Influence of Fruit Wounding on Subsequent Monilinia laxa Infection of Nectarines

Round 1

Reviewer 1 Report (Previous Reviewer 5)

The authors have fully responded to my comments raised during the first submission, and the proposed recommendations are taken into account

Sincerely.

The quality of English language style  has been improved compared to the last version

Author Response

Thank you very much for your time, for your corrections and contributions to our MS.

Best regards,

The authors

Reviewer 2 Report (New Reviewer)

The research entitled (Influence of Fruit Wounding on Subsequent Monilinia laxa Infection of Nectarines) studied the relationship between mechanical stress and infection in Nectarines. Although the research idea is good some points need handling in the manuscript.

1.       In the method part the pathogen M. laxa (accession identification: Ml3), this is not accession number in the gene bank. The authors must include the isolate sources, sequence, genetic tree, and the right accession number.

2.       What is the temperature, and humidity of the First Experiment?

3.       The stander error must include in all figures.

4.       The discussion part needs to be fortified with more recent references.

5.       The authors need to check the reference style one by one to match the journal style.

6.       The whole manuscript needs English editing and grammar correction.

The whole manuscript needs English editing and grammar correction

Author Response

Dear Reviewer,

Next, it is answered point by point, and in the new version it is marked with change control in the Word file and it is highlighted with red text.

Point 1: In the method part the pathogen M. laxa (accession identification: Ml3), this is not accession number in the gene bank. The authors must include the isolate sources, sequence, genetic tree, and the right accession number.

Response 1: The strain used is a local one, not registered in a gene bank, nor sequenced. Therefore, it has no accession number. INARE accession identification: Ml3.

It is clarified in the text that it is an INRAE identification.

Line 115: "M. laxa (accession identification: Ml3)" was changed to "M. laxa (INRAE accession ID: Ml3)"

 

Point 2: What is the temperature, and humidity of the First Experiment?

Response 2: For all experiments, the conditions are given in the paragraph 2.2 : incubation conditions; Thus, the boxes were maintained in a growth chamber at 25±1 °C, 80% relative humidity, and a photoperiod of 12 hours

 

Point 3: The stander error must include in all figures.

Response 3: In the choice we've done of figure types (boxplots, individual fruit plotting, cloud of dots) , standard errors have no place to be; Infection probability variable is integrative, by construction there is no repetition. Therefore it is not possible to add standard error on any figure of the publication. If the measured variables had been plotted directly, it is a good observation to note that the standard errors are missing.

 

Point 4: The discussion part needs to be fortified with more recent references.

Response 4: The literature is very poor concerning fruit wounding, and in particular stone fruit and link with fungal infection. The same way the compounds detected are not very common.

We dare to think that we have searched the scientific databases and selected the references that are not too far from our subject... for sure we may have missed some.

On the basis of the other reviewer advice, we added the reference of a recent review, even very generalist (Mostafa et al. 2022).

In references added: 24. Mostafa, S.; Wang, Y.; Zeng, W.; Jin, B. Plant responses to herbivory, wounding, and infection. Int. J. Mol. Sci. 2022, 7031. https://doi.org/10.3390/ijms23137031.

 

 

Point 5: The authors need to check the reference style one by one to match the journal style.

Response 5: Thanks, the references style was revised to match the journal.

 

Point 6: The whole manuscript needs English editing and grammar correction.

Response 6: Thanks, the English edition was revised twice, and a new revision by a native English speaker was recently done.

 

Many thanks! for the correction and suggestions.

The authors

 

Reviewer 3 Report (New Reviewer)

The authors investigated the relationship between mechanical wounding and Monilinia laxa infection in nectarine fruits. The materials and experimental methods were described clearly and in detail. The findings should be of high value for understanding the wounding effects on pathogen infection in peach and nectarine fruits.

In the study, the conclusion is that the wounding has no systemic effect on M. laxa infection. Multiple longitudinal wounds on the fruit were made by a razor blade. I wonder if this type of cut wound is common in the orchard. It seems to me that the common wound of fruits during harvesting is likely mechanical bumping. Maybe different types of wounding could release different VOCs thereby exerting different effects on pathogen infection. The authors should discuss such possibilities.

Did the wounding change the flavor and taste of later fruits? Since flavonoid and phenylpropanoid compounds were involved in the fruit response to wounded infection, were there any changes related to fruit taste in ripening fruits triggered by prior wounding? Although these are not the theme of the study, the results may be of interest to the audience in fruit postharvest storage.

“Fruit was disinfected for 30 seconds with 60 °C water [41].” Does 60 °C water for 30 seconds could result in an accelerated ripening of fruits?

Skin samples from all genotypes were analyzed using HPLC. Why did not include some flesh tissue in regions areas with the red reaction? Fruit skin and flesh tissue may have different specialized metabolites.

Lines 63-66, some references [20-23] of plant responses to wounds are a little out of date. Recent literature should be referenced, such as plant responses to herbivory, wounding, and infection. Int. J. Mol. Sci. 2022, 23, 7031.

Author Response

Dear Reviewer,

Next, it is answered point by point, and in the new version it is marked with change control in the Word file and it is highlighted with red text.

Point 1: In the study, the conclusion is that the wounding has no systemic effect on M. laxa infection. Multiple longitudinal wounds on the fruit were made by a razor blade. I wonder if this type of cut wound is common in the orchard. It seems to me that the common wound of fruits during harvesting is likely mechanical bumping. Maybe different types of wounding could release different VOCs thereby exerting different effects on pathogen infection. The authors should discuss such possibilities

Response 1: We chose this way of wounding because it was easily reproducible, very similar to natural micro-cracking and thin enough to allow recovery such as in case of micro-cracking, Spontaneous skin micro-cracking linked to excessively rapid fruit growth often happens before maturity. Indeed, in case of bumping after harvest, fruit are nearly unresponsive and they are no longer able to implement defense.

However, the type of wounding may actually have effect on the VOCs released but we did not find literature on this. So it’s difficult to discuss.

Added in discussion, line 493: "However, the type of wounding may have an effect on the volatile compounds released, but no literature on this was found."

 

Point 2: Did the wounding change the flavor and taste of later fruits? Since flavonoid and phenylpropanoid compounds were involved in the fruit response to wounded infection, were there any changes related to fruit taste in ripening fruits triggered by prior wounding? Although these are not the theme of the study, the results may be of interest to the audience in fruit postharvest storage.

Response 2: This is a good and interesting question, but out of the scope, indeed. We focused the study before harvest because at maturity and after, the fruit is very less reactive. Moreover it is not direct to make links between VOCs and flavor/taste. We have no data to implement this question.

 

Point 3: “Fruit was disinfected for 30 seconds with 60 °C water [41].” Does 60 °C water for 30 seconds could result in an accelerated ripening of fruits?

Response 3: We have used this protocol following the results of the tests done by the French applied experimental center for Fruit (CTIFL). They have looked to firmness, Brix (Total  soluble sugars) and acidity, and did not find any impact of this ‘bath’ compared to control.

We add the bibliographic citation in the Materials and Methods point (Reference Number 43 corresponding to Lurol et al., 2009) line 112 and in References (43. Lurol, S.; Tabaries, P.; Buffet, B.; Sobas, M .-A.; Portal, A. Lutte post récolte contre Monilia. Application d´eau chaude sur pêche. InfosCTIFL. 250. pp. 32-36.).

 

Point 4: Skin samples from all genotypes were analyzed using HPLC. Why did not include some flesh tissue in regions areas with the red reaction? Fruit skin and flesh tissue may have different specialized metabolites.

Response 4: Of course we agree that skin contains different metabolites from flesh. However, the ‘red’ coloration of the red zones was very superficial, and reduced to kin. So there was no reason to analyse flesh in this case.

 

Point 5: Lines 63-66, some references [20-23] of plant responses to wounds are a little out of date. Recent literature should be referenced, such as plant responses to herbivory, wounding, and infection. Int. J. Mol. Sci. 2022, 23, 7031.

Response 5: Thanks, the suggestion is taken into account and the bibliographic citation is added in the introduction.

Introduction line 66, the number 24 referring to the citation Mostafa et al., 2022 was added.

In references added: 24. Mostafa, S.; Wang, Y.; Zeng, W.; Jin, B. Plant responses to herbivory, wounding, and infection. Int. J. Mol. Sci. 2022, 7031. https://doi.org/10.3390/ijms23137031.

 

 

Many thanks! for the correction and suggestions.

The authors

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

Very interesting study evaluating interaction between the fruit and pathogen under various aspects. This is a very important study that provides insight in how pathogen is responding to the presence of other wounded fruit and shows reaction of the fruit to the pathogen including revealing some metabolites associated with the infection response in the fruit. Major issues I had with reading the paper is the English language and also sometime not enough information being provided. Detailed comments and suggestions are provided in the attached reviewed manuscript.

I would suggest authors use Grammarly add on to the Word software to help them analyze their manuscript text and improve the writing.

 

Comments for author File: Comments.pdf

Author Response

The MDPI English editing service was used for English Editing, after this round of revision.

Thank you very much for the review, we tried to answer point by point on the joint file.

Author Response File: Author Response.pdf

Reviewer 2 Report

Brown rot is the most important peach and nectarine disease, the link between mechanical stress and infection was explored in this study. The paper suggest the intervention of biochemical interplay between the fruit and the fungi, triggered by prior injury, and flavonoid and phenylpropanoid compounds may have an influence on the response of fruit to M. laxa infection. My concerns are as follows:

1. Whether the pathogen is representative?

2. The hardness of the fruit may affect the results of the experiment.

3. The language of the article is not authentic enough, it is recommended to rewrite it carefully.

Author Response

The MDPI English editing service was used for English Editing, after this round of revision.

Thank you very much for the review, we tried to answer point by point on the joint file.

Author Response File: Author Response.pdf

Reviewer 3 Report

In some of planting region, brown rot is the economically most popular disease affecting peach and nectarine production. The authors did excellent work to evaluate the influencing of Monilinia laxa on different nectarine cultivars and selections. Sequentially, find some phenolic and flavonoid compounds that putative related to plant-pathogen reactions. This study gave us important information of fruit defense triggered by injury. It can be accepted for publication.  

But there are some character writing errors. For example,

line 205: change ground to grounded; line 351: third its change to third, its ripening; line 361; line 368…..

Author Response

The MDPI English editing service was used for English Editing, after this round of revision.

Thank you very much for the review, we tried to answer point by point on the joint file.

Author Response File: Author Response.pdf

Reviewer 4 Report

The Writing is extremely poor, MM and Results are poorly written and need massive copy-editing and rephrasing. So confusing. Long sentences! 

The abstract/needs to be fully rephrased to improve the English flow and add more sound results (data).

Line 33: rephrase, it is confusing

42-45: Very poor wordings

Again poor English: “It is also a target of research institution groups to 50 searches for molecular markers for the resistance”.

87-91: Poor wording, please rephrase

95:INRA instead of INRAE

Line 94-96: Two sentences telling the same thing!

M and Methods

The English is very poor and is not scientific

94-102: can be rewritten. For instance, you can write: Bloom dates for X yZ cultivars grown research unit of INRA Avignon 5France, were recorded on February 10th for Zephyr and C207, February 23th for C216, February 97 Agronomy 2022, 12, x FOR PEER REVIEW 3 of 21 27th for F115 and March 8th for H165, 2018 season. Fruits with no apparent lesions or infections were selected and measurements of height, 100 equatorial diameters were recorded. Fruit disinfection was done by VERB with at 60°C hot water for 30 seconds

Line 118: Displayed: are presented

127: Being 20 fruits…. Just say (20 per each condition)

 

It is preferable to name 1^st condition, the second condition using other terms

I can not edit the whole paper, you need to have your paper edited by a native English speaker with scientific baground

Author Response

Thank you very much for the review.

The MDPI English editing service was used for English editing, after the revisions asked by the 4 reviewers. We beleive the rephrasing proposed fixed all the problems along the manuscript (the ones detailed by the reviewers, plus the problems not underlined).

INRA is no longer INRA; it has changed to INRAE.

Reviewer 5 Report

The manuscript “The Influence of fruit wounding on subsequent Monilinia laxa infection of nectarines” describes a holistic study of the influence of wounds in nectarine fruits on subsequent infection by M. laxa.

The manuscript requires extensive editing in terms of scientific and English language and style.

Why the authors have worked on M. Laxa while M. fructigena is the most considered postharvest agent. Brown rot is caused by three Monilinia species: M. fructigena, M. laxa and M. fructicola. The symptoms on fruits are difficult to identify and differentiate, The diagnostic protocols for these species should be included in the manuscript (Traditional, culture-based and microscopic methods, as well as molecular techniques). Therefore, additional information on the identification of M. laxa

Line 15 Remove peach in the abstract since the pathogen does not only attack these 2 fruits.

Line 14: The work contains numerous inconsistencies: for example, In Plant material, the author talks about nectarines, but the conclusion of the abstract is about peach.

Line 123-124: Specify the strain used and its accession number. Has the pathogen used already been proven in pathogenicity tests?

Before the experiments, sub-fungal isolates must be performed in a solid medium. The authors must specify the temperature and duration of incubation

Specify the concentration of tween used. Has the effect of Tween-80® been studied separately?

Why the authors used alcohol as a disinfectant? You have to look for the antifungal effect of alcohol! Disinfection must be carried out with sodium hypochlorite

Molecular analysis to study gene expression is needed.

 

 

Reviewer 6 Report

 

The present manuscript aims to investigate the Influence of fruit wounding on subsequent Monilinia laxa infection of nectarines. I believe that the following paper might be considered for publication in the journal of agronomy after the proposed revision.

 

In general, the research deals with an important topic and reached promising results that could help in controlling fungal diseases.

Only small English writing problems need to be modified, the authors should check the whole manuscript and correct them to fit the publication standards

 

The abstract:

 we believe that the abstract should take the following organization:

The author should cite the background of the subject and the problematic, then followed by the methodology, and end his abstract with the final results and recommendations or conclusion, for that a  new rearrangement is needed.

 

Line 27-28: “These compounds may be actively involved in plant-pathogen reactions and/or activation of metabolic pathways implicated in the susceptibility/resistance of peach to M. laxa.’

Is it peach or nectarine please clarify.

 

The Introduction:

 

1-    The sentence "The BR resistance is of quantitative inheritance [13-15], and therefore its heritability is medium to low with great environmental influence [10,16-18]." is not very clear in explaining the relationship between the inheritance and heritability.

2-    The sentence "In comparison to other plant organs/tissues, little information is available on the molecular mechanisms of wounded fruits [19]" can be made clearer and more concise by saying "Limited information is available on the molecular mechanisms of wounded fruits in comparison to other plant organs/tissues".

The novelty of the study needs to be highlighted compared to other similar studies.

The Introduction part must contain the whole background regarding the targeted problem and how to solve that problem with a comparison with the literature review; please check and revised accordingly.

 

Material and methods:

 

Please change: 2.1. Plant material to Fruit material or fruit preparation

Line 100-101: The authors report that they have only used the temperature as a way of fruit disinfection, In general, hot water treatment at 60°C may not be effective against all types of pathogens, especially those that are heat-resistant. Therefore, the use of alcohol and sodium hypochlorite or the combination of all methods may be needed to achieve more thorough and complete disinfection of the fruit.

In section: 2.2. Pathogen culture and incubation conditions

 

Is the M. laxa strain identified using Molecular means, if so please add the accession number of the fungus published in the genebank.

 

Please add the composition of the PDA media.

 

In the detailed experimental sections, several flows exist:

 

Lack of control group: There is no control group in the experiments or at least you didn’t mention them In the text

 

Small sample size: With only 20 fruits for each condition, it may be difficult to detect significant differences between the groups.

 

Lack of standardization: The procedure for making the wounds may not be standardized, which could lead to differences in the severity and nature of the wounds between the fruits.

 

 Please explain why you choose 7h of incubation

 

In the statistical analysis please explain the parameters you have used, as well as the statistical test and its harmony with the results obtained.

 

Discussion:

 

In the discussion, it could be more developed, and the authors should make their manuscript more appealing to psychopathologists and readers and they should highlight the relevance of their results compared to the literature, I feel that the discussion needs more work too.

 

 

Round 2

Reviewer 4 Report

EVen the English was improved, there is a need for deep copy-editing and rephrasing! 

The experiment design is not enough to answer the hypothesis presented in the introduction.

You stated that: The results suggest that flavonoid and phe- 659 nylpropanoid compounds may influence have an influence on the response of fruit to M. 660 laxa infection.

Your data is not enough to even state this! You didn't report the Antioxidant activity, nor measured the TFC or TPC to guide you in interpreting your results.