Differences in the Physiological Indicators of Seed Germination and Seedling Establishment of Durum Wheat (Triticum durum Desf.) Cultivars Subjected to Salinity Stress
Elena Delian
Round 1
Reviewer 1 Report
This paper analyzed the effects of salt stress on the germination of eight durum wheat genotypes seeds, and presented the results in a good way. The main suggestion are as follows.
The title needs to be improved to make it more concise and precise.
All data in the paper need to be added with significance analysis.
The horizontal and vertical coordinates of figures need to be labeled with their full names. Abbreviations should be avoided. The figure needs to be showed with self-evidence.
Try to avoid using codes or abbreviations in the table and change to the full name. The table explanation should be placed below the table.
The discussion needs to be segmented. Each paragraph should focus on a clear issue rather than being mixed together.
The schematic representation (Figure 7) is relatively simple, and it is recommended to remake or delete it.
The method needs to be written under different subheadings for clearer expression.
Author Response
REVIEWER 1
Thank you very much for accepting to review our MS and present this report.
The MS was taken up again and subjected to a total revision of its scientific content, quality, style and presentation. All recommendations are taken in account, Figures are redone to be clearer and more concise. All recommended amendments were added. They are highlighted in green in the text. Below our response to your reports
This paper analyzed the effects of salt stress on the germination of eight durum wheat genotypes seeds, and presented the results in a good way. The main suggestion are as follows.
* The recommendation regarding title was taken in consideration and title was improved as follow:
Differences in the physiological indicators of seed germination and seedling establishment of durum wheat (Triticum durum Desf.) cultivars subjected to salinity stress
* The numerous recommendations regarding significance analysis, figures, abbreviation, codes, tables, discussion, schematic fig. 7, heading and subheading are considered, and the MS was improved in style, figures, tables….
All amendments are highlighted in green in the text.
* For difference significance: Because some figures are presented as curve (not histogram), it is very difficult to add the convenient letter on each point. For that, we used horizontal discontinuous lines that mark the passage from one level to the other one of significance.
* For cultivars differences, this point was clarified in MM, section 2.1. The used cultivars are known grown in different bioclimatic stages of Tunisia, their differences in water need and their different yield.
Author Response File: 
Author Response.doc
Reviewer 2 Report
The topic of this study is one of interest considering the expansion and accentuation of the effects of saline stress on the growth and development of plants in general, and in the present case, on durum wheat. Therefore, the characterization from the physiological point of view of the response of some cultivars to this abiotic factor during the germination process and during the early growth of the seedlings can ensure the premises for the choice and use in the culture of the most suitable cultivars, which can cope with success in terms of salt stress tolerance. From the point of view of the structuring of the information and the appropriate use of the specific terminology, I appreciate that some interventions on the part of the author are necessary. In this sense, some observations and recommendations are presented below (which are also found in the manuscript reviewed by me).
1.wheat genotypes
So, genotypes or cultivars?
Eight cultivars have been used.
2. Eight cultivars have been used.
......the cultivars diversity?
Abstract
3. genotypes or cultivars?
As I understand, durum wheat cultivars have been studied.
4. ....on eight durum wheat genotypes ('Karim', 'Khia'r, 'Inrat100','Maali', 'Monastir', 'Portodur', 'Razeg', and 'Salim') ...
So, genotypes or cultivars?
5. ...salinity concentration by using sodium chloride (NaCl) (0, 2, 4, 6, and 10 g L-1).
6. I recommend:
... mean daily germination (MDG),
....mean germination time (MGT).....
7. ......cultivars differences
were revealed: 'Karim' and' Salim' are least affected, 'Razeg' and 'Maali’ are most .
8. ....osmotic effect (OE) was...
9. .......dominated the toxic effect (TE) and irreversibly inhibited the germination process.
10.
IV and SI that .....? Because SI there was not explain before.
The initial vigor and the stress index that.......studied cultivars are....
Introduction
11. ....sodium chloride (NaCl) concentration....
12. Durum wheat (Triticum durum Desf.),
Because the entire name (common and scientific) has been written earlier, now (and following), you can use only one of them: durum wheat or T. durum.
Before Results section it will be placed as
2. Materials and Methods section.
13.
3.Results?
In fact, the second section will be:
2. Materials and Methods
……
I will comment on it later.
Now, the obtained results will be presented in the same order as in Materials and Methods the different indicators have been described.
3.Results
Example:
3.1. Seeds germination capacity
....
3.2. Germination rate................
14. Germination capacity (GC, %, a) of durum wheat ....
Is this in fact the final germination percentage (FGP) (%) scientifically determined for wheat, according to the STAS rules?
15. of 100 grains (....
16. .decreased germination rate (GR) in...
Now, you can use.....germination rate or GR.
17. Data presented in Table 1 show that.....
18. Our results also show.....
19. Please,
For Y axis.....
Inside the graphs, Germination rate (%/day)?? or values after each day represent in fact the cumulative values regarding the % of seeds germinated?
The name of the cultivars need to be written as.......'Khiar'....'.Maali'....and so on
For X axis.....Germination time (days)
20.Table 1.
Germination index (GI) has not been described in Materials and Methods. Please do it.
For the control there was not determined?
Please completely write the sodium chloride concentrations....not only 2 NaCl.......but, 2 g NaCl/L...and so on
The entire cultivars name, or as......Khi.....Kar......Raz.....s
Please write in English.....as 0.98.....; NOT 0,98...and so on.....
21. The velocity coefficient (Vc) has been described in Materials and Methods section?
22. Please use the consecrated names for indicators.
23. ....'Karim' and 'Salim' remain...
24. ...by 'Inrat', 'Ihiar', 'Monastir', and 'Portodur', whereas 'Razeg' and 'Maali' are the most affected.
25. ...except 'Razeg' (GRec de-clined to 82%) and 'Maali'...
26. ..in 'Maali', 50%, and 'Razeg'.....
27. 'Razeg'
28.Table 2.
....mean germination time (MGT) and mean daily germination (MDG) ....
29. The same observation as for Table 1.
30. 73.4 .....and so on....
31. The same observation as for Table 1.
- the salt concentration explicitly
32. 67.2........and so on....
33. MGT
1.36........1.19...and so on
34. MDG
23.5....23.5....22.5.....and so on
35. Figure 3.
The non-germinated grains in salt are transferred to deionized water and the number is used to calculate the recovery percentage (%).
This phrase can be deleted. The calculation procedure should be described in Materials and Methods section.
As we can see, the statistical analysis results are not presented in the Figure 3. So, why do you say ....at α = 0.05?
36. Inside the graph, please write the cultivars’ names as I previously recommended.
37.Figure 5.
....of means of 100 replicates? or ?
- p≤ 0.05. ......but, statistical analysis has not been done
38. 2 g L-1
39.Figure 7.
DW, gmg plant -1 ?
Please, not clear here.
The indicator is....
DW (mg plant -1)?
Please pay attention to repetitions vs. replicates. Also, ...p≤ 0.05, but statistical analysis has not been done here..
I recommend writing the cultivars name as I said earlier.
40. Did you describe the initial vigor (IV) in Materials and Methods section?
41.Table 3.
Please consider my previously observations and recommendations regarding the cultivars writing and the treatments applied.
42. the stress index?
Did you describe this indicator in Materials and Methods section? If not, please do it.
43.Table 4.
Please consider my previously recommendations.
44. Table 4.
1,04c ± 0.08
Please, it is not clear.
In English
- we write for 1000.....1,000
- we write for 1,04.......1.04
So, .....
Discussion
45. ...genotypes or cultivars? as there was the presentation before
46. I appreciate that the consecrate names of the indicators should be used.
47. ..sensitive cultivars ('Razeg' and 'Maali') than in the intermediate and tolerant cultivars ('Karim' and 'Salim').
48. ...the mean daily germination (MDG)
49. ...is the osmotic effect (OE), ...
As in other cases too, if at the first apparition in text there was done the explanation for an abbreviation, later it is not necessary to repeat.
Is possible o use only the abbreviation or, if you want, only the explanation (not both).
50. ...at electrical conductivity (EC) ≥10 dS m-1
51. (potassium - K+, calcium -Ca2+, magnesium – Mg 2+, and nitrate -NO 3-)
52.
2. Materials and Methods
To be easier pursue by the reader, I recommend using subheadings here.
So, please carefully review the section.
For example:
2.1. Experimental place
......
2.2. Biological material
......
2.3. Experimental setup and design
......
2.4. Indicators regarding seed germination traits
2.4.1. Germination capacity (GC)(%)
......
2.4.2. Germination rate (GR) (%. day--1)
......
2.4.2.Germination recovery (GRec) (%)
......................
2.4.3. and so on
.....
2.5. Statistical analysis
............
53. Salim is missing.
So,
('Karim', 'Khiar', 'Razek', 'Maali', 'Inrat100', 'Portodure', and 'Monastir')
If later on you will use some abbreviations for these, please also write in the above sentence in the simple brackets, the abbreviation for each one.
54. ...per Petri dish....
55. .....imbibed with distilled water (control -0 salt) and saline solution containing 2, 4, 6, and 10 g L-1 sodium chloride (NaCl).
56. Germination capacity (GC) ?
I appreciate that in plant germination physiology it is rather used.....Germination percentage (GP) (%).......
57. Farzadfar et al., 2017 is missing from the Reference section.
58. Seedlings growth ....
59. So, As I said at the section beginning, please review the entire section and do the necessary reorganization.
60. estimates
Why do you say estimates.
Because the values have been really calculated based on concrete collecting data, I appreciate that we can say....By the various experiments there were determined the following....
61. Please consider my before observations.
62. GP (%)=
63. Germination rate (GR) (%. day-1)
64. Germination recovery (GRec) (%) ....
65. GRec (%) = .....
66. As a rule, it is named
Mean daily germination (MDG) (seeds number germinated per day)
67. As a rule it is named,
Mean germination time (MGT) (days)...
68. Statistical analysis....,,,,in the Tables.
....as we can see.
69. Please respect Instructions for authors
.https://www.mdpi.com/journal/agronomy/instructions
References should be described as follows, depending on the type of work:
Journal Articles:
1. Author 1, A.B.; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.
Example:
Wang, W. B.; Kim, Y. H.; Lee, H. S.; Kim, K. Y.; Deng, X. P.; Kwak, S. S. Analysis of antioxidant enzyme activity during germination of alfalfa under salt and drought stresses. Plant Physiology and Biochemistry, 2009, 47, 570–577.
Comments for author File: 
Comments.pdf
Author Response
RESPONSE TO REVIEWER’S REPORTS
REVIEWER 2
Thank you very much for accepting to review our MS and present this report.
The MS was taken up again and subjected to a total revision of its scientific content, quality, style and presentation. All recommendation are taken in account, Figures are redone to be clearer and more concise. All recommended amendments were added. They are highlighted in green in the text. Below our response to your reports
* After verification of scientific meaning and discussion with scientist who provided as with seeds in the National Institute of Field crops of Kef (Tunisia), the used wheat seeds in this study are cultivars. The convenient modifications are made alongside the MS.
* The cultivars names were re-adjusted with capital first letter, missed one (Salim was added)
* Mistakes, errors and missing information’s about salt stress, concentration, cultivars, abbreviation (MGT, MDG….), legends, comma in tables replaced by point … all marked remarks in the pdf you provided are taken in consideration and necessary corrected are made
* Revised Abbreviated parameters and indicators and lacking in MM are added and explained in section 2.3 (SI, IV )
* The repetition and confusion in the use of abbreviated parameters and full names are, and the use of double names (durum wheat/ triticum durum) were avoided and the overall text was revised
* MM was placed in section 2 instead of 4
* MM section was divided into subsections as follow:
- Materials and Methods
2.1. Biological material and germination experiments
2.2. Potted experiments
2.3. Experimental design and germination indicators
* Final germinationcapacity (FGC).This parameter represents the maximum the maximum germination capacity that can be achieved if the osmotic problem is solved. For this it represents the summation of the GC and GRec (germination recovery after transfert on non germinating seeds to H2O). This indicator is explained in MM, section 2.3.
* Germination index (GI). GI represent the ratio of stressed to control, for that there is no GI in control. It is usually < 1. So 1 for control
* The statistical analysis of results was made and significance added in all figures.
*. .....imbibed with distilled water (control -0 salt) and saline solution containing 2, 4, 6, and 10 g L-1 sodium chloride (NaCl), recommendation taken in account
* The germination capacity (GC) was explained and differenciated from germination percentage. They are mentioned and explained in MM
* Farzadfar et al., 2017 is missing from the Reference section. Added reference.
* Please respect Instructions for authors. The reference list was revised and rewritten according to the journal instructions.
* For difference significance: Because some figures are presented as curve (not histogram), it is very difficult to add the convenient letter on each point. For that, we used horizontal discontinuous lines that mark the passage from one level to the other one of significance.
* For cultivars differences, this point was clarified in MM, section 2.1. The used cultivars are known grown in different bioclimatic stages of Tunisia, their differences in water need and their different yield.
Author Response File: Author Response.doc
Reviewer 3 Report
The review of the paper written by Hmissi et al and submitted to Agronomy MDPI. The paper cover the aims of this journal, however based just on germination analysis and initial seedlings growth of 8 wheat durum cultivars. The results seems to be predictive however it could be useful for Tunisian growers and supplement the characteristic of the cultivars. In my opinion the results are just an observation and do not answer any scientific question. I also do not find any hypothesis stated prior to the study. This is why I have objection to published this research in journal with IF nearly 4.
I also advise to add statistical deviation to figure 2. Also in my opinion the authors should present statistic differentiation in figures. They should indicate where the difference is statistically important. The comparison should be done between different salt concentration for each of cultivars as well as for different cultivars at each of salt concentration.
I also suggest to move the last sentence from result section into conclusion.
In the M&M section I have one question. How the salt was add during seedling growth. Was it just one time at the beginning or was the plants watered with nutrient solution with salt every weeks? Do the authors monitored salt concentration in the soil? Do the authors protect the pots with plants against salt accumulation due to water evaporation?
The cited references are up to date which is very important. It is also well written and prepared. The most important doubts is about scientific level of this manuscript.
Author Response
RESPONSE TO REVIEWER’S REPORTS
REVIEWER 3
Thank you very much for accepting to review our MS and present this report.
The MS was taken up again and subjected to a total revision of its scientific content, quality, style and presentation. All recommendation are taken in account, Figures are redone to be clearer and more concise. All recommended amendments were added. They are highlighted in green in the text. Below our response to your reports
* The review of the paper written by Hmissi et al and submitted to Agronomy MDPI. The paper cover the aims of this journal, however based just on germination analysis and initial seedlings growth of 8 wheat durum cultivars. The results seems to be predictive however it could be useful for Tunisian growers and supplement the characteristic of the cultivars. In my opinion the results are just an observation and do not answer any scientific question. I also do not find any hypothesis stated prior to the study. This is why I have objection to published this research in journal with IF nearly 4.
In this paper we grant a special interest to the durum wheat response to salinity stress at the early stages of the plant cycle because this is a real problem for Tunisian Farmers. In fact, in non- irrigated agrosystems, durum wheat was usually sown after first or second precipitation of Automn, sometimes seeds and plantlets spend long times under no raining and salinized soil. Even in irrigated perimeter, the major underground water used for irrigation is saline. Thus, screening tolerant cultivars at this stage can guarantee the subsequent success of the culture. Our approach was the exploration of the variability of durum wheat response to salinity to screen cultivars with good germination and plantlet initiation under salinity stress which represent a rapid, safe, and ecofriendly approach. Our hypothesis was that a cultivar that shows some tolerance during germination and the early stages of development would certainly be tolerant at later stages of the plant cycle. We also tried to identify the most critical stage (germination or seedling establishment) in order to propose the appropriate improvement approach, such as priming if germination is more sensitive. The findings of this study can provide useful physiological indicators or traits for screening wheat cultivars for salt tolerance in future screening programs.
* I also advise to add statistical deviation to figure 2. Also in my opinion the authors should present statistic differentiation in figures. They should indicate where the difference is statistically important. The comparison should be done between different salt concentration for each of cultivars as well as for different cultivars at each of salt concentration.
Statistical analysis was done, and significance was added to all figures
* I also suggest to move the last sentence from result section into conclusion.
Recommendation taken into consideration.
* In the M&M section I have one question. How the salt was add during seedling growth. Was it just one time at the beginning or was the plants watered with nutrient solution with salt every weeks? Do the authors monitored salt concentration in the soil? Do the authors protect the pots with plants against salt accumulation due to water evaporation?
We used in this experiment a sandy soil just as a support to retain plant. Almost the conditions of strict hydroponics. Because the field capacity of the used sand is very low (7 %), plants are over irrigated every two days. The excess of solution recovered in the plate is put back into the pots next day. These explanations are added in MM
* For difference significance: Because some figures are presented as curve (not histogram), it is very difficult to add the convenient letter on each point. For that, we used horizontal discontinuous lines that mark the passage from one level to the other one of significance.
* For cultivars differences, this point was clarified in MM, section 2.1. The used cultivars are known grown in different bioclimatic stages of Tunisia, their differences in water need and their different yield.
Author Response File: Author Response.doc
