Maternal Environment and Priming Agents Effect Germination and Seedling Quality in Pitaya under Salt Stress

Round 1

Reviewer 1 Report (Previous Reviewer 2)

Comments and Suggestions for Authors

Dear Authors,

The present study evaluates the effect of salt stress on germination and emergence of pitaya seeds, priming with plant growth regulators, such as salicylic acid (SA), oxalic acid (OA) and mepiquat chloride (MC). The research subject is interesting and brings scientific important data in the field. Some changes of the manuscript should nevertheless be performed in order to improve its quality. Following specific changes should thus be performed:

Major changes

Abstract: The species are not correctly named: they need to have paternity the first time they appear in text (here and in Introduction). Please do not use expressions such as “It is thought…”.

Introduction: Similarly, species need to have paternity. You need to provide more information on the species: botanical data, traditional, culinary or industrial uses. 

All these suggested changes should be performed in order to bring further improvements to the manuscript.

Comments on the Quality of English Language

English language needs some moderate corrections.

Author Response

Suggested corrections were completed,

Kind regards.

Reviewer 2 Report (New Reviewer)

Comments and Suggestions for Authors

The authors tested the effect of salt stress on the seeds of two pitaya species with different flesh colour. Moreover, they evaluated the potential of three plant growth regulators to ameliorate the negative effects of salt stress and possibly increase important seed germination traits, including germination percentage, and seedling fresh and dry mass. The manuscript provides a sound hypothesis leading to logical scopes and objectives. Specifically, the introduction is informative and sufficiently explains the scope of the research. The experiment is well established and described, but a few clarifications must be made. Moreover, clarifications and amendments must also be made in the results and especially in all the Tables and Figure 1. The discussion and conclusions are appropriate. Overall, the manuscript has good quality and fit the scope of Horticulturae.

 

Specific comments are following.

L10. The abstract must stand-alone. An introductory paragraph should also be included before the objective.

L99. Please mention where did you obtain the fruits from.

L108. How did you choose the concentration of 150 ppm?

L117. It is important to also mention the electrical conductivity of the salt-treated water.

L119. Did you also need to re-fill with water?

L120-130. Maybe this part belongs to the following sub-sector 2.4. Please merge these parts.

L178. In Table 1, some values seem to be very close, but are significantly different. Especially in many cases the standard error is very high. For example, seedling dry matter ratio in red and white flesh colour. Also, the characteristics where PGRs show significant differences (MGT, fresh weight, GSTI). Please explain how this occurred. Are there possible errors in your analysis or presentation of the results?

L198. In Table 2, many letters of significant differences are not correct. For example, in germination %, MC in the means of flesh colour has A in 0, 2500, and 5000, but AB in 10000 ppm. In OA, only A are shown. Moreover, you should specify that the salt concentrations are measured in ppm. Please check and correct all the mistakes. The same applies for Table 3 as well.

L240. Please include letters of significant differences in Figure 1. It is difficult to read the caption in order to identify the differences among the treatments.

Comments on the Quality of English Language

Minor editing of English language required

Author Response

Suggested corrections were completed,

Kind regards.

Author Response File: Author Response.pdf

Reviewer 3 Report (New Reviewer)

Comments and Suggestions for Authors

Dear authors,

Overall, the manuscript is ok, but some points need improvement.

1- In the introduction, the last paragraph could be replaced by the formulation of a scientific hypothesis. That would be more appropriate.

2- The graphics are very dirty, remove the bottom lines.

3- All excerpts that call the figures are inappropriate. Start describing what was done, then say: it is presented in figure/table x> Please enter the data, describe it and finish with (Figure/table x).

4- In the discussion, a paragraph could be added about the physiological mechanisms affected by salinity. And how treatments can mitigate their effects.

5- In conclusion, put in the present and try to make a better parallel with the objectives

Comments for author File: Comments.pdf

Author Response

Suggested corrections were completed,

Kind regards.

Author Response File: Author Response.pdf

Reviewer 4 Report (New Reviewer)

Comments and Suggestions for Authors

In this manuscript, authors studied the effect of salt stress on germination and emergence of pitaya seeds obtained from Hyloceresu undatus and Hylocereus polyrhizus, priming with plant growth regulators, namely salicylic acid (SA), oxalic acid (OA) and mepiquat chloride (MC). However, the mechanism of effects of parent environment and initiator on germination and seedling quality of Pitaya under salt stress is unclear. More related experiments should be conducted. Moreover, the results do not make logical sense. All of these have to be confirmed one by one. My major comments are in the following:

1. In the Abstract, the study background should be introduced in brief. Additionally, the effects of plant growth regulators on seed germination under salt stress should be summarized. Authors are suggested to add these parts in the abstract.

2. In the Introduction, authors should provide more information about plant growth regulators, especially salicylic acid (SA), oxalic acid (OA) or mepiquat chloride (MC). In other words, the function and the application of SA, OA and MC should be introduced more clearly. Furthermore, in the last paragraph, authors explained the innovation of this study. However, the effects of growth regulators on seed germination in other plant species are studied widely. Authors simply switched species for the study. Therefore, the innovation of this study is not high enough.

3. In Materials and Methods, some important information was missing in 2.1. For example, the origin (brought or cultivated from some place) of different pitaya fruits needs to be added in 2.1. Are the fruits which authors selected are all fresh? Do authors have any requirements of selecting fruit size? Authors should explain these points clearly.

4. Table 1 shows the distribution of germination and emergence performances according to factors. However, some points is confusing. Authors are suggested to determine the germination related parameters with different plant growth regulators treatments and different concentration of NaCl to confirm the accuracy of ANOVA Significance levels. Moreover, in Table 2, the significance difference of MC, SA and OA under different concentration salt stress should be marked.

5. In Figure 1, the significant difference and the error bar was not shown. Additionally, each salt concentration should be shown in Figure 1.

6. In Results section, although authors determined some parameters, the evidence is not enough. More related experiments should be conducted. In addition, the results do not make logical sense. All of these have to be confirmed one by one.

7. In Discussion, authors mentioned many irrelevant content. More importantly, the explanation of your results is not shown, and the mechanism of this study is not revealed. Authors should compare your results with other similar research and give full and reasonable explanation.

Comments on the Quality of English Language

I think the English language is good and may be followed.

Author Response

Suggested corrections were completed,

Kind regards.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report (New Reviewer)

Comments and Suggestions for Authors

The authors sufficiently addressed my comments.

Therefore, I suggest the manuscript's publication in Horticulturae.

Thank you for giving me the opportunity to review it.

Author Response

Thanks one more for your valuable comments,

Kind regards.

Reviewer 4 Report (New Reviewer)

Comments and Suggestions for Authors

The English is good.

Comments on the Quality of English Language

In the abstract section, the background of the study should be included in brief.

Author Response

Suggested part was added.

Kind regards.

Author Response File: Author Response.docx

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

Comments and Suggestions for Authors

 

 

The manuscript under consideration holds significant thematic interest and relevance to both researchers and professionals in the field.

It primarily focuses on priming agents effect germination and seedling quality in pitaya under salt stress, potentially limiting its generalizability. To enhance its applicability, the authors are encouraged to include results from other varieties, verifying their investigating approach and experimental procedure.

Furthermore, the novelty of the work should be emphasized, particularly from an academic standpoint. Although the title accurately reflects the content, the introduction of other spicies would provide a more comprehensive understanding of its scope and limitations for potential future research.

The primary goal of the paper is to propose a precise and sensitive method for evaluation of the effect of salt stress on germination and emergence of pitaya seeds obtained from different fruit flesh colors, priming with plant growth regulators, namely salicylic acid (SA), oxalic acid (OA) and mepiquat chloride (MC). To strengthen the paper, a broader assessment of similar works beyond a local scale is necessary, with a focus on identifying their shortcomings.

In the 2.3. Experimental layout section, including a Data Flow Chart illustrating the steps from literature review to experimental analysis and conclusions would enhance readability.

The results and discussion demonstrate good correlation with cited literature data. However, updating the literature review with more recent references would enhance its quality. 

Regarding novelty, it should be more explicitly demonstrated.

How were the interchange terms of FC, SC and PGR were fitten into ANOVA calculation (Table 1)? They were qualitative (i.e. categorical) variables in this case?

In the "Conclusions" section, the main original scientific and research achievements should be highlighted, accompanied by a detailed plan for further research within the paper's subject.

Overall, the language and presentation are appropriate, and the research methodology is well-described, facilitating replication. The logical sequence of results and thorough analysis are commendable.

However, the paper's main drawback lies in its novelty compared to existing works. Demonstrating a substantial contribution to the research area is essential, either through further experimentation or a more comprehensive discussion of the manuscript's implications.

In conclusion, the proposed manuscript holds promise for publication in the Horticulturae journal, but it requires modifications and stronger demonstrations of novelty and contribution to the field.

 

 

 

 

Author Response

Corrections were made according to the suggestions,

Kind regards.

Author Response File: Author Response.docx

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

The present study evaluates the effect of salt stress on germination and emergence of pitaya seeds, priming with plant growth regulators. The research subject is interesting and brings scientific important data in the field. Some changes of the manuscript should nevertheless be performed in order to improve its quality. Following specific changes should thus be performed:

Major changes

Abstract: It is too long. Please shorten in order to respect the recommended 200 words in Instructions for Authors. This problem appears here for the first time, but it is valid also for the Introduction section: authors should start by introducting the species (both here and in Introduction) with its scientific name (including paternity), followed by common name. afterwards, throughout the whole manuscript, the species may be named either by using its common name, or by using its scientific abbreviation.

Introduction: It is not clear which species do you work on. Is it a single species? Is it all the Hylocereus genus? You need to provide more information on the species: botanical data, traditional, culinary or industrial uses. You present the different concepts in your study, but no similar study on the subject is presented. Authors need to clarify what their study brings in novelty, compared to these similar studies found in scientific literature. It is very important to state what exactly you bring in novelty in order to express your originality. The purpose of the study needs to be clearer in the last paragraph.

Materials and Methods: You do not offer references for all of the assays. Are all methods completely new? Because, if they are, you need to offer them a different approach.

Results and Discussions: Novelty and originality of your study is essential to be emphasized in this section once again. You need to emphasize this in terms of results, not purposes, as the Introduction should. I think that a separation between sections would be more adequate, as many results are presented, so a Discussion section appears as quite important for the manuscript. It would help for a better understanding of results. In fact, you only need to separate some of your comments to make this new section. I find it will bring improvement of the quality of your manuscript.

Conclusions: Perspectives should be added.

All these suggested changes should be performed in order to bring further improvements to the manuscript.

Comments on the Quality of English Language

English language needs some moderate corrections.

Author Response

Corrections were made according to the suggestions,

Kind regards.

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

This work aims at evaluating the effects of increased salt concentration and growth regulators on several germination parameters in two genotypes of Pitaya. The topic falls within the scope of the journal “Horticulturae”, the experiment was properly planned and conducted, and the results are interesting. Hence, I feel that the manuscript deserves to be published, provided some flaws are checked and corrected. First, I am not sure that the term “maternal”, in the title and elsewhere in the text (e.g., line 186, 237, etc.) fits the addressed meaning; I invite the Authors to check the opportunity to substitute it with some other term, such as “growing” or “germination”, according to the context. Second, I have some concerns about the statistical treatment and the presentation of results. I briefly list them in the following lines.

-        Before running an ANOVA procedure, the assumptions of normal distribution and variance homogeneity should be assessed. Since percentages based on counting (such as yours) meet very rarely these assumptions, the original data are usually transformed (cfr Steel and Torrie, “Principles and Procedures of Statistics”, but also every statistic manual), mostly by means of the square root transformation. Check this part and correct the text consequently.

-        The discussion of results should be addressed better. As shown in table 1, some of the studied variables (e.g., Germination % or Fresh seedling weight mg) experience some statistically significant interaction (SC x PGR, FC x SC, and so on). Hence, the mean effects of the corresponding main factors should be discussed with some caution, since a significant interaction effect means that the outcome of one causal variable varies based on the state of the second causal variable (and that is exactly what happens in your data). Under these circumstances, all significant interactions should be separately reported (as tables or figures) and discussed. Based on the results in table 1, the most proper data presentation should include SC x PGR and FC x SC for the germination %; FC x SC for the MGT; FC x SC, FC x PGR, SC x PGR, FC x SC x PGR for the Fresh seedling weight, and so on. Actually, the data reported in tables 2 and 3 are referred to the FC x SR x PGR interactions, that resulted non-significant for germination % and MGT; you are allowed to report all your interactions in this form even though they are not significant, but the results of statistical analysis should be clearly reported in each table (and discussed consequently).

-        It is not clear why you choose to present all data in tabular form, and only GSTI as histograms.

-        At Table 1, section “Plant Growth Regulators (PGRs)”: the indication “ns” should be reported as a separate line at the bottom of this part of the table; in the present form, it seems referred only to the MC value.

-        In all tables, the reported mean values should be accompanied with some variability index (standard deviation or standard error), even when the results of LSD (or other post-hoc tests) are reported.

-        Lines 190-194: this sentence (as well as the sentence at lines 230-231) seems suggesting that the tested genotypes (white- and red- flesh) belong to different species, rather than being different cultivars of the same species; if this is true, it should be specified in the M&M section (and some information should be better given also in the introduction). If not, please rephrase.

Line 221: proportional to what?

Author Response

Corrections were made according to the suggestions,

Kind regards.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors manage to improve the quality of the paper.

Author Response

Once again we are grateful for the valuable contributions.

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

The present study evaluates the effect of salt stress on germination and emergence of pitaya seeds, priming with plant growth regulators. The authors performed some of the suggested changes after the first round of review. However, they are not highlighted, for easy following. However, following specific changes should still be performed:

Major changes

Abstract: It is still too long. Please shorten in order to respect the recommended 200 words in Instructions for Authors. The concerned species is not mentioned. In order to respect limitations of words, there are phrases that are not needed and may be replaced in order to respect the requested structure and number of words. I did not see any changes in this part.

Introduction: I did not find the presentation of similar studies existing in scientific literature. Authors need to clarify what their study brings in novelty, compared to these similar studies found in scientific literature.

Results and Discussions: I did not find the suggested changes performed in this part. Maybe it’s because they are not highlighted. At the same time, I did not receive any explanation for changes that are not performed so I will be stating again the same comments for this part. Novelty and originality of your study is essential to be emphasized in this section once again. You need to emphasize this in terms of results, not purposes, as the Introduction should. I think that a separation between sections would be more adequate, as many results are presented, so a Discussion section appears as quite important for the manuscript. It would help for a better understanding of results. In fact, you only need to separate some of your comments to make this new section. I find it will bring improvement of the quality of your manuscript.

All these suggested changes should be performed in order to bring further improvements to the manuscript. 

Comments on the Quality of English Language

Moderate language corrections should be performed.

Author Response

Corrections were made according to the suggestions,

Kind regards.

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

I went through the revised version of this manuscript. The Authors made efforts to amend their work according to the reviewers’ indications; however, two observations of mine have been ruled out, and in my opinion some other amendments are necessary to make this work ready for publication. I list my observations in the following lines.

Line 31: “their” instead of “its”

Line 38: “organoleptic” is not a purpose

Line 61: check the opportunity to insert “In developing plants…” (Seeds have no stomas neither do photosynthesis)

Lines 89-90: check the sentence (a verb is missing)

Line 96: remove “belong …were harvested”

Line 104: Write “Petri” with capital initial.

Line 113: Remove “one”; do you mean “for each tested salt solution”?

Line 133: “with the following formula:…” ; insert formulae in separate lines

Line 135: insert the verb: “Germination stress tolerance index (GSTI, %) was obtained as follows: GSTI = …

Line 201: “undatus” with small initial

Table 1: I invite you again to modify the significance indications in the section “Plant Growth Regulators (PGRs)”: in the present form, the indication “ns” seems referred only to the MC value. Since the significance levels (*, **…) are reported in the bottom part of the table, you can remove the “ns” indication from the columns Germination, Shoot length, and Seedling dry matter ratio; also move the note “ns: not significant” to the lowest line, together with the other descriptions (*,  **, ***: means…). Furthermore, it is not clear why some values are written in bold.

Likewise, I also repeat my recommendation to insert, in all tables, the respective variability index (standard deviation or standard error).

Table 2 and table 3: indicate what is the meaning of the letters in the body of the tables.

Line 296: this is the first time you use the abbreviation SDMR; it should be better writing it in the extended form.

Figure 2: report in the caption of the figure, or as a footnote, the meaning of the abbreviations used in rows and columns.

Author Response

Corrections were made according to the suggestions,

Kind regards.

Author Response File: Author Response.docx