The Effects of Foliar Salicylic Acid and Zinc Treatments on Proline, Carotenoid, and Chlorophyll Content and Anti-Oxidant Enzyme Activity in Galanthus elwesii Hook
Round 1
Reviewer 1 Report
The manuscript presents the results of the effect of Zn and SA on biomolecules contents and enzymatic activity in samples of Galanthus elwesii. The results obtained are important for understanding the physiology of the species and contribute to its management in breeding programs.
The work is well planned and has a clear and fluent writing; however, it presents important limitations with respect to the methodologies used, the identification of the compounds and the presentation of the results.
The technique used for protein analysis only allows the determination of soluble proteins, which should be clearly stated in the abstract, results and conclusions. In case of referring to total, structural, functional, and reserve proteins, they must analyze the nitrogen content by Kjeldahl and convert it to total protein using the corresponding factor.
Table 1 is saturated with information, which makes it difficult to review and analyze the significance of the differences. Since the results of the effect of increasing concentrations of Zn and SA are shown, I suggest that they be presented in the form of graphs to better observe the effects. The same applies to Table 2.
Page 3, line 133. Indicate where the sample was obtained from.
Page 3, line 141. Change “proteine” to protein.
Page 34, Line 142. The described technique allows analysis of the presence of all free amino acids, but it is difficult for the reaction to be positive for proline because proline is a secondary amine. I suggest that this specification be made in the summary, results, and conclusions.
Page 5, Lines 185-195. I suggest that only the concentration of the solutions used be indicated and the detail of the preparation be eliminated.
Page 6, Lines 271-273. I suggest that the protein and proline concentrations be expressed with respect to fresh weight as well as the other parameters.
Page 6, Lines 271, 276. The values obtained for total protein are very low. In tissue extracts with the described extraction conditions, they should be in the order of mg.
Author Response
REVISION
Dear Editor,
All of the corrections were painted red in the text. And the corrections were given under the title of “RESPONSE to REVIEWERS”.
First of all, I would like to express my gratitude to the Reviewers for their valuable contributions. I have found the Reviewers extremely sensitive about reading and examining the article. I present the corrections as I have made taking the criticism of the Reviewers into consideration as follows.
RESPONSE TO EDITOR
The language of the article has been corrected by experts, and the species names that should be written in italics have been checked and corrected throughout the article and in the references section.
RESPOND TO REVIEWERS
REVIEWER 1
“The manuscript presents the results of the effect of Zn and SA on biomolecules contents and enzymatic activity in samples of Galanthus elwesii. The results obtained are important for understanding the physiology of the species and contribute to its management in breeding programs. The work is well planned and has a clear and fluent writing; however, it presents important limitations with respect to the methodologies used, the identification of the compounds and the presentation of the results.”
View 1
“The technique used for protein analysis only allows the determination of soluble proteins, which should be clearly stated in the abstract, results and conclusions. In case of referring to total, structural, functional, and reserve proteins, they must analyze the nitrogen content by Kjeldahl and convert it to total protein using the corresponding factor.”
Revision/Answer: Thank you for your evaluation, the Reviewer is right in his suggestion. As we mentioned in the Materials and Methods, in G. elwesii samples, the amount of soluble protein was determined by the Bradford Protein Assay Method. Total protein determination is evaluated by the Kjeldahl Method, as you emphasized. The total protein expression in the abstract, results, and conclusions sections has been changed to soluble protein.
View 2
“Table 1 is saturated with information, which makes it difficult to review and analyze the significance of the differences. Since the results of the effect of increasing concentrations of Zn and SA are shown, I suggest that they be presented in the form of graphs to better observe the effects. The same applies to Table 2.”
Revision/Answer: Some data are given as figures.
View 3
“Page 3, line 133. Indicate where the sample was obtained from.”
Revision/Answer: Fresh leaf samples were used.
View 4
“Page 3, line 141. Change “proteine” to protein.”
Revision/Answer: It was changed.
View 5
“Page 34, Line 142. The described technique allows analysis of the presence of all free amino acids, but it is difficult for the reaction to be positive for proline because proline is a secondary amine. I suggest that this specification be made in the summary, results, and conclusions.”
Revision/Answer:
View 6
“Page 5, Lines 185-195. I suggest that only the concentration of the solutions used to be indicated and the detail of the preparation be eliminated.”
Revision/Answer: That part has been corrected taking into account the Reviewer's suggestion.
View 7
“Page 6, Lines 271-273. I suggest that the protein and proline concentrations be expressed with respect to fresh weight as well as the other parameters.”
Revision/Answer: It was corrected as fresh weight.
View 8
“Page 6, Lines 271, 276. The values obtained for total protein are very low. In tissue extracts with the described extraction conditions, they should be in the order of mg.”
Revision/Answer: The amount of soluble protein in bulbous plants was checked and it was found to be at these levels.
Reviewer 2 Report
This study investigated the effect of foliar salicylic acid and zinc treatments on proline, carotenoid, chlorophyll content and anti-oxidant enzyme activity in Galanthus elwesii Hook. However, this manuscript has many issues and can not reach the level of publication.
1. In the Materials and Methods, I can not know how to spray the salicylic acid and zinc, they are mixed together? Or separate spraying? Salicylic acid first? Then zinc? Time and degree of spraying? When the fresh leaf samples were collected and which leaf in the plant? Many methods and treatments are not clearly explained.
2. In the results, there are only 2 tables. No further analysis of the data was conducted. So I can not get the clear results in this manuscript. Which treatment is suitable for Galanthus elwesii Hook.
3. There are only physiological data in this study, lack of morphological and other data.
4. There are many errors in this manuscript.
For example, Galanthus elwesii Hook should be Galanthus elwesii Hook.
In the 31 sentence, Galanthus elwesii should be italic.
Tukey's Posthoc test in the Materials and Methods, but LSD test in the results.
Six replications in the Materials and Methods, but n=3 in the tables.
Author Response
REVISION
Dear Editor,
All of the corrections were painted red in the text. And the corrections were given under the title of “RESPONSE to REVIEWERS”.
First of all, I would like to express my gratitude to the Reviewers for their valuable contributions. I have found the Reviewers extremely sensitive about reading and examining the article. I present the corrections as I have made taking the criticism of the Reviewers into consideration as follows.
RESPONSE TO EDITOR
The language of the article has been corrected by experts, and the species names that should be written in italics have been checked and corrected throughout the article and in the references section.
RESPOND TO REVIEWERS
REVIEWER 2
“This study investigated the effect of foliar salicylic acid and zinc treatments on proline, carotenoid, chlorophyll content, and anti-oxidant enzyme activity in Galanthus elwesii Hook. However, this manuscript has many issues and can not reach the level of publication.”
View 1
“In the Materials and Methods, I can not know how to spray the salicylic acid and zinc, they are mixed together. Or separate spraying? Salicylic acid first? Then zinc? Time and degree of spraying? When the fresh leaf samples were collected and which leaf in the plant? Many methods and treatments are not clearly explained.”
Revision/Answer: It was explained in detail, taking into account the Reviewer’s suggestion.
View 2
“In the results, there are only 2 tables. No further analysis of the data was performed. So I can not get clear results in this manuscript. Which treatment is suitable for Galanthus elwesii Hook.”
Revision/Answer: This study focused on enzyme activity, so other parameters will be focused on in other studies.
View 3
“There are only physiological data in this study, lack of morphological and other data.”
Revision/Answer: Only physiological parameters are taken into account, further studies will focus on considering the Reviewer’s recommendation.
View 4
“There are many errors in this manuscript.”
For example, Galanthus elwesii Hook should be Galanthus elwesii Hook.
In the 31 sentence, Galanthus elwesii should be italic.
Revision/Answer: It was checked and fixed.
View 5
“Tukey's Posthoc test in the Materials and Methods, but LSD test in the results. Six replications in the Materials and Methods, but n=3 in the tables.”
Revision/Answer: It was checked and fixed.
Reviewer 3 Report
This study delved into the effects of foliar treatments using salicylic acid and zinc on proline, carotenoid, chlorophyll content, and antioxidant enzyme activity in G. elwesii. The researchers administered three concentrations of SA and Zn, measuring enzyme activities and contents. Notably, Zn at 80 and 120 mM produced the highest APX and CAT levels. Significant increases in proline, chlorophyll b, and carotenoid content were observed with 0.5 mM SA treatments. Combining Zn at 120 mM with SA at 2 mM led to an elevation in total chlorophyll content, while Zn at 40 mM with SA at 1 mM resulted in increased chlorophyll content.
While this paper presents a commendably organized and well-written contribution to the field, a few minor suggestions should be addressed.
To enhance the study's quality, consider incorporating biomass composition measurements, including % protein, % lipids, % carbohydrates, and % polynucleic acids.
Refine the introduction to clearly highlight knowledge gaps and connect them to the research goals, emphasizing the novelty and relevance of the study.
Strengthen the discussion section by offering insights gained from the findings and their applicability to future research. Address crucial questions such as research gaps, beneficiaries of improvements, and potential future directions to enhance the paper's contribution.
In the conclusion, it's advisable to steer clear of terms like "first" or any language that might suggest exclusivity to this study. Instead, focus on the broader implications of the research findings and their significance in advancing our understanding of the subject. This will help maintain a balanced and objective tone while discussing the contribution of the study.
Ensure adherence to the "Instructions for Authors" and utilize the original draft of Horticulturae for manuscript formatting. Explicitly mention the research type above the title.
Improve the quality of language and sentence structure by addressing grammatical and typographical errors.
Author Response
REVISION
Dear Editor,
All of the corrections were painted red in the text. And the corrections were given under the title of “RESPONSE to REVIEWERS”.
First of all, I would like to express my gratitude to the Reviewers for their valuable contributions. I have found the Reviewers extremely sensitive about reading and examining the article. I present the corrections as I have made taking the criticism of the Reviewers into consideration as follows.
RESPONSE TO EDITOR
The language of the article has been corrected by experts, and the species names that should be written in italics have been checked and corrected throughout the article and in the references section.
RESPOND TO REVIEWERS
REVIEWER 3
“This study delved into the effects of foliar treatments using salicylic acid and zinc on proline, carotenoid, chlorophyll content, and antioxidant enzyme activity in G. elwesii. The researchers administered three concentrations of SA and Zn, measuring enzyme activities and contents. Notably, Zn at 80 and 120 mM produced the highest APX and CAT levels. Significant increases in proline, chlorophyll b, and carotenoid content were observed with 0.5 mM SA treatments. Combining Zn at 120 mM with SA at 2 mM led to an elevation in total chlorophyll content, while Zn at 40 mM with SA at 1 mM resulted in increased chlorophyll content. While this paper presents a commendably organized and well-written contribution to the field, a few minor suggestions should be addressed.”
View 1
“ To enhance the study's quality, consider incorporating biomass composition measurements, including % protein, % lipids, % carbohydrates, and % polynucleic acids. ”
Revision/Answer: Considering the Reviewer's recommendation, these parameters will be focused on in future studies.
View 2
“Refine the introduction to clearly highlight knowledge gaps and connect them to the research goals, emphasizing the novelty and relevance of the study.”
Revision/Answer: The purpose of the study was emphasized in the introduction, and the gaps were removed.
View 3
“Strengthen the discussion section by offering insights gained from the findings and their applicability to future research. Address crucial questions such as research gaps, beneficiaries of improvements, and potential future directions to enhance the paper's contribution.”
Revision/Answer: The conclusion part has been changed taking into account the Reviewer's suggestion.
View 4
“ In the conclusion, it's advisable to steer clear of terms like "first" or any language that might suggest exclusivity to this study. Instead, focus on the broader implications of the research findings and their significance in advancing our understanding of the subject. This will help maintain a balanced and objective tone while discussing the contribution of the study. ”
Revision/Answer: Words like "first" and "only" were omitted from the conclusion.
View 5
“Ensure adherence to the "Instructions for Authors" and utilize the original draft of Horticulturae for manuscript formatting. Explicitly mention the research type above the title. ”
Revision/Answer: Necessary corrections were made taking into account the Reviewer’s instructions.
Round 2
Reviewer 1 Report
The manuscript has partially improved with the modifications made, however, the expression of the concentration of soluble protein and proline in table 1, is now expressed as a (ug/ml FW), which is difficult to understand, they refer to ml of fresh weight, when It must be a unit of weight and not of volume.
Regarding the protein concentration, it is mentioned that the values were checked, however, the very low amount of protein conflicts with the activity values of the enzymes, since very low activity results would be obtained with the concentrations that report. I suggest that the amount of extract used in enzyme activity assays be indicated in order to correlate activity values with protein concentration.
Additionally, I suggest describing the protein quantification in more detail, since a 4 ul cuvette was used from a 5 ml extract. Probably, the length of the optical step of the cuvette or the conversion of the data causes deviation of the results.
Author Response
REVISION 2
Dear Editor,
All of the corrections were painted red in the text. And the corrections were given under the title of ‘RESPOND to REVIEWERS’.
First of all, I would like to express my gratitude to the referees for their valuable contributions. I have found the referees extremely sensitive about reading and examining the article. I present the corrections as I have made taking the criticism of the referees into consideration as follows.
RESPOND TO REVIEWERS
REVIEWER 1
View 1
“The manuscript has partially improved with the modifications made, however, the expression of the concentration of soluble protein and proline in table 1, is now expressed as a (ug/ml FW), which is difficult to understand, they refer to ml of fresh weight, when It must be a unit of weight and not of volume”.
Revision/Answer: The µg/mL values expressed for soluble protein and proline in Table 1 were used for the concentrations prepared to obtain the y = 0.0887x + 0.2703 R² = 0.9851 graph. Since the analyzes were made from fresh leaves, the results in g were corrected in Table 1 and article.
View 2
“Regarding the protein concentration, it is mentioned that the values were checked, however, the very low amount of protein conflicts with the activity values of the enzymes, since very low activity results would be obtained with the concentrations that report. I suggest that the amount of extract used in enzyme activity assays be indicated in order to correlate activity values with protein concentration”.
Revision/Answer: 2.4. under the heading of the article. The first three sentences of the Bradford protein assay section describe the preparation of homogenants for the determination of enzyme activity. In the relevant section, it was stated that homogenant was prepared from 1 g of fresh leaf sample.
View 3
“Additionally, I suggest describing the protein quantification in more detail, since a 4 ul cuvette was used from a 5 ml extract. Probably, the length of the optical step of the cuvette or the conversion of the data causes deviation of the results.”
Reviewer 2 Report
The authors have provided a revised version along with a cover letter, but the manuscript has not been sufficiently improved to warrant publication in Horticulturae. Because the methods in this manuscript were not adequately described. And the results were not further analysized.Author Response
REVISION 2
Dear Editor,
All of the corrections were painted red in the text. And the corrections were given under the title of ‘RESPOND to REVIEWERS’.
First of all, I would like to express my gratitude to the referees for their valuable contributions. I have found the referees extremely sensitive about reading and examining the article. I present the corrections as I have made taking the criticism of the referees into consideration as follows.
RESPOND TO REVIEWERS
REVIEWER 2
“The authors have provided a revised version along with a cover letter, but the manuscript has not been sufficiently improved to warrant publication in Horticulturae. Because the methods in this manuscript were not adequately described. And the results were not further analysized.”
Revision/Answer:
In revision 1, the material and method were adequately explained. Whether the article in preprint that the referee objected to here is accepted as final. This is not a process related to us, it was offered to us by the Horticulturae and we approved the preprint.
Reviewer 3 Report
The manuscript has undergone significant enhancements and is now suitable for acceptance in its present form.
The manuscript has undergone significant enhancements and is now suitable for acceptance in its present form.
Author Response
REVISION 2
Dear Editor,
All of the corrections were painted red in the text. And the corrections were given under the title of ‘RESPOND to REVIEWERS’.
First of all, I would like to express my gratitude to the referees for their valuable contributions. I have found the referees extremely sensitive about reading and examining the article. I present the corrections as I have made taking the criticism of the referees into consideration as follows.
REVIEWER 3
“The manuscript has undergone significant enhancements and is now suitable for acceptance in its present form.”
Revision/Answer: Thank you
