Iron Sulfate (FeSO4) Improved Physiological Attributes and Antioxidant Capacity by Reducing Oxidative Stress of Oryza sativa L. Cultivars in Alkaline Soil
, Manal Ahmed Naseeb 3, Arwa Saad Almasaudi 3 and Steve Harakeh 4
Round 1
Reviewer 1 Report
sustainability-2063350: This manuscript is interesting. But, some issues must be improved.
1) Line 30: What is “ROS” stands for?
2) Lines 66-67: Paddy field also plays an important role for soil organic carbon, which can mitigate CO2 emission. Please see these papers. [Soil organic carbon in sandy paddy fields of Northeast Thailand: A Review. Agronomy 2020, 10(8), 1061.] [Rice paddy soils are a quantitatively important carbon store according to a global synthesis. Commun Earth Environ 2021. 2, 154.]
3) Lines 79-80: “…it is essential to find techniques that can increase the concentration of Fe in vegetables.” This statement is not make sense because you focused on rice.
4) Lines 92-92: Why four different rice cultivars were considered? Do they have any difference potential? The reason is needed.
5) In the Experimental Design, the management practices are needed to explain in details. For example, how many kg of rice seeds were sown? How about type and amount of chemical fertilizers?, What kind of water management in the plot? and so on. Some photos of experiment are also useful.
6) Results: The relationship between FeSO4 and soil physical and chemical properties (e.g. pH, ECe, OM, Avail. P, K, Ca, Mg, and so on) should be analyzed, reported and discussed.
Author Response
REVIEWER: 01
Line 30: What is “ROS” stands for?
Response: ROS stands for reactive oxygen species and have added in manuscript.
2) Lines 66-67: Paddy field also plays an important role for soil organic carbon, which can mitigate CO2 emission. Please see these papers. [Soil organic carbon in sandy paddy fields of Northeast Thailand: A Review. Agronomy 2020, 10(8), 1061.] [Rice paddy soils are a quantitatively important carbon store according to a global synthesis. Commun Earth Environ 2021. 2, 154.]
Response: checked and added in revised version.
3) Lines 79-80: “…it is essential to find techniques that can increase the concentration of Fe in vegetables.” This statement is not make sense because you focused on rice
Response: This statement is corrected as “it is essential to find techniques that can increase the concentration of Fe in cereal crops”.
4) Lines 92-92: Why four different rice cultivars were considered? Do they have any difference potential? The reason is needed.
Response: Four rice varieties are used to prevent duplicating the same information as others writing in our field, allowing us to find our own, unique approach to your topic. But by using multiple varieties, we can check whether the effects of all the different treatments are the same or different for all the varieties.
5) In the Experimental Design, the management practices are needed to explain in details. For example, how many kg of rice seeds were sown? How about type and amount of chemical fertilizers?, What kind of water management in the plot? And so on. Some photos of experiment are also useful.
Response: We planted 5-6 seeds in plastic pots 1/2 inches below the soil, with the water depth set at 2-3 cm. Further detailed have been added in material and methods.
6) Results: The relationship between FeSO4 and soil physical and chemical properties (e.g. pH, ECe, OM, Avail. P, K, Ca, Mg, and so on) should be analyzed, reported and discussed.
Response: Added in material and methods. FeSO4 application increased the iron content while P and K decreased.
Reviewer 2 Report
This topic is interesting, but the manuscript is poorly written and requires revision. I've got some suggestions to improve this manuscript.
Abstract: It is unclear why this study was important. The authors should add some numerical data. How much of an increase or decrease in all parameters should be expressed by using percentages?
Furthermore, authors need to rewrite the keywords they should be more descriptive.
Authors first have to create an abbreviation of Oryza sativa as O. sativa, then use the abbreviation instead of the full form throughout the manuscript.
Methodology: In the experimental design, what was available in terms of phosphorus and potassium? The author should mention the value.
Did the authors check the soil properties after iron sulphate treatment? If so, how much iron increased as a result of the treatment?
In material and methods for how many minutes does the author centrifuge the sample for H2O2 determination?
Results are well written but some words need to change. It is more accurate to write "normal growth of rice" in place of "rice's normal growth" and "plant growth or growth of a plant" in place of "plant's growth."
Authors used the value of "standard deviation" or "standard error" for error bars should mention it in the figure's caption.
In the discussion section: authors mentioned that "stressed plants exhibit dysregulated reactive oxygen metabolism and an imbalance of free radicals." What does the author mean by "stressed plants"? Vague sentence.
"In addition to antioxidant enzymes non-enzymatic activity, we determined antioxidant non-enzymatic activity" unclear sentence need to rewrite this sentence.
Avoid beginning a sentence with an acronym or an abbreviation.
In conclusion: fine but with many grammatical issues, and the author should add some recommendations (which variety showed better response as well as which concentration).
In conclusion, always use may be; avoid using should be.
Author Response
REVIEWER: 02
Abstract: It is unclear why this study was important. The authors should add some numerical data. How much of an increase or decrease in all parameters should be expressed by using percentages?
Response: The importance of the present research work has been added to L18–21. In addition, some numerical data and percentage increases are included L33-35.
Furthermore, authors need to rewrite the keywords they should be more descriptive.
Response: keywords are replaced by “biofortication, micronutrient, iron, Oryza sativa L., antioxidants, oxidative stress markers”.
Ion of Oryza sativa as O. sativa, then use the abbreviation instead of the full form throughout the manuscript.
Response: It is corrected throughout the manuscript in the revised version.
Did the authors check the soil properties after iron sulphate treatment? If so, how much iron increased as a result of the treatment?
Response: Soil properties were checked after the treatment, but the pH of the soil remained alkaline throughout the experiment, indicating low iron solubility. After the application of FeSO4, soil pH slightly changed and caused an increase in iron content to 4.4 mg/kg.
In material and methods for how many minutes does the author centrifuge the sample for H2O2 determination?
Response: centrifugation time has been added for 15 minutes
Authors used the value of "standard deviation" or "standard error" for error bars should mention it in the figure's caption.
Response: standard deviation used for this line regarding to this added in each caption “Error bars represent the standard deviation (SD) of six replicates”.
In the discussion section: authors mentioned that "stressed plants exhibit dysregulated reactive oxygen metabolism and an imbalance of free radicals." What does the author mean by "stressed plants"? Vague sentence.
Response: Plants in stressful conditions display an imbalance of free radicals and a dysregulated reactive oxygen metabolism, which leads to the buildup of ROS, which in turn causes and accelerates lipid peroxidation and culminate in the breakdown of membrane integrity
In addition to antioxidant enzymes non-enzymatic activity, we determined antioxidant non-enzymatic activity" unclear sentence need to rewrite this sentence.
Response: In this study, an integrative overview of antioxidant enzymes and non-enzymatic activity showed that iron deficiency causes an increase in oxidative damage. To deal with this situation, the use of FeSO4 increased antioxidant enzymatic and non-enzymatic activity (Figures 3, 4, and 5).
Avoid beginning a sentence with an acronym or an abbreviation.
Response: we have accordingly revised
In conclusion: fine but with many grammatical issues, and the author should add some recommendations (which variety showed better response as well as which concentration).
Response: The variety name and concentration that showed the best results have been added in conclusion with a recommendation.
In conclusion, always use may be; avoid using should be.
Response: It is corrected as per your advice.
Reviewer 3 Report
Iron is a limiting element for rice growth and development in alkaline soils. Also, aerobic cultivation usually faces the same problem in rice plants. It has an important value to establish some useful approaches alleviating the adverse effects of iron deficiency on rice plants. In this study, authors focused on the theme of Iron sulfate (FeSO4) improved physiological attributes and anti-oxidant capacity by reducing oxidative stress of Oryza sativa L. cultivars in alkaline soil. Pot experiment was conducted with four cultivars and five FeSO4 concentration treatments. Some useful results were presented in this study. However, there were still many issues to be solved for publication the manuscript in Sustainability.
Abstract: The conclusion should be presented clearly around FeSO4 concentration treatments.
Introduction: (1)The studying theme was the effects of FeSO4 on physiological characteristics in rice plants. Therefore, I suggest that the background should focus on the rice production and Iron deficiency firstly instead of malnutrition. Malnutrition was only one of the reasons we conducted this study. (2) There have many reports on the theme (the effects of Fe on growth and development in rice plants), author must point out previous results and put forward scientific hypotheses in here.
Material and Methods: More details must be wrote in here.
(1) Why you chose the four cultivars whether they had different tolerances when response to iron-deficiency. Thus, you’d better introduction the relevant characteristics of cultivars.
(2) Whether there is a reference for setting different Fe concentration treatments.
(3) Line 111, How to calculate the water efficiency (%). In addition, water efficiency should be separated from the section because water efficiency is not belong to morphological parameter.
(4) Line 118, Wf, Wd, and Wt should be explained in here.
Results：There were many unclear expression.
(1) The title of 3.2 is error. Please correct it.
(2) Lines 206 to 207, it is unclear, which Fe treatments were compared in here. Please re-wrote it.
(3) Line 211, move (P<0.05) to the end of the sentence. A significantly decrease should be corrected as a significant decrease.
(4)Lines 227 to 229,Lines 236 to 239, The sentences were also unclear, which treatments? Please re-wrote it.
Discussion: Your results must be compared with previous results. However, the information is little lack, especially around the point how the Fe affect growth and development in rice plants.
Conclusion: Iron is important for a variety of processes, including DNA synthesis, respiration, 368
and photosynthesis. In plants, crop quality and productivity are affected by iron defi- 369
ciency. The sentence should be deleted from here.
Author Response
REVIEWER: 03
Abstract: The conclusion should be presented clearly around FeSO4 concentration treatments.
Response: The conclusive remarks and concentration, which showed the significant results, has been added to the abstract
Introduction: (1)The studying theme was the effects of FeSO4 on physiological characteristics in rice plants. Therefore, I suggest that the background should focus on the rice production and Iron deficiency firstly instead of malnutrition. Malnutrition was only one of the reasons we conducted this study. (2) There have many reports on the theme (the effects of Fe on growth and development in rice plants), author must point out previous results and put forward scientific hypotheses in here.
Response: Introduction section has been revised.
Material and Methods: More details must be wrote in here.
- Why you chose the four cultivars whether they had different tolerances when response to iron-deficiency. Thus, you’d better introduction the relevant characteristics of cultivars.
Response: Two Basmati varieties were chosen for this study based on their iron content. According to our previous study, two Basmati varieties, Basmati-515 and Basmati-198, have the highest and lowest iron contents of 22.0 and 14.1 ppm, respectively, while PK-386 had 19.0 ppm. KSK-133, a coarse variety with high yields and extra-long grains, was chosen.
- Whether there is a reference for setting different Fe concentration treatments
Response: These concentrations are selected after checking different concentrations in screening test in soil and agar as well.
- Line 111, How to calculate the water efficiency (%). In addition, water efficiency should be separated from the section because water efficiency is not belong to morphological parameter
Response: The formula to calculate water efficiency has been added as well as separated from the morphological section in results and in caption also.
- Line 118, Wf, Wd, and Wt should be explained in here
Response: RWC (%) = [Wf (fresh weight) - Wd (dry weight)) / (Wt (turgid weight) - Wd (dry weight)] * 100
Results：There were many unclear expression.
(1) The title of 3.2 is error. Please correct it.
Response: Title of 3.2is corrected now as “Effect of FeSO4 on leaf area, relative water content and photosynthtic pigments”
(2) Lines 206 to 207, it is unclear, which Fe treatments were compared in here. Please re-wrote it.
Response: We've rewritten this in result section, and no concentrations have been added.
(3) Line 211, move (P<0.05) to the end of the sentence. A significantly decrease should be corrected as a significant decrease.
Response: (P<0.05) has been corrected throughout the manuscript
- Lines 227 to 229, Lines 236 to 239, The sentences were also unclear, which treatments? Please re-wrote it.
Response: We've rewritten this in result section, and no concentrations have been added.
Discussion: Your results must be compared with previous results. However, the information is little lack, especially around the point how the Fe affect growth and development in rice plants.
Response: More study have been added
Conclusion: Iron is important for a variety of processes, including DNA synthesis, respiration, and photosynthesis. In plants, crop quality and productivity are affected by iron deficiency. The sentence should be deleted from here.
Response: These sentence has been removed from conclusion section.
Reviewer 4 Report
The authors test the iron sulfate effect on 4 rice genotypes in alkali soil conditions at the seedling stage. Their experimental design and conducted analysis were well selected and performed.
I have some minor comments for this manuscript.
1-L20: please correct photosynthetic content with "photosynthetic pigment"
2-L24-25: please arrange text punto
3-L27: please remove "present study found that"
4-L56: please remove dot (.) before citation.
5-L60, 64, 76: Merge the citations
6-L67: "Significant" S has to be a small letter
7-L101,116, 130: Please correct celcius degree abbreviation and seperated from number in all text.
8-L144, 152: Please follow same pattern for abbreviation, keep one blank gap between number and abbreviations and check all in text.
9-L159: for statittic (p > 0.05) p has to be ithalic and has to be lower symbol (<), please check all text and correct them.
10-L188: Those are not photosynthetic parameters, photosynthetic pigments. Please correct the phrase.
11-L194, 209: please use "cultivars" after the "O.sativa"
12-L382: Please correct text punto.
Author Response
REVIEWER: 04
1-L20: please correct photosynthetic content with "photosynthetic pigment"
Response: it is corrected throughout the manuscript
2-L24-25: please arrange text punto
Response: These sentences are corrected.
3-L27: please remove "present study found that"
Response: it is corrected
4-L56: please remove dot (.) before citation
Response: dot has been removed before citation
5-L60, 64, 76: Merge the citations
Response: All the citations have now been combined
6-L67: "Significant" S has to be a small letter
Response: S has taken the place of s
7-L101,116, 130: Please correct celcius degree abbreviation and separated from number in all text.
Response: Celcius degree abbreviation has been corrected (°C) and separated from all numbers throughout the manuscript.
8-L144, 152: Please follow same pattern for abbreviation, keep one blank gap between number and abbreviations and check all in text.
Response: All are corrected throughout the manuscript.
9-L159: for statittic (p > 0.05) p has to be italic and has to be lower symbol (<), please check all text and correct them.
Response: (p > 0.05) has to be changed to (p < 0.05) and italics throughout the manuscript.
10-L188: Those are not photosynthetic parameters, photosynthetic pigments. Please correct the phrase.
Response: This line has been corrected, and photosynthetic parameters are now photosynthetic pigments.
11-L194, 209: please use "cultivars" after the "O.sativa"
Response: Cultivars word has been added after the O. sativa and corrected throughout the manuscript
12-L382: Please correct text punto
Response: already corrected as per your advice
Round 2
Reviewer 1 Report
sustainability-2063350-V2: Thank you very much for your revision. Although the manuscript was improved, some issues are remaining and need to be revised.
1) The revised manuscript is not provided the Line number, making difficult to add the specific suggestions.
2) Page 2: “Rice paddy soils are a quantitatively important carbon store according to a global synthesis [2].” This sentence must be explained more detail. Paddy field also plays an important role for soil organic carbon, and rice yield. Please see this paper. [Soil organic carbon in sandy paddy fields of Northeast Thailand: A Review. Agronomy 2020, 10(8), 1061.].
3) Some photos of experiment plots must be added.
4) “ES (0.60 dS m-1 )” What is “ES”?
5) Which version of SPSS was used?
6) In conclusion: Which FeSO4 concentration is recommended to apply?
Author Response
Comments and Suggestions for Authors
sustainability-2063350-V2: Thank you very much for your revision. Although the manuscript was improved, some issues are remaining and need to be revised.
1) The revised manuscript is not provided the Line number, making it difficult to add the specific suggestions.
Response: The line Number has been added in the revised version
2) Page 2: “Rice paddy soils are a quantitatively important carbon store according to a global synthesis [2].” This sentence must be explained more detail. Paddy field also plays an important role for soil organic carbon, and rice yield. Please see this paper. [Soil organic carbon in sandy paddy fields of Northeast Thailand: A Review. Agronomy 2020, 10(8), 1061.].
Response: More information regarding this has been added in the L 46-49.
3) Some photos of experiment plots must be added.
Response: photos of all cultivars have been added at the end of the paper L 708-713
4) “ES (0.60 dS m-1 )” What is “ES”?
Response: It is corrected and ES is replaced with EC L120
5) Which version of SPSS was used?
Response: The version of SPSS 22.0 is mentioned in statistical analysis L179-181.
6) In conclusion: Which FeSO4 concentration is recommended to apply?
Response: The concentration that showed significantly highest results (500mM) has been discussed in the conclusion L408-411
Reviewer 3 Report
Authors solved all issues raised by me. The quality of the manuscript was significantly improved after revising. There are no additional comments in this time.
Author Response
Thank you very much for your precious time to review and improve our research work.
Regards
Round 3
Reviewer 1 Report
Accept in present form.
Author Response
Thank you so much.
Regards
Ali
