Reducing Postharvest Losses in Organic Apples: The Role of Yeast Consortia Against Botrytis cinerea
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors
I congratulate the uthors for their valuable work!
Reducing food loss trough proper storage conditions is very current topic in this overpopulated world especially when it comes with chemical residue free practices.
- I recommend to reshape the first part of the introduction (~row 25-37) trough this viewpoint.This study gives potential answers for a dominantly postharvest problem trough only postharvest solutions. Therfore i hhyink the authors should not describe the wider problems of the global food demands and in-field agricultural practices, but focus on food loss and prevention trough (especially biological) postharvest treatments. The viewpoint in row 301-306 in the discussion is way better tot he introduction as well, it feels this part belong tot he introduction.
- Row 46: mentioning the teleomorph is a good practice in education, but modern molecular identification-based taxonomy works with one current name in case of fungi. If the authors insit to include this name it should be referred as synonym.
- As it seems the yeast strains were not properly identified in this study. If this identification was done in a previous work, it should be clearly cited, or if the strains are originated from a gene bank acession numbers should be referred to ensure tracking of a proper identification.
- Identification of the pathogen is also not desribed. Description of morphological identification is minimally expected, however species level identificaton has to be supported with molecular genetic identification in the Botrytis genus (e.g. differentiation of cinerea and B. pseudocinerea).
- Row 92: Why the yeast cell suspension is adjusted exactly to 2x107 CFU/ml? If it is from a previous method please cite it! Cite all method from previous sources!
- Please give the type, (+producer, country, city) f hemacytometers!
- Row 105: Surface disinfection usually conducted with Neomagnol. Please cite this sodium-hypoclorite method from earlyer studies!
- Row 117-118: Please clarify number of replications! As i understood: The experiment was conducted in 3-3 reps started from two different mother colonies (2 separate experiments), then these reps were used as 6 reps in statistical analysis.
- Row 122: Country and city of procucer should be added.
- Part 2.5: Volume and type of the flasks and volume of the suspension should be added.
- Row 134: 1 replication consist from 10 apples. As it seems the authors used mean of the means (mean 10 apples then 5 reps). Please clarify! In this case the errors of the means of 10 apples are lost data, or occur only in better sampling. Therefore I recommend nested design ANOVA for statistical analysis of this part, however this method is also acceptable.
- Part 2.6 and 2.7 should be merged to eliminate redundancy. Please focus ont he differences!
- In the description of statistical analysis the methods of normality and homoscedasticity testing should be present. If these was not done please ensure, that the assumptions of ANOVA are fulfilled. If not use nonparametric tests!
- Fig 1 and 2: Compact letter display should be added (e.g.:letters) based on pairwise or post hoc tests. The sentence „Bars represent SE at 170 a confidence level of 99%.” is not clear. a) this is not a bar diagram, errors are represented on „whiskers” b) represented error can be SE or confidence interval but not both, c) I assume 99% confidence means alpha set as 1%, but this belongs to the compact letter display differences (e.g.: different if p<0,01).
- Fig 3: Same as in case of Fig 1-2. In the last sentence of figure decription should be added, that the differences are based on Fisher LSD tests (or Mann-Whitney if nonparametric). I assume LSD from the Materials and methods, but the figure should be fully understandable on its own.
- All mean comparisons: sincet he tests were evaluated in good number of repetitions and the errors are not too big, I recommend to use a more conservative test (e.g.:Bonferroni test) for post-hoc tests. It suggests a more strict evaluation, however LSD is acceptable as well.
- Fig 4: Same as fig 1-2.
- In case of tables should be added, that the differences (CLD) are based on Fisher LSD tests (or Mann-Whitney if nonparametric).
- Row 256-258: please clarify that the populations can be stable for a wider interval, but the experiments lasted only 7 and 14 days.
- Row 261: I think 116/67 is a typo, the authors mean 117/67.
Author Response
Thank you for your valuable review. All suggestions were taken into consideration and the whole paper was revised. We added missing information and clarified experimental design where necessary.
Response: The introduction have been rephrased according to the suggestions.
Row 46: mentioning the teleomorph is a good practice in education, but modern molecular identification-based taxonomy works with one current name in case of fungi. If the authors insit to include this name it should be referred as synonym
Response: Corrected.
As it seems the yeast strains were not properly identified in this study. If this identification was done in a previous work, it should be clearly cited, or if the strains are originated from a gene bank acession numbers should be referred to ensure tracking of a proper identification.
Response: The information has been added.
Identification of the pathogen is also not desribed. Description of morphological identification is minimally expected, however species level identificaton has to be supported with molecular genetic identification in the Botrytis genus (e.g. differentiation of cinerea and B. pseudocinerea).
Response: The information on B. cinerea source and identification was added.
Row 92: Why the yeast cell suspension is adjusted exactly to 2x107 CFU/ml? If it is from a previous method please cite it! Cite all method from previous sources!
Response: Explanation was added.
Please give the type, (+producer, country, city) f hemacytometers!
Response: Added.
Row 105: Surface disinfection usually conducted with Neomagnol. Please cite this sodium-hypoclorite method from earlyer studies!
Response: Addded.
Row 117-118: Please clarify number of replications! As i understood: The experiment was conducted in 3-3 reps started from two different mother colonies (2 separate experiments), then these reps were used as 6 reps in statistical analysis.
Response: Clarified.
Row 122: Country and city of procucer should be added.
Response: Added.
Part 2.5: Volume and type of the flasks and volume of the suspension should be added.
Response: Added.
Row 134: 1 replication consist from 10 apples. As it seems the authors used mean of the means (mean 10 apples then 5 reps). Please clarify! In this case the errors of the means of 10 apples are lost data, or occur only in better sampling. Therefore I recommend nested design ANOVA for statistical analysis of this part, however this method is also acceptable.
Response: Clarified.
Part 2.6 and 2.7 should be merged to eliminate redundancy. Please focus ont he differences!
Response: Merged.
In the description of statistical analysis the methods of normality and homoscedasticity testing should be present. If these was not done please ensure, that the assumptions of ANOVA are fulfilled. If not use nonparametric tests!
Response: Clarified.
Fig 1 and 2: Compact letter display should be added (e.g.:letters) based on pairwise or post hoc tests. The sentence „Bars represent SE at 170 a confidence level of 99%.” is not clear. a) this is not a bar diagram, errors are represented on „whiskers” b) represented error can be SE or confidence interval but not both, c) I assume 99% confidence means alpha set as 1%, but this belongs to the compact letter display differences (e.g.: different if p<0,01).
Fig 3: Same as in case of Fig 1-2. In the last sentence of figure decription should be added, that the differences are based on Fisher LSD tests (or Mann-Whitney if nonparametric). I assume LSD from the Materials and methods, but the figure should be fully understandable on its own.
All mean comparisons: sincet he tests were evaluated in good number of repetitions and the errors are not too big, I recommend to use a more conservative test (e.g.:Bonferroni test) for post-hoc tests. It suggests a more strict evaluation, however LSD is acceptable as well.
Fig 4: Same as fig 1-2.
Response: All figures were checked and corrected.
In case of tables should be added, that the differences (CLD) are based on Fisher LSD tests (or Mann-Whitney if nonparametric).
Response: Added.
Row 256-258: please clarify that the populations can be stable for a wider interval, but the experiments lasted only 7 and 14 days.
Response: Clarified.
Row 261: I think 116/67 is a typo, the authors mean 117/67.
Response: Yes, thank you.
Reviewer 2 Report
Comments and Suggestions for Authors
The manuscript entitled ' Reducing postharvest losses in organic apples: the role of yeast consortia against Botrytis cinerea' represents the utility of yeast consortia for managing Botrytis rot in apple. The research is meticulously performed, but it needs some additional information, like
- Molecular identification of yeast culture (accession number)
- In vitro compatibility assay for the yeast cultures for checking the suitability of developing consortia
- Different typo errors.Best of luck
Comments for author File: 
Comments.pdf
Comments on the Quality of English Language
The manuscript needs minor modification
Author Response
Thank you very much for taking the time to review our article. We appreciate your valuable comments. We reviewed the article for typos and had a native speaker check it.
The manuscript entitled ' Reducing postharvest losses in organic apples: the role of yeast consortia against Botrytis cinerea' represents the utility of yeast consortia for managing Botrytis rot in apple. The research is meticulously performed, but it needs some additional information, like
- Molecular identification of yeast culture (accession number)
Response: We added the information on yeast identification to the article. They will be molecularly assessed in the future.
- In vitro compatibility assay for the yeast cultures for checking the suitability of developing consortia
Response: The information was added.
- Different typo errors. Best of luck.
Response: We carefully checked the article. Thank you very much once more.
Reviewer 3 Report
Comments and Suggestions for Authors
The manuscript “Reducing postharvest losses in organic apples: the role of yeast consortia against Botrytis cinerea,” authored by Krzyminska and Kowalska, is an important study, and for organic apple cultivation, this kind of option should be available. The present study's main focus is on the use of yeast consortia in biocontrol against Botrytis cinerea, but it should be mentioned that its findings advance existing research. Have similar yeast combinations been tested before? If so, what distinguishes this study?
The methodology lacks details on the experimental setup, including inoculum concentration, duration of storage trials, and methods used to assess colonization and disease suppression.
It is unclear whether the interactions between yeast strains were analyzed at the molecular or metabolic level to understand potential synergistic effects.
The storage conditions (4°C and 23°C) are relevant, but additional intermediate conditions may provide better insights into the real-world applicability of these consortia.
There is no mention of statistical methods used to validate the efficacy of the yeast consortia. Were appropriate controls and replicates included? How were significance levels determined?
The study suggests effective suppression of B. cinerea but does not elaborate on whether the yeasts act via competition, antibiosis, or induced resistance in apples. A more mechanistic explanation would strengthen the findings.
The potential application in commercial apple storage is not discussed. Would these consortia be compatible with existing postharvest handling practices? Are there any constraints regarding formulation or shelf life?
The abstract is clear but could briefly mention the statistical significance of the findings.
Since organic farming is a focus of the study, the study could discuss regulatory approval and consumer acceptance of yeast-based biocontrol agents.
Author Response
Thank you very much for taking the time to review our manuscript. Please find the responses to each of your comments below.
The manuscript “Reducing postharvest losses in organic apples: the role of yeast consortia against Botrytis cinerea,” authored by Krzyminska and Kowalska, is an important study, and for organic apple cultivation, this kind of option should be available. The present study's main focus is on the use of yeast consortia in biocontrol against Botrytis cinerea, but it should be mentioned that its findings advance existing research. Have similar yeast combinations been tested before? If so, what distinguishes this study?
Response: We added the information.
The methodology lacks details on the experimental setup, including inoculum concentration, duration of storage trials, and methods used to assess colonization and disease suppression.
Response: Missing information was added.
It is unclear whether the interactions between yeast strains were analyzed at the molecular or metabolic level to understand potential synergistic effects.
Response: It was clarified.
The storage conditions (4°C and 23°C) are relevant, but additional intermediate conditions may provide better insights into the real-world applicability of these consortia.
Response: Thank you. The temperatures were picked for storage conditions, so the results can apply to real world usage. More temperatures can be picked in future experiments.
There is no mention of statistical methods used to validate the efficacy of the yeast consortia. Were appropriate controls and replicates included? How were significance levels determined?
Response: Missing information was added.
The study suggests effective suppression of B. cinerea but does not elaborate on whether the yeasts act via competition, antibiosis, or induced resistance in apples. A more mechanistic explanation would strengthen the findings.
Response: Some mechanisms were discussed, it will be studied further in the following experiments.
The potential application in commercial apple storage is not discussed. Would these consortia be compatible with existing postharvest handling practices? Are there any constraints regarding formulation or shelf life?
Response: Thank you, we will include such information in our future study.
The abstract is clear but could briefly mention the statistical significance of the findings.
Response: Added.
Since organic farming is a focus of the study, the study could discuss regulatory approval and consumer acceptance of yeast-based biocontrol agents.
Response: Thank you, we will include such information in our future study.
Round 2
Reviewer 1 Report
Comments and Suggestions for Authors
I congratulate the authors, the manuscript improved a lot.
The introduction now serves as a good base for the study.
My earlier concerns regarding the statistical analysis are partly solved (important for repeatability of the study). The authors try to clarify the number of samplings and repetitions with different wording or semantics, however these could be simply communicated witht exact numbers: e.g.: xy test was conducted in two independent experiments in three repetitions and the results of the two experiments were used as six replications (two experiments treated as one in the process of evaluation) in the statistical analysis.
In case of part 2.6. I think they used mean of the means method: mean of 10 apple as samplings mean of the 3 or 5 repetitions (with errors) wich are the mean of 10 apple as samplings.
Testing of normality and homoscedasticity is now mentioned, but the exat conducted tests (e.g. Shapiro-Wilk and Levene or other ones) are not described.
Language and minor recommendations:
row 12-13: statistically ipmortant should be statistically proven or this can be deleted.
row 31: "/h" in the end of the sentence should be a typo or an unhyperlinked citation.
Producer of API ID 32 C system is mentioned, this lacks country and city.
Part 2.6. and 2.7. is now merged, so statistical analysis can be 2.7. not "2.8.".
All of the above require minor changes to the manuscript, after these will be revised, I recommend the manusript to publish.
Author Response
Thank you very much once again for taking the time to review our paper. I’ve incorporated all your recommendations and hopefully now the article is improved.
The introduction now serves as a good base for the study.
Response: Thank you.
My earlier concerns regarding the statistical analysis are partly solved (important for repeatability of the study). The authors try to clarify the number of samplings and repetitions with different wording or semantics, however these could be simply communicated witht exact numbers: e.g.: xy test was conducted in two independent experiments in three repetitions and the results of the two experiments were used as six replications (two experiments treated as one in the process of evaluation) in the statistical analysis.
In case of part 2.6. I think they used mean of the means method: mean of 10 apple as samplings mean of the 3 or 5 repetitions (with errors) wich are the mean of 10 apple as samplings.
Testing of normality and homoscedasticity is now mentioned, but the exat conducted tests (e.g. Shapiro-Wilk and Levene or other ones) are not described.
Response: I’ve completed missing information.
Language and minor recommendations:
row 12-13: statistically ipmortant should be statistically proven or this can be deleted.
Response: Changed.
row 31: "/h" in the end of the sentence should be a typo or an unhyperlinked citation.
Response: Removed.
Producer of API ID 32 C system is mentioned, this lacks country and city.
Response: Added.
Part 2.6. and 2.7. is now merged, so statistical analysis can be 2.7. not "2.8.".
Response: Corrected.
All of the above require minor changes to the manuscript, after these will be revised, I recommend the manusript to publish.
Response: Thank you.
Reviewer 3 Report
Comments and Suggestions for Authors
The authors have revised manuscript as per suggested and now it acceptable in its current state.
Author Response
The authors have revised manuscript as per suggested and now it acceptable in its current state.
Response: Thank you very much once again for taking the time to review our manuscript.
