Germination and Emergence Responses of Alfalfa, Triticale and Quinoa Irrigated with Brackish Groundwater and Desalination Concentrate

Round 1

Reviewer 1 Report

Brief summary: The re-submitted manuscript described how irrigation with brackish groundwater and desalinization concentrate can influence germination and emergence of alfalfa, triticale (forage crop) and quinoa (food crop). I appreciated the authors effort to modify the manuscript in order to answer the comment raised after the first submission. Some minor changes are recommended.

Comment #1: In the material and methods section, row 148-149, I suggest to add the reference for 30% threshold of irrigation, as you answerend in the response form ( George H. Hargreaves...) or either to explain the method.

Comment #2: I suggest to add in the manuscript the table (and so relative reference) with EC of the sand soils planted with triticale and alfalfa, as you answered in the response form (Response 12).

Comment #3: Why don't you want to add data on gas exchange and transpiration (with eventual discussion) as you included in the response 13? They complete the manuscript, in my opinion. 

Author Response

Response to Reviewer 1 Comments

MS Title: “Germination and Emergence responses of alfalfa, triticale and quinoa irrigated with brackish groundwater and desalination concentrate”

Overall Comments:

Point 1. In the material and methods section, row 148-149, I suggest to add the reference for 30% threshold of irrigation, as you answered in the response form (George H. Hargreaves...) or either to explain the method.

Response 1. In the material and methods section, row 148-149, now Line 153-157, we have added the reference for using 30% as threshold of irrigation and explained the method as well.

Point 2. I suggest to add in the manuscript the table (and so relative reference) with EC of the sand soils planted with triticale and alfalfa, as you answered in the response form (Response 12).

Response 2. It is a good suggestion, but we also think it would be better to add a small paragraph explaining how the saline irrigation impacted the soil salinity at end of the experiment in the discussion section, without much deviating the focus of the results which is more on the germination and emergence parameters. Hope you agree with us. Please see lines 456-462 in discussion section.

Point 3. Why don't you want to add data on gas exchange and transpiration (with eventual discussion) as you included in the response 13? They complete the manuscript, in my opinion. 

Response 3. I really appreciate your suggestion on adding the data on gas exchange and transpiration rates in this manuscript, however, we would like to clarify that the gas exchange and transpiration measurements were recorded in a completely different experiment on evapotranspiration. This data were not included in the earlier manuscript because it was collected towards the end (10 days before the harvest) of the experiment, when the plants were already showing chlorosis symptoms and the readings were very low.

Reviewer 2 Report

The manuscript with the title “Germination and emergence responses of alfalfa, triticale and quinoa irrigated with brackish groundwater and desalination concentrate”  was evaluated following reviewer instructions from the journal of Agronomy. I found this paper very interesting; it provided a piece of valuable information which producer can consider before establishing crops under poor or saline soils. I found this manuscript suitable for a publication in Agronomy, and I believe this work can attract potential readers working on this topic. All sections were well written, and scientifically sounded. For this manuscript to move forward, authors should take into consideration a few comments placed below.

In the introduction section, authors should mention the biological differences between crops; in this study, authors are comparing three different crops, such as grass, herbaceous and forage legume. In alignment with differences in plant species, I was wondering if such differences between crops were associated with the root system type. Grasses and legumes crops have a different root system; at some point, tolerance to stressor conditions can be related to the root architecture. It would be good if authors can provide some background in the introduction of biological differences between crops.

Line 74, need space between oC and the next word.

Line 76 to 79, please provide some references here.

Figure 2, add a legend in the Y-axis (days)

Table 6, make a more esthetic table, place soil in the first column and crop specie in the second. Please, arrange numbers to the left side of each cell

Figure 4, add a legend in Y-axis (days)

References should have the same style, some references are provided with journals names abbreviation, while others have full name, the same for italic and no italics names.

Author Response

Response to Reviewer 2 Comments

MS Title: “Germination and Emergence responses of alfalfa, triticale and quinoa irrigated with brackish groundwater and desalination concentrate”

Overall Comments:

Point 1. In the introduction section, authors should mention the biological differences between crops; in this study, authors are comparing three different crops, such as grass, herbaceous and forage legume. In alignment with differences in plant species, I was wondering if such differences between crops were associated with the root system type. Grasses and legumes crops have a different root system; at some point, tolerance to stressor conditions can be related to the root architecture. It would be good if authors can provide some background in the introduction of biological differences between crops.

Response 1. We really appreciate your suggestion to add biological differences between the three crops. We have added relevant biological information for each species. Please see lines 85-95 in the introduction section.

• Line 74, need space between oC and the next word.
• We inserted the space between oC and the next word in line 74 of the manuscript.
• Line 76 to 79, please provide some references here.
• We have added the reference in line 77.
• Figure 2, add a legend in the Y-axis (days)
• We added the legend ‘MGT in days’ on Y-axis in Figure 2.
• Table 6, make a more esthetic table, place soil in the first column and crop species in the second. Please, arrange numbers to the left side of each cell
• We edited Table 6 as suggested.
• Figure 4, add a legend in Y-axis (days)
• We added the legend ‘MET in days’ on Y-axis in Figure 4.
• References should have the same style, some references are provided with journals names abbreviation, while others have full name, the same for italic and no italics names.
• We corrected citations with complete journal names and the titles are italicized in the references now.

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

Thanks for the authors' efforts on the irrigation science.  This paper demonstrates the germination and emergence responses of three species to saline and treated irrigation water. The results are novel and the experiment design is appropriate. However, there are too many problems with writing a scientific paper. I would provide a few examples as follows:

1. There are too many editorial faults. Look at figure 1. Which one is figure 1? and the letter "B" in "Fig. 1". Also, there are too many double spaces between words. How about the sentence "The results were in total agreement with [36,37].Salinity tolerance differed with various stages of plant growth especially in alfalfa and quinoa." in the discussion section? Again, these are only a few examples and these mistakes greatly undermine the reliability of this paper. So, please carefully revise the manuscript again and again.
2. The way of referring the references in the text is inappropriate. "In general, according to [68,69] most species either....", "[8] in their germination study reported..." these are not the correct way to referring a reference in the text. Again, these are a few examples of referencing mistakes.
3. Last but not least, the way of writing a scientific paper is too short to be published in this journal. A few examples. There are too many paragraphs in the manuscript. There should be less than 6 paragraphs in the introduction section, especially in an experiment paper. The fact that there are many paragraphs is a testament to the author's lack of a clear understanding of what this study means. Please refer to the commentary of Plaxco (2010), "The art of writing science", to improve the current article.  

I am really sorry that I must be critical of your paper. My critics are not about your experiments or findings, but the way or your presentations. I hope the authors keep improving this article so that many readers would enjoy reading the interesting findings of this work. Many thanks again!

Reviewer 2 Report

Brief summary: The manuscript investigate how irrigation with brackish groundwater and desalinization concentrate can influence germination and emergence of alfalfa, triticale (forage crop) and quinoa (food crop). The work provide results taking into account several germination/emergence variables, and by them authors suggested triticale as a promising salt tolerante forage crop.

Broad comments and Specific comments:

Comment # 1. In the abstract, I suggest authors to add some values concerning germinability and emergence % to improve it a bit.

Comment # 2. I recommend authors to check tables and figures formatting: almost all have problems like misalignement.

Comment # 3. Introduction in my opinion is too long: I suggest to authors to shorten it between 10-20%. Readers could feel a bit lost about the focus of the topic.

Comment # 4. Still in the Introduction, I suggest to authors to change place (and reduce) the description of the three crops considered for the experiment: I would put the descriptions from row 111 to row 124 just after row 82, integrating it with the current paragraphs. In my opinion than the description should be more linear to read.

Comment # 5. The presented methods are detailed, and statistical analisys are sound (mixed model is a good approach). However, I have a couple of major criticisms concerning experiment concepts:

1) Being an experiment on saline condition effects on forage and food crop germination/emergence, in my opinion is strictly necessary to have ions elemental analysis on processed seedlings/plantlets (at least Na⁺ and K⁺ concentration). Germination and emergence are influenced by ions balance, and you also state that in the discussion (from row 84 to row 86 of the discussion section). Moreover, besides the ionic factors, the osmotic component are really important to assess total effects on germinability experiments under saline conditions (according to Munns et al., 1995); So I ask to authours if they have this data. In case not, I suggest to perform those analysis and I strongly recommend to add it in the manuscript, describeing also the methods.

2) Authors did not give any information about biomass (Dry Weight, growth parameters such us elongation of the seedlings from time zero and/or relative growth rate) production and water content of the seedling/plantlets under different saline irrigation regime: I really suggest to add those information and their methods if they have, since also those parameters really contribute to consider a crop as salt tolerant or salt sensitive. Even if you state in the discussion section (row 8 and 9 of discussion) that some "studies indicated there is no relation between germination, emergence and plants ability to produce biomass", in my opinion is still an important parameter to show, especially when you wanna provide ideas on salt tolerance or sensitivity.

Comment # 6. In the Table 1: do you also have K concentration for the physiochemical properties of soils used for the experiment? Potassium play an important role in Na homeostasis, especially in the case of a saline condition experiment.

Comment # 7. Table 2: you should add the treatment acronim near the dsm^-1 description for each saline condition, like you did for control; I suggest also to eliminate the last column (EC): it's redundant, you already stated it in the first one.

Comment # 8. Still in the Material and Methods section: from the end of row 149 to row 160, in my opinion is a bit redundant; I suggest to shorten and simply refer to Table 2.

Comment # 9. In the Germination experiments description, at row 166, you state "replicated germination experiments"? but I suggest to cite the number of time you repeated the experiments; the same for paragraph 2.2. Moreover, I am not sure I understood the number of the biological replicates: are them 25? (see row 169 of mat and methods, in wich you state "twenty five seeds were placed..."). Or 25 seeds for different blocks? Could you please clraify that? Thank you

Comment # 10. Could you please explain how and why you decided to rewater the pots when soil water content was depleted about to 30% of the saturated water content? (see row 189-190 of "emergence experiments", still in material and methods section). It's really important to understand how and why you decided 30% as a thrashold for rewatering, otherwise you are going to apply also a drought stress.

Comment # 11. When you describe the variables computations and the relative formulas, I suggest to write it twice: with "G" when describes germinability, or with "E" when they assess Emergence; otherwise can be confusing.

Comment # 12. I am asking to authors if they measured EC in the soil at the end of the experiment. It would be nice to have an idea of the total salinization of the environment.

Comment # 13. I am asking if authors did some physiological measurements like gas exchanges or transpiration rate or chlorophyll content.

Comment # 14. In the results section, the number progression of figure caption is not correct: after fig 1 for germinability % (Soil main effects), you still use "Figure 1" for the germinability % (Species main effects). Please correct it (and also in the text).

Comment # 15. In the results section, page 13, "Emergence Index (EI), Timson's Index (TI) and Timson modified Index (Tmod)", there is no reminder to EI table (the 7) when you speak about EI. Please add it.

Comment # 16. In the discussion section, page 16 row 2, I am not sure that it's needed to add the title "Germination and Emergence".

Comment # 17. From row 42 to row 44 of the discussion: you cannot coclude that, if you don't add ions data it's just a speculation or a suggestion.

Comment # 18. In the conclusion section, I ask to authors to stress more new findings of their work respect to other recent publication in this area.