to the Bt toxin) and although part of the larvae feed on transgenic maize diet, another considerable part survives and pupates. Rewritten in the new version were collected from maize or alfalfa crops in Lleida Changed, line 108 16:8 light: dark (h) photoperiod Corrected relative humidity (>60%) Corrected the pupae were placed in the adult-emerging conditions at 25ºC Rephrase this “(cylindrical methacrylate tubes of 25 cm Ø and 40 cm high, closed at the base with an expanded polystyrene lid and at the top with a white gauze) with a drinker with honey solution and a cotton ball for egg laying.” All M&M have been rewritten 80⁰C until use. -80 º C Corrected diet made with non-Bt leaves (non-Bt), M&M have been rewritten rewrite in scientific way (5.5 cm ø x 3cm high) and use X from symbols not from alphabets Done correct the space between degree and Celsius throughout the text of manuscript (e.g., 25 º C) Done Better to use symbolic multiplication instead of others……for (6 cages* 6 females/cage) and 30 males (6 cages* 5 males/cage) for each condition. & ANOVA was used (the factors were, diet * sex) Maintain space with * for two-way (parentsdiet) ANOVA on development duration Done Maintain space (Bt β-carotene). Done Space between fig and number (e.g., Fig1 = Fig 1) Done Results Percent mortality of the Helicoverpa Corrected Discussion Please update with latest references We have used the latest references we have found Overall, please carefully crosscheck the reference mentioned in the text and list. ________________________________________ 6. 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 #1: No Reviewer #2: Yes: Abid Ali Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0269585.r002
25 May 2022 Decision Letter - Patrizia Falabella, Editor Parental effects of Bt toxin and vitamin A on Helicoverpa armigera PONE-D-22-04499R1 Dear Dr. Eizaguirre, 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, Patrizia Falabella Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0269585.r003
Formally Accepted
23 Jun 2022 Acceptance Letter - Patrizia Falabella, Editor PONE-D-22-04499R1 Parental effects of Bt toxin and vitamin A on Helicoverpa armigera Dear Dr. Eizaguirre: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. 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. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Prof. Patrizia Falabella Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0269585.r004

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