Monitoring Rose Black Spot Disease Using Electrical Impedance Spectroscopy

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Please find the attached file for the comments.

Comments for author File: Comments.pdf

Author Response

Response to Reviewer 1 Comments

 

 

 

Thank you very much for the valuable comments on our manuscript “Identification of Rose Black Spot Resistance Using Electrical Impedance Spectroscopy” from MDPI in Agronomy - agronomy-3732463. We have considered your comments and revised the manuscript accordingly. In this revised version, changes to our manuscript were all highlighted within the document in yellow.

Our responses towards your valuable comments and suggestions are attached herewith.

 

Yours sincerely

 

Tianyi Ma

 

 

College of Horticulture,

 

Hebei Agricultural University,

Baoding, Hebei Province, China

 

Email: mty2802620991@163.com

 

 

Comment 1: The manuscript does not clearly delineate what is novel in this work relative to prior studies that applied EIS to plant disease diagnostics. Explicitly state whether the novelty lies in the specific cultivar comparisons, identification of extracellular resistivity (r_e) as a key indicator, the time-course of detection, or integration with ultrastructural findings. Include a paragraph in the Introduction to highlight this contribution.

Response 1:

Thank you very much for your valuable suggestions. We have added a discussion in the Introduction section regarding the limitations of previous applications of EIS (Please refer to lines 36 to 44.). Meanwhile, we have also highlighted the innovation of applying EIS technology to plant disease diagnosis in the Introduction (Please refer to lines 58 to 70).

Comment 2: The manuscript is verbose in some sections, especially the Results and Discussion, with frequent restatement of findings already evident in figures/tables. Revise these sections for conciseness, focusing on interpretation and significance rather than repetition of descriptive details. Several sentences use awkward or imprecise phrasing. Example: “the resistant variance consistently showing higher impedance”, “the resistant cultivar consistently exhibited higher impedance.”

Response 2:

Thank you very much for your valuable comments. We have revised the "Results" and "Discussion" sections to make them more concise and clearer for easier interpretation by readers (Please refer to lines 159 to 165、195 to 199、205 to 210、241 to 244、252 to 254、263 to 272、297 to 302、330 to 334、338 to 340、347 to 349、360 to 362 and 378 to 393). In addition, we have corrected the previously inaccurate statements; (Please refer to lines 394 to 411).

Comment 3: The statistical methods are inadequately described. The manuscript mentions significance testing and correlation analysis but does not specify the statistical tests employed (e.g., ANOVA, post hoc comparisons, correlation type). Provide full details of the statistical analyses, including tests used, confidence level, and software. Figures and tables do not consistently report measures of variability (e.g., standard error bars) or exact p-values. Ensure all figures and tables include these where appropriate

 

 

Response 3:

Thank you very much for your valuable suggestions. We have revised the Statistical Analysis section to clearly describe the statistical methods used (Please refer to lines 129 to 155). In addition, we have added figure/table captions below each image and table to help readers better understand the experiments and study design (Please refer to lines182 to 183、192 to 193、202 to 203、214 to 215、226 to 227、237 to 238、249 to 250、260 to 261、295 to 296 and 305 to 306).

Comment 4: Figures: Axis labels in several figures lack units; figure captions are terse and do not stand alone. Revise captions to clearly explain what is shown, including experimental conditions and time points.

Tables: The regression models are presented in text-heavy format that is hard to follow.

Consider formatting them more clearly with proper notation and explanatory footnotes.

Response  4:

Thank you very much for your valuable suggestions. We have revised all the figures in the manuscript by adding the missing units and updating the figure captions to facilitate clearer interpretation by readers (Please refer to lines 166 to 171、180 to 183、190 to 193、200 to 203、211 to 215、224 to 227、235 to 238、247 to 250 and 258 to 261). In addition, we have rephrased the regression models presented in the figures and supplemented the captions accordingly to improve reader understanding (Please refer to lines 295 to 302 and 305 to 306).

Comment 5: The Introduction is lengthy and sometimes redundant, with more focus on general background than on the specific problem and gap addressed in this study. Shorten the general background, and expand the discussion on why existing methods are insufficient, and why EIS is particularly suited for this purpose.

Response 5:

Thank you very much for your valuable comments. We have revised and streamlined the redundant parts in the Introduction, and added a discussion on the limitations of existing methods as well as the advantages of EIS (Please refer to lines 30 to 44 and 58 to 70).

Comment 6: The Discussion largely reiterates results and lacks critical evaluation of limitations. For example, the influence of environmental factors, cultivar-specific physiology, and field applicability are not discussed. Add a paragraph discussing limitations and potential confounding factors, and propose directions for future research (e.g., field validation, broader cultivar testing).

Response  6:

Thank you very much for your valuable comments. We have added a discussion of the current limitations, potential interfering factors, and future research directions in the Discussion section (Please refer to lines 330 to 334、338 to 340、347 to 349、
360 to 362 and 378 to 393).

Comment 7：The Conclusion section is descriptive and does not effectively synthesize broader implications or practical applications. Frame the conclusion to emphasize how the findings could guide breeding programs, disease monitoring strategies, and future methodological developments

Response 7：

Thank you very much for your valuable suggestion. We have revised the Conclusion section to highlight the practical implications of the findings for future development ( Please refer to lines 395 to 411).

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

I have been carefully reviewing the manuscript entitled “Identification of Rose Black Spot Resistance Using Electrical Impedance Spectroscopy” and I have found that it very interesting and sound. Abstract is clear, Introduction is well written and organized, Materials and methods can be easily followed and understood and Results are well structured, showed and discussed. Additionally, your paper proposal adds novelty to the field. So that, it is suitable for publication.

Nevertheless, there are some minor aspects that should be reviewed prior acceptance. Please let me list them point by point.

1. Please specify the type or paper in the first line, first page.
2. Appart of providing the coordenates (Longitude and Latitude) please provide, city and province when referring to your institution in line 98, page 3.
3. Please review figures 1 and 2 as size titles in axis X and Y are unreadable and set its size in the way they are in figure s3 and 4.
4. Finally, I have some concerns regarding statitstics and mathematical models:
   1. Could authors please justify the low R2 values obtained in the models? I expected R2 equal to 0.80 or higher. Understanding that significance values are good, are these R2 values acceptable by the authors? Please explain and/or discuss.
   2. In the same line of argumentation, have the authors considered using machine learning or ANNs? Fittings would notably increase by these methods. Also explain and/or discuss.

In my point of view, the paper would be suitable for publication after attending to the above-mentioned aspects. So that, congratulations to the authors!

Best regards,

Author Response

Response to Reviewer 2 Comments

 

 

 

Thank you very much for the valuable comments on our manuscript “Identification of Rose Black Spot Resistance Using Electrical Impedance Spectroscopy” from MDPI in Agronomy - agronomy-3732463. We have considered your comments and revised the manuscript accordingly. In this revised version, changes to our manuscript were all highlighted within the document in yellow.

Our responses towards your valuable comments and suggestions are attached herewith.

 

Yours sincerely

 

Tianyi Ma

 

 

College of Horticulture,

 

Hebei Agricultural University,

Baoding, Hebei Province, China

 

Email: mty2802620991@163.com

 

 

Comment 1: Please specify the type or paper in the first line, first page.

Response 1:

Thank you very much for your suggestion. We have specified the article type (line 1).

 

Comment 2: Appart of providing the coordenates (Longitude and Latitude) please provide, city and province when referring to your institution in line 98, page 3.

Response 2:

Thank you very much for your valuable suggestions. We have already added the cities and provinces in the article (line 88).

 

Comment 3: Please review figures 1 and 2 as size titles in axis X and Y are unreadable and set its size in the way they are in figure s3 and 4.

Response 3:

Thank you very much for your valuable comments. We have adjusted the sizes of Figures 1 and 2 to improve readability for the readers(line 166 to 171).

 

Comment 4.1: Finally, I have some concerns regarding statitstics and mathematical models:Could authors please justify the low R² values obtained in the models? I expected R² equal to 0.80 or higher. Understanding that significance values are good, are these R² values acceptable by the authors? Please explain and/or discuss.

 

Response 4.1:

Thank you for your valuable comments. During the seven sampling periods, environmental variations led to fluctuations in certain physiological indicators (e.g., reducing sugar), which resulted in a relatively low coefficient of determination (R²) when constructing regression models with r_e. However, correlation analysis showed that r_e is highly correlated with physiological parameters such as reducing sugar, indicating its potential to reflect the incidence of disease in roses. In future studies, we will consider strictly controlling environmental conditions to minimize such fluctuations and improve the accuracy of the regression models.

 

Comment 4.2: In the same line of argumentation, have the authors considered using machine learning or ANNs? Fittings would notably increase by these methods. Also explain and/or discuss.

 

Response 4.2:

Thank you for your valuable comments. We used the least squares method to construct a linear regression model due to its simplicity, strong interpretability, and good alignment with the knowledge structure of current practitioners in China’s rose industry. In future studies, we will consider applying machine learning or artificial neural network approaches to improve model accuracy and better meet the needs of practical applications in future production.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

Dear Authors,

You can find my comments in attached file.

Comments for author File: Comments.pdf

Author Response

Response to Reviewer 3 Comments

 

 

 

Thankyou very much for the valuable comments on our manuscript “Identification of Rose Black Spot Resistance Using Electrical Impedance Spectroscopy” from MDPI in Agronomy - agronomy-3732463. We have considered your comments and revised the manuscript accordingly. In this revised version, changes to our manuscript were all highlighted within the document in yellow.

Our responses towards your valuable comments and suggestions are attached herewith.

 

Yours sincerely

Tianyi Ma

College of Horticulture,

Hebei Agricultural University,

Baoding, Hebei Province, China

Email: mty2802620991@163.com

 

 

Comment 1:

The study presents interesting data as a result of the applied methods and protocols. The results obtained shed light on biotic stress caused by Diplocapron rosae. The data obtained are useful in assessing the biotic stress caused by the pathogen, the physiological and biochemical changes that occur in resistant and susceptible genotypes as a result of the infectious process. These findings can also be used in breeding of resistant rose genotypes to Diplocapron rosae. However, the methods conducted in the research cannot be used to determine the exact time to start control under real production conditions in the field or in greenhouses. The reasons for this are: that the experiments were carried out with plants that are known to be infected with the pathogen; -by the methods used in the study pathogen(s) cannot be identified, therefore, it cannot be proved an infection and accordingly predict the exact time for treatment with the appropriate fungicides. For these reasons, I recommend that concept be changed.

 

Response 1:

Thank you very much for your suggestion. We believe that the environmental conditions and plant growth status were consistent throughout the experiment, which allows us to rule out the influence of environmental stress on the plants. Therefore, this method is considered feasible for monitoring disease occurrence. However, we fully acknowledge that the method detects the presence of disease but cannot identify the specific type of pathogen. We fully accept your suggestion regarding the need to change the concept, and we have revised the title accordingly to: Monitoring Rose Black Spot Disease Using Electrical Imped-ance Spectroscopy. (Please refer to line 2.)

 

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

the context of the issue, highlighting the agronomic significance of the disease and the shortcomings of traditional control methods. I found the selection of two contrasting cultivars, ‘Carefree Wonder’ and ‘Red Cap,’ to be particularly insightful, as well as the well-considered temporal sampling design, which facilitated a comprehensive assessment of the physiological and structural responses to infection.

While the experimental procedure is generally well described, I believe the manuscript could be further improved by incorporating a few additional technical details. For instance, it would be beneficial to specify the environmental conditions during inoculation, provide a brief overview of the Colocolo model used for electrochemical impedance spectroscopy (EIS), and elucidate the physiological rationale behind the selection of biochemical parameters. These enhancements would contribute to greater clarity and reproducibility.

Nonetheless, I found that the integration of EIS with biochemical analyses—such as assessments of sugar content, proline levels, defense-related enzymes, and ultrastructural observations—represents one of the manuscript’s key strengths. Collectively, these approaches yield robust and comprehensive evidence.

The clarity and logical flow of the results facilitate the interpretation of cultivar-specific responses and the dynamics of electrical parameters as the infection advances. The figures and tables effectively support the findings; however, I recommend reviewing the titles and legends for consistency, especially to avoid errors like “FigureFig.” Additionally, I believe that Table 2 would benefit from improved formatting to enhance the interpretability of the regression models.

The conclusions are strongly backed by the data, especially the clear correlations of re and τ with physiological indicators of defense. These findings highlight their potential as early indicators of resistance. The authors’ interpretations align well with existing literature, and I did not identify any issues related to self-citation or ethical concerns.

Comments on the Quality of English Language

From a writing perspective, the manuscript is coherent and informative. However, I believe it could benefit from a round of stylistic editing to enhance linguistic precision, eliminate redundancies, and standardize technical terminology. A few minor, recurring errors could be resolved with a careful review of the text.

It provides valuable insights for researchers in plant physiology, crop protection, and disease diagnostics. The proposed non-destructive approach is particularly relevant within the framework of sustainable and technology-driven agriculture. With some adjustments in presentation and writing, I believe this manuscript warrants publication.

Author Response

Response to Reviewer 4 Comments

 

Thank you very much for the valuable comments on our manuscript “Identification of Rose Black Spot Resistance Using Electrical Impedance Spectroscopy” from MDPI in Agronomy - agronomy-3732463. We have considered your comments and revised the manuscript accordingly. In this revised version, changes to our manuscript were all highlighted within the document in yellow.

 

Our responses towards your valuable comments and suggestions are attached herewith.

 

Yours sincerely

Tianyi Ma

 

College of Horticulture,

Hebei Agricultural University,

Baoding, Hebei Province, China

Email: mty2802620991@163.com

 

 

Comment 1: While the experimental procedure is generally well described, I believe the manuscript could be further improved by incorporating a few additional technical details. For instance, it would be beneficial to specify the environmental conditions during inoculation, provide a brief overview of the Colocolo model used for electrochemical impedance spectroscopy (EIS), and elucidate the physiological rationale behind the selection of biochemical parameters. These enhancements would contribute to greater clarity and reproducibility.

 

Response 1:

Thank you for your valuable suggestions. We have added the environmental conditions of the experiment in the Materials and Methods section ( Please refer to lines 88 to 90). A description of the Cole-Cole model has been added in the Statistical Analysis section (Please refer to lines 129 to 155). In the Introduction, we have added an explanation of the principles behind the selected physiological parameters to help readers better understand the study (Please refer to lines 45 to 57).

 

Comment 2: The clarity and logical flow of the results facilitate the interpretation of cultivar-specific responses and the dynamics of electrical parameters as the infection advances. The figures and tables effectively support the findings; however, I recommend reviewing the titles and legends for consistency, especially to avoid errors like “FigureFig.” Additionally, I believe that Table 2 would benefit from improved formatting to enhance the interpretability of the regression models.

 

Response 2:

Thank you very much for your valuable comments. We have standardized the titles and figure legends throughout the manuscript. In addition, we have added a caption to Table 2 to help readers better understand the content (Please refer to lines 295 to 306).

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript is revised according to the recommendations.