Integrative Effect of UV-B and Some Organic Amendments on Growth, Phenolic and Flavonoid Compounds, and Antioxidant Activity of Basil (Ocimum basilicum L.) Plants

Round 1

Reviewer 1 Report

General Summary:

Thank you for giving me an opportunity to review the manuscript “Biometric and biochemical changes of basil (Ocimum basilicum L.) plants in response to UV-B exposure and organic amendments application ” by Santin et al. In this paper, the author tried to combine the effects of B (2% w/w), WD (1:100), and their combination (BWD) on some biometric and biochemical parameters of basil plants (Ocimum basilicum L.) exposed to different doses of UV-B radiation (0, 1, 2 h d-1 ; UV-B irradiance of 1.36 W m−2) in controlled conditions. Overall, the paper presents valuable findings that contribute to the understanding of plant responses to environmental stressors and the potential benefits of organic amendments. The experimental design appears robust, allowing for a clear assessment of the impact of UV-B radiation and organic amendments. However, I have observed some minor issues which should be addressed as minor revisions.

Specific comments:

Please refer to the following comments:

1.      Please add numerical values of mentioned parameters in the abstract text.

2.      Ensure consistent formatting of units throughout the paper.

3.      The authors used B and C abbreviations. B is for biochar right? But, it has puzzled the text. The general reader will think if it is UV-B or biochar. Please correct this issue in the entire manuscript.

4.      At their first mention, please write the full names of the element names.

5.      Table 1: Please provide the SD of at least three replications. Also, a t-test is a must to derive a significant difference between soil and soil/biochar mixture.

6.      How do authors justify: the long-term effects of UV-B exposure and organic amendments on basil plants?

7.      In discussion: Address potential confounding factors and acknowledge any uncertainties in the data interpretation. This will help readers understand the scope and boundaries of the research.

8.      Conclusion: commendable.

Author Response

REVIEWER 1

Thank you for giving me an opportunity to review the manuscript “Biometric and biochemical changes of basil (Ocimum basilicum L.) plants in response to UV-B exposure and organic amendments application ” by Santin et al. In this paper, the author tried to combine the effects of B (2% w/w), WD (1:100), and their combination (BWD) on some biometric and biochemical parameters of basil plants (Ocimum basilicum L.) exposed to different doses of UV-B radiation (0, 1, 2 h d-1 ; UV-B irradiance of 1.36 W m−2) in controlled conditions. Overall, the paper presents valuable findings that contribute to the understanding of plant responses to environmental stressors and the potential benefits of organic amendments. The experimental design appears robust, allowing for a clear assessment of the impact of UV-B radiation and organic amendments. However, I have observed some minor issues which should be addressed as minor revisions.

Specific comments:

Please refer to the following comments:

1. Please add numerical values of mentioned parameters in the abstract text.

Reply: Numerical values were added in the abstract text as requested.

2. Ensure consistent formatting of units throughout the paper.

Reply: We made our best to check the units throughout the paper.

3. The authors used B and C abbreviations. B is for biochar right? But, it has puzzled the text. The general reader will think if it is UV-B or biochar. Please correct this issue in the entire manuscript.

Reply: As suggested, B was removed from the text and replaced with biochar.

4. At their first mention, please write the full names of the element names.

Reply: As suggested, we wrote the full name of the element.

5. Table 1: Please provide the SD of at least three replications. Also, a t-test is a must to derive a significant difference between soil and soil/biochar mixture.

Reply: We thank the reviewer for his/her suggestion. The SD of each values was reported in table 1. Student t-test of three replicates for each treatment was performed. As reported, TOC, K exc. and CEC were significantly different between the two groups, as showed in the table.

6. How do authors justify: the long-term effects of UV-B exposure and organic amendments on basil plants?

Reply: The aim of the experiment was to verify the influence of different amendments and UV-B exposures during the growth of basil plants in pre-flowering stage. We did not evaluate the effect of these treatments during the complete life cycle of plants (pre-flowering, full-flowering and post-flowering), which could be an interesting point to address in future investigations. Indeed, it is likely that plants in a different growth stage might have a similar or opposite behavior to the same soil and light conditions. We thank the reviewer for his/her observation.

7. In discussion: Address potential confounding factors and acknowledge any uncertainties in the data interpretation. This will help readers understand the scope and boundaries of the research.

Reply: We thank the reviewer for his/her valuable comment. A paragraph was added at the end of the discussion to address the reviewer comments regarding the uncertainties that might affect our study.

8. Conclusion: commendable.

Reply: We thank the reviewer this comment.

Author Response File: Author Response.docx

Reviewer 2 Report

I have had the pleasure of reviewing your manuscript titled " Biometric and biochemical changes of basil (Ocimum basilicum  L.) plants in response to UV-B exposure and organic amendments application." I must commend your comprehensive and robust approach to this pertinent issue in the realm of agricultural science.

The issue of soil fertility is indeed a pressing one, and your research not only highlights the problem but provides a tangible and innovative solution. Your study design, especially the integration of using some novel soil amendments (i.e., biochar (B) and wood distillate) with the exposure to UV-B radiation is appropriately rigorous, enabling a holistic understanding of the variables at play. Your focus on some measurements, including growth characteristics, phenolics and flavonoids as well as antioxidants activity is highly commendable. However, there are some crucial amendments required as follow:

Point 1: The suggested title of the MS should represent accurately the performed experiments without unnecessary information. I would suggest the title to be “Integrative effect of UV-B and some organic amendments on growth, phenolic and flavonoid compounds, and antioxidant activity of basil (Ocimum basilicum  L.) plants.”

Point 2: My second comment pertains to the term “wood distillate (WD)” used as one of the treatments. From the agricultural point of view this term is not common. I would rather prefer to use the term wood vinegar instead in the whole MS.

Point 3: Line 104-110 where you illustrated the main objective of the study. The hypothesis of your work needs to be more justified. Please rephrase.

Point 4: My next comment pertains to the photoreceptor’s measurements. One of the main objects of your study is to evaluate the response of basil plants to UV-B. by which should have been evaluated through the measurement of the main photoreceptor’s such as:-

ü Chlorophyll, absorbing PAR (Photosynthetic Active Radiation) and transferring energy to the reaction centers.

ü Phytochrome , absorbing mainly in the red (R) and far-red (FR).

ü Cryptochrome, absorbing blue and UV-A region (320-400nm).

ü UV-B photoreceptors, absorbing UV-B region (280 and 320 nm).

However, you did not present any photoreceptor’s measurements by which are essential for your results justification. You should have done these parameters while performing the experiment. If YES, please include this data in the results. Measurements.

Point 5: My second comment pertains to the largely descriptive nature of your results. While the data you present offer valuable insights, there could be additional analyses to deepen the interpretation and implications of your findings. Specifically, I recommend complementing your trait-by-trait analyses with correlation analysis and Principal Component Analysis (PCA).

Incorporating these statistical tools would not only strengthen your results but also provide a more nuanced understanding of the data. It would contribute to a richer and more comprehensive discussion of the interplay between selenium application, water stress, and the physiological and biochemical responses of the common bean plants.

Point 6: While your manuscript clearly reflects a sound understanding of the existing literature, I would like to emphasize the importance of including more recent publications in your references. Scholarly conversations are continually evolving and including citations from the last three years (especially from 2023) will ensure your work is positioned within the most current state of the field.

Point 7: While your manuscript displays a strong command of the topic and presents compelling findings, I noticed some minor language issues and inconsistencies throughout the text. These could potentially hinder the clarity of your message and disrupt the reader's engagement with your work.

I noticed some minor language issues and inconsistencies throughout the text. These could potentially hinder the clarity of your message and disrupt the reader's engagement with your work.

Author Response

REVIEWER 2

Comments and Suggestions for Authors

I have had the pleasure of reviewing your manuscript titled " Biometric and biochemical changes of basil (Ocimum basilicum  L.) plants in response to UV-B exposure and organic amendments application." I must commend your comprehensive and robust approach to this pertinent issue in the realm of agricultural science.

The issue of soil fertility is indeed a pressing one, and your research not only highlights the problem but provides a tangible and innovative solution. Your study design, especially the integration of using some novel soil amendments (i.e., biochar (B) and wood distillate) with the exposure to UV-B radiation is appropriately rigorous, enabling a holistic understanding of the variables at play. Your focus on some measurements, including growth characteristics, phenolics and flavonoids as well as antioxidants activity is highly commendable. However, there are some crucial amendments required as follow:

Point 1: The suggested title of the MS should represent accurately the performed experiments without unnecessary information. I would suggest the title to be “Integrative effect of UV-B and some organic amendments on growth, phenolic and flavonoid compounds, and antioxidant activity of basil (Ocimum basilicum  L.) plants.”

Reply: We thank the reviewer for his/her suggestion, the title was modified as suggested.

Point 2: My second comment pertains to the term “wood distillate (WD)” used as one of the treatments. From the agricultural point of view this term is not common. I would rather prefer to use the term wood vinegar instead in the whole MS.

Reply: We thank the reviewer for his/her suggestion. The term “wood distillate (WD)” was reported in several recent publications concerning agricultural topics. Anyway, we added the terms “pyroligneous acid” and “wood vinegar” to the keywords because are generally considered synonyms. We think that this can make the "wood distillate" term more common and facilitate its use for future research. 

Point 3: Line 104-110 where you illustrated the main objective of the study. The hypothesis of your work needs to be more justified. Please rephrase.

Reply: We thank the reviewer for his/her suggestion, we rephrased the aim of our work to make it more understandable.

Point 4: My next comment pertains to the photoreceptor’s measurements. One of the main objects of your study is to evaluate the response of basil plants to UV-B. by which should have been evaluated through the measurement of the main photoreceptor’s such as:-

ü Chlorophyll, absorbing PAR (Photosynthetic Active Radiation) and transferring energy to the reaction centers.

ü Phytochrome , absorbing mainly in the red (R) and far-red (FR).

ü Cryptochrome, absorbing blue and UV-A region (320-400nm).

ü UV-B photoreceptors, absorbing UV-B region (280 and 320 nm).

However, you did not present any photoreceptor’s measurements by which are essential for your results justification. You should have done these parameters while performing the experiment. If YES, please include this data in the results. Measurements.

Reply: We do not think the chlorophyll, the phytochrome and the cryptochrome investigations are pertinent to evaluate the plant response to UV-B radiation, since they are not the photoreceptors for the UV-B range. In addition, PAR intensity and quality was the same in both control and UV-B chambers.

For what concerns the UV-B photoreceptors, the UV RESISTANCE LOCUS 8 (UVR8), it is constitutively expressed within the cell, therefore its concentration does not vary in response to the UV-B exposure. When hit by UV-B, it rapidly monomerize and trigger a specific signaling pathway. However, it is highly challenging to detect the UVR8 monomer, since the re-dimerization kinetics occur in a very short time, requires specific UVR8 antibodies, and needs a dedicated experiment.

Besides, the aim of this study was to evaluate the possible enrichment in nutraceuticals (in terms of phenolics, flavonoids, and antioxidant capacity) and the changes in biometric parameters of basil plants in response to a co-application of above- and below-ground treatments, not a deep investigation on mechanisms of light perception and transduction.

Point 5: My second comment pertains to the largely descriptive nature of your results. While the data you present offer valuable insights, there could be additional analyses to deepen the interpretation and implications of your findings. Specifically, I recommend complementing your trait-by-trait analyses with correlation analysis and Principal Component Analysis (PCA).

Incorporating these statistical tools would not only strengthen your results but also provide a more nuanced understanding of the data. It would contribute to a richer and more comprehensive discussion of the interplay between selenium application, water stress, and the physiological and biochemical responses of the common bean plants.

Reply: We thank the reviewer for pointing it out. Accordingly, we have added a paragraph in the Results section on a multivariate statistical analysis, a canonical discriminant analysis, considering the biochemical traits analyzed (total phenolics and flavonoid content, and antioxidant capacity) and separately for the vegetative shoot apices and the fully developed leaves. It was not possible to also combine the biometric parameters, since they have been measured on roots and aerial parts, therefore on different tissues/parts of the plant in respect of the biochemical data. Besides, we do not think it could be helpful to include two more CDAs on the biometric variables, since it would consider just the length and weight variables.

In addition, we think the reviewer got confused when referring to the selenium application and water stress on bean plants.

Point 6: While your manuscript clearly reflects a sound understanding of the existing literature, I would like to emphasize the importance of including more recent publications in your references. Scholarly conversations are continually evolving and including citations from the last three years (especially from 2023) will ensure your work is positioned within the most current state of the field.

Reply: We thank the reviewer for his/her suggestion, we tried to update the literature with more recent publications.

Point 7: While your manuscript displays a strong command of the topic and presents compelling findings, I noticed some minor language issues and inconsistencies throughout the text. These could potentially hinder the clarity of your message and disrupt the reader's engagement with your work.

Reply: We thank the reviewer for his/her comment, we tried to make our message more understandable particularly in the aim paragraph.

Comments on the Quality of English Language

I noticed some minor language issues and inconsistencies throughout the text. These could potentially hinder the clarity of your message and disrupt the reader's engagement with your work.

Reply: We thank the reviewer for his/her comment, we tried to solve the language issues.

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

The required amendments have been performed, good luck