PONE-D-21-17085

The antagonistic mechanism of Bacillus velezensis ZW10 against rice blast disease and its evaluation as a potential biopesticide

PLOS ONE

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Based on the comments of the reviewers and based on my own review of the manuscript, I recommend it for a major revision

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Reviewers' comments:

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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: Partly

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

Reviewer #1: No

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?

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Reviewer #1: No

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: Comments on MS No. PONE-D-21-17085

MS title: The antagonistic mechanism of Bacillus velezensis ZW10 against rice blast disease and its evaluation as a potential biopesticide

Rice blast caused by Magnaporthe oryzae is one of the most serious diseases of rice causing significant yield loss every year. Extensive work has been done on genetics of blast resistance and several blast resistant rice cultivars are available for commercial cultivation. A large number of highly effective chemicals are also available to manage the disease in the standing crop. However, identification and use of bio-control agents to manage rice blast disease is also equally important.

The authors may note the following points

Line No. 19: it causes the rice grain yield decreases

The author can make: it causes the rice grain yield reduction

Line 25: In the in vivo tests most of M. oryzae could not infect

The sentence should be modified properly

Lines 59-60: In recent years, the research on B. velezensis mainly focused on promoting the growth of animals and plants

The authors may provide reference please

Line 64: Dai Zhang et al.

Should it be only Zhang et al.?

Lines 68-70: Previous studies characterized the physical and chemical properties of B. velezensis ZW10 and elucidated its antagonistic activity against M. oryzae.

Please provide the reference

Line 70: In the paper

Authors can make it ‘in this paper’

Line 76: B. velezensis ZW10 was inoculated to Landy medium (10 L) to be cultured at 35 ℃

The authors are requested to make the sentence proper. The authors should also give the composition of Landy medium or any reference which details the Landy medium composition

Lines 80-84: The authors mentioned that the ethyl acetate extract had the maximum inhibitory effect against the blast fungus and this extract was then further separated using column chromatography and the running phase CH2Cl2 /MeOH at ratio of 40:60 yielded maximum elution of inhibitory compounds. What is the actual compound which gave the inhibitory activity against blast fungus?

Line 91: What was the logic in using GFP-tagged M. oryzae isolate for conidial germination study? Whether the same GFP-tagged M. oryzae isolate was used for field inoculation studies?

Line 96: the germination degree of conidia

The authors can modify the sentence

Lines 107-108: Please give the composition of ‘complete medium’ and any suitable reference. The authors also can mention the objective of this experiment. Is it to know, whether the antagonist produces any cell wall degrading enzymes?

Line 115: Infection of rice leaf sheaths by M. oryzae

The objective of this experiment is not clear

Lines 122-124: Was this experiment was done under glass house condition or inside a plant growth chamber? Whether experiment was done under a particular temperature and RH condition? Whether any treatment was kept with only spraying with CFB (no blast pathogen inoculation)?

Line 172: The fermentation broth was extracted by different polar organic solvents

But hexane is not a polar solvent

Line 189-190: In the treatment group, the germination rate of conidia was only 3.87 ±1.33% after 2 h, and higher than 90% after 8 h.

But Figure 2b shows that germination rate did not cross beyond 20% even after 48 hpi. Please check and clarify. Why in Figure 1b Y-axis it is written formation rate? Should it be germination rate? Overall quality of the figure is not good

Line 216: The antibacterial substances of ZW10 …..

Should it be antifungal?

Figure 5: Pictures/graphs are not clear

Lines 224-228: Upregulation of these genes involved in defense was only due to CFB or pathogen inoculation also has some role? The authors could have taken one treatment with only CFB application.

Lines 235-243: Application of CFB has drastically reduced the leaf and neck blast severity under field condition and equal to carbendazim. Most of the literatures suggest comparatively lower level of protection by biocontrol agents compared to recommended pesticide. How many sprays of CFB were given? The authors could have repeated the field experiment to confirm exceptionally high level of protection by CFB. Graph showing the neck blast control is not clear. Whether any field trial was taken using the antagonist formulation?

Reference # 26 is incompletely cited

Line # 414-415: Ref # 33: 33. Ou SH. Pathogen Variability and Host Resistance in Rice Blast Disease. Ann rev phytopathol. 2014; 18(1):167-187.

Please check the year. It should be 1980

The manuscript requires improvement in its presentation style and language. Also the authors should suggest practical use of this antagonist.

Reviewer #2: Bacillus velezensis is harmless to human being so it should be an excellent candidate as bioagent controlling plant pathogens. The most interesting is that it is widely distributed so will be available easily. But in the present MS there are few issues which are needed for modification. In line no. 44 of introduction you have written which leaf blast and panicle blast are the most common and most harmful: In many places neck blast is emerging in severe form causing massive yield loss. Change the line no 47 as problems in humans and livestock, as well as pathogens are developing resistance to the fungicides instead of resistance to pathogens. Sentence no. 48 & 49 modify the sentence as Therefore, there is a growing interest in the discovery and development of new and improved fungicides based on natural products which are environment friendly as well as the...

Pseudomonas aeruginosa magna has sustained: Please check the species name. besides this is not a good example as Pseudomonas aeruginosa can cause infections in the blood, lungs (pneumonia), or other parts of the body after surgery in human. In line no. 57 B. velezensis, a new species of Bacillus, was discovered in 2005: please give reference. the growth of animals and plants , induction of systemic resistance: Please give reference. 92.07 ± 0.53% after 12 h. In the treatment group, the germination rate of conidia was only 3.87 ± 1.33% after 2 h:In case of blast more than 10,000 spores are formed from a single lesion of a susceptible variety and a susceptible variety contain more than 100 lesions in it’s leaf blades so 1.33% also is not less: How do you justify that? normal appressorium structure due to the abnormal germ tube formation (Fig. 2c):How normal appressorium can be formed due to abnormal germ tube formation? Please rectify the sentence. Please modify all the bibliography in similar style like Annual review of microbiology should be written as Annual Review of Microbiology. Journal of Cereal ence or Journal of Cereal Science?? Frontiers in plant science: please change it to maintain uniform style.

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Reviewer #1: No

Reviewer #2: Yes: Dr. Arup Kumar Mukherjee

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The antagonistic mechanism of Bacillus velezensis ZW10 against rice blast disease：evaluation of ZW10 as a potential biopesticide

PONE-D-21-17085R1

Dear Dr. Xu,

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.

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Kind regards,

R M Sundaram, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

I understand that the authors have addressed the concerns/suggestions of the reviewers. In view of this, I recommend the manuscript for publication in PLoS one

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 #1: All comments have been addressed

Reviewer #2: 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 #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

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 #1: Yes

Reviewer #2: 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 #1: Yes

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 #1: MS # PONE-D-21-17085R1

MS Title: The antagonistic mechanism of Bacillus velezensis ZW10 against rice blast disease：evaluation of ZW10 as a potential biopesticide

Comments

The authors have addressed all the queries and incorporated all the corrections and clarifications raised by the reviewers

Line # 59: was discovered in 2005[13].

Please give a space between 2005 and [13]

Line # 66: antifungal effects of LPs and VOCs released by B. velezensis

Please put the full form of LP and VOC

Line 68: on Xanthomonas oryzae pv. oryzae

pv. will not be in italics

Line # 206: broken, and; moreover

Please make necessary changes

The manuscript may be accepted for publication

Reviewer #2: The article is now modified as per the suggestions. The reference also modified. So, it may be considered for acceptance.

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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 #1: No

Reviewer #2: Yes: Dr. Arup Kumar Mukherjee