Effective Applications of Bacillus subtilis and B. amyloliquefaciens as Biocontrol Agents of Damping-Off Disease and Biostimulation of Tomato Plants
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis study is aimed to investigate the effective applications of Bacillus subtilis and B. amyloliquefaciens as biocontrol agents of damping-off disease and biostimulation of tomato plants.
The introduction provides adequate details on previous knowledge of biological control related to the given Bacillus species. The experimental approaches and data analyses are appropriate, employing suitable methods and analyses to achieve the stated objectives.
The results include significant findings, such as i) two Bacillus species were isolated from the rhizosphere of tomato plants offering as possible biocontrol agents; ii) in a greenhouse experiment, tomato plants treated with each of these antagonistic Bacillus strains significantly suppressed fungal disease, improved plant growth parameters, increased the content of photosynthetic pigments, antioxidants enzymes and total phenols and increased macronutrients content and yield during the two growing seasons.
The discussion section is offering acceptable comparisons of the obtained results with previous literature.
In the conclusion is quite short and weak and need improvements. In addition, this section should also highlight the future implications of the study and also discuss its limitations.
The study contains interesting elements that could be worthy of publication in an international journal after appropriate revisions.
Other suggestions and comments:
L52-54: Alternaria solani, Septoria lycopersici, Botrytis cinerea, Fusarium oxysporum, Pythium debaryanum, Verticillium dahliae, Rhizoctonia solani, and Phytophthora infestans – should be in italic
L61-62: P. aphanidermatum, P. debaryanum, P. spinosum, P. myriotylum and P. echinogynum, R. solani - should be in italic
L113: F. oxysporum KT224063, P. debaryanum OP823136 and R. solani – full name is needed in the title.
L123: B. subtilis – full name is needed in the title.
L130: B. subtilis and B. amyloliquefaciens against F. oxysporum, P. 130
debaryanum, and R. solani.- should be in italic and full name is needed in the title
L133: In vitro B. subtilis F. oxysporum, P. debaryanum, and R. solani. - should be in italic and full name is needed in the title
L149: Latin names should be in full in the title.
L162: Latin names should be in full in the title.
L200: Latin names should be in full in the title.
L212: Latin names should be in full in the title.
L244: Latin names should be in full in the title.
L554: J Med Plants Stud – should be in italic
In many cases, points are missing after page numbers.
Author Response
Please see the attachment
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe publication submitted to me for review on the use of Bacillus subtilis and B. amyloliquefaciens as biological control agents against tomato diseases caused by Fusarium oxysporum, Pythium debaryanum and Rhizoctonia solani is valuable and meets the current needs of sustainable agriculture. The work is solid and based on reliable experimental studies. Minor changes in the clarity of the presentation of the results, a deepening of the interpretation and a clearer emphasis on the practical significance of the research would increase its scientific value.
Below I provide comments on possible weaknesses, ambiguities and suggestions that will strengthen the character of the scientific work and adapt it to the requirements of publication.
• Abstract: It is short and concise, but can be improved by highlighting:
o Why is the use of Bacillus spp. important in the context of sustainable agriculture?
o Possible practical applications for agriculture.
• Introduction:
o Explain in more detail why these three pathogens were chosen (F. oxysporum, P. debaryanum and R. solani). Are they particularly common in tomato crops?
o Avoid repeating general information about Bacillus spp. For example, the mechanism of action of Bacillus as a PGPR is repeated several times.
• Materials and Methods:
o There is no information on why the dual culture method was chosen as the primary method for evaluating antagonism. Were other methods considered, e.g. testing for volatile organic compounds (VOCs)?
o Explain why the variety “Super Strain B” was selected. Is it particularly susceptible to pathogens?
o It is worth adding a brief description of why certain enzymes were selected (PO, PAL, PPO). What role do they play in plant resistance?
• Results:
o When discussing the results, there is no evidence of correlation between enzyme activity, plant growth and disease reduction. Does higher PAL and PPO activity correlate with pathogen reduction?
• Discussion:
o It would be worth discussing in more detail why B. subtilis is more effective against F. oxysporum and R. solani, while B. amyloliquefaciens is more effective against P. debaryanum. Could the differences be due to the production of specific metabolites?
o Limitations of the study: it might be worth adding a section describing possible limitations, e.g. lack of volatile metabolite analysis or field trials.
• Conclusions o It might be worth suggesting future studies, e.g. testing these Bacillus strains under field conditions or analysing their mechanisms of action at the molecular level.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
