Using Plant-Based Preparations to Protect Common Bean against Halo Blight Disease: The Potential of Nettle to Trigger the Immune System

Round 1

Reviewer 1 Report

In the manuscript entitled, “Using plants to protect common bean against halo blight disease: the potential of nettle to trigger the immune system”, authors have evaluated the effects of

 Plant-based preparations on   pathogen infections mediated by   Pseudomonas syringae pv. Phaseolicola. It appears interesting; however, I have a quarry to be addressed before the acceptance of manuscript.

Herein, authors evaluated the expression profiles of various genes such as PR1, WRKY33, MAPKK, RIN4, PR1, PAL1 along with an unknown gene, however, since in the defense related mechanisms plant cell wall play a key role in altering resistance, why authors have largely ignored (except PAL1) observing the expression profiles of genes directly or indirectly involved in cell wall biosynthesis, for instance, cellulose synthases?

Author Response

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Reviewer 2 Report

The manuscript entitled "Using plants to protect common bean against halo blight disease: the potential of nettle to trigger the immune system" studied the effect of plant-based preparations (PBPs) on managing the disease caused by  Pseudomonas syringae pv. phaseolicola (Pph). The authors included four plant-based preparations in the study and investigated these PBPs' ability to inhibit the growth of Pph and induce host defence response. The authors selected two PBPs based on higher inhibitory effects on the growth of Pph. The authors tested them by measuring the total H2O2, lipid peroxidation and antioxidant enzymatic activities, and the expression of six defence-related genes.

The introduction is well written and easy to follow. The experimental design and data analysis appeared to be reasonable and easy to follow.  Unfortunately, the reviewer found that results and discussion are confusing and difficult to interpret. Authors showed that the production of H2O2 was reduced by 50% after PBP treatment, compared to the mock treatment. The PBP treatments resulted in the up-regulation of  SOD and lipid POX, but down-regulation of CAT. These results are contradictory.  The authors stated that RG is a resveratrol-rich PBP, a well-known ROS-scavenger molecule and lipid peroxidation protector. However, the reviewer found that it is not very convincing. Also, it is hard t believe that the nearly 50% reduction in the production of H2O2 is due to the mobilization of H2O2 to the signalling process. Is there any evidence supporting this statement?

Authors claimed that Urtica dioica plant-based preparations could be useful to diminish halo blight disease effects. The reviewer found there was not sufficient data to support this statement.

Author Response

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Reviewer 3 Report

1. This article is not intended as a review，so the part of induction of this paper was  too long, should be shorten to 3-4 paragraphs.
2. the name of Figure A1 and A2, should be Reorder it to 1, 2, 3 etc.
3. In Figure 1, different letters such as a, b ,c etc. need to be marked to indicate significant differences.
4. It also exists in Figure 2, and different letters need to be marked（su ch as : A, E, F), even if the difference between different treatments is not significant.
5. qRT-PCR analysis was performed on 5 genes in this paper, but the expression analysis of these 5 genes was not found in section 3.3
6. In section 3.4 of this paper, although there is no significant difference in yield indicators among different treatments, further analysis is still needed. For example, you can find out which treatment has the highest yield and how much it increases the yield compared to the control.
7. inspection 3.5, I can see from Figure 6 that different treatments have significant effects on common bean length and fresh weight, but there is no significant difference in final yield in this paper. What is the reason?

Author Response

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Reviewer 4 Report

Summary: 

Thank you so much for allowing me to review the manuscript. The manuscript dealt with Using plants to protect the common bean against halo blight disease: the potential of nettle to trigger the immune system. The author has written the manuscript clearly. The manuscript has simple; however, it is enough to provide some insight into the role of PBPs against halo blight. 

Minor comments:

1. Manuscript looks wordy in all the sections, which can address with clarity.

2. All the graphs must change into black and greyscale to understand by color blind people. 

3. All the references and abbreviations of journals should be corrected as per journal guidelines. For instance, Ref#2, 8, 9, and 114.    

4. This gene expression analysis demonstrated that plants treated with U-PBP showed the overexpression of all studied defensive genes. The word overexpression is not appropriate that can change from “overexpression to upregulation. 

5. The fold changes values (Example 2-fold) for defensive genes by qRT-PCR analysis can be given after subtracting the control.

6. Author should perform the experiment based on HPLC or GC-MS methods to quantify the presence of the secondary metabolites in the plant-based preparations (PBPs). Therefore, the result will be meaningful to convince the readers to correlate defense mechanisms by applying PBPs rather than citing the previous references. 

 7. The Common bean roots phenotype in the treated PBPs showed a reduced roots phenotype (Fig.5). Have authors recorded any data related to root phenotype? And which can be given. 

  8. May this title be changed: Using plants-based extract to protect the common bean against halo blight disease: the potential of nettle to trigger the immune system. 

Author Response

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Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Thanks for authors response and revises, and The new version of this paper has met the requirements for publication in this journal.

Author Response

Thank you very much for your revision.

Best regards

Reviewer 4 Report

No comments. 

Author Response

Thank you very much for your revision.

Best regards