Multilocus Sequence Analysis and Detection of Copper Ion Resistance of Xanthomonas phaseoli pv. manihotis Causing Bacterial Blight in Cassava

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

In this article author use Cassava (Manihot esculenta Crantz.) to analyze the multilocus sequences and copper ion resistance detection in Xanthomonas phaseoli pv. manihotis that causes bacterial blight in host plant. The article presents previously unpublished data and overall research, and the findings given by the authors are good. Authors used molecular and bioinformatics approaches that could be given a weightage to the interpretation of data. In my view the article could be accepted for publication in this journal after some minor revision, rectifying typo-errors, and formatting in its current form.

 

Some comments and recommendations are listed below:

1)      Title looks incomplete, the authors should add the name of host plant i.e., cassava. I suggested the title “Multilocus sequence analysis and detection of copper ion resistance of Xanthomonas phaseoli pv. Manihotis causing Bacterial Blight in Manihot esculenta Crantz. (Cassava)” for more clear understanding.

2)      In abstract and in introduction, italicize ‘Manihot esculenta’ Crantz, and wherever it appears.

3)      Authors should cite reference (FAO) in the reference list.

4)      Avoid the overlapping between text and ref. cited in the manuscript.

5)      Format table 2 as per journal guidelines authors should check for font format, size, etc.

6)      In section 2.3, spell check for sub-heading.

7)      In line 115,137,144,150, ‘CuSO4 solution’ valency should be subscripted.

8)      In line 143, 149, and 151, it would be better if authors could use the subscript for wavelength as in OD₆₀₀ = 0.5.

9)      In line 171, ‘4210 nucleotides(Appendix A)’ give the space between overlapping words. Also find the same typo-errors throughout the manuscript.

10)  The authors should have to identify the typo-errors by themselves and make thorough revision before re-submitting.

 

Review comments:

 The manuscript could be accepted for publication after some minor changes and corrections in its current form.

 

 

Comments on the Quality of English Language

The language is OK

Author Response

Dear reviewer

Thank you for your warm suggestion. For your suggestions, the response is as follows.

 

1)  Title looks incomplete, the authors should add the name of host plant i.e., cassava. I suggested the title “Multilocus sequence analysis and detection of copper ion resistance of Xanthomonas phaseoli pv. Manihotis causing Bacterial Blight in Manihot esculenta Crantz. (Cassava)” for more clear understanding.

I accept this suggestion.
2)  In abstract and in introduction, italicize ‘Manihot esculenta’ Crantz, and wherever it appears.

I accept this suggestion.

3)  Authors should cite reference (FAO) in the reference list.

The statistical data (FAO) link was cited in the reference list. By the way, the data of 2021 was adopted in the revision manuscript.

4)      Avoid the overlapping between text and ref. cited in the manuscript.

The order of all the ref. Cited was checked and modified.

5)      Format table 2 as per journal guidelines authors should check for font format, size, etc.

The Format of table 2 was checked and modified.

6)      In section 2.3, spell check for sub-heading.

The wrong word ‘assayis’ was amend as ‘assays’.

7)      In line 115,137,144,150, ‘CuSO4 solution’ valency should be subscripted.

All the valency was subscripted in the revision manuscript.

8)      In line 143, 149, and 151, it would be better if authors could use the subscript for wavelength as in OD600 = 0.5.

All the number 600 was subscripted in the revision manuscript.

9)      In line 171, ‘4210 nucleotides(Appendix A)’ give the space between overlapping words. Also find the same typo-errors throughout the manuscript.

The manuscript was checked, and all the typo-errors were amend.

10)  The authors should have to identify the typo-errors by themselves and make thorough revision before re-submitting.

 The manuscript was checked, and all the typo-errors was amended.

 

We think your suggestion is very good, and the revision manuscript has been prepared. If you think further modifications are necessary, please let me know.

 

Very sincerely yours.

Shi Tao

 

Environment and Plant Protection Institute

Chinese Academy of Tropical Agricultural Sciences

E-mail：[email protected]

 

 

 

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Dr

Happy day

The paper is fine but need some more work

1-      More details are needed through the paper, for example, how did you make the sequencing process?

2-      Kindly show us the phylogenic tree which is fare better than the tree. I suggest for you to try to remove the region where there are similarities within sequences. And re-build your phylogenetic tree to get better result.

3-      I highly recommended to put a future opinion based on your result in the conclusion part. For example, you can recommend using antimicrobial agents other than that based on the copper toxicity.

4-      Some technical points should be put in concern such as using ddwater and bacterial filter before the PCR which did not remove all the existing DNA (as natural contaminants). Only sterilization can get rid from any DNA contamination.

with my pleasure

Comments for author File: Comments.pdf

Author Response

Dear reviewer

Thank you for your warm suggestion. For your suggestions, the response is as follows.

1-More details are needed through the paper, for example, how did you make the sequencing process?

    The primer sequences related with amplicon sequencing, and the sequencing of amplicon were supplemented in part ‘Materials and Methods’ of the revision manuscript. Each amplicon was sequenced with two or more primers, the short and partly overlapped sequences were merged, and the full sequence was obtained. Primer synthesis and amplicon sequencing were performed by BGI Co., Ltd (Shenzhen, Guangdong, China).

2-  Kindly show us the phylogenic tree which is fare better than the tree. I suggest for you to try to remove the region where there are similarities within sequences. And re-build your phylogenetic tree to get better result.

   In this paper, the partial sequences of five genes were used for genetic diversity analysis. All the strains had the same sequence of dnaK, gyrB, and efp, only a few polymorphisms were observed in atpD and rpoD. So, I know this phylogenic tree is not very satisfactory, but the Circular tree is a worse choice. By the way, one paragraph introducing the analysis result was supplement in the revision manuscript.

3- I highly recommended to put a future opinion based on your result in the conclusion part. For example, you can recommend using antimicrobial agents other than that based on the copper toxicity.

   I accept this suggestion. The preliminary research results indicate that ethylicin(chinese name is 乙蒜素) and 2-amino-5-sulfydryl-1,3,4-thiadiazole zinc(chinese name is 噻唑锌) were effective for CBB.

4- Some technical points should be put in concern such as using ddwater and bacterial filter before the PCR which did not remove all the existing DNA (as natural contaminants). Only sterilization can get rid from any DNA contamination.

   The reagents for PCR amplification reaction (including sterilized distilled water), PCR amplification system were supplemented in part ‘Materials and Methods’ of the revision manuscript.

By the way, the suggestion in peer-review-30063585.v1 is very good. The corresponding modifications had been completed in the revision manuscript.

We think your suggestion is very helpful, and the revision manuscript has been prepared. If you think further modifications are necessary, please let me know.

 

Very sincerely yours.

Shi Tao

 

Environment and Plant Protection Institute

Chinese Academy of Tropical Agricultural Sciences

E-mail：[email protected]