Jatropha curcas Seed Germination: Effect of Seed Imbibition, Aging, Storage, and Salinity

Round 1

Reviewer 1 Report (Previous Reviewer 3)

Comments and Suggestions for Authors

After revision, this manuscript has improved. However, I strongly recommend that the authors carefully examine the data described in the results. Such as in lines 343-344, the r = -0.72, but in figure 2A, the data showed r2 = 0.721.

Author Response

Dear reviewer, the authors thank you for your valuable time and inform you that the text is correct, as is the corresponding figure. However, the negative sign was omitted in the figure. We apologize for the lapse and ask that you consider the new corrected version.

Reviewer 2 Report (Previous Reviewer 4)

Comments and Suggestions for Authors

The authors have done an excellent job in revising and shortening the manuscript. The writing quality has greatly improved. I know it was difficult to delete the ontogeny data set, but the manuscript has been improved by focusing on the remaining large data set. 

When authors change journals, the authors carry 100% of the responsibility in re-formatting the manuscript. In the future please try to do the appropriate re-formatting to adhere to the new journal's rules. Concerning the lack of "–" on your keyboard, it is easy to copy this from the MDPI manuscript template then paste it into your manuscript. 

I still believe this manuscript is much too long and more complicated than necessary. This is a straightforward study and there is no need for 100+ citations and 40 citations in the Introduction. I suggest minor revision to shorten the manuscript, focusing on reducing the length of the Introduction and Discussion sections by 20% to 30%. If the editors do not feel this is necessary, then I am ok with acceptance. 

Full disclosure, I am not qualified to review Section 3.6. 

Author Response

In the attachment

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.

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Comments to the manuscript by Pompelli et al.

 

This manuscript explores key aspects of Jatropha curcas seeds, including their ontogeny, water permeability, germination, and the effects of imbibition, aging, storage, and salinity. It thoroughly analyzes factors influencing seed traits of J. curcas, with well-structured data presentation with clear tables and figures. The study is relevant to biofuel production, offering valuable insights into optimizing seed storage and germination to support sustainable energy and agricultural practices.

Below are general comments on the manuscript:

1-     The language is clear and acceptable. However, little sections, such as anatomical descriptions, can be improved.

2-     The manuscript includes overly detailed sections, such as the anatomical descriptions of J. curcas that can have summarized.

3-     The discussion needs additional citations (references) to strengthen the scientific context and comprehensively compare results.

4-     In the abstract, the authors mention the economic feasibility of the storage method. It would also be helpful to explain this aspect in the discussion.

5-     In the introduction, adding morphological traits such as the plant size, leaf characteristics, and life form of J. curcas is very important.

 

6-     To enhance understanding, some figures and tables must be better integrated into the text, with clear explanations in the manuscript.

Author Response

see attached file

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The research objectives outlined in this paper exhibit clear directions and possess significant practical implications within the fields of environment and energy. The investigation into the quality retention, water permeability, germination ability, and salt stress response of Jatropha curcas seeds under storage conditions demonstrates a degree of originality and offers novel insights and data for this area of study. Notably, the experiment involving the controlled storage of seeds through temperature and humidity regulation enhances our understanding of effective seed quality preservation in practical production settings, thereby contributing valuable support to production practices. However, there are areas that could benefit from further development: the paper lacks a quantitative comparison of specific seed quality parameters (such as oil content) and germination efficiency, both of which are critical to seed development and storage. It is advisable to incorporate additional experimental data to more directly assess the influence of storage conditions on biodiesel yield potential. Additionally, the research findings regarding seed germination under salt stress warrant further elaboration. While the germination performance of seeds exposed to varying concentrations of NaCl is addressed, the physiological changes occurring in seeds under salt stress have not been thoroughly investigated. It is recommended to include measurements such as internal ion content, variations in osmotic regulating substances, and the expression of seed resistance genes under salt stress to provide a more comprehensive evaluation of the impact of salt stress on seeds. It is recommended that the authors include practical application suggestions in the conclusion section. For instance, they could propose the development of new seed treatment protocols informed by the research findings, as well as the integration of other bioengineering methods to enhance the production efficiency of biodiesel crops.

Author Response

see attached letter

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

1. In the Abstract part, it is recommended to simplify the background description to highlight the methods, results, and conclusions.

2. The Introduction part is too long. I suggest the author reprepare this part as 4 or 5 paragraphs to describe the importance of J. curcas for biodiesel production, point out the shortcomings or limitations of existing research on seed storage and aging, provide popular seed storage methods, clarify the scientific questions of this study, and highlight its innovation.

3. This study evaluated the salt tolerance of J. curcas seeds, but the correlation between the research topic and salt tolerance evaluation is weak. The necessity of salt tolerance evaluation should be fully established in the Introduction part to make the salt tolerance experiment more logical and scientifically significant.

4. In the Results part, there are multiple obvious errors in the description of the trends and significance of germination rate and mean germination time (MGT), and the mechanism analysis behind the data should be appropriately deepened. Suggest systematizing the structure of the results section to ensure natural transitions between each part of the results.

5. The discussion part of this study is relatively complete and detailed, but the depth of the discussion should be increased to highlight the practical application background and significance of the results.

6. In the Methods part, on page 30, lines 1061-1062, how about the biological replicate design for SFW and SDW analysis? On page 31, line 1107, formulas for seed germination rate and MGT calculation should be provided.

7. On page 32, line 1167, it is mentioned that all experiments in this study were conducted using five replicates, but this is not the case. For example, on page 31, line 1098, it is explicitly mentioned that four replicates were used.

8. In the conclusions and future perspectives part, the main conclusions should be highlighted and the order of paragraphs should be adjusted. I don't understand why the conclusion section first describes the effect of salt stress on seed germination.

Comments on the Quality of English Language

This article studied the viability and physiological characteristics of long-term storage of Jatropha curcas seeds at 4℃, and briefly assessed the salt tolerance of the seeds, providing a useful reference for seed storage management in biodiesel production. However, this article is too lengthy, making it difficult for readers to grasp the key points, and some issues also need to be addressed.

Author Response

see attached letter

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

This is a very long manuscript based on a very large data set. The experimental protocols are detailed and the number of seeds that were exploited was impressive. The work by these authors can certainly be packaged in a manner that is publishable. However, in my opinion this manuscript is poorly organized, the methods are not consistent among the experiments, the statistical protocols are not explained, and there are too many experimental issues for a single manuscript. The manuscript is very confusing throughout.

The manuscript needs to be rewritten by a native English speaker. There are sentences throughout that say nothing of value. Just general information that does not really belong in a science article. For example lines 34-36. This sentence makes no sense.

For citations within the text, the dash should not be used in line 43 and throughout the manuscript. “1-3” should be replace with “1–3”.

Line 60 and every other time that “et al” is used, the style needs to be standardized. No comma. No italics.

The manuscript suffers from inconsistencies.

-For example, in lines 1063-1065 seed dry weight is referred to as SDW, Sdw, and Sdw. The abbreviation should be the same throughout the manuscript.

-A second example is the definition of germination, which is different among the experiments. For example in line 1074 germination is defined as when the seedling is 1 cm above the medium surface, line 1104 germination is when the radicle is 0.5 mm in length, in line 1165 germination is defined as when the seedling emerges from the surface of the medium. This is three different ways to quantify the same term in the methods. This is very confusing for the reader. Moreover, germination is not the same as emergence of the seedling from the soil surface. So use of the term germination for the studies in lines 1074 and 1165 is not accurate.

The greatest concern with the manuscript is the variety of issues that are studied. I believe the ontogeny data should be one manuscript and the storage and aging data should be a separate manuscript. This would remove much of the confusion and difficulty in absorbing the information.

Abstract

The Abstract needs to be completely rewritten. There is repetition in lines 23-26. The abstract must stand alone. The reader has no idea what the authors mean by controlled humidity, oil quality, or reduced metabolic activity or reduced deterioration. All of these terms must be explained in the abstract. The concluding remarks in lines 34-37 are very confusing. The conclusions mention storage but do not mention the ontogeny data.

Introduction

The Introduction needs to be completely rewritten. It is much too long and contains more citations than are needed. It reads like a literature review with a flow that is confusing. The Introduction should just introduce the subject matter, explain what is known, then explain why the experiments were conducted and how the new information will add to the subject matter. After reading the long paragraphs the reader is left with the feeling that the subject is well-understood. Then when reading lines 180-183 there is no way to evaluate why this will add anything to what is already known. The Introduction needs to be shortened and focused on a clear depiction of how the findings add to the subject. Entire paragraphs such as lines 55-105 should be moved to the Discussion. Some paragraphs are about storage of seeds, then a paragraph on salinity, then back to a paragraph on storage. These types of confusing flow of the subject matter just confuses the reader.

Materials and Methods

This section is after the Introduction section for Horticulturae. The Methods section should begin on line 184 as Section 2. These kinds of mistakes by the authors indicate a lack of concern for obeying the template that is provided.

The methods must be completely rewritten. There are many statements that do not make sense. The reader must be able to repeat the methods.

-Use of the terms 2020 and 2021 cause the reader to think that the same protocols were used for the two years. In fact, one experiment was conducted in 2021 with fresh seeds (defined as 2021) and one year old seeds (defined as 2020). Referring to the two batches of seeds should not use 2020 and 2021. A different term should be used such as fresh versus one-year-old seeds.

-In line 1073 the seeds are covered with 200g of sand. The area of the trays is never mentioned, so the reader cannot ascertain how deep the seeds are sown. Depth of sowing is what is important, not the weight of sand put in each tray.

-In lines 1125-1127 the seeds were selected according to size and physiological appearance. What does this mean? How can a reader repeat these methods that make no sense?

-In lines 1134-1151 the details of equipment are lacking. What is the model and manufacturer of each piece of equipment? What is the chamber size for the gas exchange protocols? These are mandatory details when pieces of equipment are used.

-In line 1157 there are five dose levels for the salinity study which range from 0 to 150. But in Figure 14 there are six dose levels which range from 0 to 100. These kinds of mistakes are indefensible. The reader is left with only confusion.

-Section 4.7 is incomplete. Example 1, nothing is mentioned about the PCA protocols, but Section 2.5.4 has an entire figure and text about the multivariate results. Example 2, the results shown in Figure 8 and the text associated with this figure indicate a factorial. The means of storage temperature are compared, but the means of storage time are also compared. This indicates a two-way factorial ANOVA was conducted. But Section 4.7 contains no mention of a factorial analysis. Example 3 pertains to the results shown in 2.5.3. The methods section has no information of any type to explain the methods used to generate Figure 12. There is no way for the reader to understand what statistical approaches were used. Therefore, there is no way for the reader to repeat the study.

Results

There are too many general introduction types of content in the Results subsections before the text begins to convey the findings. For example, lines 186-188, lines 285-287, lines 375-378, lines 458-464, lines 501-507, lines 611-617. The Results section should just state the findings, it should not have general introductory statements about what will be conveyed. There are also too many summary types of content in the Results subsections after the findings have been conveyed. For example, lines 360-367, lines 450-456, lines 482-498, lines 528-534, lines 646-668. These types of general interpretation statements belong in the Discussion section.

The flow of the Results section is very confusing.

-Figure 4 and Table 1 are cited in section 2.1. But Figure 4 and Table 1 appear much later after section 2.2.2.

- Figure 7 is cited in section 2.3.1. But it does not appear until section 2.3.3.

-Line 193. How can fruit weight be measured in mm?

-Line 196. The color “citrus” is not really a color. This needs to be defined.

-Line 218. The term “funicle” is not a normal botany term. Please define or use a different term.

-Lines 269-270. This is not a result. Delete.

-Line 294 and throughout. The minus sign “−“ needs to be used rather than a dash.

-Line 359. Figure 5 caption needs to be reworded. Panels A and B are not seed traits, they are solution characteristics.

-Figures 6, 9, 10, and 14. The SE values are shown, letters to indicate means separation are used, and regressions are shown. This is three ways of comparing the treatment means. This is redundant. If pairwise means separation is used then regressions should not be used. If regressions are used then means separation should not be used. Also, the methods section does not mention how the regressions were calculated. Also, some of the regressions do not look like they would be significant. The equation and r-square values are shown but the P values are not shown for the regressions. For example, I do not believe the line in Figure 6B is significant.

-Line 417. The reader has no idea what “staggered germination” means.

-Figure 10 and 11. What does the “MS” that is in some of the units? I have never seen this before.

-Section 2.5.3 is pure speculation. It does not belong in a Results section.

Discussion

The entire Discussion section should be deleted and rewritten. The current format is (1) detailed literature reviews on the subject, (2) general statements about the subject being important, and (3) repeated some of the results. There is no attempt to interpret the results. There is no attempt to explain how the new findings expand on what is already known.

Literature cited

Line 1232 and every reference thereafter. The dash is not appropriate for page ranges. Please replace with “–“.

Comments on the Quality of English Language

The general writing style is very difficult to understand. It is repetitive and the flow of the sequential paragraphs appears to have not been considered.

Author Response

see attached letter

Author Response File: Author Response.pdf