Management of spot disease of Indian mustard caused by Alternaria brassicae using botanical fungicide

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1. 

   Access Microbiology

   Nov 10, 2023

   Comments to Author

   The manuscript analyses the incidence of spot disease in a field of Indian mustard and explores the inhibitory effect of different botanical extracts on conidial germination of the pathogenic fungus Alternaria brassicae. The title of the manuscript does not reflect the actual content of the research. The research objectives are not clearly stated and both parts of the research (field study and antifungal assay) are not well linked, at least by the way it is explained. The provided methods need a deep revision and complementation of the information. Further, identification of Alternaria isolates by molecular methods would be required and data from re-infection assays must be provided. Statistical analysis was appropriate; however, the interpretation of the results was not consistent. Conclusions …

   Comments to Author

   The manuscript analyses the incidence of spot disease in a field of Indian mustard and explores the inhibitory effect of different botanical extracts on conidial germination of the pathogenic fungus Alternaria brassicae. The title of the manuscript does not reflect the actual content of the research. The research objectives are not clearly stated and both parts of the research (field study and antifungal assay) are not well linked, at least by the way it is explained. The provided methods need a deep revision and complementation of the information. Further, identification of Alternaria isolates by molecular methods would be required and data from re-infection assays must be provided. Statistical analysis was appropriate; however, the interpretation of the results was not consistent. Conclusions sometimes are not supported either by the presented results or references. The article requires major revision for publication. Below, my suggestions for improvement of the manuscript are found: 1. Abstract - Line 25: "As the garlic extract is a botanical fungicide, this can be applied without causing any significant hazards to the environment." Please, rewrite this sentence. The fact that an extract comes from a plant does not mean that it is not negative for the environment. 2. Introduction - Line 30: "Indian mustard [Brassica juncea (L.) Czern and Coss.] is a herb of the Brassicaceae (Cruciferae) family and is the third most important oilseed crop in the world in terms of production." Please, add a reference. - Line 31: "It is the source of edible oil which cannot be replaced with other substitutes". Please rewrite this sentence or remove. There are many alternative edible oils, such as sunflower or olive oil. - Line 36: The unit lakh is not a unit from the International System, please convert to Million ha or just ha. - Line 38: Please, add a reference for the information given in the paragraph. - Line 56: Please, replace the "@" by the proper symbol for registered trademark "®". 3. Materials and methods - Line 70: Please, explain this better. I don't understand the fact that this area has Kharif, Rabi and Zayad, but then mustard is a chief crop. - Line 71: You explain that the infection was assessed during January, but exactly when? I think this is important because you then discuss that a secondary infection was due to the rainfall of the 22nd of January, however you don't explain when your samples were collected. - Line 77: You explain that RH749 variety was cultivated by "most" of the farmers. This is not specific enough. Belonged all your sampled plants to this variety or did you sample different varieties? - Line 82: Please, replace the term "samples" by "sites" and rewrite the sentence. Your samples are the plants taken at different sites. Do so in Table 1. - Line 85: You explain that you used the software ImageJ to measure the leaf area, but you don't explain how you measured the infected leaf area. Was it done with the same software? If so, please explain the parameters considered for the analysis in ImageJ. - Line 88: Please, rewrite the last sentence. - Line 89: Please, replace "inoculated" by "incubated". - Identification of pathogen: Nowadays, molecular methods, such as sequencing of gene markers, are required for the proper identification of microorganisms. Please, provide molecular evidence about the identification of your isolates. - Re-inoculation of new healthy plants with the isolates obtained from the prospection is required to verify that those were actually the causal agent of the observed symptoms, following Koch postulates. The authors state that they have conducted these experiments, although these data are not shown. Please, provide the results. - Antifungal bioassay: The experimental design and procedure is not properly described. Please, include number of replicates, the origin of the A. brassicae isolate used for the experiment and the positive and negative controls included. I also miss information about the concentration of the extracts and how the application was done. Additionally, germination rates or frequencies should be provided, as a quantification of the inhibition. Qualitative (+, -) evaluation is not sufficient. - Line 113: Remove the word "for". - Line 114: Please, indicate the concentration of conidia used for the experiment. - Line 122: This is actually not explained in previous sections. 4. Results and discussion - Table 1: Please, replace "No. sample" by "Site". - Lines 134 - 139: The statistical analysis conducted of leaf area and infected leaf area add low value to the analysis, because the relevant analysis is the disease severity. Anyway, the authors state that the areas of the studied leaves were significantly different, but the infected area among leaves was homogeneous. However, the statistical analysis showed that there were significant differences among sites in both parameters, so that statement is not supported by the results. - Line 136 - 137: This discussion is not supported by your results. - Line 138 - 139: This statement is not supported by your investigation since there is not a monitoring of the disease along time. The authors don't provide data about when could take place a primary infection to say that the rain on the 22nd of January was the responsible for the secondary infection. - Line 176: Please, remove "significantly". - Line 177: "when cultured conidia were tested for infectivity (not included in this result) these were again very similar to the conidia described in Fig. 2 d." This information must be included. Please, provide the results of the experiment. - Line 200-201: "These findings may be attributed to the fact that neem is a good insecticide but poor fungicide." That finding can be attributed to many facts (concentration, method of extraction, among others), please discuss further and add references. - Antifungal bioassay: Germination rates or frequencies should be provided as a quantification of the inhibition. Qualitative (+, -) evaluation is not sufficient. - Line 213: Please, rewrite this sentence in order to make clear that this statement is only based on previous literature. The way it is written makes one think that the activity found in your extract is due to allicin, however the research does not tackle extract composition or investigation of the antifungal compound. - Line 2018 - 220: Please, explain and discuss the results obtained by Yadav et al. (2023) in their study. 5. Conclusions - Line 222: Why is the disease underestimated? Please, explain or provide a reference. - Line 225: "Rain and high humidity in the field promoted the secondary infection very rapidly and all the plants were infected within a few days and 100% disease incidence was recorded." The authors don't provide evidence supporting this conclusion. - Line 239 - 241: Again, the authors write about allicin as if it was the active compound of their garlic extract, however they do not provide evidence on this.

   Please rate the manuscript for methodological rigour

   Poor

   Please rate the quality of the presentation and structure of the manuscript

   Poor

   To what extent are the conclusions supported by the data?

   Partially support

   Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?

   No

   Is there a potential financial or other conflict of interest between yourself and the author(s)?

   No

   If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?

   Yes

   Read the original source
2. 

   Access Microbiology

   Nov 10, 2023

   We have now received comments from two reviewers who recommend revision to your manuscript. In particular, I would encourage you to address your title, consistency in result interpretation and clarity of methods description.

   Read the original source
3. 

   Access Microbiology

   Sep 27, 2023

   Comments to Author

   Overall, the manuscript lacks the correct in-text referencing format with insufficient references used throughout, with many references too old to be used without addition of further references. This limits how your research can fit into the broader context of it's field. I also have concerns regarding the identification of isolates as there is no secondary confirmation method for species identification. Introduction - Why do most paragraphs have a single reference at the end of the paragraph? In-text references should be next to the information they're referring to, not a list at the end of a paragraph. Line 41 - Why use a reference this old? There are more recent articles that show this and more. Line 47 - 'shrivel' is not descriptive enough. Line 48 - 'most of the times' is not scientifically …

   Comments to Author

   Overall, the manuscript lacks the correct in-text referencing format with insufficient references used throughout, with many references too old to be used without addition of further references. This limits how your research can fit into the broader context of it's field. I also have concerns regarding the identification of isolates as there is no secondary confirmation method for species identification. Introduction - Why do most paragraphs have a single reference at the end of the paragraph? In-text references should be next to the information they're referring to, not a list at the end of a paragraph. Line 41 - Why use a reference this old? There are more recent articles that show this and more. Line 47 - 'shrivel' is not descriptive enough. Line 48 - 'most of the times' is not scientifically acceptable language. Line 50 - 'Hence' is used wrong. You haven't made a strong enough justification to use 'hence'. Line 52 - you say 'various researcher' but only provide a single reference. Is it various, or is it just one article? Line 62 - never refer to anything as a 'fact' in an article. Line 88 - you don't need to say agar and medium for the same thing. Line 93 - Did you run a PCR to confirm identification? Any sequencing? Identification based on morphology and microscopy has a lot of error associated with it. The reference for this identification is far too old. Many updates have been made since then in morphological identifications. Line 106 - If centrifuge speeds are given in rpm, the make of the centrifuge and rotor size is needed. Centrifuge speeds should always be given in g. Data analysis is very weak and is purely descriptive. A more critical and comparative approach is needed. This is also true of the discussion points. References from the 1980s should not be included in a discussion unless it is critically necessary.

   Please rate the manuscript for methodological rigour

   Poor

   Please rate the quality of the presentation and structure of the manuscript

   Satisfactory

   To what extent are the conclusions supported by the data?

   Partially support

   Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?

   No

   Is there a potential financial or other conflict of interest between yourself and the author(s)?

   No

   If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?

   Yes

   Read the original source
4. 

   Version published to 10.1099/acmi.0.000692.v1 on Access Microbiology

   Aug 29, 2023