Biocontrol of Cercospora leaf spot in sugar beet by a novel Bacillus velezensis KT27 strain: Enhanced antifungal activity and growth promotion in laboratory and field conditions

Agnieszka Wita; Wojciech Białas; Katarzyna Czaczyk; Agnieszka Drożdżyńska; Łukasz Sobiech; Monika Grzanka; Jakub Danielewicz; Ewa Jajor; Joanna Horoszkiewicz; Roman Marecik

doi:10.1371/journal.pone.0323889

Peer Review History

Original SubmissionFebruary 13, 2025
13 Feb 2025 Author Response https://doi.org/10.1371/journal.pone.0323889.r001
26 Feb 2025 Decision Letter - Estibaliz Sansinenea, Editor PONE-D-25-07815Use of Bacillus valezensis KT 27 induced by fungi in the protection of sugar beets against Cercospora Leaf Spot (CLS)PLOS ONE Dear Dr. Marecik, 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. Please submit your revised manuscript by Apr 12 2025 11:59PM. 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 are unable to adhere to our open data policy, please kindly revise your statement to explain your reasoning and we will seek the editor's input on an exemption. Please be assured that, once you have provided your new statement, the assessment of your exemption will not hold up the peer review process. Additional Editor Comments: The reviewers have commented about the Ms and reccommended major revision before its acceptation. I invite the authors to respond carefully all reviewers comments doing the neccessary changes . [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: Partly Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: No 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: No 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: No ****** 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: 1-The submitted manuscripts has significant technical weaknesses and lacks novelty. The writing quality is poor, and the ideas are repetitive. The researchers used a single bacterial strain alongside three pesticides, which creates an unfair comparison. It would have been more appropriate to include multiple bacterial strains for a more balanced evaluation. Additionally, the methodology requires substantial revision to ensure clarity and robustness. Must of the results i can't believe it .2-The images in Figure 1 indicate the absence of a proper control treatment. Although the researcher claims that one of the wells contains a control treatment, a thorough review reveals that the so-called control treatment actually impacts the fungus. This is both unacceptable and ethically inappropriate.3-Please revise the research and simplify the sentences for clarity. The section on materials and methods needs to be rewritten in a clear and organized manner. The results are not presented in a logical sequence and should be restructured accordingly. Additionally, the discussion is weak and requires improvement. It is recommended to incorporate the suggested sources identified in the research to strengthen the discussion.4-The discussion section requires the addition of relevant sources and the removal of those that are not directly applicable to the topic.5- No images of molecular analysis, field experiment, images of treated and untreated plants, results of MALDI-TOF MS . Reviewer #2: There are some suggestions and recommendations to improve the study impact. • Further Studies on Mechanisms: Future research should focus on elucidating the precise mechanisms of action of Bacillus velezensis KT27, including the roles of specific metabolites in antagonism. • Broader Field Trials: Expanding the field trials to different geographical locations and crop types could help validate the versatility of the strain as a biocontrol agent under diverse environmental conditions. • Integrated Pest Management (IPM): Research should investigate the combination of Bacillus velezensis KT27 with other biocontrol agents or cultural practices in Integrated Pest Management (IPM) systems to assess its compatibility and potential synergistic effects. • Ecotoxicological Assessments: More comprehensive studies on the environmental impact of the strain, including its effects on non-target organisms and soil health, would be beneficial for its eventual commercial application. ****** 6. 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Revision 1
14 Apr 2025 Author Response Dear Reviewers, We would like to express our deepest gratitude for your thoughtful and constructive comments regarding our manuscript. Your insightful suggestions have been of great value to us, and we sincerely appreciate the time and effort you invested in reviewing our work. We are confident that the revisions made in response to your remarks have considerably improved the scientific rigor, clarity, and overall quality of the manuscript. During the revision process, we also devoted careful attention to refining the language, as well as enhancing the technical and formal aspects of the manuscript. To facilitate your review of the revised version, we have enabled the “Track Changes” feature to clearly indicate all substantial modifications. We have also included responses to your comments directly in the form of annotations. Nevertheless, we would kindly point out that the final, clean version of the manuscript best represents the full extent of the improvements made. As part of the revision process, we have amended the manuscript title, which we believe now more accurately reflects the scope and essence of the study. Additionally, we have revised the titles of several subsections to improve structure and readability. Following your valuable suggestions, we have also updated the reference list and included photographs from the sugar beet field trials in which our bacterial biopreparation was applied. We would also like to take this opportunity to clarify that the illustrative images provided reflect fungistatic activity observed under in vitro conditions. The well on the left side of the photograph demonstrates the activity of the bacterial biomass of Bacillus velezensis KT27, whereas the well on the right side corresponds to the post-culture fluid devoid of bacterial cells. We did not include images from the control treatment, as the application of sterile saline into wells on control plates did not result in any inhibition of fungal growth (i.e., no visible clearing zones were present). In our view, these would not contribute additional informative value. We fully acknowledge that this is a broad and multifaceted topic, and that there are various possible approaches to its investigation. At the same time, we wish to emphasize that verifying laboratory results under field conditions—though often complex—is critical for confirming the efficacy of a biological preparation in practical agricultural settings. We stand by the reliability and validity of the results we have obtained. While numerous studies are focused on identifying novel microbial strains with unique metabolic capacities for plant protection, we would like to highlight that the induction of fungistatic activity in microorganisms—such as the one we demonstrated—is still relatively underexplored. We sincerely hope that the publication of our findings will encourage further research others into this promising and important area. We wholeheartedly agree with your recommendation to continue investigations along the lines outlined in the manuscript. Indeed, we are actively pursuing further research with the ultimate goal of formally registering our biopreparation, based on the B. velezensis KT27 strain, for protection against CLS, and eventually making it commercially available. In our opinion, the subject of our research is well aligned with the increasingly urgent need to reduce the reliance on conventional chemical plant protection agents, as well as to mitigate the growing problem of pathogen resistance. These efforts, alongside a reduction in mineral fertilization, may substantially contribute to the improvement of environmental conditions impacted by modern agricultural practices. Once again, we extend our sincere appreciation for your kind attention, careful review, and valuable input. We are truly grateful for the opportunity to improve our manuscript with your guidance. With the highest respect and kind regards, The Authors Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0323889.r003
16 Apr 2025 Decision Letter - Estibaliz Sansinenea, Editor Biocontrol of Cercospora Leaf Spot in sugar beet by a novel Bacillus velezensis KT27 strain: Enhanced antifungal activity and growth promotion in laboratory and field conditions PONE-D-25-07815R1 Dear Dr. Marecik, 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 will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, 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, Estibaliz Sansinenea Academic Editor PLOS ONE Additional Editor Comments (optional): The authors have followed all comments suggested by the reviewers therefore the MS has been improved and it can be accepted in the current form. Reviewers' comments: https://doi.org/10.1371/journal.pone.0323889.r004
Formally Accepted
Acceptance Letter - Estibaliz Sansinenea, Editor PONE-D-25-07815R1 PLOS ONE Dear Dr. Marecik, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, 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 customercare@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 Dr. Estibaliz Sansinenea Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0323889.r005

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