Microbial Biopesticides: Diversity, Scope, and Mechanisms Involved in Plant Disease Control

Round 1

Reviewer 1 Report

Review of Vero, et al 2023 “Microbial biopesticides: diversity, etc.”

 

This is an exceptionally comprehensive, sophisticated, and up-to-date literature review on the subject of microbial biopesticides for plant diseases control. Citations number 235 and most are very recent publications. The review contains many original ideas and comments and would be valuable for anyone interested in non-chemical disease control approaches.

 

The review is acceptable for publication after some minor revisions, mostly regarding word selection, tense, comma use, and excessive and unneeded words.

 

With 25 pages of single-space prose and tables, with many subject headings, a table of contents would facilitate retrieving information from the review if one is acceptable to this journal.

 

L28 Microbial food safety? This term applies to microbial pathogens or harm to humans they may cause and this subject is not raised in this review.  Replace “Microbial damages…” with “Losses caused by plant pathogens are among the most significant.”

 

L30 Insert after “have” the following: “…encouraged investigators to develop alternatives.”

 

L30-31. Replace this statement with “Alternatives based on microbial biopesticides tend to be safer and more environmentally benign than conventional pesticides. In recent years, formulations based on biopesticides have progressively increased in number and diversity and have attracted commercial interest.”

 

L35 Insert a comma after “factors”

 

L38 Insert a comma after “bacteriophages”

 

L40 Replace “like” with “such as”, insert a comma after “competition”

 

L49 Why is citation 4 about mycoparasitism related genes called out here – should citation 6 be used instead, since it is about food losses?

 

L51 Replace “work” with “effort”, delete the comma

 

L53 Insert a comma after “damage”

 

L54 Insert a comma after “…viruses)”

 

L54 State “cause”

 

L58-61 It would be a good idea to include a reference for these statements from the EPA. https://www.epa.gov/ingredients-used-pesticide-products/what-are-biopesticides

 

L78 Replace the very general term “microbial spoilage” with “plant pathogens”

 

L81 Replace “against” with “to inhibit”

 

L91 Insert a space after “Table”

 

L92-93 State “Modes of action that involve direct interaction and physical contact of the biological control agent and pathogen on different crops.”

 

L117 Replace “have been” with “were”

 

L127 Replace “being” and “and are”

 

L137 State “…reported, such as the control of …”

 

L141 Replace “have also be possible” with “were accomplished”

 

L156 Insert “that” after “therapy”

 

L158 Insert “light” after “UV”

 

L160 State “…bacteria. These…” Replace “accurate” with “optimal” or “effective”

 

L165 State “contents”

 

L172 Replace “contain” with “contents”

 

L174 Replace “formed by the” with “composed of”

 

L175 Replace “content” with “contents”

 

L176 Replace “is” with “are”. “Progeny” is usually assumed to be plural.

 

L191 Replace “approximate” with “approach”

 

L193 Replace “content is” with “contents are”

 

L196 Insert “population” before “density”

 

L200 Insert “environmental” before “stresses”

 

L201 Delete “We can mention” - it is not needed.

 

L203 Unclear statement – “…controlling the appearance of…”? Replace with this with “controlled” or “inhibited”

 

L207 Replace “grow at the expense of living fungi” with “use living fungi as a nutritional source”

 

L215 Replace “cultivated” with “cultured”

 

L217 Replace “the” with “this” before “fungal”

 

L231 Replace “first ones” with “former”

 

L236 Insert “upon” after “preyed”

 

L240 Insert “occurs” after “yeasts”

 

L247 Replace “establish parasitic interactions with” with “parasitize”

 

L253 Delete the comma

 

L267 Insert a comma before “have”

 

L274 Insert a comma after “Clonostachys”

 

L279 Is the “C.” here a Clonostachys sp.?

 

L286 Spell out genus of “G.”

 

L288 Eliminate excess spaces 

 

L291 Replace comma with a period after “[58]” Delete “The” – it is not needed.

 

L303 Would “absorbs” be a better word than “drains”?

 

L377 Replace “Up to date” with “Currently” or “At this time”

 

L383 State “sustainable”

 

L385 Replace “the” with “their”

 

L406 Insert a period after “[96]”, then insert “this” after “however”

 

L407 “the regulation is complex and time-consuming”

 

L410 What does “easier to handle” refer to?

 

L418 Replace “guarantee” with “ensure” or “enhance”

 

L421 Insert a space after “nanosheets”

 

L424 Delete comma. “indicate” is a better word than “suggest”

 

L441 Space needed after “other”

 

L460 Delete “In fact,” it is not needed

 

L477 Insert a comma after “genera”

 

L478 Delete “have demonstrated their ability to” this statement is not needed

 

L484 “this kind of” – it is not needed

 

L487 State “contents”

 

L488 Replace “against” with “of”

 

L449 Add a space after “2022”: “Balthazar et al. 2022 [109]” See similar correction needed on “Chung et al. 2016”

 

L503 Replace “in the inhibitory activity of the fungus against” with  “to inhibit the fungus B. cinerea”

 

L504 “confirmed”

 

L521 Delete “it has been demonstrated that” – it is not needed.

 

L525 State “contents” and “are”

 

L 549 Spell out and expand description of “SOS stress response”

 

L552 Rather than “interfere”, a negative term, “influence” is a better term here.

 

L563 Replace “was” with “is”, since the product is available now.

 

L602 The “a” is not needed. “broad-spectrum” is a term used more often than “wide-spectrum”

 

L620 State “Since that time, yeasts  have been classified into three killer toxin mode of action classes: killer, sensitive, and neutral.”

 

L624 Replace “this kind of” with “these”, and does “has been acknowledged” mean “reported”? Use “reported”

 

L625 Delete “In fact”

 

L643 Insert “the occupation of”

 

L646 Replace “has been proven that could act by inhibiting” with “was shown to inhibit” or “was reported to inhibit”

 

L648 Replace from “beating…” with “controlling human diseases and fungi in plants and foods”

 

L650 State “Santos et al [179] isolated and purified a killer toxin (CYC 1106) from Pichia membranifaciens that could be used to control B. cinerea in preharvest applications.”

 

L653 Space needed after “collaborators”, italics needed on “S. cervisiae”

 

L657 Replace ‘have” with “had”, state “from plant infection studies.”

 

L663 Sate “which cause severe losses of stone fruit”

 

L679 Replace “have shown antimicrobial activity against” with “inhibit”

 

L680 Replace “have demonstrated their ability to” with “inhibit”

 

L684 Replace “against” with “from”

 

L685 Insert “such” before “as”, state “greenhouses”

 

L686 Delete “These data suggest that”

 

L690 Delete the comma – it is not needed

 

L697 State “bull’s eye rot”

 

L705 Replace “tried to determine” with “examined” or “studied”

 

L731 Replace “evidencing” with “evidence” or “indicating”

 

L736 Replace “promptly” with “readily”

 

L792 Replace “bacteria” with “bacterium”, and “cause” with “causes”

 

L813-814 The use of “effect” is unclear here – virulence was already stated, so what “effect” does this refer to?

 

L824 Could a numbered reference be called out here?

 

L826 Replace “In the last decades” with “Recently” or “In recent years”

 

L830 & L832 Replace “scope of action” with “spectrum of activity”

 

L839-842 Safety is mentioned in L839-842 and only briefly elsewhere in the manuscript. Why is it recommended here that only the potential risks of secondary modes of action be evaluated?  The mode of action, whether primary or secondary, could possibly have little to do with human or environmental risk, especially with the application of viable microorganisms. The US EPA regulates the “Use of an antimicrobial for pre- and/or post-harvest field use on crops.” on a case by case basis (https://www.epa.gov/pesticide-registration/pesticide-registration-manual-chapter-18-other-federal-or-state-agency). In some cases, regulators denied approval of microorganism for biological control purposes if it was capable of growth at human body temperature and present on a food product. This concern was raised when postharvest decay control required large and viable antagonistic populations to be present to effectively protect harvested products during marketing and in consumers possession. 

 

References.

 

Titles of peer-reviewed journal papers should have consistent style – typically only the first word in the title is capitalized, while other words in the title are in lower case. Book chapter titles usually are in title case. 

 

The use of bold font is also inconsistent, especially when years are stated.

 

L1096 Citation is incomplete.

 

L1162 Remove excessive spaces – spacing inconsistencies occur elsewhere as well.

 

L1198 Italics needed on Botrytis cinerea

 

L1347 Italics needed on journal title

Author Response

Response to Reviewer 1 comments

We revised and improved the paper because of your valuable feedback and would like to express our appreciation for the extremely thoughtful suggestions. All grammar and language suggestions were considered. Language was also revised and improved to get a more readable manuscript. Other suggestions were evaluated and point-by-point responses to each one are listed below:

1. With 25 pages of single-space prose and tables, with many subject headings, a table of contents would facilitate retrieving information from the review if one is acceptable to this journal.

Response 1: The reviewer is right. We will include a table of contents in the final version

2. L28 Microbial food safety? This term applies to microbial pathogens or harm to humans they may cause and this subject is not raised in this review.  Replace “Microbial damages…” with “Losses caused by plant pathogens are among the most significant.”

Response 2: We referred to food safety because there are plant pathogens that produce mycotoxins (in the field such as F. graminearum or F. verticillioides or during storage such as Aspergillus flavus or Penicillium expansum) that could be dangerous for consumers.

3. L30 Insert after “have” the following: “…encouraged investigators to develop alternatives.”
4. L30-31. Replace this statement with “Alternatives based on microbial biopesticides tend to be safer and more environmentally benign than conventional pesticides. In recent years, formulations based on biopesticides have progressively increased in number and diversity and have attracted commercial interest.”
5. L35 Insert a comma after “factors”
6. L38 Insert a comma after “bacteriophages”
7. L40 Replace “like” with “such as”, insert a comma after “competition”
8. L49 Why is citation 4 about mycoparasitism related genes called out here – should citation 6 be used instead, since it is about food losses?
9. L51 Replace “work” with “effort”, delete the comma
10. L53 Insert a comma after “damage”
11. L54 Insert a comma after “…viruses)”
12. L54 State “cause”

Response 3 to 12: All suggestions have been accepted.

13. L58-61 It would be a good idea to include a reference for these statements from the EPA. https://www.epa.gov/ingredients-used-pesticide-products/what-are-biopesticides

Response 13: This reference was included. 

14. L78 Replace the very general term “microbial spoilage” with “plant pathogens”
15. L81 Replace “against” with “to inhibit”
16. L91 Insert a space after “Table”
17. L92-93 State “Modes of action that involve direct interaction and physical contact of the biological control agent and pathogen on different crops.”
18. L117 Replace “have been” with “were”
19. L127 Replace “being” and “and are”
20. L137 State “…reported, such as the control of …”
21. L141 Replace “have also be possible” with “were accomplished”
22. L156 Insert “that” after “therapy”
23. L158 Insert “light” after “UV”
24. L160 State “…bacteria. These…” Replace “accurate” with “optimal” or “effective”
25. L165 State “contents”
26. L172 Replace “contain” with “contents”
27. L174 Replace “formed by the” with “composed of”
28. L175 Replace “content” with “contents”
29. L176 Replace “is” with “are”. “Progeny” is usually assumed to be plural.
30. L191 Replace “approximate” with “approach”
31. L193 Replace “content is” with “contents are”
32. L196 Insert “population” before “density”
33. L200 Insert “environmental” before “stresses”
34. L201 Delete “We can mention” - it is not needed.
35. L203 Unclear statement – “…controlling the appearance of…”? Replace with this with “controlled” or “inhibited”
36. L207 Replace “grow at the expense of living fungi” with “use living fungi as a nutritional source”
37. L215 Replace “cultivated” with “cultured”
38. L217 Replace “the” with “this” before “fungal”
39. L231 Replace “first ones” with “former”
40. L236 Insert “upon” after “preyed”
41. L240 Insert “occurs” after “yeasts”
42. L247 Replace “establish parasitic interactions with” with “parasitize”
43. L253 Delete the comma
44. L267 Insert a comma before “have”
45. L274 Insert a comma after “Clonostachys”
46. L279 Is the “C.” here a Clonostachys sp.?
47. L286 Spell out genus of “G.”
48. L288 Eliminate excess spaces 
49. L291 Replace comma with a period after “[58]” Delete “The” – it is not needed.
50. L303 Would “absorbs” be a better word than “drains”?
51. L377 Replace “Up to date” with “Currently” or “At this time”

Response 14 to 51: All suggestions have been accepted.

52. L383 State “sustainable”
53. L385 Replace “the” with “their”
54. L406 Insert a period after “[96]”, then insert “this” after “however” 
55. L407 “the regulation is complex and time-consuming”
56. L410 What does “easier to handle” refer to?
57. L418 Replace “guarantee” with “ensure” or “enhance”
58. L421 Insert a space after “nanosheets”
59. L424 Delete comma. “indicate” is a better word than “suggest”
60. L441 Space needed after “other”

Response 52 to 60: Section 2.5 was rewritten following the editor´s recommendation. Suggestions of reviewer 1 were also incorporated.

61. L460 Delete “In fact,” it is not needed
62. L477 Insert a comma after “genera”
63. L478 Delete “have demonstrated their ability to” this statement is not needed
64. L484 “this kind of” – it is not needed
65. L487 State “contents”
66. L488 Replace “against” with “of”
67. L449 Add a space after “2022”: “Balthazar et al. 2022 [109]” See similar correction needed on “Chung et al. 2016”
68. L503 Replace “in the inhibitory activity of the fungus against” with  “to inhibit the fungus B. cinerea”
69. L504 “confirmed”
70. L521 Delete “it has been demonstrated that” – it is not needed.
71. L525 State “contents” and “are”
72. L 549 Spell out and expand description of “SOS stress response”
73. L552 Rather than “interfere”, a negative term, “influence” is a better term here.
74. L563 Replace “was” with “is”, since the product is available now.
75. L602 The “a” is not needed. “broad-spectrum” is a term used more often than “wide-spectrum”
76. L620 State “Since that time, yeasts  have been classified into three killer toxin mode of action classes: killer, sensitive, and neutral.” 
77. L624 Replace “this kind of” with “these”, and does “has been acknowledged” mean “reported”? Use “reported”
78. L625 Delete “In fact”
79. L643 Insert “the occupation of”
80. L646 Replace “has been proven that could act by inhibiting” with “was shown to inhibit” or “was reported to inhibit”
81. L648 Replace from “beating…” with “controlling human diseases and fungi in plants and foods”
82. L650 State “Santos et al [179] isolated and purified a killer toxin (CYC 1106) from Pichia membranifaciens that could be used to control B. cinerea in preharvest applications.”
83. L653 Space needed after “collaborators”, italics needed on “S. cervisiae”
84. L657 Replace ‘have” with “had”, state “from plant infection studies.”
85. L663 Sate “which cause severe losses of stone fruit”
86. L679 Replace “have shown antimicrobial activity against” with “inhibit”
87. L680 Replace “have demonstrated their ability to” with “inhibit”
88. L684 Replace “against” with “from”
89. L685 Insert “such” before “as”, state “greenhouses”
90. L686 Delete “These data suggest that”
91. L690 Delete the comma – it is not needed
92. L697 State “bull’s eye rot”
93. L705 Replace “tried to determine” with “examined” or “studied”
94. L731 Replace “evidencing” with “evidence” or “indicating”
95. L736 Replace “promptly” with “readily”
96. L792 Replace “bacteria” with “bacterium”, and “cause” with “causes”

Response 61 to 96: All suggestions have been accepted.

97. L813-814 The use of “effect” is unclear here – virulence was already stated, so what “effect” does this refer to?

Response 97: “Effect” was deleted and sentence rewritten.

98. L824 Could a numbered reference be called out here?

Response 98: A numbered reference was included.

99. L826 Replace “In the last decades” with “Recently” or “In recent years”
100. L830 & L832 Replace “scope of action” with “spectrum of activity”

Response 99 to 100: All suggestions have been accepted.

101. L839-842 Safety is mentioned in L839-842 and only briefly elsewhere in the manuscript. Why is it recommended here that only the potential risks of secondary modes of action be evaluated?  The mode of action, whether primary or secondary, could possibly have little to do with human or environmental risk, especially with the application of viable microorganisms. The US EPA regulates the “Use of an antimicrobial for pre- and/or post-harvest field use on crops.” on a case by case basis (https://www.epa.gov/pesticide-registration/pesticide-registration-manual-chapter-18-other-federal-or-state-agency). In some cases, regulators denied approval of microorganism for biological control purposes if it was capable of growth at human body temperature and present on a food product. This concern was raised when postharvest decay control required large and viable antagonistic populations to be present to effectively protect harvested products during marketing and in consumers possession. 

 Response 101: The reviewer is right. The paragraph was rewritten. “Thus, both primary and secondary modes of action should be assessed to identify any potentially harmful impact on human health (producers, harvesters, processors, and consumers), the environment, other crops, and beneficial microorganisms.”

102. Titles of peer-reviewed journal papers should have consistent style – typically only the first word in the title is capitalized, while other words in the title are in lower case. Book chapter titles usually are in title case. 

Response 102: References style was revised carefully.

103. The use of bold font is also inconsistent, especially when years are stated.

 Response 103:  References style was revised carefully.

104. L1096 Citation is incomplete.

Response 104: Citation was completed.

105. L1162 Remove excessive spaces – spacing inconsistencies occur elsewhere as well.

 Response 105: Excessive spaces were removed.

106. L1198 Italics needed on Botrytis cinerea

 Response 106:  Done

107. L1347 Italics needed on journal title

Response 107:  Done

Reviewer 2 Report

Dear authors,

Some minor corrections are needed.

Please verify automatic correction (spaces, comas, italics, ...).

Please consider the comments included in the text.

 

Comments for author File: Comments.pdf

Author Response

Response to Reviewer 2

We would like to express our appreciation for the extremely thoughtful suggestions. We revised and improved the paper as a result of your valuable feedback. Language was  corrected and improved to get a more readable manuscript. Each comment was evaluated and point-by-point responses to each one are below. 

1. L 44 This sentence “ Food losses can be described as the reduction in the quantity and quality of food during the production and storage stages” is equal to the first sentence of the abstract. Please rephrase one of them.

Response 1: The sentence was rephrased

2. L92 Title of the table must be on the same page.

Response 2:  In the final version it will be corrected

3. Table 1 Pseudomonas in italic

Response 3: Done

4. L229 Include authors' name

Response 4:  The sentence was rewritten

5. L 246 Mycoparasitism, please expand

Response 5:  This paragraph is introductory to this section. Further explanations of the different kinds of mycoparasitism are exposed in the following sections.

6. L291 Please, rewrite. This sentence doesn't make sense. “mycoparasite has been developed [58], The 291 mycotrophic behaviour is characteristic of Trichoderma and rosea [63].”

Response 6: The sentence was rewritten

7. L294 (64, 65]

Response 7: The curved bracket was substituted by a right bracket.

8. L351 was caused reduced

Response 8: The sentence was rewritten

9. L472 Pleease review the definition of Antibiotics

Response 9:  The definition of antibiotics used in this review is a conservative one. Could you please guide us in order to actualize it?