Effect of Salinity and Nitrogen Fertilization Levels on Growth Parameters of Sarcocornia fruticosa, Salicornia brachiata, and Arthrocnemum macrostachyum
, Zhadyrassyn Nurbekova 1, Dinara Oshanova 1, Arvind Kumar Dubey 2, Kusum Khatri 2, Varsha Mudgal 3, Anurag Mudgal 3, Amir Neori 4,5, Muki Shpigel 4, Rajeev Kumar Srivastava 6, Luísa Margarida Batista Custódio 7, Dominic Standing 8,* and Moshe Sagi 8,*Round 1
Reviewer 1 Report
The work may have some interesting aspects, but my main concern is the lack of control samples that would give a more precise indication of the plants' response to the various treatments. I recommend the inclusion of controls.
Also, some parts of the manuscript need to be rewritten: the introduction should be shortened and better focused on the objectives, and the discussion less repetitive of the results and more focused on their discussion.
Author Response
Agronomy - 1797302
We thank the reviewers for their time and effort. In the light of their suggestions, we have made extensive revisions to the manuscript. In particular the Results figures have been completely reformatted and the legends rewritten. The Discussion has also been rewritten with better focus and the deletion of distracting details. We think these revisions greatly enhance the readability of the manuscript. Once again, we thank the reviewers for their suggestions.
The number of revisions (more than 500) is such that detailing them one by one would be impractical. Please find the track edited pdf version attached.
Our responses to specific comments are written below.
Reply to Reviewer 1
Comment: The work may have some interesting aspects, but my main concern is the lack of control samples that would give a more precise indication of the plants' response to the various treatments. I recommend the inclusion of controls.
Response:
With regard to control plants in case of a study that includes halophytic plants such as Salicornia and/or Sarcocornia, it should be considered that ‘zero salinity’ as the control treatment would in itself, be a stressor. Accordingly, it is entirely appropriate to include a salt treatment as the control, when the plant material is Salicornia and/or Sarcocornia and compare other salinity level treatments to it, as done before by Ventura et al., (18, 24, 25), and Kurmanbayeva et al., (50).
The above paragraph is now written at the end of the discussion section.
Comment: Also, some parts of the manuscript need to be rewritten: the introduction should be shortened and better focused on the objectives, and the discussion less repetitive of the results and more focused on their discussion.
Response: The Discussion has been extensively rewritten with the reviewer’s comments in mind. For example, specific mention of treatment results has been deleted unless it specifically strengthens the argument. Discussion of the methods has also been deleted. The second reviewer found no fault with the Introduction except that he/she would like to see some recent literature citations. Additional references have been added to address this.
Reply to Reviewer 2
Authors showed an interesting study in which they are looking for potential crop species between halophytes using an already halophyte crop species (S. fruticosa) as a reference. They obtained interesting results not only for the candidates but aslo for S. fruticosa.
Introduction its well structure, it includes the main topic and the aim of the manuscript is clearly indicated and material and methods are explained in detail. However there is some consideration I would like to make.
Comment: Introduction: There are some references talking about the innovation in halophytes crops but they are ten-year-olds. I would want to know the most recent literature about the topic.
Response: Additional references have been included.
Comment: Material and methods: Temperature between 30 and 35ºC were used for the germination and I think that it could be too high. Why did the authors this range of temperature?
Response: Although we have not performed a ‘germination x temperature’ experiment on these species/ecoptypes, our extensive experience in this lab has shown that these conditions give excellent germination.
Comment: It is said that a parallel trial was setup for plants to flower, however it is not established the importance that flowering have in this experiment.
Response: Now it is written in lines 142 to 150 as follows: " It was noted that S. brachiata did not flower under the 14-hour day length and thus biomass accumulation was not interrupted by flowering. To examine if under natural day length conditions the successive harvest will prevent flowering, a parallel trial was set up in a greenhouse under natural day length for 30.8523° N, 34.7834° E, the temperature was partially controlled (~10°C min to 40°C max), and midday PAR reached 650-700 Mol/m2/s. Two nine L pots of ten plants per pot were planted, using the same potting medium as for the growth room experiments, for S. brachiata and S. fruticosa VM in June 2021. Plants received the LN+LS treatment and were subject to a repetitive harvest regime (same as the growth room plants) over one year.".
Supplementary photo S2 shows a flowering example of S. brachiata grown for seed production.
Comment: Why did the author not use and ANOVA or a statistical analysis that allow you to compare multiple treatments instead of comparing one-one each treatment?
Response: We used one-way ANOVA (fit y by x) followed by Tukey’s HSD for the majority of the analyses. The Data Analysis text has been corrected to reflect this.
Comment: Results: This section is hard to follow as it is shown. Species and ecotypes are sometimes referred as two letters (For instance, AM) and in other occasion it use the full name. It does not help to read it fluently. Each section is divided in two and I think that it is not needed.
Response: the text has been edited so that the two-letter identifier is used through this section. With respect to dividing each result into two parts (within ecotype and between ecotypes), the authors deliberately chose this style to separate the different analyses and avoid confusion.
Comment: Figures must be changed. They are not intuitive and the abbreviation of treatment should be replaced by salinity and N concentrations. There are plenty of papers in the literature where multiple combinations of treatments are showed clearer than this. In addition, letters used to represent significant differences are also hard to interpret.
Response: The Figures have been significantly altered. Now, each species/ecotype is represented by a different shaded bar and each treatment is separated by a dashed line. Comparisons within each treatment are still in lower case but now in red. The legends have been altered to reflect this.
Comment: Results are too specific, like a description of the figures; it would be more helpful if they just summarize the global changes that treatments showed pointing specific values that are interesting for the discussion.
Response: This section has been re-edited. The average paragraph length for each result is two to three sentences. Often it is only a single sentence. This section was written in accordance with the journal instructions: ‘concise and precise description of the experimental results’.
Comment: In Table 2: Why did not show the harvest for all ecotypes and species?
Response: Only two contrasting species were tested in the greenhouse, SB and VM. This is mentioned in the text.
Comment: Discussion: It is too descriptive. The results are again displayed in a more summarized way and it can make the manuscript repetitive. The discussion is poor in general and it sometimes shows a lack of connection between the parameters measures.
Response: This section has been significantly re-written to take account of the reviewer’s comments, i.e. removal of directly mentioned results, as well as description of the methods.
Comment: Line 420: “with those of [6] at a similar salt level” [6] must show the surname of the authors.
Response: This has been corrected to show the surnames.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Authors showed an interesting study in which they are looking for potential crop species between halophytes using an already halophyte crop species (S. fruticosa) as a reference. They obtained interesting results not only for the candidates but aslo for S. fruticosa.
Introduction its well structure, it includes the main topic and the aim of the manuscript is clearly indicated and material and methods are explained in detail. However there is some consideration I would like to make.
Introduction: There are some references talking about the innovation in halophytes crops but they are ten-year-olds. I would want to know the most recent literature about the topic.
Material and methods: Temperature between 30 and 35ºC were used for the germination and I think that it could be too high. Why did the authors this range of temperature?
It is said that a parallel trial was setup for plants to flower, however it is not established the importance that flowering have in this experiment.
Why did the author not use and ANOVA or a statistical analysis that allow you to compare multiple treatments instead of comparing one-one each treatment?
Results: This section is hard to follow as it is shown. Species and ecotypes are sometimes referred as two letters (For instance, AM) and in other occasion it use the full name. It does not help to read it fluently. Each section is divided in two and I think that it is not needed.
Figures must be changed. They are not intuitive and the abbreviation of treatment should be replaced by salinity and N concentrations. There are plenty of papers in the literature where multiple combinations of treatments are showed clearer than this. In addition, letters used to represent significant differences are also hard to interpret.
Results are too specific, like a description of the figures; it would be more helpful if they just summarize the global changes that treatments showed pointing specific values that are interesting for the discussion.
In Table 2: Why did not show the harvest for all ecotypes and species?
Discussion: It is too descriptive. The results are again displayed in a more summarized way and it can make the manuscript repetitive. The discussion is poor in general and it sometimes shows a lack of connection between the parameters measures.
Line 420: “with those of [6] at a similar salt level” [6] must show the surname of the authors.
Author Response
Agronomy - 1797302
We thank the reviewers for their time and effort. In the light of their suggestions, we have made extensive revisions to the manuscript. In particular the Results figures have been completely reformatted and the legends rewritten. The Discussion has also been rewritten with better focus and the deletion of distracting details. We think these revisions greatly enhance the readability of the manuscript. Once again, we thank the reviewers for their suggestions.
The number of revisions (more than 500) is such that detailing them one by one would be impractical. Please find the track edited pdf version attached.
Our responses to specific comments are written below.
Reply to Reviewer 1
Comment: The work may have some interesting aspects, but my main concern is the lack of control samples that would give a more precise indication of the plants' response to the various treatments. I recommend the inclusion of controls.
Response:
With regard to control plants in case of a study that includes halophytic plants such as Salicornia and/or Sarcocornia, it should be considered that ‘zero salinity’ as the control treatment would in itself, be a stressor. Accordingly, it is entirely appropriate to include a salt treatment as the control, when the plant material is Salicornia and/or Sarcocornia and compare other salinity level treatments to it, as done before by Ventura et al., (18, 24, 25), and Kurmanbayeva et al., (50).
The above paragraph is now written at the end of the discussion section.
Comment: Also, some parts of the manuscript need to be rewritten: the introduction should be shortened and better focused on the objectives, and the discussion less repetitive of the results and more focused on their discussion.
Response: The Discussion has been extensively rewritten with the reviewer’s comments in mind. For example, specific mention of treatment results has been deleted unless it specifically strengthens the argument. Discussion of the methods has also been deleted. The second reviewer found no fault with the Introduction except that he/she would like to see some recent literature citations. Additional references have been added to address this.
Reply to Reviewer 2
Authors showed an interesting study in which they are looking for potential crop species between halophytes using an already halophyte crop species (S. fruticosa) as a reference. They obtained interesting results not only for the candidates but aslo for S. fruticosa.
Introduction its well structure, it includes the main topic and the aim of the manuscript is clearly indicated and material and methods are explained in detail. However there is some consideration I would like to make.
Comment: Introduction: There are some references talking about the innovation in halophytes crops but they are ten-year-olds. I would want to know the most recent literature about the topic.
Response: Additional references have been included.
Comment: Material and methods: Temperature between 30 and 35ºC were used for the germination and I think that it could be too high. Why did the authors this range of temperature?
Response: Although we have not performed a ‘germination x temperature’ experiment on these species/ecoptypes, our extensive experience in this lab has shown that these conditions give excellent germination.
Comment: It is said that a parallel trial was setup for plants to flower, however it is not established the importance that flowering have in this experiment.
Response: Now it is written in lines 142 to 150 as follows: " It was noted that S. brachiata did not flower under the 14-hour day length and thus biomass accumulation was not interrupted by flowering. To examine if under natural day length conditions the successive harvest will prevent flowering, a parallel trial was set up in a greenhouse under natural day length for 30.8523° N, 34.7834° E, the temperature was partially controlled (~10°C min to 40°C max), and midday PAR reached 650-700 Mol/m2/s. Two nine L pots of ten plants per pot were planted, using the same potting medium as for the growth room experiments, for S. brachiata and S. fruticosa VM in June 2021. Plants received the LN+LS treatment and were subject to a repetitive harvest regime (same as the growth room plants) over one year.".
Supplementary photo S2 shows a flowering example of S. brachiata grown for seed production.
Comment: Why did the author not use and ANOVA or a statistical analysis that allow you to compare multiple treatments instead of comparing one-one each treatment?
Response: We used one-way ANOVA (fit y by x) followed by Tukey’s HSD for the majority of the analyses. The Data Analysis text has been corrected to reflect this.
Comment: Results: This section is hard to follow as it is shown. Species and ecotypes are sometimes referred as two letters (For instance, AM) and in other occasion it use the full name. It does not help to read it fluently. Each section is divided in two and I think that it is not needed.
Response: the text has been edited so that the two-letter identifier is used through this section. With respect to dividing each result into two parts (within ecotype and between ecotypes), the authors deliberately chose this style to separate the different analyses and avoid confusion.
Comment: Figures must be changed. They are not intuitive and the abbreviation of treatment should be replaced by salinity and N concentrations. There are plenty of papers in the literature where multiple combinations of treatments are showed clearer than this. In addition, letters used to represent significant differences are also hard to interpret.
Response: The Figures have been significantly altered. Now, each species/ecotype is represented by a different shaded bar and each treatment is separated by a dashed line. Comparisons within each treatment are still in lower case but now in red. The legends have been altered to reflect this.
Comment: Results are too specific, like a description of the figures; it would be more helpful if they just summarize the global changes that treatments showed pointing specific values that are interesting for the discussion.
Response: This section has been re-edited. The average paragraph length for each result is two to three sentences. Often it is only a single sentence. This section was written in accordance with the journal instructions: ‘concise and precise description of the experimental results’.
Comment: In Table 2: Why did not show the harvest for all ecotypes and species?
Response: Only two contrasting species were tested in the greenhouse, SB and VM. This is mentioned in the text.
Comment: Discussion: It is too descriptive. The results are again displayed in a more summarized way and it can make the manuscript repetitive. The discussion is poor in general and it sometimes shows a lack of connection between the parameters measures.
Response: This section has been significantly re-written to take account of the reviewer’s comments, i.e. removal of directly mentioned results, as well as description of the methods.
Comment: Line 420: “with those of [6] at a similar salt level” [6] must show the surname of the authors.
Response: This has been corrected to show the surnames.
Author Response File: Author Response.pdf
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.
