Expression of the Protein Phosphatase Gene SlPP2C28 Confers Enhanced Tolerance to Bacterial Wilt in Tobacco

Round 1

Reviewer 1 Report (Previous Reviewer 1)

Comments and Suggestions for Authors

The term POD is showed disconnectedly, with both italic and normal fonts used in different places (e.g., lines 451 and 453). Please make sure consistent formatting throughout the manuscript.

In line 433, 'gene o' is unclear. Please clarify whether this.

Comments for author File: Comments.pdf

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report (New Reviewer)

Comments and Suggestions for Authors

Article Expression of the Protein Phosphatase Gene SlPP2C28 Enhanced Tolerance to Bacterial Wilt in Tobacco by authors
Lei Ni, Yafei Qin, Mei Wang, Jianfang Qiu, Daodao Tang, Liantian Chen, Lang Wu, Jinhua Li, Yu Pan, Xingguo Zhang considers the possibility of using the SlPP2C28 gene to increase resistance to bacterial wilt.
The manuscript is formatted according to the journal rules and contains all the necessary sections.
The article reflects all the necessary methodological approaches and relevant results.
Minor comments.
There should be no conclusions at the end of the introduction - they should be moved to the conclusion. In the last paragraph of the introduction, it is necessary to formulate the hypotheses being tested, goals and objectives, but not the results.
In the results section, all the photos do not contain a scale.
In Figure 4, photo b is of very poor quality - it is not clear what can be seen there. Figure 4a also lacks a scale bar, the names of the fabrics are not labeled, the image is too small,
Figure 5 - again no scale bars.
After adding materials and making edits, the article can be published

Comments on the Quality of English Language

The language of writing does not require correction

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report (New Reviewer)

Comments and Suggestions for Authors

The peer-reviewed article “Expression of the Protein Phosphatase Gene SlPP2C28 Confers Enhanced Tolerance to Bacterial Wilt in Tobacco” is devoted to the producing transgenic Nicotiana benthamiana plants with expression SlPP2C28 gene from tomato for improving resistance to Ralstonia solanacearum. The authors found that overexpression of SlPP2C28 gene in transgenic tobacco plants induced ROS scavenging and expression different genes encoding PR-protein. This investigation is interesting and relevant, since it has both fundamental and applied significance. The article is well organized and illustrated. There are remarks that should be taken into account.

1. Add of the author-classifier to the genus and species names plants.
2. L100, L437. CaMV 35S → CaMV 35S; The author used genetic construction for Agrobacterium-mediated transformation containing target gene driven by the CaMV 35S promotor (L. 98-100 “We generated transgenic tobacco lines overexpressing SlPP2C28 under the control of the constitutive CaMV 35S promoter”) or double CaMV 35S promoter (Fig. 2a. 2x35S). Please, unified CaMV 35S (text of manuscript) and 2x35S (Fig. 2a).
3. L46-47. Under pathogen infection, plants rapidly produce excessive reactive oxygen species 46 (ROS), such as superoxide (O₂^-) and hydrogen peroxide (H₂O₂)… This phrase is not correctly, because not only superoxide (O₂^-) and hydrogen peroxide (H₂O₂) produced under pathogen damage.
4. L118-120. Please, unified cultivar or variety.
5. The authors do not indicate number of control and treated plants which used in each biological replication for all experiments (n=?). How many control and treated plants (or leaves?) were used for RNA isolation for each experiment?
6. The conclusion section is missing.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This manuscript describes the overexpression of SlPP2C28 in transgenic tobacco plants confer resistance to the R. solanacearum-led bacterial wilt. This study may help reveal the molecular mechanism underlying SlPP2C28-led genetic resistance to tomato bacterial wilt.

Please refer the following comments

Lines 112 – 115: Given that all three tomato cultivars are obtained from Professor Jinhua Li, it is desirable to cite the source once together rather than citing it individually for each tomato cultivar.

Line 129: Figs. S1 and S2 not found

Lines 219 – 224: Fig 1b & Fig 1c – Bars filled with different pattern along with the current colour different would be better to differentiate individual bar. Presently, it is difficult to compare and understand.

Line 247: Fig 2c – PCR products appear in different sizes; this is evident when compare the CK+ with the OE-2 and OE-3. Any explanation?

Line 248: Fig 2d – Rotating the x-axis labels would improve visualisation.

Line 282: It should be “column” not “row”.  i.e. The first column shows the effects of sterile water, and the second column shows the inoculation treatment effects.

Line 284: It would be better to keep all black arrow in the same size in the Figs.

Discussion part needs to be improved. Lines 372 to 424 appears as Introduction format, while Lines 425 to 439 appears as summary format of results section. It is better to combine these two formats appropriately. For example, Line 436, states that “which is consistent with previous studies”. A briefly mention what are those studies with citations in this line to make it like a discussion part.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

The manuscript demonstrated that the SlPP2C28 overexpression improved tolerance to Bacterial Wilt in Tobacco.

1. There are fundamental questions regarding why authors selected SlPP2C28 over the other 8 genes that were reported in Qiu et al, 2022.
2. Why do authors decide to overexpress SlPP2C28 from AC cultivar rather than the resistant HC.
3. As you are explaining, the overall response is resistant, the results showed that the disease damage index, even in OE-3, reached similar to as WT disease index at the end of day 9. I would call this response either a delayed damage in OE-3 or enhanced tolerance rather than resistance.
4. The discussion format is not aligned with the peer-review process. 
5. Please improve Fig .1 a and b with either colors or fill patterns. It is hard to comprehend the various shared of grey for clear understanding.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

The English language is above average, but in many instances, the statement structure is confusing and does not convey the correct message. I have highlighted a few of such statements in PDF.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

See annotated pdf.

Comments up to results are shown since the text requires a great improvement to be reviewed further.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

English needs to be greatly improved. 

Author Response

Please see the attachment

Author Response File: Author Response.pdf