Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Manuscript titled “Mechanism of Intermittent Hypobaric Affecting Post-harvest Quality of Cassava Roots: An Integrated Analysis Based on Respiration, Energy Metabolism, and Transcriptomics” reports a series of experiments aimed at determining the effects of hypobaric storage to preserve cassava roots, which deteriorate rapidly under atmospheric storage. There are some comments and suggestions for the authors:

 

1. The introduction mentions the use of intermittent hypobaric storage, however, a clear definition is not provided. Can you please provide a brief description of the general strategy used for IHS?
2. “notable progress” is mentioned in line 49, can you please provide specific data about the benefits of utilizing IHS on cassava? For example, on its shelf-life, quality parameters, nutritional composition, etc., additional information about this method may better support your study.
3. In section 2.1, can you please provide additional information about your experimental setup? For example, the specific vacuum pump used, the volume of the container, its material (plastic, glass, etc.), number of samples stored per container, etc. If you have images to illustrate this process, please consider including them if possible.
4. Also in section 2.1, since your document’s title mentions an “intermittent” treatment, it could be assumed that the samples are periodically stored at low and atmospheric pressure, however, this appears to not be the case since only a continuous low-pressure storage is specified. Thus, is there any information missing from this section, or can the treatment be simply referred to as “hypobaric” and omitting the term “intermittent”? If it is the latter, then then document’s title and main text should also be modified accordingly.
5. Section 2.2.3 refers the reader to various methods published elsewhere. Although it may not be necessary to repeat a method in great detail, mentioning at least its name and/or the apparatus used could be informative to the reader. For example, “crude fiber was quantified according to the XYZ method, using an XYZ apparatus/kit”. Please consider providing this basic information on this section.
6. In section 2.2.5, please specify the full names of the ADH and L-LDH enzymes, also, please specify if the activities are per gram of fresh or dry tissue.
7. Section 3.4.1 mentions results of a PCA analysis, however, this is not described in materials and methods; please include it there.
8. Lines 251 – 252 mention “The AS shows deficiencies in the gene expression of these core proteins, leading to abnormal ribosome assembly”. Since your work did not actually analyze ribosomal abnormalities, the phrasing should be modified in order to make it clear to the reader that this information is from other organisms, and not from the present document. As currently written, it can be understood that you analyzed cassava’s ribosomes and found these anomalies.
9. Related to the previous comment, figure 7 shows the ribosomes of Haloarculata marismortui and Thermus aquaticus, two unrelated organisms. These images could be separated from the heatmaps and shown in an independent figure, since they appear to be from other works; they could also be omitted and the original papers cited and discussed (these references also appear to be missing).

Author Response

Dear Reviewer 1,

We are sincerely glad to get your professional suggestions. We’ve detailed responses to each of them below.

 

Comment 1: The introduction mentions the use of intermittent hypobaric storage, however, a clear definition is not provided. Can you please provide a brief description of the general strategy used for IHS?

Response 1: Thank you for your comments. Intermittent Hypobaric Storage (IHS) means that every 12 hours, the storage conditions (including air pressure, temperature, humility and CO₂ concentration) are restored to room conditions to maintain the air balance in the chamber. The details description are shown in ‘2.1 Materials’.

 

Comment 2: "notable progress" is mentioned in line 49, can you please provide specific data about the benefits of utilizing IHS on cassava? For example, on its shelf-life, quality parameters, nutritional composition, etc., additional information about this method may better support your study.

Response 2: Agree. We sincerely appreciate your attention and valuable suggestions. Based on the previous research findings of our team, cassava treated with the IHS storage technology exhibited no significant PPD within 20 days of storage. Meanwhile, no statistically significant differences were observed in terms of its nutritional quality and appearance characteristics when compared with freshly cassava roots. These findings have been documented and published in relevant peer-reviewed papers, which tittle is: ‘The effect of postharvest water migration on metabolism of cassava root by hypobaric storage’. 

 

Comment 3: In section 2.1, can you please provide additional information about your experimental setup? For example, the specific vacuum pump used, the volume of the container, its material (plastic, glass, etc.), number of samples stored per container, etc. If you have images to illustrate this process, please consider including them if possible.

Response 3: We agree with this comment. For the present experiment, a traditional household aluminum rice storage chamber equipped with an exhaust function was employed, the picture is provided below (Fig. 1). The volume of samples inside the chamber generally does not exceed two-thirds of the total volume of the chamber. These have been modified in the corresponding text as required. Fig1. aluminum rice storage chamber is provided in the File of Fig 1.

Comment 4: Also in section 2.1, since your document's title mentions an "intermittent" treatment, it could be assumed that the samples are periodically stored at low and atmospheric pressure, however, this appears to not be the case since only a continuous low-pressure storage is specified. Thus, is there any information missing from this section, or can the treatment be simply referred to as "hypobaric" and omitting the term "intermittent"? If it is the latter, then then document's title and main text should also be modified accordingly.

Response 4: We agree with this comment. We will refined the methods and conditions regarding intermittent in 2.1 Materials and Methods. We hope to retain the term ‘intermittent' in order to distinguish it from continuous hypobaric conditions.

 

Comment 5: Section 2.2.3 refers the reader to various methods published elsewhere. Although it may not be necessary to repeat a method in great detail, mentioning at least its name and/or the apparatus used could be informative to the reader. For example, "crude fiber was quantified according to the XYZ method, using an XYZ apparatus/kit". Please consider providing this basic information on this section.

Response 5: We agree with this comment. The methods had been resolved in the main text of ‘2.2.1 Determination of Weight Loss and Moisture Content’.

 

Comment 6: In section 2.2.5, please specify the full names of the ADH and L-LDH enzymes, also, please specify if the activities are per gram of fresh or dry tissue.

Response 6: Thank you very much for your reminder. The full names of ‘ADH’, ‘L-LDH’ and ‘ATP’ are ‘Alcohol dehydrogenase’, ‘L-Lactic Dehydrogenase’ and ‘AdenosineTriphosphate’ respectively, which had been supplemented in ‘2.2.4’. Meanwhile, in the measure,we were used the fresh tissues.

 

Comment 7: Section 3.4.1 mentions results of a PCA analysis, however, this is not described in materials and methods; please include it there.

Response 7: We agree with this comment. PCA analysis had been provided in 2.3 Statistice.

 

Comment 8: Lines 251-252 mention "The AS shows deficiencies in the gene expression of these core proteins, leading to abnormal ribosome assembly". Since your work did not actually analyze ribosomal abnormalities, the phrasing should be modified in order to make it clear to the reader that this information is from other organisms, and not from the present document. As currently written, it can be understood that you analyzed cassava's ribosomes and found these anomalies.

Response 8: We agree with this comment. We have removed the statement about protein expression affecting ribosome structure.

 

Comment 9: Related to the previous comment, figure 7 shows the ribosomes of Haloarculata marismortui and Thermus aquaticus, two unrelated organisms. These images could be separated from the heatmaps and shown in an independent figure, since they appear to be from other works; they could also be omitted and the original papers cited and discussed (these references also appear to be missing).

Response 9: We agree with this comment. In Figure 7, the images of the ribosome structure have been removed. The new composite figure contains only heatmap information, describing the genes that are significantly different in the ribosome pathway.

Thank you for taking the time to read our manuscript and for providing systematic guidance and efficient assistance. If you have any questions, please let’s know.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This manuscript investigates the effects of Intermittent Hypobaric Storage (IHS) on postharvest quality of cassava roots, with particular emphasis on respiratory physiology, energy metabolism, and transcriptomic regulation. The topic is relevant and potentially valuable for improving cassava postharvest management. However, the manuscript requires substantial revision before it can be considered for publication. Major concerns relate to insufficient methodological details, unclear descriptions of IHS treatment, and widespread issues with writing quality (grammar, spelling, and terminology).

Below are my specific comments and suggestions:

General Comments

1. The description of the intermittent hypobaric treatment is insufficient. Critical parameters (duration at the pressure level, frequency, recovery conditions, and total treatment time) must be clearly and quantitatively described to ensure reproducibility.
2. The manuscript contains numerous grammatical errors, misspellings, awkward sentence structures, and inconsistent terminology, which substantially affect readability. A thorough language revision by a fluent English speaker is strongly recommended.
3. Terminology should be used consistently. For example, “postharvest” should be used instead of “post-harvest” throughout the manuscript.

Specific Comments

• Lines 33–34

Please rewrite the sentence:

“PPD significantly reduces the quality and limits the development of cassava’s economic value.”

The sentence is vague and awkwardly phrased.

• Lines 67–72 (Materials and Methods)

Please provide the following details:

- Average fresh weight (g) and size of individual cassava roots

- Number of roots per replicate and total sample size

- Mass of each replicate

• Line 68

All abbreviations should be spelled out at first mention, for example, “AS.” Using “Control” instead of “AS” may improve clarity.

• Lines 75–76

Please rewrite:

“The moisture content of cassava roots was evaluated on days across the Wang’s Method.”

This sentence is grammatically incorrect and unclear.

Similar revision is needed for Lines 78–79.

 

• Line 77

The term “weight rate” is not appropriate. Please use a standard term such as “weight loss” or “weight loss percentage.”

 

• Lines 81–83

Each analytical method should be briefly introduced, including its basic principle, without excessive procedural details.

 

• Lines 85–90 (Respiration Measurement)

Please clarify the headspace atmosphere conditions, especially for IHS-treated roots.

- How long were samples incubated before gas measurement?

- Was a closed or flow-through system used?

- How was CO₂ accumulation controlled?

It is difficult to assess respiration differences without knowing whether CO₂ reached levels high enough to affect root metabolism.

 

• Figure 2

Please specify:

- Storage conditions for the control group

- Whether roots were covered or exposed

- Relative humidity during storage

 

• Lines 328–330

Please rewrite this section to improve clarity and grammatical correctness.

 

• Lines 332–335

Please rewrite this section; the current sentence structure is confusing and difficult to follow.

 

• Line 336

The term “Patents” appears to be inappropriate or misplaced in this context. Please clarify or remove.

Author Response

Dear Reviewer,

We are very honored to receive your professional advice. Regarding your suggestions and questions, we have provided answers to each one below.

General Comments 1: The description of the intermittent hypobaric treatment is insufficient. Critical parameters (duration at the pressure level, frequency, recovery conditions, and total treatment time) must be clearly and quantitatively described to ensure reproducibility.

Response: Agree. We sincerely appreciate your attention and valuable suggestions. The description of intermittent hypobaric condition has already been provided in the main text, located in section 2.1 Materials.

General Comments 2: The manuscript contains numerous grammatical errors, misspellings, awkward sentence structures, and inconsistent terminology, which substantially affect readability. A thorough language revision by a fluent English speaker is strongly recommended.

Response: We have carefully reviewed the entire text and revised any sentences, structures, or grammar that might cause misunderstandings, ensuring that the readability of the manuscript is not affected.

General Comments 3: Terminology should be used consistently. For example, "postharvest" should be used instead of "post-harvest" throughout the manuscript.

Response: We agree with this comment. These have been modified in the manuscript as required.

 

Specific Comments 1: Lines 33-34. Please rewrite the sentence "PPD significantly reduces the quality and limits the development of cassava's economic value". The sentence is vague and awkwardly phrased.

Response: Agree. Thank you for your comments. We have modified obscure expressions; the wordings are provided in the revised manuscript, highlighted in red font (Lines 32-34).

Specific Comments 2: Lines 67-72 (Materials and Methods). Please provide the following details: Average fresh weight (g) and size of individual cassava roots. Number of roots per replicate and total sample size. Mass of each replicate

Response: We agree with this comment. In 2. Materials and Methods, we provided the average sample weight of individual cassava roots, including the total sample size (Lines 65-66).

Specific Comments 3: Line 68.All abbreviations should be spelled out at first mention, for example, "AS." Using "Control" instead of "AS" may improve clarity.

Response: We agree with this comment. These have been modified in the manuscript as required.

Specific Comments 4: Lines 75-76. Please rewrite:"The moisture content of cassava roots was evaluated on days across the Wang's Method."This sentence is grammatically incorrect and unclear. Similar revision is needed for Lines 78-79.

Response: We agree with this comment. The description of the methods for moisture content and water loss rate has been revised (Lines 77-84).

Specific Comments 5: Line 77. The term "weight rate" is not appropriate. Please use a standard term such as "weight loss" or "weight loss percentage."

Response: We agree with this comment. We have provided the term ‘weight loss' to replace the inappropriate term ‘weight rate' (Lines 80).

Specific Comments 6: Lines 81-83. Each analytical method should be briefly introduced, including its basic principle, without excessive procedural details.

Response: Agree. We have streamlined the description of procedural details in 2.3 Statistics (Lines 134-140).

Specific Comments 7: Lines 85-90 (Respiration Measurement). Please clarify the headspace atmosphere conditions, especially for IHS-treated roots. How long were samples incubated before gas measurement? Was a closed or flow-through system used? How was CO₂ accumulation controlled? It is difficult to assess respiration differences without knowing whether CO₂ reached levels high enough to affect root metabolism.

Response: We agree with this comment. Breathing detection is conducted in a sealed chamber. After the vacuum pump reduces the chamber pressure to 0.05 MPa, the air detector readings are taken once the values stabilize for about 10 minutes, then read again after an hour. Multiple measurements are averaged to obtain accurate air detection results. The description of method has already been provided in the main text, located in section 2.2.3 Respiration Rate.

Specific Comments 8: Figure 2. Please specify: Storage conditions for the control group. Whether roots were covered or exposed. Relative humidity during storage.

Response: We agree with this comment. The storage activities of the control were carried out in the chambers under atmosphere conditions. The description of relative humidity has been added in Section 2.1 Materials (Lines 70-72). These have been modified in the corresponding text as required.

Specific Comments 9: Lines 328-330.Please rewrite this section to improve clarity and grammatical correctness.

Response: We agree with this comment. The conclusion section has been rewritten using a modified language expression (Lines 331-335).

Specific Comments 10: Lines 332-335. Please rewrite this section; the current sentence structure is confusing and difficult to follow.

Response: We agree with this comment. The conclusion section has been rewritten using a modified language expression (Lines 337).

Specific Comments 11: Line 336. The term "Patents" appears to be inappropriate or misplaced in this context. Please clarify or remove.

Response: We agree with this comment. Due to a misunderstanding of the template format, the term 'Patents' appears to be inappropriate. We have removed this term.

Thank you for providing constructive feedback and clear guidance for our experiments and paper. If there are any remaining issues, we hope you will contact us promptly.

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

 

Overall Assessment:
The manuscript addresses an important topic related to postharvest preservation of cassava roots using Intermittent Hypobaric Storage (IHS). However, the writing quality and clarity of presentation require substantial improvement before the manuscript can be considered for publication. At present, the text is difficult to follow, and several methodological details are unclear. I strongly recommend a thorough revision to improve readability, organization, and precision.

Comments

1. Writing and Readability:
   The manuscript is hard to read due to awkward sentence structures and lack of flow. Please revise the entire text for clarity and conciseness. Consider professional language editing or assistance from a native English speaker.

2. Methodological Details (Lines 65–75):

   • The description of IHS procedures is incomplete. Please specify:
     • Duration of the “on” and “off” hypobaric cycles.
     • Storage conditions for the control roots: Were they kept open without packaging or covered?
     • Number of hypobaric chambers used and whether treatments were replicated across chambers.
   • These details are essential for reproducibility.

3. Terminology (Line 78):
   The phrase “at each samples point” is unclear. Please rephrase to something like “at each sampling point” or “at each time point.”

4. Materials and Methods Style (Lines 81–82):
   The current style repeatedly uses “xx was determined as described by [citation].” This forces readers to consult multiple references for basic principles. Instead, briefly describe the principle or method in the text, for example:
   “Weight loss was determined based on reduction in root mass during storage [citation].”
   This approach improves readability and transparency.

5. Figures and Data Presentation (Lines 145–147):
   Figures 1 and 2 do not include temperature data.

 

 

Author Response

Dear Reviewer,

We are very honored to receive your professional advice. Regarding your suggestions and questions, we have provided answers to each one below.

Comments 1: Writing and Readability: The manuscript is hard to read due to awkward sentence structures and lack of flow. Please revise the entire text for clarity and conciseness. Consider professional language editing or assistance from a native English speaker.

Response: Thank you for your professional comment. These have been modified in the manuscript as required.

Comments 2: Methodological Details (Lines 65–75):

The description of IHS procedures is incomplete. Please specify: Duration of the “on” and “off” hypobaric cycles. Storage conditions for the control roots: Were they kept open without packaging or covered? Number of hypobaric chambers used and whether treatments were replicated across chambers. These details are essential for reproducibility.

Response: A hypobaric cycle consists of 12 hours of 'on' and 0.5 hours of 'off,' with two hypobaric cycles making up one day. The entire hypobaric storage lasts 35 days. The control roots were kept in chambers without packaging or covered. We have previously demonstrated the stability of storage experiments through multiple tests. In this experiments, just two chambers were used in two storage groups. All of details will be provided in manuscript.(Line 65-75)

Comments 3: Terminology (Line 78):
The phrase “at each samples point” is unclear. Please rephrase to something like “at each sampling point” or “at each time point.”

Response: We agree with this comment. These have been modified in the manuscript as required (Line 78).

Comments 4: Materials and Methods Style (Lines 81–82):
The current style repeatedly uses “xx was determined as described by [citation].” This forces readers to consult multiple references for basic principles. Instead, briefly describe the principle or method in the text, for example: “Weight loss was determined based on reduction in root mass during storage [citation].”

Response: Thank you for your systematic comment. These have been modified in the manuscript as required (Line 79-105).

 

Comments 5: Figures and Data Presentation (Lines 145–147): Figures 1 and 2 do not include temperature data.

Response: We agree with this comment. These have been modified in the manuscript as required (Line 73-75, and supplement).

Thank you for taking the time to read our manuscript and for providing systematic guidance and efficient assistance. If there are any remaining issues, we hope you will contact us promptly.