How Different Na⁺ Concentrations Affect Anatomical, Nutritional Physiological, Biochemical, and Morphological Aspects in Soybean Plants: A Multidisciplinary and Comparative Approach

Round 1

Reviewer 1 Report

The manuscript “How different Na+ concentrations affect anatomical, nutritional physiological, biochemical, and morphological aspects in soybean plants: A multidisciplinary and comparative approach” (agronomy-2024543) showed impact fo the Na concentration at nutritional, biochemical, physiological and morphological parameters in soybean. This manuscript is interesting to readers, especially, based a Special Issue “Antioxidant Defences in Crop Plants”

The experiment is well-design, and the manuscript was well-written.

The authors have done a large amount of work, employing various references and critical analysis based on a scientific method and structure. The introduction, M&M, the results and discussion topic is good and minor points its necessary by adjusting in English syntax. In general, tables and figures are good qualities and major adjust are necessary. Tables and figures displayed great significance to interpretation of the data.

My conception, the manuscript is suitable for publication, after minor corrections. The structure is adequate, and the information is new and of great significance for comprehension to Agronomy journal readers.

Please, use to template “Microsoft or Latex Template” of manuscript to figures, tables and references need adjust following “Author Instructions in Agronomy”.

-Figures and tables need correction following “Author Instructions” and displayed after first citation.

#1: There is a scope for improvement in the introduction section: a) additional emphasis on the significance of the study, b) scientific and economic contribution of the paper; c) prospectively to other plants to agronomic interest. Maybe a one paragraphs with potential economic by important this manuscript to science crop;

#2. Your hypothesis is not clear in last paragraph to introduction. Pease write “Our hypothesis was…”

#3: Please. All standardization of nomenclature equipment/reagents/software when necessary. Example: Fabricant, City, State, Country (three-letter). Check all manuscript.

#5. I’m suggesting to authors an Abbreviations list after conclusion.

#6. Please check for “Author Instruction” and standardization to manuscript, example, Figure/Figures, not Fig; References;

#7. Alphabetic order keywords;

#8. Check, al references. Many data of the last 15 years old!

#9. Activated in initial experiment but, how many samples were analysed per period (time)? Antioxidant enzyme assays; what is E.C. (Enzyme Commission number) to enzymes analysed?

Material and methods;

Topic 2.9: chlorophyll and carotenoids

“in gs (negative oscillations of 52% to 85%).”, what did you mean by that? de

Check author contribution following “Author Instructions”.

Conclusion: What are future perspectives for production on farms? What is the impact of the temperate climate or soil with higher Na+/K+ in Cerrado and other Brazilian soils with increased salinity in recent years?

Author Response

Dear reviewers and editor linked to manuscript # agronomy-2024543: We are submitting the revised manuscript to the journal submission site for your review. As instructed in your e-mail on 18 Nov 2022, we have carefully considered all the reviewer’s comments and fully addressed them in the revised manuscript. Our responses to each specific reviewer comments are as follows (red into manuscript): General comments from Managing Editor: Your manuscript has now been reviewed by experts in the field. Please find your manuscript with the referee reports at this link: https://susy.mdpi.com/user/manuscripts/resubmit/fe8b282269cc9f510fcb6ef6e36b5c41 Please revise the manuscript according to the referees' comments and upload the revised file within 4 days. Decision: Minor revisions Diana Otvos Assistant Editor Agronomy General comments from Reviewer 1: The manuscript “How different Na+ concentrations affect anatomical, nutritional physiological, biochemical, and morphological aspects in soybean plants: A multidisciplinary and comparative approach” (agronomy-2024543) showed impact fo the Na concentration at nutritional, biochemical, physiological and morphological parameters in soybean. This manuscript is interesting to readers, especially, based a Special Issue “Antioxidant Defences in Crop Plants”. The experiment is well-design, and the manuscript was well-written. The authors have done a large amount of work, employing various references and critical analysis based on a scientific method and structure. The introduction, M&M, the results and discussion topic is good and minor points its necessary by adjusting in English syntax. In general, tables and figures are good qualities and major adjust are necessary. Tables and figures displayed great significance to interpretation of the data. My conception, the manuscript is suitable for publication, after minor corrections. The structure is adequate, and the information is new and of great significance for comprehension to Agronomy journal readers. Please, use to template “Microsoft or Latex Template” of manuscript to figures, tables and references need adjust following “Author Instructions in Agronomy”. Figures and tables need correction following “Author Instructions” and displayed after first citation. Reviewer 1: There is a scope for improvement in the introduction section: a) additional emphasis on the significance of the study, b) scientific and economic contribution of the paper; c) prospectively to other plants to agronomic interest. Maybe a one paragraphs with potential economic by important this manuscript to science crop. Authors: One paragraph elaborated and inserted in introduction section (below): “Soybean (Glycine max (L.) Merr.) is one of the most important crops in the world because its seeds are rich in oils and proteins [1] that can be used in foods, animal feed [2] and as an energy source in biofuels [3]. Global production is estimated at approximately 338 million tons, with the United States being the main producer, followed by Brazil and Argentina. However, one of the main factors limiting soybean cultivation is soil salinity [4], a matter of increasing importance since approximately 800 million hectares of land are affected by this environmental problem.” Reviewer 1: Your hypothesis is not clear in last paragraph to introduction. Pease write “Our hypothesis was…” Authors: This sentence was added (below): “Our hypothesis investigated the problems caused by different Na+ concentrations on morphological aspects in soybean plants, we considered that a multidisciplinary, broad and comparative approach was essential to explain the deleterious effects. On this basis, our research measured anatomical, nutritional, physiological, and biochemical interferences, detailing possible consequences on morphological responses in soybean plants subjected to progressive salt stress.” Reviewer 1: Please. All standardization of nomenclature equipment/reagents/software when necessary. Example: Fabricant, City, State, Country (three-letter). Check all manuscript. Authors: These modifications were implemented (below): “inductively coupled plasma mass spectrometer (model ICP-MS 7900, Agilent, United States).” “spectrophotometer (model UV-M51; Bel Photonics, Italy).” Reviewer 1: I’m suggesting to authors an Abbreviations list after conclusion. Authors: Abbreviation list was removed, being applied and described into manuscript. Reviewer 1: Please check for “Author Instruction” and standardization to manuscript, example, Figure/Figures, not Fig; References; Authors: Figures and reference styles were checked and adjusted, in agreement author instruction. Reviewer 1: Alphabetic order keywords; Authors: Keywords were adjusted. Reviewer 1: Check, al references. Many data of the last 15 years old! Authors: References were corrected. Old references remained only in the materials and methods or/and specific sentences. Reviewer 1: Activated in initial experiment but, how many samples were analysed per period (time)? Antioxidant enzyme assays; what is E.C. (Enzyme Commission number) to enzymes analysed? Authors: These informations were included (below): “Antioxidant enzymes [SOD (EC 1.15.1.1), CAT (EC 1.11.1.6), APX (EC 1.11.1.11), and POX (EC 1.11.1.7)]” “Fig. 5. Activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POX) in soybean plants submitted to salt stress. Bars with different letters indicate significant differences from the Scott-Knott test (P

Reviewer 2 Report

Comment-1

I think it is more intuitive to replace the tables with column charts.

Comment-2

Fig.2 All of the pictures are somewhat fuzzy. The stomata should be enlarged to have a better definition.

Comment-3

Fig.2 The cell in pictures C and D are smaller than others, but all of the bars are same. Please check it. And the magnification of pictures in Fig.3 are different. You need point out the wax with arrows in the pictures.

Comment-4

Fig.3 It seems like that the gap between cells in C, E, G are bigger than A and I. Is this caused by Na⁺ treatment?

Comment-5

Why does the most of wax concentrat in the cell gaps in Fig.3 G, H, I, J ?

Comment-6

Is the white fluff in Fig. 3B wax? If Yes, Why dose the surface of H and J become smooth?

Comment-7

I find the cell walls were stained by toluidine blue O in the reference but not the text. You should note it in the legend.

Comment-8

Can you provide the pictures of thichome under different treatment?

Comment-9

Why is the spaces in the palisade parenchyma under 200 mM Na⁺ (Fig. 4J) treatment smaller than that of 150 mM (Fig. 4H) and 100 mM (Fig. 4F) ? And the spongy parenchyma disorders degree of Fig. 4J is less than Fig. 4H and F?

Comment-10

Table 6 Formats of 9.65 ±0.87b and 9.86 ±0.63b need to be adjusted.

Author Response

Dear reviewers and editor linked to manuscript # agronomy-2024543: We are submitting the revised manuscript to the journal submission site for your review. As instructed in your e-mail on 18 Nov 2022, we have carefully considered all the reviewer’s comments and fully addressed them in the revised manuscript. Our responses to each specific reviewer comments are as follows (red into manuscript): General comments from Managing Editor: Your manuscript has now been reviewed by experts in the field. Please find your manuscript with the referee reports at this link: https://susy.mdpi.com/user/manuscripts/resubmit/fe8b282269cc9f510fcb6ef6e36b5c41 Please revise the manuscript according to the referees' comments and upload the revised file within 4 days. Decision: Minor revisions Diana Otvos Assistant Editor Agronomy General comments from Reviewer 1: The manuscript “How different Na+ concentrations affect anatomical, nutritional physiological, biochemical, and morphological aspects in soybean plants: A multidisciplinary and comparative approach” (agronomy-2024543) showed impact fo the Na concentration at nutritional, biochemical, physiological and morphological parameters in soybean. This manuscript is interesting to readers, especially, based a Special Issue “Antioxidant Defences in Crop Plants”. The experiment is well-design, and the manuscript was well-written. The authors have done a large amount of work, employing various references and critical analysis based on a scientific method and structure. The introduction, M&M, the results and discussion topic is good and minor points its necessary by adjusting in English syntax. In general, tables and figures are good qualities and major adjust are necessary. Tables and figures displayed great significance to interpretation of the data. My conception, the manuscript is suitable for publication, after minor corrections. The structure is adequate, and the information is new and of great significance for comprehension to Agronomy journal readers. Please, use to template “Microsoft or Latex Template” of manuscript to figures, tables and references need adjust following “Author Instructions in Agronomy”. Figures and tables need correction following “Author Instructions” and displayed after first citation. Reviewer 1: There is a scope for improvement in the introduction section: a) additional emphasis on the significance of the study, b) scientific and economic contribution of the paper; c) prospectively to other plants to agronomic interest. Maybe a one paragraphs with potential economic by important this manuscript to science crop. Authors: One paragraph elaborated and inserted in introduction section (below): “Soybean (Glycine max (L.) Merr.) is one of the most important crops in the world because its seeds are rich in oils and proteins [1] that can be used in foods, animal feed [2] and as an energy source in biofuels [3]. Global production is estimated at approximately 338 million tons, with the United States being the main producer, followed by Brazil and Argentina. However, one of the main factors limiting soybean cultivation is soil salinity [4], a matter of increasing importance since approximately 800 million hectares of land are affected by this environmental problem.” Reviewer 1: Your hypothesis is not clear in last paragraph to introduction. Pease write “Our hypothesis was…” Authors: This sentence was added (below): “Our hypothesis investigated the problems caused by different Na+ concentrations on morphological aspects in soybean plants, we considered that a multidisciplinary, broad and comparative approach was essential to explain the deleterious effects. On this basis, our research measured anatomical, nutritional, physiological, and biochemical interferences, detailing possible consequences on morphological responses in soybean plants subjected to progressive salt stress.” Reviewer 1: Please. All standardization of nomenclature equipment/reagents/software when necessary. Example: Fabricant, City, State, Country (three-letter). Check all manuscript. Authors: These modifications were implemented (below): “inductively coupled plasma mass spectrometer (model ICP-MS 7900, Agilent, United States).” “spectrophotometer (model UV-M51; Bel Photonics, Italy).” Reviewer 1: I’m suggesting to authors an Abbreviations list after conclusion. Authors: Abbreviation list was removed, being applied and described into manuscript. Reviewer 1: Please check for “Author Instruction” and standardization to manuscript, example, Figure/Figures, not Fig; References; Authors: Figures and reference styles were checked and adjusted, in agreement author instruction. Reviewer 1: Alphabetic order keywords; Authors: Keywords were adjusted. Reviewer 1: Check, al references. Many data of the last 15 years old! Authors: References were corrected. Old references remained only in the materials and methods or/and specific sentences. Reviewer 1: Activated in initial experiment but, how many samples were analysed per period (time)? Antioxidant enzyme assays; what is E.C. (Enzyme Commission number) to enzymes analysed? Authors: These informations were included (below): “Antioxidant enzymes [SOD (EC 1.15.1.1), CAT (EC 1.11.1.6), APX (EC 1.11.1.11), and POX (EC 1.11.1.7)]” “Fig. 5. Activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POX) in soybean plants submitted to salt stress. Bars with different letters indicate significant differences from the Scott-Knott test (P