Comparative Transcriptome Analysis Reveals the Mechanism by Ethylene and 1-Methylcyclopropene Treatment on the Vase Life of Cut Oncidium hybridum ‘Honey Angle’

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The article entitled "Comparative transcriptome analysis reveals the mechanism by ethylene and 1-methylcyclopropene (1-MCP) treatment on the vase life of cut Oncidium hybridum ‘Honey Angle’" deals with the investigation of the molecular mechanisms of this cultivar's cut flowers senescence, the identification of key regularity genes and the analysis of gene expression patterns, while 1-MCP was evaluated as an effective postharvest treatment for vase life extending.

It is distinguished by originality and novelty, since no other study about postharvest treatments for cut flowers of Oncidium hybridum was found in literature. The quality of presentation is high and is supported by suitable references, making it interesting and understandable to readers.

Only the following minor corrections are proposed:

1. Regarding the keywords, they are all found in the title as well. Since the maximum of 10 keywords hasn't been reached, the authors could also use some more general words, such as orchids, cut flower, postharvest handling etc.
2. Lines 80-84: These lines should be moved to results or/ and conclusions. However, the authors could write in the aims of this study that they were also to provide critical insights into the molecular Oncidium senescence and evaluate 1-MCP as an effective postharvest treatment for extending its vase life.
3. Lines 230-231:

- The phrase "(E) average percentage weight loss" doesn't fit to this figure and should be erased.

- A comma should be added after the description of each subfigure, meaning (A) vase life, (B)...etc.

- In line 231 "and" should be before (F)

4. Line 246: in the phrase "treated with of" the word "of" should be erased.
5. Lines 593-595: the letters from (A) to (H), that characterize each subfigure, should be added in the caption of figure 8 like that: (A) APX:...; (B) XZH:.... etc.
6. Line 680: Better write "Nine" instead of number 9 in the beginning of the sentence.
7. Line 986: "2015" should be in bold.

Comments for author File: Comments.pdf

Author Response

The changes and responses to reviewers’ comments are listed as below. The line (L) numbers refer to our revised manuscript submitted 3/18/2025.

Thank you for your review of our paper. We have answered each of your points below.

Response to Reviewer 1:

The article entitled "Comparative transcriptome analysis reveals the mechanism by ethylene and 1-methylcyclopropene (1-MCP) treatment on the vase life of cut Oncidium hybridum ‘Honey Angle’ deals with the investigation of the molecular mechanisms of this cultivar's cut flowers senescence, the identification of key regularity genes and the analysis of gene expression patterns, while 1-MCP was evaluated as an effective postharvest treatment for vase life extending.

It is distinguished by originality and novelty, since no other study about postharvest treatments for cut flowers of Oncidium hybridum was found in literature. The quality of presentation is high and is supported by suitable references, making it interesting and understandable to readers.

Only the following minor corrections are proposed:

 

Regarding the keywords, they are all found in the title as well. Since the maximum of 10 keywords hasn't been reached, the authors could also use some more general words, such as orchids, cut flower, postharvest handling etc.

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the manuscript. As you suggested, we have changed the keywords as: Orchids; cut flower; postharvest handling; Ethylene; 1-methylcyclopropene (1-MCP); Vase life; Transcriptome on Page 1, Line 30-31 of the revised manuscript.

 

Lines 80-84: These lines should be moved to results or/ and conclusions. However, the authors could write in the aims of this study that they were also to provide critical insights into the molecular Oncidium senescence and evaluate 1-MCP as an effective postharvest treatment for extending its vase life.

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the manuscript. As you suggested, we have rewritten in the ‘Introduction’ part on Page 2, Line 81-84 of the revised manuscript.

 

Lines 230-231:

- The phrase "(E) average percentage weight loss" doesn't fit to this figure and should be erased.

- A comma should be added after the description of each subfigure, meaning (A) vase life, (B)...etc.

- In line 231 "and" should be before (F)

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the figure captions. We have corrected on Page 5, Line 233-234 of the revised manuscript.

 

Line 246: in the phrase "treated with of" the word "of" should be erased.

• RESPONSE: Thank you for noting this. We have corrected on Page 5, Line 248 of the revised manuscript.

 

Lines 593-595: the letters from (A) to (H), that characterize each subfigure, should be added in the caption of figure 8 like that: (A) APX:...; (B) XZH:.... etc.

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the figure captions. We have corrected on Page 13, Line 606-609 of the revised manuscript.

 

Line 680: Better write "Nine" instead of number 9 in the beginning of the sentence.

• RESPONSE: Thank you for noting this. We have corrected on Page 15, Line 693 of the revised manuscript.

 

Line 986: "2015" should be in bold.

• RESPONSE: Thank you for noting this. We have corrected on Page 21, Line 998 of the revised manuscript.

 

Thanks very much to editor and reviewers for your precious suggestions!

 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This manuscript examined the role of ethylene in maintaining the viability of Oncidium hybridum cultivar 'Honey Angle' flowers in vases. It is a good and logical work with many methods. However, there are some questions and minor comments.

The introduction is well written, contains the necessary references, briefly but informatively describes the object of study, the role of ethylene and ethylene inhibitor in the processes under study.

The methods are described in detail, although there are some questions about them:
1) It is not entirely clear, was the treatment with ethylene and inhibitor carried out throughout the study? When various measurements were taken, how did the gas composition in the boxes change?
2) How were the plants weighed? Were the plants taken out of the vases to remove water? How could this affect the vital signs of the plants? If they were weighed in vases, how did they achieve the accuracy of the measurements?
3) A number of measurements required temporary removal of individual samples. Were they then returned to the remaining flowers? Could specific conditions (placing in a bottle or separate smaller boxes) affect the indicators of the samples?

The results and discussion section contains a large amount of information. All measurements presented form a complete picture of the processes under study. There are some minor technical comments on this section.
1) It is necessary to indicate in the caption to Figure 2 what the arrows and highlighted areas point to.
2) Figures 4-8 are impossible to see properly, and if you zoom in on the pictures, everything becomes blurry.
3) Error rates merge with diagrams (Figure 1), it would be better to make them in a different color. The statistical difference rates at the top of Figure 1C are unclear.
4) 3.12 (line 651) - one cannot talk about enzyme activity if the section is about genes.
5) It is not entirely clear why these genes were chosen for PCR validation (Figure S1).

The conclusion reflects the essence of the work. The list of references contains a sufficient number of references.

Author Response

The changes and responses to reviewers’ comments are listed as below. The line (L) numbers refer to our revised manuscript submitted 3/18/2025.

Thank you for your review of our paper. We have answered each of your points below.

Response to Reviewer 2:

This manuscript examined the role of ethylene in maintaining the viability of Oncidium hybridum cultivar 'Honey Angle' flowers in vases. It is a good and logical work with many methods. However, there are some questions and minor comments.

The introduction is well written, contains the necessary references, briefly but informatively describes the object of study, the role of ethylene and ethylene inhibitor in the processes under study.

The methods are described in detail, although there are some questions about them:

 

1) It is not entirely clear, was the treatment with ethylene and inhibitor carried out throughout the study? When various measurements were taken, how did the gas composition in the boxes change?

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the manuscript. In this study, cut flowers were subjected to a short-term treatment with ethylene and 1-MCP for 12 hours in an 80 L airtight plastic boxes. During the treatment, water retention cotton was placed at the ends of the cut flowers to prevent water loss and wilting. After treatment, the cut flowers were removed from the plastic boxes, and the water retention cotton was discarded. Three cut flowers were then placed in a vase containing 150 ml of tap water. During the postharvest measurements, the cut flowers had already been removed from the container, ensuring that the measurement of the indicators did not affect the gas composition inside the container. As you suggested, we have rewritten this part in the methods section on Page 3, Line 97-107 of the revised manuscript.

 

2) How were the plants weighed? Were the plants taken out of the vases to remove water? How could this affect the vital signs of the plants? If they were weighed in vases, how did they achieve the accuracy of the measurements?

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the manuscript. To weigh the cut flowers, we removed them from the vase and quickly dried the moisture on the stems with absorbent paper. Then promptly placed the flowers on the scale. The weighing process was completed within 1 minute, minimizing any impact on the state of the cut flowers. As you suggested, we have rewritten this part in the methods section on Page 3, Line 117-120 of the revised manuscript.

 

3) A number of measurements required temporary removal of individual samples. Were they then returned to the remaining flowers? Could specific conditions (placing in a bottle or separate smaller boxes) affect the indicators of the samples?

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the manuscript. Measurement indicators such as vase life, flowering ratio, petal brightness, and maximum petal diameter were directly measured in the vase without removing the cut flowers. Samples for odor measurement and RNA extraction were consumed after measurement and did not need to be returned to the vase. The main indicators requiring temporary removal of individual samples for measurement and subsequent return to the vase were the weight loss rate and respiration rate. The measurement of the weight loss rate was completed rapidly, typically within 1 minute, minimizing any impact on the cut flowers. For respiration rate measurements, the cut flowers were placed in a box for approximately one hour. Water retention cotton was wrapped around the ends of the cut flower stems to reduce water loss, wilting, and changes in other indicators. Preliminary experiments indicated that cut flowers typically exhibited petal wilting and other affected indicators when placed in a non-moisturized environment for 3-4 hours. One hour in the box with water retention cotton did not affect the physiological indicators of the cut flowers.

 

The results and discussion section contains a large amount of information. All measurements presented form a complete picture of the processes under study. There are some minor technical comments on this section.

1) It is necessary to indicate in the caption to Figure 2 what the arrows and highlighted areas point to.

• RESPONSE: Thank you for noting this. The blue marks indicated curled or wilted petals; the red circles showed areas where the tissues have faded and turned whitish. We have corrected on Page 6, Line 293-294 of the revised manuscript.

 

2) Figures 4-8 are impossible to see properly, and if you zoom in on the pictures, everything becomes blurry.

• RESPONSE: Thank you for noting this. We have addressed the issues with Figures 4-8 to ensure better visibility and clarity of the revised manuscript. In addition, we have also uploaded the original picture file as an attachment for Figures 4-8.

 

3) Error rates merge with diagrams (Figure 1), it would be better to make them in a different color. The statistical difference rates at the top of Figure 1C are unclear.

• RESPONSE: Thank you for noting this. We have corrected on Figure 1 of the revised manuscript.

 

4) 3.12 (line 651) - one cannot talk about enzyme activity if the section is about genes.

• RESPONSE: Thank you for noting this. We have corrected on Page 14, Line 664 of the revised manuscript.

 

5) It is not entirely clear why these genes were chosen for PCR validation (Figure S1).

The conclusion reflects the essence of the work. The list of references contains a sufficient number of references.

• RESPONSE: We sincerely appreciate your insightful comments and suggestions for revising the manuscript. After RNA transcriptome sequencing, PCR validation was conducted to avoid technical errors, biological sample variation, and data analysis discrepancies. The selected genes were primarily differentially expressed and functionally significant following the treatment of Oncidium cut flowers with ethylene and 1-MCP. Additionally, these genes exhibited relatively high expression levels and were believed to play a crucial role in regulating the extension of the postharvest vase life of Oncidium. They were identified as key targets for further investigation in subsequent studies.

Thanks very much to editor and reviewers for your precious suggestions!

 

Author Response File: Author Response.pdf