Current Progress in Microbial Biocontrol of Banana Fusarium Wilt: A Systematic Review

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

All my comments are available in the pdf document provided.

Comments for author File: Comments.pdf

Author Response

Comment 1: The abstract does not highlight anything new about this work compared to previously published ones.

Response 1: the summary was improved by indicating the contributions and novelties of this new revision

Comment 2: This term has just been used a little higher systematic

Response 2: The term "systematic" is part of the type of study conducted (systematic review); we cannot use a synonym.

Comment 3: This phrase is too generic, be a little more specific.

Response 3: the phrase was changed to be more specific

Comment 4: These words are very repetitive

Response 4: the term was changed to avoid repetitiveness

Comment 5: Use synonyms to refer to this expression.

Response 5: synonyms have been used for this expression

Comment 6: Already described before, put Foc

Response 6: from here on Foc is used

Comment 7: This has also been described above.

Response 7: as the term was amended

Comment 8: R4T or TR4?

Response 8: TR4

Comment 9: Indicate the acronym the first time it is used, not now.

Response 9: acronym was modified

Comment 10: The quality of this image needs to be improved.

Response 10: the image quality has been improved

Comment 11: Use Foc

Response 11: Use Foc

Comment 12: Use the acronym.

Response 12: acronym was used

Comment 13: I think it was already described

Response 13: acronym was used

Comment 14: Clarify this sentence.

Response 14: sentence clarified

Comment 15: Specify which race 4.

Response 15: race was specified

Comment 16: Has it already been mentioned in the text?

Response 16: yes, the acronym was used

Comment 17: More quality in the image and these words placed horizontally like the other raids.
Use more colors, each country a different one,
On light colored panels the white letters do not look good.

Response 17: improved colors by country and image quality

Comment 18: Add sp. cubense

Response 18: cubense was added

Comment 19: Why does it have a hyphen? It has been used before in the text and it did not have one.

Response 19: modified to standardize the term throughout the text

Comment 20: Clarify these two sentences, it is not clear what the percentages are for China.

Response 20: percentages for China were clarified

Comment 21: Remove the italics from "and".

Response 21: italics removed

Comment 22: The figure is not understandable, it is not self-explanatory and the figure caption does not clarify anything. I would eliminate it or replace it or explain it well in the text and in the footer or with a legend.

Response 22: the figure was improved, also the legend was improved to explain it better

Comment 23: Indicate what "NA" means. Why are the references indicated with a number and the name of the first author in the table and only with a number in the main text? is this correct?

Response 23: clarified the use of both terms ND and NA in the table footnote and left only the number of the citation for clarity

Comment 24: In this column, start the word with a capital letter. Apply to entire column

Response 24: capitalization was modified as suggested

Comment 25: Why is it written in Spanish? put it in english

Response 25: all the terms that were in Spanish in the tables have been modified

Comment 26: sp. without italics.

Response 26: italics removed

Comment 27: Remove the dashes or put them in all similar cases of the table

Response 27: terms were standardized

Comment 28: Lowercase for plot and trial

Response 28: were changed to lowercase

Comment 29: Add the genotype as in the rest of the cultivars

Response 29: only in those cases where the author of the article consulted indicates the genotype is shown, in the other cases it was decided not to add it. 

Comment 30: Add a dot after sp (sp.)

Response 30: the period was added in all abbreviations

Comment 31: Adjust the table so that the s is not alone on the line. Take this into account for the entire table.

Response 31: adjustments were made in all cases

Comment 32: Add a dot after et al (et al.)

Response 32: the point was added in all cases

Comment 30: Is something missing here?

Response 30: the species was added correctly

Comment 31: remove 1 after prigigallo

Response 31: the number was removed

Comment 32: This table footer is not well understood. Rewrite it please.

Response 32: corrected table footer

Comment 33: Does all this follow from figure 4? Rewrite and direct correctly to the source of information

Response 33: the source of the information was rewritten and correctly cited.

Comment 34: Use the acronym BCA throughou the text

Response 34: the acronym was used

Comment 35: This sentence is not understood, clarify it, please

Response 35: the sentence was clarified

Comment 36: Remove Fusarium. Foc is already described

Response 36: was used Foc

Comment 37: use an acronym for fusarium wilt of banana, for instance, FWB or BFW.

Response 37: acronym was used

Comment 38: In this section reference should be made to Figure 4. Comment on it in the appropriate places.

Response 38: Figure 4 is discussed in this section

Comment 39: add references for these statements

Response 39: reference was added

Comment 40: This is already described above

Response 40: was modified

Comment 41: It is better to talk about concentration or total number of conidia. Indicating only the volume does not clarify anything

Response 41: indicated as suggested

Comment 42: The information in this section should be expanded.

Response 42: this section has been expanded and indicated others varieties

Comment 43: Are you referring to subtropical race 4 here? Please clarify.

Response 43: reference clarified

Comment 44: Foc and FWB or BFW instead of Fusarium oxysporum and Fusarium wilt.

Response 44: the term was clarified

Comment 45: check the italics of scientific names

Response 45: italics in references have been corrected

Comment 46: The authors name is duplicated at least in references 98, 99, 100. Check all the references in this regard

Response 46: the reference was corrected

 

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript "Current Progress in Microbial Biocontrol of Banana Fusarium Wilt: A Systematic Review" by Richard Solórzano et. al. reports on a systematic review on the use of antagonists against the causative agent of fusarium of banana plant roots.

Biological protection of plants, especially in the cultivation of tropical plants, is important for maintaining the ecological stability of agroecosystems and obtaining a safe crop. The work is generally logical, but it is carelessly designed, there are many errors and ambiguities. In addition, there are a lot of inaccuracies in the methods that should not be in an article applying for publication in a highly rated journal.  In addition, there are significant errors and issues in the work that, in my opinion, require serious adjustments.

Main remarks:

- The word "Progress" given in the title of the article suggests the use of something new, however, the article contains only precedents for the use of antagonists and an assessment of their effectiveness, and therefore the authors need to either change the title or add new information and sections. For example, methods of antagonist delivery, the use of metabolites, encapsulation in formulations, etc.

- There is no clear research hypothesis in the paper, and the "introduction" section presents a problem, but does not formulate a specific research question or hypothesis. The authors should define this point more clearly.

- The Methods section describes the selection process for articles, but the text itself does not clearly analyze which sources were excluded and for what reasons.

- The question arises whether the "quality" parameter of the articles was taken into account when selecting the papers, but the quality criteria of the articles themselves are not specified.

- Why don't the authors, noting the dominance of Asian studies, discuss the possible consequences of this on a global scale?

-The article describes the use of a variety of microorganisms, but does not provide a detailed comparison of the analysis of their effectiveness in different conditions.

Minor remarks:

- The authors, having provided so much information about antagonist microorganisms, do not discuss the mechanisms of their action. It must be added to the text.

- The use of antagonists may have side effects, such as adverse effects on local soil microbiomes, but this aspect is not discussed.

- Although the article discusses the potential of biocontrol and the use of these microorganisms in commercial crop production, it provides little information about current commercial products and the prospects for their use.

- The article contains a lot of data on the geographical distribution of research and the effectiveness of different microorganisms, but few graphs or tables to facilitate the perception of information. I suggest that the authors add a global map and indicate on it where the studies were performed and where the strains were isolated.

- The conclusion talks about the prospects of biocontrol, but clear recommendations on its use in the field are not given. It is necessary to add to the text.

-In some places, the text is overloaded with complex structures, which makes it difficult to understand.

I urge the authors to improve the quality of the text and thereby increase the importance of the publication.

Author Response

Main remarks:

Comments 1: The word "Progress" given in the title of the article suggests the use of something new, however, the article contains only precedents for the use of antagonists and an assessment of their effectiveness, and therefore the authors need to either change the title or add new information and sections. For example, methods of antagonist delivery, the use of metabolites, encapsulation in formulations, etc.

Response 1: We appreciate the suggestion on the use of the term Progress in the title. We consider that progress in microbial biocontrol of Fusarium wilt in banana is relevant since the emergence of new races of the pathogen, especially Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4), which has prompted the search for more effective management strategies.
The article discusses recent advances in the application of antagonistic microorganisms, including the use of consortia, mixtures with organic matter and, to a lesser extent, synthetic communities. However, studies on new application and formulation methods, such as encapsulation or the use of metabolites, are still limited and lack sufficient evidence to be extensively discussed in this review. Despite this, we mention these emerging approaches as developing lines of research.
Since the article already highlights recent trends and the use of novel strategies in some cases, we believe that the term Progress is still appropriate. However, we are open to adjusting the title if deemed necessary to more accurately reflect the scope of the study.

Comments 2: There is no clear research hypothesis in the paper, and the "introduction" section presents a problem, but does not formulate a specific research question or hypothesis. The authors should define this point more clearly.

Response 2: The hypothesis “Biological control based on microorganisms represents an effective and sustainable alternative for the management of Fusarium wilt in banana, and its efficacy varies according to the type of microorganism, mechanism of action and application conditions” was added.

Comments 3: The Methods section describes the selection process for articles, but the text itself does not clearly analyze which sources were excluded and for what reasons.

Response 3: In addition to the inclusion and exclusion criteria already mentioned in the methodology, “Studies with incomplete data, lack of replicability, absence of adequate controls in biocontrol trials or that did not specify the characterization of the microorganisms used” were excluded.

Comments 4: The question arises whether the "quality" parameter of the articles was taken into account when selecting the papers, but the quality criteria of the articles themselves are not specified.

Response 4: To guarantee the rigorousness of this systematic review, quality parameters based on the following were taken into account:
1. methodological rigor: studies with detailed methodologies, adequate experimental design (appropriate controls and replicability) and validated statistical analyses were prioritized.
Scientific relevance: We selected articles that addressed the biocontrol of Fusarium oxysporum f. sp. cubense in banana, excluding studies with no direct application in this crop.
3.    Verifiable sources: Publications in indexed scientific journals and articles with access to complete and replicable data were considered.
4.    Impact and soundness of results: Priority was given to studies that presented statistically significant results, with adequate interpretation and discussion of their findings.

Comments 5: Why don't the authors, noting the dominance of Asian studies, discuss the possible consequences of this on a global scale?

Response 5: was added in the country results section:
The predominance of studies of Asian origin in the literature on Fusarium biocontrol in banana may influence the global applicability of the findings due to agroecological, genetic, and crop management differences in other producing regions. Asia is a key epicenter of banana production and the impact of Fusarium oxysporum f. sp. cubense Race 4 Tropical (Foc R4T), which has prompted further research in this region. However, the limited representation of studies from Latin America and Africa-also important producing areas-may restrict the extrapolation of biocontrol strategies to other soil and climatic conditions, production systems, and locally grown banana varieties.

Comments 6: The article describes the use of a variety of microorganisms, but does not provide a detailed comparison of the analysis of their effectiveness in different conditions.

Response 6: We appreciate the observation that a more detailed comparative analysis of the effectiveness of microorganisms under different conditions is needed. However, in the review we found that many of the studies do not uniformly specify soil conditions, such as pH, electrical conductivity (EC) or climatic characteristics, which makes direct comparison between them difficult.
Despite this limitation, it was observed that, among the species of microorganisms analyzed, the responses to biocontrol were consistent and robust. For this reason, it was not considered appropriate to delve into these parameters as determinants of the biological control response in this study. Nevertheless, we recognize the importance of these factors and suggest that future research incorporate them in greater detail to improve understanding of their influence on the efficacy of microbial agents.

Minor remarks:

Comments 7: The authors, having provided so much information about antagonist microorganisms, do not discuss the mechanisms of their action. It must be added to the text.

Response 7: The mechanisms of action of antagonistic microorganisms are multiple and non-exclusive, since being biological organisms, it is not possible to completely suppress or isolate all their interactions and abilities in the soil ecosystem. In the manuscript, mention is made of the most commonly recognized mechanisms in the literature, such as competition for nutrients and space, production of antifungal metabolites, induction of systemic resistance, and mycoparasitisation.
However, it is important to note that, although some studies report the expression of several of these mechanisms, in most of the reviewed papers they were not measured directly, but were assumed to be a function of the type of microorganism used. Since this review focuses on the efficacy of biocontrollers and not on descriptive studies of individual mechanisms, detailed inclusion of these aspects could skew the interpretation of the body of evidence collected. However, a mention of this limitation and the need for further research along these lines will be included in the revised version.

Comments 8: The use of antagonists may have side effects, such as adverse effects on local soil microbiomes, but this aspect is not discussed.

Response 8: according to the literature reviewed has been added: 
The use of antagonistic microorganisms may generate collateral effects on soil microbiomes, an issue that has been addressed in some studies, although with still limited and variable results. In general, the introduction of biocontrol agents can modify the local microbial composition, either through competition with native populations or by altering soil ecological balances.
However, the literature reviewed does not provide conclusive data on significant negative impacts on soil microbiomes following the application of antagonists for the control of Fusarium oxysporum f. sp. cubense. In fact, some research suggests that certain microorganisms may even enhance microbial biodiversity and favor soil resilience by promoting more functionally balanced communities.

Comments 9: Although the article discusses the potential of biocontrol and the use of these microorganisms in commercial crop production, it provides little information about current commercial products and the prospects for their use.

Response 9: according to the literature reviewed has been added: 
We recognize the importance of addressing the current status of commercial products based on antagonistic microorganisms and their applicability in banana production on a commercial scale. However, the literature reviewed focuses on efficacy studies under experimental conditions, while information on commercial formulations and their field performance is still limited and scattered. Some products based on Trichoderma spp., Bacillus spp. and Pseudomonas spp. are already available on the market and have been reported in efficacy studies, but none on Foc. 

Comments 10: The article contains a lot of data on the geographical distribution of research and the effectiveness of different microorganisms, but few graphs or tables to facilitate the perception of information. I suggest that the authors add a global map and indicate on it where the studies were performed and where the strains were isolated.

Response 10: is indicated in the chord chart, it is not necessary to use a world map because there will be many empty and unreadable spaces in the figure.

Comments 11: The conclusion talks about the prospects of biocontrol, but clear recommendations on its use in the field are not given. It is necessary to add to the text.

Response 11: has been added:
The review has identified strategies that have demonstrated greater efficacy in improving the establishment of microorganisms in soil. To strengthen the applicability of these findings, it is recommended to apply in order of importance Trichoderma spp, Bacillus spp, Streptomyces spp. and Pseudomonas spp. It is recommended to adopt practices based on available evidence, such as:
1. Selection of microorganisms: Prioritize the use of microbial consortia rather than pure cultures, especially those that include Pseudomonas spp. due to their greater stability and consistency in Foc control.
Timing of application: Inoculate seedlings before transplanting or in early stages of the crop to favor rhizosphere colonization.
3.    Soil conditions: Incorporate organic matter together with the microorganisms to improve their persistence and activity in the soil.
4.    Monitoring and dose adjustment: Conduct trials under local conditions before large-scale adoption, since efficacy varies according to soil type, climate and variety of banana cultivated. Doses are highly variable, so it is suggested that average concentrations can be a minimum of 1x105 and adjust them according to whether they are applied in solid form combined with organic matter or liquid directly per plant. 

Comments 12: In some places, the text is overloaded with complex structures, which makes it difficult to understand.

Response 12: the wording was improved and several points were clarified according to the reviewers' observations

Comments 13: I urge the authors to improve the quality of the text and thereby increase the importance of the publication.

Response 13: We appreciate the suggestion and recognize the importance of improving the clarity and accuracy of the text to strengthen the impact of the publication. As part of this process, we revised the manuscript to optimize the writing, ensuring a smoother structure and greater coherence in the presentation of information.

Reviewer 3 Report

Comments and Suggestions for Authors

The article presents a good quality of the review research. The authors analyzed the important problem showing the direction for future research and development of the biocontrolling products. 

After reading the text I found some points, which need to be checked and corrected. 

line 160 - add to the equation x100%

lines 16, 104, 174, 185,192, 202, 215, 248, 253, 258, 522,  - please correct (exclude partly italics typing) "focusing", "focused" "focus"

line 260 - and without italics

Table 1 - sp.; spp. - without italics

Table 1 - ND; NA - explain what they mean; "Tallos de banano" - what does it mean?

R4T - should be explained as race 4 tropical, it is mentioned in the text but no additional data is provided. 

line 312 - and without italics

line 362 - Streptomyces with italics

line 363 - I am unsure if the author properly used "red dots"; please check it because there are black dots on the graph. 

line 526 - before "Williams" add cv.

line 530v - R4ST - it is mentioned only once in the text; what it rferes to? Please explain, correct.

Another suggestion is to reorganize Table 1 by grouping cases according to the type of the experiment and the organism. This means that first should be presented Dual Culture Trials, next Seedling trial, and Experimental Plot Trial. Within each type of experiment, the organisms should be grouped, one by one. This will make the table 1 easier to read. 

Comments for author File: Comments.pdf

Author Response

Comments 1: The article presents a good quality of the review research. The authors analyzed the important problem showing the direction for future research and development of the biocontrolling products. 

Response 1: thanks you

Comments 2: After reading the text I found some points, which need to be checked and corrected. 

Response 2: have been corrected according to your observations

Comments 3: line 160 - add to the equation x100%

Response 3: was added thanks

Comments 4: lines 16, 104, 174, 185,192, 202, 215, 248, 253, 258, 522,  - please correct (exclude partly italics typing) "focusing", "focused" "focus"

Response 4: corrected as observed

Comments 5: line 260 - and without italics

Response 5: italics removed

Comments 6: Table 1 - sp.; spp. - without italics

Response 6:  italics removed

Comments 7: Table 1 - ND; NA - explain what they mean; "Tallos de banano" - what does it mean?

Response 7: failed to translate was corrected

Comments 8: R4T - should be explained as race 4 tropical, it is mentioned in the text but no additional data is provided. 

Response 8: explained as tropical breed 4 for clarity

Comments 9: line 312 - and without italics

Response 9: italics removed

Comments 10: line 362 - Streptomyces with italics

Response 10:  italics added

Comments 11: line 363 - I am unsure if the author properly used "red dots"; please check it because there are black dots on the graph. 

Response 11: was reviewed and modified

Comments 12: line 526 - before "Williams" add cv.

Response 12: cv. added

Comments 13: line 530v - R4ST - it is mentioned only once in the text; what it rferes to? Please explain, correct.

Response 13: has been corrected

Comments 14: Another suggestion is to reorganize Table 1 by grouping cases according to the type of the experiment and the organism. This means that first should be presented Dual Culture Trials, next Seedling trial, and Experimental Plot Trial. Within each type of experiment, the organisms should be grouped, one by one. This will make the table 1 easier to read. 

Response 14: the table has been reorganized according to the indications

Reviewer 4 Report

Comments and Suggestions for Authors

(1) The major concerns: (i) the study did not analyze the cost of different microoorganisms as biocontrol agents to control Foc; (ii) no inforamtion about registering and commercilation of the biocontrol agents; (ii) no imformation about mechanisms of the biocontrol agents.

(2) Minor concerns: (i) editorial mistakes (see my labels in reviewed version); (ii) references, format should be carefully checked.

Comments for author File: Comments.pdf

Author Response

Comentarios 1: (1) Las principales preocupaciones: (i) el estudio no analizó el costo de los diferentes microorganismos como agentes de biocontrol para controlar Foc; (ii) no hay información sobre el registro y comercialización de los agentes de biocontrol; (ii) no hay información sobre los mecanismos de los agentes de biocontrol.

Respuesta 2: (i) los estudios revisados ​​no detallan costos e inferirlos puede ser impreciso; por otro lado, muchos de los estudios son experimentales y no comerciales, nuestro estudio se enfoca en eficacia y no en costos que podrían agregarse pero sin fundamento suficiente en los artículos revisados ​​(ii) los estudios son experimentales y no sobre cepas comerciales; por lo tanto, no hemos revisado los costos ni los aspectos regulatorios que podrían analizarse en otro estudio (ii) agregamos y discutimos los mecanismos de biocontrol.

Comentarios 2: (2) Preocupaciones menores: (i) errores editoriales (ver mis etiquetas en la versión revisada); (ii) referencias y el formato deben revisarse cuidadosamente.

Respuesta 2: (i) se corrigieron los errores de redacción señalados en el documento adjunto (ii) se corrigió el formato de las referencias indicadas en el documento adjunto por el revisor.

 

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

Thanks to the authors for the adjustments. Now the work has acquired a higher quality