Combined Effects of Magnetized Irrigation and Water Source on Italian Lettuce (Lactuca sativa L. var. ramosa Hort.) Growth and Gene Expression

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Irrigation water quality is vital for good crop growth and development. Magnetization technique for improving quality of irrigation water is relatively new technique. Present study describes the effects of magnetized irrigation using different water sources on lettuce growth and gene expression, providing theoretical support for the application of magnetized water technology in agriculture. The study concludes that magnetization treatment increased the EC, pH, and DO of irrigation water sources. The EC of saline water and the pH of recycled water were significantly higher than other sources. Magnetized irrigation increased the content of N, K, Ca, and Mg in lettuce, improved quality, increased fresh and dry weight, and water productivity, while reducing water consumption. For magnetized fresh water irrigation, the content of N, K Ca and Mg in lettuce increased significantly increased. Soluble sugar, VitC, fresh and dry weight as well as WPc increased showing the best overall improvement in quality, fresh weight and WPc. Compared to fresh water irrigation, magnetized recycled water irrigation led to higher levels of N content, soluble protein, soluble sugar, VitC, fresh weight, and WPc, and lower water consumption, with fresh weight and WPc reaching significant. Overall the study is presented well, however, few queries need to be addressed.

-In controlled environments, treatments should be repeated five times at least.

-A sketch showing the magnatization process may be presented.

-Results are interpreted well, however, need more discussion.

-Conclusion needs to be brief and to the point according to the findings.

-There are errors in english and format.

Author Response

Please refer to the attached document

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The general theme of the manuscript is sound, although there are serious doubts about it. The manuscript has major flaws, including: how to magnetize water, the quality of recycled water, and salinity treatment. The prepared water was not saline in salinity treatment, because the electrical conductivity was below 4 dS/m.

Detailed comments:

The scientific name should be italicized in the title (and elsewhere).

How strong was the magnetization of water? Mentioned in the abstract.

There is no need to number the results in the abstract.

When magnetizing water increases the electrical conductivity of water, how does saltwater increase growth and yield?

Line 121: randomized complete block design, not randomized block design

Your salinity treatment is not correct, 2.46 g·L−1 is not considered salinity. This amount of salt creates an EC of less than 4 dS/m (about 3.8), which is not salt.

 In a table, the full specifications of the two water used, especially recycled water, should be provided. This is essential.

No details of how water is magnetized are given.

Author Response

Please refer to the attached document

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript entitled, "Effects of Magnetized Irrigation with Different Water Sources on Italian Lettuce (Lactuca sativa L. var. ramose) Growth and Gene Expression Analysis" is a good two-year study. However, I found some errors that will improve this manuscript.

 

Title

·         Do italic the scientific name

Abstract

·         Results are not clearly explained, write in a mechanistic way. “Magnetized irrigation improved lettuce quality, increased fresh and dry 16 weight and WPc, and reduced water consumption” This is not the way to explain quality, then growth and then others parameters

Introduction

·         “Luo et al. also found that magnetized water” “Zhao et al. reported that magnetized irrigation” “Zhang J.H. et al. observed significant” cite the references according to journal requirement.

·         Cite new studies and remove old studies

Materials and Methods

·         “Italian lettuce (Lactuca sativa L. var. ramosa Hort.) was used as the experimental material” From where have you got the seeds?

·         “2.3.2. Mineral content of lettuce The dried lettuce leaves were ground using a grinder, and the total nitrogen content in the lettuce leaves was determined by the Kjeldahl method. The contents of P, K, Ca, Mg, and Na were measured using an inductively coupled plasma optical emission spectrometer (715-ES, Agilent Technologies, USA)”. Not clearly explained. Write the references…

·         Use the full form and then abbreviations “P, K, Ca, Mg, and Na”

Results

·         It is clearly explained

·         What are the units of these parameters? “P, K, Ca, Mg, and Na”

Discussion

·         It is very poorly written. Add more comparative and recent work to explain the mechanism of how these treatments affect the soil and plant physiology, yield and quality parameters etc..

References

·         These are according to journal requirements. Please do

Comments on the Quality of English Language

Minor editing is required

Author Response

Please refer to the attached document

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

          The manuscript entitled “Effects of magnetized irrigation with different water sources on Italian lettuce (Lactuca sativa L. var. ramose) growth and gene expression analysis” by X. Yao, X. Wang, M. Qu, Y. Wei, F. Shan, and Y. Li (Ref. agronomy-3257061) presents the results from a two-season experiment in which the effects of irrigation treatments with different water sources have been combined with magnetization of those waters on lettuce growth and gene expresion were compared. The authors observed that magnetizing fresh water or recycled water improved the performance of lettuce when compared to their respective controls, while magnetizing saline water did not produce observable enhancements. The manuscript is interesting and the experiment has been carried out with care. Moreover, the manuscript fits within the scope of Agronomy.

          Despite its interest, this manuscript has several flaws that prevent its publication in the journal in its present form. The main drawback of this work is that the Discussion section is weak and needs considerable improvements. In addition, the Conclusions section needs to be re-written, as it is just a repetition of results. Moreover, the figures presented need improvements. Furthermore, additional information must be included in the Materials and Methods section. Finally, English language should be checked by a native speaker, although in general the manuscript is understandable.

          In view of these drawbacks and the specific comments below, I suggest a major revision of this manuscript so it reaches the standards for being published in Agronomy.

         

          Specific comments:

Title:

The title is confusing. Did you magnetized water or the irrigation system? I suggest re-phrasing it to: “Combined effects of magnetized irrigation and water source on Italian lettuce (Lactuca sativa L. var. ‘ramose’) growth and gene expression”.

 

Abstract:

The abstract is too long and needs to be reduced.

Lines 8-9: I suggest moving “and molecular mechanisms” to the end of the sentence instead of “transcriptomic analysis”.

Line 11: I would remove “The results showed that: (1)”.

Lines 13-14: “with the increases of pH and DO reaching the highly significant levels (P<0.01)” can be removed.

Lines 16-17: This sentence repeats the contents of the previous and following ones. I suggest removing it.

Lines 21-22: “with fresh weight […] Transcriptome results showed that”, I would remove this.

Lines 26-27: There are no drawbacks of using recycled water?

 

Keywords:

Please, do not use words that already appear in the title of your manuscript, such as “magnetized water irrigation” and “lettuce growth”.

 

Introduction:

This section explains clearly the motivation and aims of the current study.

Line 32: I would remove “for farmland irrigation”.

Line 33: “This is critical”, what is “this”?

Line 35: “domestic life” instead of “daily life”.

Line 36: Citations are needed here.

Line 44: “magnetized water irrigation treated water”. This does not make sense. Please, re-phrase this.

Line 52: Remove “J.H.” after “Zhang”.

Line 59: Reference number 10 is devoted to eggplants and not to cotton, as implied in this sentence.

Line 61: “relatively few studies”, why not citing some of them?

Lines 64-67: Citations are needed here.

Line 71: “Existing studies mainly focused on” instead of “Most existing studies focus on”.

Line 78: Remove “the” before “alternating”.

Line 79: Remove “regime”.

Line 82: “ERF, WRKY, MYB, and bHLH” are not defined. What do these abbreviations mean?

Line 90: Please, define “WPc”.

Lines 94-99: This is repetitive and redundant with respect to lines 84-93.

 

Materials and Methods:

This section requires additional information describing some aspects of the study that I detail below:

Line 104: Please, check the editing of the geographical coordinates.

Line 105: “L. var.” and "Hort." should not be written in italics.

Line 107: “air-dried soil". Where was this soil collected? What are its main characteristics (texture, pH, organic matter…)

Line 117: Units should not be written in italics.

Line 126: “three replications". How many plants per replication?

Lines 130-131; Units should not be written in italics.

Line 141: How many emitters per pot? What was the flow rate of those emitters?

Line 144: Please, define “EC" and "DO" when first used.

Line 149: “using a grinder”, either remove or provide brand and model.

Lines 161-162: Improve, as not only the wheighing of pots is included in the calculation of water productivity.

Lines 163-164: Please, be consistent with the way in which you present the units. It should be either kg/m³ or kg m^-3, but do not use both forms in the manuscript.

Line 166: How many lettuce plants were used for transcriptome analysis?

Line 167: Do you mean that you used plants from all replications in the experiment?

Line 173: Please, define “FPKM”.

Line 175: A citation is needed here.

Line 178: “GO” and "KEGG" must be defined when first used.

Line 180: Citations are needed here.

Line 184: Did you check whether data met the assumptions for this test?

 

Analysis of results:

This section should be called, simply, “Results".

Line 187: Remove “The".

Lines 189-190: “of fresh, recycled, and saline water" instead of "of fresh water, recycled water, and saline water”.

Line 196: Include "those from” before "other water sources" and remove "Further".

Line 197: “produced" instead of "obtained".

Figure 1: In the caption, indicate the meaning of the abbreviations used in the figure.

Table 1: In the title, indicate the meaning of the abbreviations used in the table. The first sentence in the footnote should be included in the title of the table.

Line 200: Remove “The”.

Line 211: Include “lettuce plants irrigated with” before “different water sources”.

Figure 2: The note should be moved to the caption of the figure. Indicate the meaning of the abbreviations used in the figure. Besides, panel B refers to 2019 and not 2018 as it is indicated in the caption.

Table 2: Indicate the meaning of “M”, “T”, asterisks and “ns”.

Line 213: Remove “The”.

Lines 225-226: To which source of water are you referring to here?

Line 229: Remove “Further”.

Lines 238-239: Please, re-phrase this sentence because it is unclear.

Lines 243-245: I suggest removing this sentence.

Table 3: Indicate the meaning of the abbreviations used and that of significance letters. Besides, in this table, some units are shown in format with slash (g/plant) while others are shown with superscripts (mg g^-1). Please, be consistent with the format in which you present the units all over the text.

Table 4: Please, indicate the meaning of the abbreviations and the asterisks. Each table should be self-explanatory.

Line 255: Please, re-phrase the title of this sub-section.

Line 256: “PCA” has not been defined.

Lines 257-260: These are not results.

Line 263: Remove “and” before “allowing”.

Figure 3: “FPKM” and “PCA” have not been defined. Moreover, the abbreviations used in the graphs should be defined.

Line 268: “screened” instead of “creened”.

Figure 4: In the Y-axis, remove “The”. “DEG” has not been defined. Include the meaning of the abbreviations for the treatments.

Line 275: Remove “water” after “fresh”.

Line 277: I suggest removing “after magnetized water irrigation”.

Figure 5: The legends in these graphs are very difficult to read. Define “DEGs”, please.

Line 278: What is “GO”?

Lines 279-280: “irrigated with magnetized water from three sources” instead of “irrigated after magnetized water irrigation with three water sources”.

Lines 286-299: These sentences are confusing. Please, re-phrase them.

Figure 6: The letters in the right of the graphs are impossible to read. Include (a), (b) and (c) for each of the panels and re-phrase the caption accordingly. Each caption should be self-explanatory.

Line 307: “water sources” instead of “irrigation sources”.

Lines 308-309: Remove “comparison group”. Besides, this is not clear in the figure.

Lines 309-316: These are not results, but some sort of discussion.

Line 317: “improves water quality by regulating cell membrane functions”. This does not make sense, as cell membrane functions do not regulate water quality, as implied in this sentence.

Lines 320-325: These are not results, but some sort of discussion.

Line 326: Remove “comparison groups”.

Lines 327-328: Not clear in the figure.

Lines 328-335: These are not results. Most of these two paragraphs are discussion and not results.

Figure 7: Please, improve this figure. It is unclear and difficult to interpret. Symbols and fonts in the legends are very small. The caption is not self-explanatory. I suggest splitting it in two or three figures.

 

Discussion:

This section is rather weak. English should be improved.

Lines 338-339: “yield, WPc, and gene expression of lettuce” instead of “yield, and WPc of lettuce, and gene expression analysis”.

Lines 341-342: Remove “and the EC, pH, and DO of irrigation water are vital to crop growth”.

Line 342: Remove “treatment” after “magnetization”.

Lines 343: “and the effects of different water sources were different”, what do you mean?

Line 352: Remove “we found that”.

Line 353: Remove “treatment”.

Line 354: Remove “altering the water quality of these irrigation sources”.

Lines 354-355: In which sense?

Lines 355-359: This is messy. It needs re-phrasing.

Line 360: Remove “the results of most”.

Lines 361-367: Please, condense information and re-phrase.

Line 370: “in plant cells” instead of “by plant cells”.

Line 374: Remove “treatment”.

Lines 381-384: Check English and re-phrase this sentence.

Lines 386-392: This is repetitive with respect to lines 337-340.

 

Conclusion:

The conclusions are interesting, but the section is too long and it is mostly a repetition of results. Please, condense information and re-write this section.

 

Data availability statement:

This is not true. Data are not included in the article, just a description and interpretation of the results generated with a dataset which is not included in the article. Please, re-phrase.

 

References:

Please, edit the references according to the journal style. For instance, use abbreviated titles of the journals. Some examples of mistakes are listed below:

Lines 438-439: Why is this reference in capital letters? What are the page numbers?

Lines 443-444: Include the page or article numbers, please.

Line 446: “Festuca arundinacea” should be written in italics. Include the page or article numbers, please.

Lines 448-449: “Triticum aestivum” should be written in italics. Include the page or article numbers, please.

Line 455: Please, include the article number.

Line 460: Please, include the article number.

Line 466: Please, include the article number.

Line 471: “Meloidogyne incognita” should be written in italics.

Line 472: This reference is incomplete. Please, check it and correct it.

Line 492: “Phaseolus vulgaris” should be written in italics.

Line 495: “Haloxylon ammodendron” should be written in italics.

Line 498: “Melissa officinalis” should be written in italics.

Lines 500-502: Why is this reference in capital letters? “Lactuca sativa” should be written in italics.

Lines 506-508: Why is this reference in capital letters? “Gossypium hirsutum” should be written in italics.

Lines 511-513: “GhCIPK6a” should be written in italics. Please, include the article number.

Comments on the Quality of English Language

English must be revised all over the manuscript for correcting some mistakes and confusing sentences. In general, language is correct, but some passages are unclear.

Author Response

Please refer to the attached document

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The authors paid good attention to my comments and presented some reasons that satisfied me and made me think their manuscript is acceptable.

 

Author Response

Dear reviewer:

Thank you for your decision and constructivecomments on my manuscript.

Reviewer 3 Report

Comments and Suggestions for Authors

Manuscript Title: Combined Effects of Magnetized Irrigation and Water Source on Italian Lettuce (Lactuca sativa L. var. ramosa Hort.) Growth and Gene Expression

Authors have addressed all the comments according to suggestion. Now, it is recommended for publication.

Comments on the Quality of English Language

Minor changes are needed which can be rectified in galley proof. 

Author Response

Dear reviewer:

Thank you for your decision and constructivecomments on my manuscript.We have touched up the English of the article.

Reviewer 4 Report

Comments and Suggestions for Authors

          The revised version of the manuscript entitled “Combined effects of magnetized irrigation and water source on Italian lettuce (Lactuca sativa L. var. ramosa Hort.) growth and gene expression” (Ref. agronomy-3257061-v2) represents a considerable improvement from the original submission. The authors have considered all of my comments and suggestions on the previous version, as well as those from other reviewers, and performed changes in the text accordingly.

          However, I suggest English being revised by a native speaker as writing is too verbose (especially in the Results section). Additionally, figure captions should be improved (sometimes, not even panels are referred to in the caption), the abbreviations of the treatments to be compared are not defined in any of the figure captions and table titles.

          In view of the enhancements that authors have performed, I suggest a minor revision of this manuscript before being accepted for publication in Agronomy.

         

          Specific comments:

Abstract:

The abstract is informative but it is too long and needs to be reduced to meet the requirements specified in the instructions for authors of the journal, as they indicate that abstract should not be longer than 300 words.

For instance, I suggest removing portions of text in lines 9-11, 13-15, 24-25, and 28-29.

Line 4: “lettuce was taken” instead of “lettuce taken”.

 

Keywords:

“Lettuce” is already included in the title.

 

Introduction:

This section explains clearly the motivation and aims of the current study.

Line 47: “Additionally” instead of “Wang, et al. also found that”.

Line 49: “Moreover” instead of “Zhao, et al. reported that”.

Lines 51-52: Remove “Zhang, et al. Observed”.

Line 55: “For instance” instead of “Zhou, et al. demonstrated that”.

Line 58: Remove “Sun, et al. also reported that”.

Line 89: “WPc” must be defined here, not only in the abstract.

Line 92: “mechanisms” instead of “mechanism”.

 

Materials and Methods:

Table 1: “Particle size” instead of “Soil size”.

Table 2: Modify the title to “Nutrient contents in the soil where the experiment was conducted”.

Line 143: “EC” and “DO” must be defined when they first appear in the text, not only in the abstract.

Line 148: “were collected” instead of “are collected”.

Line 149: Remove “in the lettuce leaves”.

Line 151: “were measured” instead of “are measured”.

Line 183: “Kyoto” instead of “kyoto”.

 

Results:

Line 193: I would remove “of different irrigation water sources”.

Table 3: “of different sources” instead of “under different irrigation water sources”. I suggest indicating the mean of the abbreviations of the treatments shown in the table.

Line 210: Increased with respect to what?

Lines 216-218: Please, check English and re-phrase this sentence.

Lines 265-266: I would remove “and allowing further analysis to proceed”.

Figure 5: There are three panels in this figure, but this fact is not considered in the caption.

 

Discussion:

Line 338: Remove “Hosseini, et al found that”.

Line 342: Remove “de Oliveira, et al found that”.

Line 347: Remove one of the two “water”.

Lines 349-356: Please, remove the name of the authors, as the reference is already included in numbers. Check the way in which you express the reports from other researchers, as it is here is also everywhere in the manuscript.

Lines 365-368: Same comment as above.

Line 391: “multiple molecular and metabolic pathways” instead of “multiple molecular pathways and metabolic pathways”.

Lines 397-402: I suggest removing this sentence, which is mainly a conclusion.

 

Conclusion:

Line 416: Remove “which provides a theoretical basis for the molecular mechanism”.

 

Data availability statement:

This is not true. Data are not included in the article, just a description and interpretation of the results generated with a dataset which is not included in the article. Please, re-phrase.

 

Author Response

Please see the attachment

Author Response File: Author Response.pdf