Application of Fluorescence Spectroscopy for Early Detection of Fungal Infection of Winter Wheat Grains
, Olga K. Persidskaya 1, Veronika M. Andreevskaya 2, Natalia A. Semenova 1 and Sergey V. Gudkov 1Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe work presented in the Manuscript, entitled „ Application of fluorescence spectroscopy for early detection of fungal infection of winter wheat grains„. There are shortcomings and modifications that should be included in order to enhance the final manuscript for the readers.
1- An introduction sentence should be added at the beginning of the abstract to explain the importance of this work.
2- In line 13. Fluorescence Spectrometer should be changed to fluorescence spectrometer. Please check it at all manuscript.
3- The sentence in line 19 to line 21 should be rephrased to be clear for the reader.
4- Line 35. Please change volumes to scale?
5- In introduction section, what is the novelty (originality) of the work? and what is new in your work that makes a difference in the body of knowledge? The introduction should be presented the previous studies, which have the same objective of this study.
6- Line 71. The goal of this paper should be changed to the objective of this work.
7- The numbers of Subsection under Materials and Methods should be changed to 2.1., 2.2. , 2.3. …………els.
8- Line 85. Please add the citation for the first sentence?
9- The results and Discussion sections are well written.
10- Please, write the practical applications of your work in a separate section, before the conclusions and provide your good perspectives?
11- Please write the limitation of this work in conclusion section?
Comments on the Quality of English LanguageMinor editing of English language required
Author Response
Comments 1: An introduction sentence should be added at the beginning of the abstract to explain the importance of this work.
Response 1: We agree. The introduction sentence is added (see red text).
Comments 2: In line 13. Fluorescence Spectrometer should be changed to fluorescence spectrometer. Please check it at all manuscript.
Response 2: We agree. Change made (red).
Comments 3: The sentence in line 19 to line 21 should be rephrased to be clear for the reader.
Response 3: The sentence is rephrased (red).
Comments 4: Please change volumes to scale?
Response 4: We agree. Change made (red).
Comments 5: In introduction section, what is the novelty (originality) of the work? and what is new in your work that makes a difference in the body of knowledge? The introduction should be presented the previous studies, which have the same objective of this study.
Response 5: We agree. Additions have been made to the introduction (see red text).
Comments 6: Line 71. The goal of this paper should be changed to the objective of this work.
Response 6: We agree. Change made (red).
Comments 7: The numbers of Subsection under Materials and Methods should be changed to 2.1., 2.2. , 2.3. …………els.
Response 7: We agree. Changes made (red).
Comments 8: Please add the citation for the first sentence?
Response 8: We have added the citation and slightly modified the text (red).
Comments 9: The results and Discussion sections are well written.
Response 9: Dear reviewer, thank you for your kind words, as well as for your valuable comments and questions, thanks to you our manuscript becomes better and of higher quality.
Comments 10: Please, write the practical applications of your work in a separate section, before the conclusions and provide your good perspectives?
Response 10: We agree. We have added this item at the end of Discussion (red).
Comments 11: Please write the limitation of this work in conclusion section?
Response 11: We have touched on this issue in the Discussion and Conclusions sections (red text).
Reviewer 2 Report
Comments and Suggestions for AuthorsThe reviewed manuscript is devoted to the urgent problem of early detection of fungal damage to plant grains.
The authors conducted a large amount of research on modern equipment and obtained encouraging results.
The overview of the current state of the problem is good, the Manuscript contains many modern sources of information. At the same time, when analyzing the methods, it should be discussed how much the proposed EEM method is more difficult to implement and more expensive than the traditional fluorescence method (lines 61-64).
The Materials and Methods section below shows the measurement of wet grain (Figure 1), but in real conditions, the infected grain may be dry. How important is Figure 2 in the Manuscript?
In conclusion, it is necessary to indicate how the results obtained can be used in practical agricultural engineering.
The figures should be placed immediately after their first mention, and not on the next page (for example, Figure 3). This is extremely inconvenient for readers.
Author Response
Comments 1: The reviewed manuscript is devoted to the urgent problem of early detection of fungal damage to plant grains.
The authors conducted a large amount of research on modern equipment and obtained encouraging results.
Response 1: Dear reviewer, thank you for your kind words, as well as for your valuable comments and questions, thanks to you our manuscript becomes better and of higher quality.
Comments 2: The overview of the current state of the problem is good, the Manuscript contains many modern sources of information. At the same time, when analyzing the methods, it should be discussed how much the proposed EEM method is more difficult to implement and more expensive than the traditional fluorescence method (lines 61-64).
Response 2: We have touched on this issue at several points in the text (red).
Comments 3: The Materials and Methods section below shows the measurement of wet grain (Figure 1), but in real conditions, the infected grain may be dry. How important is Figure 2 in the Manuscript?
Response 3: Thank you for pointing this out. Really, figure 2 does not carry valuable information and therefore it was decided to delete it. Accordingly, the numbering of figures and their mentioning in the text has been changed. The figure was replaced in “Supplementary materials” file.
We have slightly changed the text (We've done a word substitution “cuvette for dry samples” to “cell for solid and powder samples”. (Line 138-139)
Comments 4: In conclusion, it is necessary to indicate how the results obtained can be used in practical agricultural engineering.
Response 4: We agree. We have added this item at the end of Discussion (red).
Comments 5: The figures should be placed immediately after their first mention, and not on the next page (for example, Figure 3). This is extremely inconvenient for readers
Response 5: Thank you for pointing this out. We have moved the figures to the places where they are first mentioned in the text.
Reviewer 3 Report
Comments and Suggestions for Authors[1] Fluorescence spectroscopy technology demonstrates significant potential for early detection (1-2 days) of wheat fungal infections, highlighting its high sensitivity, non-destructive nature, and diverse analytical capabilities. Detailed experimental design, multiple repetitions, comparison of different fungi, and analysis of fluorescence peak shifts further enhance the reliability and reproducibility of the research results (see Section 3 Results and Section 4 Discussion).
[2] The fluorescence peaks of different compounds may overlap in the spectrum, particularly in complex samples, affecting the accuracy and resolution of the results. It is suggested to discuss how to address this issue in the conclusion, such as combining with other detection techniques or methods (see Section 4 Discussion).
[3] Fluorescence spectroscopy data is complex and requires specialized knowledge and software for analysis and interpretation, which can be challenging for non-specialists. It is recommended to provide more detailed data analysis steps and methods in the article (see Section 3.4).
[4] Although fluorescence spectroscopy itself is non-destructive, the preparation and handling of samples may require certain techniques and time, especially in large-scale detection. It is suggested to discuss the specific requirements and improvement methods for sample handling in the article (see Section 3.2).
[5] High-quality fluorescence spectrometers and related equipment can be expensive, requiring significant financial investment for laboratories. It is recommended to mention equipment costs and economic feasibility in the discussion section, particularly for large-scale agricultural applications (see Section 5 Conclusion).
[6] Although fluorescence spectroscopy can detect fungal infections, it may not be specific enough for different fungal species. It is suggested to mention the possible combination with other techniques (such as PCR or ELISA) to improve specificity (see Section 4 Discussion).
[7] ï‚·Many citations are more than 5 years old. Kindly add new citations to ensure that more than 50% of citations are from the last 5 years. The keywords in the paper are inconsistently formatted. The numbering of subheadings in Part 2 is incorrect, e.g., 3.1, 3.2, etc. In Part 3, all subheadings are not numbered, e.g., Principal Component Analysis, etc. In Part 3, the structure of the “Analysis of Peaks” section is unclear; subheadings and serial numbers should be added. In Part 3, the descriptions of some figures and their references are inconsistent, such as “we also applied PCA (Figure 6)” and “Because the peaks in the spectra (Figure 4) varied in both intensity and location, we decided to analyze these parameters separately.”
[8] References are not cited in accordance with the standard, e.g., [18][19], [24][25][26][28][29][30] are not cited in the text. Please explain the innovation of the article and discuss its current deficiencies and future research directions. The structure of the experimental section is unclear. The discussion section is not clearly described.
[9] Kindly explain in detail why this particular method is chosen and why it has not been studied yet. And if it has been studied, what were the limitations you found that the manuscript addresses. The manuscript contains many abbreviations; it is advisable to add a list of abbreviations. The experiments in the article are mainly conducted in laboratory settings, lacking data support for field applications, which may face different challenges in actual agricultural use.
Author Response
Comments 1: Fluorescence spectroscopy technology demonstrates significant potential for early detection (1-2 days) of wheat fungal infections, highlighting its high sensitivity, non-destructive nature, and diverse analytical capabilities. Detailed experimental design, multiple repetitions, comparison of different fungi, and analysis of fluorescence peak shifts further enhance the reliability and reproducibility of the research results (see Section 3 Results and Section 4 Discussion).
Response 1: Dear reviewer, thank you for your kind words, as well as for your valuable comments and questions, thanks to you our manuscript becomes better and of higher quality.
Comments 2: The fluorescence peaks of different compounds may overlap in the spectrum, particularly in complex samples, affecting the accuracy and resolution of the results. It is suggested to discuss how to address this issue in the conclusion, such as combining with other detection techniques or methods (see Section 4 Discussion).
Response 2: Thank you for pointing this out. We have added the remark at the end of Discussion (red text).
Comments 3: Fluorescence spectroscopy data is complex and requires specialized knowledge and software for analysis and interpretation, which can be challenging for non-specialists. It is recommended to provide more detailed data analysis steps and methods in the article (see Section 3.4).
Response 3: We have added Section 2.5 “Finding the peaks of the emission excitation matrix” to Materials and Methods.
Comments 4: Although fluorescence spectroscopy itself is non-destructive, the preparation and handling of samples may require certain techniques and time, especially in large-scale detection. It is suggested to discuss the specific requirements and improvement methods for sample handling in the article (see Section 3.2).
Response 4: The advantage of fluorescence spectroscopy is that the samples do not require any special preparation.
Comments 5: High-quality fluorescence spectrometers and related equipment can be expensive, requiring significant financial investment for laboratories. It is recommended to mention equipment costs and economic feasibility in the discussion section, particularly for large-scale agricultural applications (see Section 5 Conclusion).
Response 5: We have touched on this issue at several points in the text (red).
Comments 6: Although fluorescence spectroscopy can detect fungal infections, it may not be specific enough for different fungal species. It is suggested to mention the possible combination with other techniques (such as PCR or ELISA) to improve specificity (see Section 4 Discussion).
Response 6: Thank you for pointing this out. We have we mentioned this moment at the end of Discussion (red text).
Comments 7: Many citations are more than 5 years old. Kindly add new citations to ensure that more than 50% of citations are from the last 5 years. The keywords in the paper are inconsistently formatted. The numbering of subheadings in Part 2 is incorrect, e.g., 3.1, 3.2, etc. In Part 3, all subheadings are not numbered, e.g., Principal Component Analysis, etc. In Part 3, the structure of the “Analysis of Peaks” section is unclear; subheadings and serial numbers should be added. In Part 3, the descriptions of some figures and their references are inconsistent, such as “we also applied PCA (Figure 6)” and “Because the peaks in the spectra (Figure 4) varied in both intensity and location, we decided to analyze these parameters separately.”
Response 7: We have added 36 new citations from the last 5 years. We have corrected keywords. We have changed the subsection numbers in the Materials and Methods section to the correct ones. We've corrected the structure of the Results. We have aligned the figures and their references.
Comments 8: References are not cited in accordance with the standard, e.g., [18][19], [24][25][26][28][29][30] are not cited in the text. Please explain the innovation of the article and discuss its current deficiencies and future research directions. The structure of the experimental section is unclear. The discussion section is not clearly described.
Response 8: We have put the references in the required form, cited indicated references in the text. We have discussed stated specified issues in the text (red). We hope that our revisions were able to better understand our study.
Comments 9: Kindly explain in detail why this particular method is chosen and why it has not been studied yet. And if it has been studied, what were the limitations you found that the manuscript addresses. The manuscript contains many abbreviations; it is advisable to add a list of abbreviations. The experiments in the article are mainly conducted in laboratory settings, lacking data support for field applications, which may face different challenges in actual agricultural use
Response 9: We have explained the choice of the method in the Introduction. We have added a list of abbreviations in “Supplementary materials”. Yes, of course this is a laboratory experiment, but we consider this step is necessary to develop a device for practical applications.
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe authors improved the manuscript according to my comments. It can be accepted for the publication.
