Vulnerability of Walnut Pruning Wounds to Fungal Trunk Pathogens and Seasonal Conidial Dynamics of Botryosphaeriaceae in the Maule Region, Chile

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors,

This paper provides interesting and novel information about the susceptibility of wounds to fungal infection, in depends on wound age.

This is crucial for prevention and protection methods against diseases, not only in walnut orchards. The authors also confirmed the spore dispersal of Botryosphaeriaceae and Diaporthaceae trends through the year in correlation with rainfall, temperature, and humidity.

The experiment was well designed, the results are clearly presented, and the discussion is very interesting. I have only a few comments.

Materials and Methods

line 159: Field experiments were carried out during the 2023-24 and 2024-25 dormant seasons.

Rev: In my opinion, the exact term of experiment establishment should be added. The dormant season is too wide term. All data were collected during 2023 and 2024, so exactly when was the experiment assumed? All results refer to 2023 and 2024. Could you clarify this?

Results

line 258: Control treatments remained unaffected throughout the experimental period, confirming that lesion development was specific to the fungal inoculations.

Rev.: Results on Fig. 1C and D showed that the lesion caused by some pathogens didn’t differ from the length of the control lesion, e.g., Fig. 1C and D –  Di. patogenica.

line 282 - Diaporthe patagonica maintained low pathogenicity across both years, producing 6.3 mm and 5.8 mm lesions on 30- and 45-day-old wounds, respectively, in 2024-25 (Fig. 2C, D).

Rev.: In Fig. 2 the 2023 is only mentioned. Why do you use 2024-25?

line 416: Dothiorella sarmentorum exhibited intermediate virulence, with lesion lengths ranging from 26.5 mm on fresh 417 wounds to 4.0 mm at 45 days in 2023–24, and a slightly higher persistence in 2024–25 (8.0 418 mm at 45 days). 

Rv.: See above

Discussion:

line 434: This discovery has practical implications, showing that a wound age is more susceptible to some pathogens than others.

Rev.: Wound age can’t be susceptible, but fresh wounds or older wounds can, probably an aged wound (?). Furthermore, the wound age plays a role in susceptibility….

 

Author Response

Reviewer 1 comments

Comments and Suggestions for Authors

Dear Authors,

This paper provides interesting and novel information about the susceptibility of wounds to fungal infection, in depends on wound age. This is crucial for prevention and protection methods against diseases, not only in walnut orchards. The authors also confirmed the spore dispersal of Botryosphaeriaceae and Diaporthaceae trends through the year in correlation with rainfall, temperature, and humidity. The experiment was well designed, the results are clearly presented, and the discussion is very interesting. I have only a few comments.

Materials and Methods

line 159: Field experiments were carried out during the 2023-24 and 2024-25 dormant seasons.

Rev: In my opinion, the exact term of experiment establishment should be added. The dormant season is too wide term. All data were collected during 2023 and 2024, so exactly when was the experiment assumed? All results refer to 2023 and 2024. Could you clarify this?

Response: Thank you for this valuable comment. We agree that specifying the exact timing of the experiment is crucial for clarity and reproducibility. As suggested, we have now clarified the months during which the pruning and inoculation were performed.

Results

line 258: Control treatments remained unaffected throughout the experimental period, confirming that lesion development was specific to the fungal inoculations.

Rev.: Results on Fig. 1C and D showed that the lesion caused by some pathogens didn’t differ from the length of the control lesion, e.g., Fig. 1C and D –  Di. patogenica.

Response: Thank you for this astute observation regarding the description of the control treatments. We agree that the original statement was an overgeneralization and did not accurately reflect the data for Di. patagonica presented in Figure 1C and D. The correction has been made in the last sentence of the paragraph as highlighted.

 

line 282 - Diaporthe patagonica maintained low pathogenicity across both years, producing 6.3 mm and 5.8 mm lesions on 30- and 45-day-old wounds, respectively, in 2024-25 (Fig. 2C, D).

Rev.: In Fig. 2 the 2023 is only mentioned. Why do you use 2024-25?

Response: Thank you for pointing out this inconsistency. You are correct; the text mistakenly referred to the season as "2024-25" while the corresponding figure uses "2024" for the same data set. We have corrected the text in line 282.

line 416: Dothiorella sarmentorum exhibited intermediate virulence, with lesion lengths ranging from 26.5 mm on fresh 417 wounds to 4.0 mm at 45 days in 2023–24, and a slightly higher persistence in 2024–25 (8.0 418 mm at 45 days). 

Rv.: See above

Response: Thank you for highlighting this further instance of inconsistent terminology. We apologize for the oversight. We have now corrected the text in line 416 and have performed a global search to ensure that all references to the experimental years now consistently use the calendar years (2023 and 2024) to match the labels used in all figures. The corrected sentence is now on line 416.

 

Discussion:

line 434: This discovery has practical implications, showing that a wound age is more susceptible to some pathogens than others.

Rev.: Wound age can’t be susceptible, but fresh wounds or older wounds can, probably an aged wound (?). Furthermore, the wound age plays a role in susceptibility….

Response: Thank you for this precise correction regarding the scientific language. We agree that the original phrasing was inaccurate. We have revised the sentence to correctly reflect that susceptibility is a property of the wound, which is influenced by its age. The sentence in line 434 has been changed to

These investigations has practical implications, showing that fresh wounds are more susceptible to some fungal pathogens while older wounds are more susceptible to others.

 

 

Reviewer 2 Report

Comments and Suggestions for Authors

Submission ID: microorganisms-3926278

Title of the manuscript: "Vulnerability of Walnut Pruning Wounds to Fungal Trunk Pathogens and Seasonal Conidial Dynamics of Botryosphaeriaceae in Maule region, Chile ".

The authors presented their work on the evaluation of the age of pruning wounds on the lesion development on lignified walnut twigs inoculated with species from the Botryosphaeriaceae and Diaporthaceae families. The subject is very interesting, and the authors did a lot of work, and the methodology used is adequate for the objectives of the study. The results are of interest and support the conclusions. However, several points should be addressed by the authors before acceptance as follows:

Comments

Point 1: Line 42-43, Rearrange alphabetically.

Point 2: Line 145, What did the authors mean by 2% potato dextrose agar (PDA)? Is it the agar percentage? Not clear.

Point 3: Line 177, What is APDA?

Point 4: Line 229, was used

Point 5: Figure 1&2, Error bars???

Point 6: In the results, authors should add one sentence at the end of each paragraph to conclude the whole paragraph to make it easy for the reader.

Point 7: In the discussion section, please cite the figures to keep the readers in contact with the results.

Respectfully, reviewer

Author Response

Reviewer 2, point by point response to comments

 

Title of the manuscript: "Vulnerability of Walnut Pruning Wounds to Fungal Trunk Pathogens and Seasonal Conidial Dynamics of Botryosphaeriaceae in Maule region, Chile ". The authors presented their work on the evaluation of the age of pruning wounds on the lesion development on lignified walnut twigs inoculated with species from the Botryosphaeriaceae and Diaporthaceae families. The subject is very interesting, and the authors did a lot of work, and the methodology used is adequate for the objectives of the study. The results are of interest and support the conclusions. However, several points should be addressed by the authors before acceptance as follows:

Comments

Point 1: Line 42-43, Rearrange alphabetically.

Response: Thank you for this note. We have confirmed that the keywords are now listed in alphabetical order as requested.

Airborne inoculum; Botryosphaeriaceae; Diaporthe; Neofusicoccum nonquaesitum.

Point 2: Line 145, What did the authors mean by 2% potato dextrose agar (PDA)? Is it the agar percentage? Not clear.

Response: Thank you for pointing out the ambiguity in the text, the authors have now corrected the mistake and the 2% has been deleted from the sentence in track changes and highlted.

Point 3: Line 177, What is APDA?

Response: Acidified Potato Dextrose Agar (APDA). The correction has been made in the  sentence “Each isolate was reactivated for inoculation by transferring a 5-mm mycelium plug to fresh Acidified Potato Dextrose Agar (APDA) [prepared by adding 0.5 mL L⁻¹ of 92% lactic acid to PDA] and incubated at 25 ± 2°C for 10 days”.

Point 4: Line 229, was used

Response: The correction has been made in the main file.

Point 5: Figure 1&2, Error bars???

Response: Thank you for your valuable comment, the correction has been made in the main file by adding the revised images showing the error bars on the columns.

Point 6: In the results, authors should add one sentence at the end of each paragraph to conclude the whole paragraph to make it easy for the reader.

Response: Thank you for your valuable comment, one sentence at the end of each paragraph to conclude the whole paragraph has been added in the result section.

Point 7: In the discussion section, please cite the figures to keep the readers in contact with the results.

Response: The correction has been made in the discussion section.

Respectfully, reviewer