Postharvest Application of Bacillus amyloliquefaciens PMB04 Fermentation Broth Reduces Anthracnose Occurrence in Mango Fruit
and Yi-Hsien Lin 2,*Round 1
Reviewer 1 Report
The manuscript deals with a very interesting topic, however, it requires extensiveEnglish language proofreading and the results and discussion section has to be substiantially improved and expanded.
I strongly suggest that authors revise the manuscript and resubmit.
Author Response
Dear reviewer:
Thank you for your careful review and kindly suggestions. We revised the manuscript based on the suggestions and responded as follows:
Response:
We thank the reviewer’s suggestion. We have sent our manuscript to a certified English editing center for extensive language proofreading and editing. Please see the tracked edits in the revised manuscript.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The manuscript “Postharvest application of Bacillus amyloliquefaciens PMB04 fermentation broth reduces the occurrence of anthracnose in mango fruit” showed the B. a. PMB04 can control the anthracnose of mango, however some conclusions were not provided because of middle writing level.
Take the abstract as an example.
1. Although we previously study demonstrated 14 that B. amyloliquefaciens (PMB04) had strong antagonistic activity against anthracnose in strawberry 15 fruit, whether PMB04 could control anthracose in mango fruit remains unexplored. This is an unbelievable hypothesis, strawberry anthracnose is caused by Colletotrichum, as well as mango anthracnose. So, PMB04 is speculated to control the mango anthracnose.
2. The inhibitory activity of PMB04 against Cg mycelium growth was comparable to fungicides pyraclostrobin and difenoconazole. This is a confused conclusion.
3. To further explore the application of PMB04, a PMB04 fermentation broth (PMB04FB) was prepared which showed superior efficacy in the reduction of anthracnose disease severity on mangoes compared to PMB04 suspension, difenoconazole, or pyraclostrobin. This is also a confused conclusion.
We found PMB04FB increased the cell death of Cg conidia. lethality rate?
Figure 1A. needs to revies, standard error a and b?
In shor, the manuscript needs a lot of writing before considering for acceptation
Author Response
Dear reviewer:
Thank you for your careful review and kindly suggestions. We revised the manuscript based on the suggestions and responded as follows:
- Although we previously study demonstrated that B. amyloliquefaciens (PMB04) had strong antagonistic activity against anthracnose in strawberry fruit, whether PMB04 could control anthracose in mango fruit remains unexplored. This is an unbelievable hypothesis, strawberry anthracnose is caused by Colletotrichum, as well as mango anthracnose. So, PMB04 is speculated to control the mango anthracnose.
Response:
We thank the reviewer’s comment and have edited the text in our revised manuscript: “B. amyloliquefaciens (PMB04) exhibits strong antagonistic activity against anthracnose in strawberry fruit, yet has not been explored for anthracnose control in mango fruit.” In Abstract P1 L14-16.
- The inhibitory activity of PMB04 against Cg mycelium growth was comparable to fungicides pyraclostrobin and difenoconazole. This is a confused conclusion.
Response:
We thank the reviewer’s comment and have made edits to clarify the conclusion in our revised manuscript: “PMB04 demonstrated better inhibitory activity against Cg mycelial growth than the fungicides pyraclostrobin and difenoconazole.” P1 L18-19
- To further explore the application of PMB04, a PMB04 fermentation broth (PMB04FB) was prepared which showed superior efficacy in the reduction of anthracnose disease severity on mangoes compared to PMB04 suspension, difenoconazole, or pyraclostrobin. This is also a confused conclusion.
Response:
We thank the reviewer’s comment and have edited the text in our revised manuscript: “A PMB04 fermentation broth (PMB04FB) was prepared and tested, and showed superior efficacy in the reduction of anthracnose disease severity in mangoes compared to a PMB04 suspension, difenoconazole, or pyraclostrobin.” P1 L19-21
- We found PMB04FB increased the cell death of Cg conidia. lethality rate?
Response:
We thank the reviewer’s comment and have edited the text in our revised manuscript: “We further found PMB04FB increased the mortality ratio of Cg conidia.” P1 L26-27
- Figure 1A. needs to revies, standard error a and b?
Response:
We thank the reviewer’s comment. The bars represent standard deviation. We have revised the figure legend of Figure 1A and other figures accordingly.
- In shor, the manuscript needs a lot of writing before considering for acceptation
Response:
We thank the reviewer’s suggestion. We have sent our manuscript to a certified English editing center for extensive language proofreading and editing. Please see the tracked edits in the revised manuscript.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments to the Author
The findings of the research are innovative achieved through well planned experimentation. There are some queries to be addressed before considering for the publication?
As the anthracnose disease of mango is pre as well as post harvest problem, that are mainly managed during pre-harvest in order to avoid the health concern due to the chemical fungicides/ pesticides.
1. Different antagonistic Bacillus spp. strains were used in this study, is there any health concern due to these strains or not? As the ultimate result would have direct impact on human health while the treatments being used for the management of post-harvest mango disease due to perishable nature of mango fruits.
2. As I learned from the manuscript, mango anthracnose in this research work was managed during pos-harvest; however, at many places it seems that research work represents both; pre- and post-harvest anthracnose.
Thus, it must be clarified throughout the manuscript by using the post-harvest anthracnose rather than anthracnose only.
Subsequently, the spore suspension of C. gloeosporioides at 1 × 105 conidia mL-1 was spotted onto the fruit, which was incubated in a plastic box at 30 oC for 7 days.
3. What method was used to apply the spore suspension? Need to be clarified.
Was that injecting? Dipping? With or without mechanical injury? As all these has impact on mango fruit as well as on disease development.
In the conclusion as explained:
To maintain crop quality and avoid postharvest losses due to anthracnose disease
4. How the quality of the crop be maintained through this research? As the work done only for post-harvest anthracnose disease of mango fruits
5. Some suggestion are give in the text to be followed throughout the manuscript (file is attached).
Comments for author File: Comments.pdf
Author Response
Dear reviewer:
Thank you for your careful review and kindly suggestions. We revised the manuscript based on the suggestions and responded as follows:
- Different antagonistic Bacillus spp. strains were used in this study, is there any health concern due to these strains or not? As the ultimate result would have direct impact on human health while the treatments being used for the management of post-harvest mango disease due to perishable nature of mango fruits.
Response:
We thank the reviewer’s comment and question. Bacillus amyloliquefaciens is a bacterial species identified as GRAS (Generally recognized as safe) in FDA of U.S. And, PMB04 has been evaluated to pose no risk of oral acute toxicity and pathogenicity to rats according to U.S. EPA OCSPP harmonized Test Guidelines 885.3050.
Reference:
US Food and Drug Administration (1999). Carbohydrase and Protease Enzyme Preparations Derived from Bacillus Subtilis or Bacillus amyloliquefaciens; Affirmation of GRAS Status as Direct Food Ingredients. Fed. Reg. 64 (78), 19887–18895
- As I learned from the manuscript, mango anthracnose in this research work was managed during pos-harvest; however, at many places it seems that research work represents both; pre- and post-harvest anthracnose. Thus, it must be clarified throughout the manuscript by using the post-harvest anthracnose rather than anthracnose only.
Response:
We thank the reviewer’s suggestion. Indeed, the control of mango anthracnose does need to be combined with pre-harvest and post-harvest control. At present, the main control strategy adopted in Taiwan is still chemical control, so the fruits used in our experiments are fresh fruits that have stopped spraying for more than 2 weeks. That's why we do not recommend pre-harvest practices in our manuscript. We have made edits throughout the revised manuscript to clarify the use in post-harvest anthracnose only
Subsequently, the spore suspension of C. gloeosporioides at 1 × 105 conidia mL-1 was spotted onto the fruit, which was incubated in a plastic box at 30 oC for 7 days.
- What method was used to apply the spore suspension? Need to be clarified. Was that injecting? Dipping? With or without mechanical injury? As all these has impact on mango fruit as well as on disease development.
Response:
We thank the reviewer’s comment. We applied C. gloeosporioides spores by dipping the spore suspension without mechanical injury. We have edited the Materials and methods section to better clarify our method.
In the conclusion as explained:
To maintain crop quality and avoid postharvest losses due to anthracnose disease
- How the quality of the crop be maintained through this research? As the work done only for post-harvest anthracnose disease of mango fruits
Response:
In our manuscript, we only focused on the control of anthracnose on post-harvest mango fruits. Since the assay on fruits can only be carried out for 3 months a year, we thank the reviewer for the great suggestion to add quality control data in future research.
- Some suggestion are give in the text to be followed throughout the manuscript (file is attached).
Response:
We sincerely thank all the reviewer’s suggestions. We have edited the text in the revised manuscript according to the reviewer’s comments given in the attached file.
Round 2
Reviewer 2 Report
I don't have any questions.
