Secondary Metabolites from Streptomyces araujoniae S-03 Show Biocontrol Potential against Rhododendron Root Rot Caused by Phytophthora cinnamomi
and Yinjuan Zhao 1,3,*
Beatriz Pérez
Round 1
Reviewer 1 Report
-Put in italics: Rhododrendon, Phytophthora cinnamomi and Streptomyces araujoniae title and throughout the text.
-Reduce the introduction (lines 1 to 16) to a single sentence.
-Continue: 'In this study...
-Add methodology: how you isolated and identified the S-03 strain of Streptomyces araujoniae
-Reorganize the text, first methodology (number of repetitions, control, variables evaluated, statistical analysis of the data).
-Add methodology. How enzymes, secondary metabolites and volatile substances were obtained?
-Add methodology: How biocontrol/nhibitory effect were evaluated?
-Add methodology and results of pathogenicity/virulence test of Phytophthora cinnamomi
-Rewrite conclusions (last four lines). Such as: Streptomyces araujoniae reduced.......in rhododrendon plants. Therefore it may be used to evaluate control of Phytopthora cinnamomi in others hosts.
-Add references
-Add photos, tables, etc.
average
Author Response
For research article
|
Response to Reviewer X Comments
|
||
|
1. Summary |
|
|
|
Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted/in track changes in the re-submitted files.
|
||
|
2. Questions for General Evaluation |
Reviewer’s Evaluation |
Response and Revisions |
|
Does the introduction provide sufficient background and include all relevant references? |
Must be improved |
] |
|
Are all the cited references relevant to the research? |
Not applicable |
|
|
Is the research design appropriate? |
Must be improved |
|
|
Are the methods adequately described? |
Must be improved |
|
|
Are the results clearly presented? |
Must be improved |
|
|
Are the conclusions supported by the results? |
Must be improved |
|
|
3. Point-by-point response to Comments and Suggestions for Authors |
||
|
Comments 1: [-Put in italics: Rhododrendon, Phytophthora cinnamomi and Streptomyces araujoniae title and throughout the text.] |
||
|
Response 1: Thank you for pointing this out. We agree with this comment. Therefore, We have corrected the italicized Latin names in the full text.
|
||
|
Comments 2: -Reduce the introduction (lines 1 to 16) to a single sentence.] |
||
|
Response 2: This passage introduces the harm of Phytophthora cinnamomi from China and abroad, showing an increasing trend year by year. It's progressive. We don't think it's repetitive.
Comments 3: - Add methodology: how you isolated and identified the S-03 strain of Streptomyces araujoniae
Response 3: Thank you for pointing this out. We agree with this comment. We have added the method of strain screening.line104~110.
Comments4: Reorganize the text, first methodology (number of repetitions, control, variables evaluated, statistical analysis of the data).
|
||
Response 4: We have deleted the jumbled sentences.line 127-135
Comments5: Add methodology. How enzymes, secondary metabolites and volatile substances were obtained?
Response 5: From line 146 to line 153, the mycelium of Phytophthora cinnamomi filament in the treatment group and the control group was ground with Tris-HCl buffer (0.05 mol/L, pH 7.5, 4℃) at a ratio of 1 to 5 (V: W). Centrifuge the homogenate at 4℃ for 15 minutes at the speed of 10,000 g/min, and the obtained supernatant is the crude enzyme solution, which is used to detect the activity of β -galacturonidase and polygalacturonase.
The acquisition of secondary metabolites is in title 2.6. Preliminary separation and purification of secondary metabolites by organic solvent extraction and silica gel column chromatography.
In the plate antagonism experiment with partition, it was found that S-03 strain could not produce volatile substances with antibacterial effect. Therefore, the volatile substances of strain S-03 were not obtained in this paper.
Comments6: Add methodology: How biocontrol/nhibitory effect were evaluated?
Response 6: The inhibition rate of mycelium growth was calculated and the inhibition effect was evaluated by plate antagonism experiment.
Comments7: Add methodology and results of pathogenicity/virulence test of Phytophthora cinnamomi
Response 7: The pathogenicity of Phytophthora cinnamomi can be evaluated by observing the growth status of inoculated plants. The leaves of the plants wilted, the fibrous roots of the roots decreased, and the roots rotted.
Comments8: Rewrite conclusions (last four lines). Such as: Streptomyces araujoniae reduced.......in rhododrendon plants. Therefore it may be used to evaluate control of Phytopthora cinnamomi in others hosts.
Response 8: We have rewritten the conclusion.line 359
Author Response File: 
Author Response.docx
Reviewer 2 Report
The study is interesting and reports for the first time that Streptomyces araujoniae can be used for the biological control of P. cinnamomi. I think this study will add more information to those working in the same field.
Although of that, there are some comments are listed below:
The first letters of title words should be in capital according to the style of the journal.
In Materials and Methods, many experiments lack supported references.
Line 100: The authors should mention the place and country of Nanjing Forestry.
Line 127: Streptomyces araujoniae should be in italic.
Line 134: The procedure for SEM examination should be written in more detail.
Line 138: The rpm of centrifugation should be updated to g.
Line 145: Sterile water should be corrected to sterile distilled water.
Line 153: Correct medium to media.
Line 164: What is the type of seed used?
Line 167: The rpm of centrifugation should be updated to g.
Line 182: Rhododendron should be not in italic, since it is in italic title.
Line 183: What do the authors mean by Splendid?
Line 185: P. cinnamomi should be in italic.
Line 187: P. cinnamomi should be in italic.
Line 203: Correct The microscopy to The SEM microscopy.
Line 220: The Legend of Figure 2 should start with SEM micrographs showing the effects.
Line 225: Correct in the control group to that of the control group.
Line 224: MDA should be written in detail when mentioned at first.
Line 228: Phytophthora should be written in italic.
Line 242: Phytophthora should be written in italic.
Line 261: Rhododendron should be not in italic.
Line 266: Phytophthora should be written in italic.
Line 281: Streptomyces should be written in italic.
Line 283: Streptomyces should be written in italic.
Lines 285, and 286: The mentioned organisms should be written in italics.
Line 310: Streptomyces should be written in italic.
Line 314: Botrytis cinerea should be written in italic.
Line 318: Correct hyphae to hyphal.
Line 328: The mentioned organisms should be written in italics.
The authors should write a conclusion
The authors should follow the style of the journal in writing the reference list.
Few language mistakes are found
Author Response
|
Response to Reviewer X Comments
|
||
|
1. Summary |
|
|
|
Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted/in track changes in the re-submitted files. |
||
|
2. Questions for General Evaluation |
Reviewer’s Evaluation |
Response and Revisions |
|
Does the introduction provide sufficient background and include all relevant references? |
Yes |
[] |
|
Are all the cited references relevant to the research? |
Yes |
|
|
Is the research design appropriate? |
Yes |
|
|
Are the methods adequately described? |
Can be improved |
|
|
Are the results clearly presented? |
Yes |
|
|
Are the conclusions supported by the results? |
Not applicable |
|
|
3. Point-by-point response to Comments and Suggestions for Authors |
||
|
Comments 1: [The first letters of title words should be in capital according to the style of the journal.] |
||
|
Response 1: Thank you for pointing this out. We agree with this comment. Therefore, We have made the first letter of the title is capitalized..
|
||
|
Comments 2: - In Materials and Methods, many experiments lack supported references.] |
||
|
Response 2: Some methods are references, and some things are experimental methods we do ourselves, so we add references..
Comments 3: - The authors should mention the place and country of Nanjing Forestry. Response 3: We added (china, Jiangsu) to it.line104
Comments4: Streptomyces araujoniae should be in italic.
|
||
Response 4: Has been italicized.line 139
Comments5: The procedure for SEM examination should be written in more detail.
Response 5: We add some operating conditions and methods of scanning electron microscope.line 146-162
Comments6: The rpm of centrifugation should be updated to g
Response 6: We have changed the centrifugal speed rpm to g. line167 line198
Comments7: Sterile water should be corrected to sterile distilled water.
Response 7: We have changed Sterile water to sterile distilled water.line 175
Comments8: Correct medium to media.
Response 8: We have changed medium to media
Comments9: What is the type of seed used?
Response9: the fermented seed liquid mean Pre-fermentation medium.
Comments10: Rhododendron should be not in italic, since it is in italic title. Line 185: P. cinnamomi should be in italic. Line 187: P. cinnamomi should be in italic. Line 228: Phytophthora should be written in italic. Line 242: Phytophthora should be written in italic. Line 261: Rhododendron should be not in italic. Line 266: Phytophthora should be written in italic. Line 281: Streptomyces should be written in italic. Line 283: Streptomyces should be written in italic. Lines 285, and 286: The mentioned organisms should be written in italics. Line 310: Streptomyces should be written in italic. Line 314: Botrytis cinerea should be written in italic. Line 328: The mentioned organisms should be written in italics.
Response10: All the above changes about italics have been completed.
Comments11: Line 183: What do the authors mean by Splendid?
Response11: Rhododendron plants are healthy. Repeat with the back “health”, I choose to delete.
Comments12: Line 203: Correct The microscopy to The SEM microscopy.
Response12: We have changed The microscopy to The SEM microscopy
Comments13: Line 220: The Legend of Figure 2 should start with SEM micrographs showing the effects.
Response13: The picture order has been adjusted.
Comments14: Line 225: Correct in the control group to that of the control group.
Response14: We removed in.
Comments15: MDA should be written in detail when mentioned at first.
Response15: We add the full name of malondialdehyde.line 180
Comments16: Line 318: Correct hyphae to hyphal.
Response16: We have changed hyphae to hyphal line 353
Comments17: The authors should write a conclusion
Response17: We have rewritten the conclusion.line 359
Comments18: The authors should follow the style of the journal in writing the reference list.
Response18: References are revised according to the periodical template.
Author Response File: Author Response.docx
Reviewer 3 Report
Please see the attached file of comments and suggestions.
Comments for author File: 
Comments.pdf
Please note the common microbiology terminology as presented in the comment file.
Also, the use of nonscientific and no-precise terminology.
Author Response
|
Response to Reviewer X Comments
|
||
|
1. Summary |
|
|
|
Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted/in track changes in the re-submitted files. |
||
|
2. Questions for General Evaluation |
Reviewer’s Evaluation |
Response and Revisions |
|
Does the introduction provide sufficient background and include all relevant references? |
Yes |
[] |
|
Are all the cited references relevant to the research? |
Not applicable |
|
|
Is the research design appropriate? |
Can be improved |
|
|
Are the methods adequately described? |
Yes |
|
|
Are the results clearly presented? |
Yes |
|
|
Are the conclusions supported by the results? |
Yes |
|
|
3. Point-by-point response to Comments and Suggestions for Authors |
||
|
Comments 1: [The present presentation of this work went directly to a single isolation of a soil inhabiting microorganism. Logically such research work would have gone through an intensive process of isolation targeting certain group(s) and or kind or genera or species of soil microorganisms. Or, it may have been obtained from an authenticated resource(es). So, the researchers need to clarify this point according to the above stated concept.] |
||
|
Response 1: Thank you for pointing this out. We agree with this comment. Indeed, some specific bacteria in the soil have been reported, but there are still some undeveloped excellent strains in the soil. From the perspective of the development of resource microorganisms, we should screen more new functional microorganisms. Fortunately, we have screened an excellent actinomycete.
|
||
|
Comments 2: - The Title; Streptomyces araujoniae S-03 acts as a biocontrol agent against the rhododendron pathogen Phytophthora cinnamomi by producing secondary metabolites. This plant disease case is a Root Rot disease. It is known as, Phytophthora root rot. It is caused by several species of the genus Phytophthora such as P. catorum and P. cinnamomic.
|
||
|
Response 2: Thanks for your comment .We have changed the title.Secondary metabolites from the bacterium isolate Streptomyces araujoniae S-03 show biocontrol potential against rhododendron Phytophthora root rot caused by Phytophthora cinnamomic.
Comments 3: - Line # 99. The statement “S-03 bacteria were isolated from healthy rhododendron soil in the diseased rhododendron cluster at…” It is confusning to say, ‘isolated from healthy rhododendron soil in the diseased rhododendron cluster’. What is ‘healthy soil’?? And how come ‘healthy soil’ in diseased..?? Needs to be rephrased showing the full name of the bacterial isolate. Correct the grammatic case ‘were’? Clarify ‘healthy rhododendron soil in the diseased rhododendron’?
Response 3: We have changed the description.line 103
Comments4: Line # 150. Give the full name of MDA..
|
||
Response 4: We add the full name of malondialdehyde.line 180
Comments5: Line # 152. ‘2.5 Detection of the antibacterial capacity of S-03’. Or do you mean ‘Antifungal’?? Or do you mean
Response 5: We have changed the antibacterial activity to antagonistic activity. Do you think it is reasonable?
Comments6: Fig 1. Line # 211. Give full name of the isolate; S. araujoniae S-03
Response 6: We have changed the full name of the isolate; S. araujoniae S-03 In the legend.
Comments7: Fig 2. The SCM pictures of the treated culture need to be clearer and in focus.
Response 7: We have changed the picture.
Comments8: Line # 254. Fig 4, Same comment as above, Detection of Antibacterial. Also, B in this figure, it is the ‘liquid culture
extract’. It does not sound right saying ‘Fermented culture broth’. It is the Liquid extract of a liquid media or ‘broth’ that was fermented by the microorganism, your bacterial isolate.
Response 8: We have changed Fermented culture broth to Fermented Filtrate
Comments9: Also, C in this Fig 4. Need not to publish all those pictures with no effect. The whole figure could be replaced by
figures of fractions; 6-11. Then this does not describe the elution time fragment which will indicate some
concentration effect from beginning to tapering end??
Response9: We believe that the completed process can reflect the separation time of the active substance, thus proving its chemical polarity.
Comments10: Lie # 216. ‘Inhibitory 216 effect of fermentation broth from strain S-03’. The use of ‘fermentation broth’ indicate using all of the culture including the microorganism. While checking extracellular activity of a microorganism in culture needs to separate
the organism from the culture. That is to extract the liquid only without the organism and use as liquid culture extract. Note: It
This paragraph is very much of the introduction is more precise and scientifically correct to use ‘liquid culture extract’ here and wherever it is meant to be describing the
extracted liquid from a liquid culture of the microorganism. In this treatment and results in Fig 1, the tested material is a ‘Raw liquid culture extract’ to differentiate this from the use of purified fractionated part(s) of that extract.
Response10: In method 2.3, the fermentation broth used later has been removed bacteria with a 0.22 µm filter.
Comments11: Line 183: Line# 278-288. This paragraph is very much of the introduction. It needs to be rewritten directly starting for instance with line 289.
Response11: We removed the redundant introduction of Streptomyces.
Comments12: Line 203: Line # 300. ‘cinnamom blight’ OR ‘Phytophthora root rot’ !!??
Response12: We have changed cinnamom blight to Phytophthora root rot.line 335
Comments13: Line # 309. ‘Herein, the strain S-03 isolated and identified as S. araujoniae’. Needs to be rephrased. Herein an isolate of S. araujoniae and characterized as S. araujoniae S-03..
Response13: This sentence has been modified according to your request.line 344
Comments14: Line 311. ‘This strain was named after Janet Magali de Arrau to 311 recognize her contribution to the study
of actinomycetes in Brazil’ This is not clear was this isolate previously obtained by them? Or it may connotate that it may show what indicate the name of that person, which is currently is not !!
Response14: We removed the description about Janet Magali de Arrau.line 347
Comments15: Line# 317. ‘the suspension’ this is not a scientifically precise description.
Response15: We have changed ‘the suspension’ to fermented liquid
Comments16: Line 324. Do you mean the ANTIBIOTIC activity not ANTIBACTERIAL. Here you are investigating this
bacterial isolate (Actimycetous) for its activity against another fungus so you are dealing with ANTI FUNGAL activity in specific or antibiotic in a general sense.
Response16: We have changed the antibacterial activity to antagonistic activity. Do you think it is reasonable?
Comments17: Lines 323-343. This paragraph is a rehash of the results and it needs to be rewritten is discussion phormat
Response17: We have rewritten the conclusion.line 359
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
-TITLE: the word SHOW in the title should be in past tense (SHOWED)
-TITLE: delete the C in cinnamomi
-TITLE: rewrite the title such as:
Secondary metabolites from Streptomyces araujoniae S-03 showed biocontrol potential against rhododrendon root rot caused by Phytophthora cinnamomi.
-ABSTRACT:
Line 18: replace phytophthora for Phythophthora rot diseases
Line 22: replace In this study, a strain...for In this study, a bacterial strain...
-Line 23: replace diseased rhododrondron for diseased rhododendron plants
-Line 36: replace
average
Author Response
Dear reviewer
Thank you very much for your letter and advice.We have revised the paper,and would like to re-submit it for your consideration. We have addressed the comments raised by the reviewers, and the amendments are highlighted in red in the revised manuscript. Change show in title to shown and delete the C in cinnamomi. The revised title is Secondary Metabolites from Streptomyces Araujoniae S-03 Showed Biocontrol Potential Against Rhododendron Root Rot Caused by Phytophthora Cinnamomi. In the abstract, in line 18, change phytophthora to Phythophthora rot diseases. In line 22, add bacterial before "strain". Finally, change the disease Rhododendron to diseased rhododendron plants.We hope that the revision is acceptable,and I look forward to hearing from you soon.
With Best wishes
Your sincerely
Sun zhimin
Author Response File: Author Response.docx
