Responses of Aroma Related Metabolic Attributes of Opisthopappus longilobus Flowers to Environmental Changes
, Weili Hao, Shan He, Li Liu, Xiaolong Feng, Qiyang Qie, Min Chai * and Yiling Wang *Round 1
Reviewer 1 Report
The manuscript titled “Responses of aroma related metabolic attributes of Opisthopappus longilobus flowers to environmental changes” reports data regarding the difference in metabolites extracted from flowers of two Opisthopappus species: O. longilobus wild and transplanted flowers, and O. taihangensis wild flowers. The work is interesting and result are convincing, however, they have to be better described. English should be carefully revised.
Revisions:
Please, explain abbreviation the first time they appear (for example CLW, CLT, TH).
“KEGG enrichment analysis revealed that the differential metabolites were enriched
into 47 pathways in CLW/CLT, 59 metabolic pathways in TH/CLW, and 61 pathways in
TH/CLT. Meanwhile, the significantly different metabolic pathways were all involved in
secondary plant metabolite biosynthesis, phenylpropanoid biosynthesis, and arginine
biosynthesis between TH/CLT, TH/CLW, and CLW/CLT.”: the results with statistical parameters have to be shown.
Aromatic compound expression: which is the unit utilized?
Is the Variation coefficient expressed as %?
Data reported in the tables should not be repeated in the text. Therefore, sentences could result more readable. I.e.:
For CLW, the most highly expressed metabolite was phenyl acetate with a content of 148542.13, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate, with relative contents of 120830.08, 56960.99, 39622.58, and 26437.30, respectively. The least expressed metabolite was p-Cresol with a relative content of 514.12.
Became:
For CLW, the most highly expressed metabolite was phenyl acetate, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate. The least expressed metabolite was p-Cresol. Etc.
In the Network diagram, authors write: Most of the metabolites were relatively dispersed. The plot shows that most of the metabolites were well connected. Otherwise, please explain better.
Methods
Authors should better explain what reciprocal transplantation is. We expect that O. longilobus individuals from Xiangtang Mountains were transplanted to Shennong Mountains and vice versa. Otherwise, the transplantation is not reciprocal. Differently, the reader understand that plants were transplanted to a garden that is not the environment of the other species. Moreover, table A3 is not informative. It seems that CLW and CLT are from the same location. What does CL stand for? Why CLW1-3 are separated from CLW4? Please, also add a column with species name.
The Extraction of total metabolite method is not clear. It seems that collected samples were liquid. Please explain better.
Results and/or Table A2: Could authors report at least the other most important environmental factors considered?
Table 2. What does asterisk mean?
Figure 2 and Table 1: add the measure unit
Figure 3: substance names are not readable.
Figure A1: : PCA diagram is not readable
Figure 4 is too small.
Author Response
Dear Editor and reviewers，
We deeply appreciate the editor and reviewers taking your valuable time to review our manuscript entitled ‘Responses of aroma related metabolic attributes of Opisthopappus longilobus flowers to environmental changes (plants-2261211)’. Those comments are valuable and very helpful. We have read through comments carefully and have made corrections. Based on the instructions provided in your letter, we uploaded the file of the revised manuscript. The responses to the reviewer's comments are presented following:
Response to the reviewer 1’s comments
The manuscript titled “Responses of aroma related metabolic attributes of Opisthopappus longilobus flowers to environmental changes” reports data regarding the difference in metabolites extracted from flowers of two Opisthopappus species: O. longilobus wild and transplanted flowers, and O. taihangensis wild flowers. The work is interesting and result are convincing, however, they have to be better described. English should be carefully revised.
Answer: Thank you much. We carefully revised our manuscript based on your comments.
- Please, explain abbreviation the first time they appear (for example CLW, CLT, TH).
Answer: The abbreviation of CLW, CLT, TH was explained in the manuscript when first time presented. Thank you much.
- “KEGG enrichment analysis revealed that the differential metabolites were enriched
into 47 pathways in CLW/CLT, 59 metabolic pathways in TH/CLW, and 61 pathways in
TH/CLT. Meanwhile, the significantly different metabolic pathways were all involved in
secondary plant metabolite biosynthesis, phenylpropanoid biosynthesis, and arginine
biosynthesis between TH/CLT, TH/CLW, and CLW/CLT.”: the results with statistical parameters have to be shown.
Answer: Dear, according your comments, we added an affiliated table A1. In this table, the detailed information about the pathways and statistical parameters were displayed.
- Aromatic compound expression: which is the unit utilized?
Answer: Based on the analyzed methods of metabonomics, the expression of aromatic compounds is only a relative value which obtained by calculating the peak area of the characteristic ion for each substance. Consequently, there are no units for these metabolites’ expression.
Is the Variation coefficient expressed as %?
Answer: The relative sentences were reviewed, thank you.
- Data reported in the tables should not be repeated in the text. Therefore, sentences could result more readable. I.e.:
For CLW, the most highly expressed metabolite was phenyl acetate with a content of 148542.13, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate, with relative contents of 120830.08, 56960.99, 39622.58, and 26437.30, respectively. The least expressed metabolite was p-Cresol with a relative content of 514.12.
Became:
For CLW, the most highly expressed metabolite was phenyl acetate, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate. The least expressed metabolite was p-Cresol. Etc.
Answer: According your suggests, the sentences have been rewritten.
- In the Network diagram, authors write: Most of the metabolites were relatively dispersed. The plot shows that most of the metabolites were well connected. Otherwise, please explain better.
Answer: The sentences were carefully reviewed, thank you much.
- Methods
Authors should better explain what reciprocal transplantation is. We expect that O. longilobus individuals from Xiangtang Mountains were transplanted to Shennong Mountains and vice versa. Otherwise, the transplantation is not reciprocal. Differently, the reader understand that plants were transplanted to a garden that is not the environment of the other species.
Moreover, table A3 is not informative. It seems that CLW and CLT are from the same location. What does CL stand for? Why CLW1-3 are separated from CLW4? Please, also add a column with species name.
Answer: Thank your comments much. Yes, our transplantation was not reciprocal only transplanting. The accurate transplant site should be Xiangtangshan National Park of China. Generally, the environmental conditions are different between lands and mountains. As know, O. longilobus grows on the cliffs of Taihang Mountains. When transplanted into the Xiangtangshan National Park, it also faces to the changed surroundings, such as average temperature, maximum temperature, and average rainfall. And the detailed descriptions have been displayed in the section 4.1. In addition, the contents of table A3 was also reviewed.
References:
① Li, W.Q. Monitoring of vegetation change in mining area based on multi-source remote sensing data: a case study of Fengfeng mining area. Hebei University of Engineering 2021.
② Ye, H., et al. Localized environmental heterogeneity drives the population differentiation of two endangered and endemic Opisthopappus Shih species. BMC Ecology and Evolution 2021, 21, 56.
③ Liu, L., et al. Geographic distribution pattern and ecological niche differentiation of endangered Opisthopappus in Taihang Mountains. Brazilian Journal of Botany 2023, 46, 217-226.
7.The Extraction of total metabolite method is not clear. It seems that collected samples were liquid. Please explain better.
Answer: This section was rewrote, thank you.
- Results and/or Table A2: Could authors report at least the other most important environmental factors considered?
Answer: We reviewed the contents of the table and added the results of PCA.
Table 2. What does asterisk mean?
Answer: The note of asterisk was added, thank you.
Figure 2 and Table 1: add the measure unit
Answer: The relative parts were reviewed.
Figure 3: substance names are not readable.
Answer: Thank you, this figure was reviewed.
Figure A1: PCA diagram is not readable
Answer: Figure A1 was reedited.
Figure 4 is too small.
Answer: Figure 4 was reviewed, thank you.
Sincerely,
Zhixia Liu and Yiling Wang
Author Response File: 
Author Response.docx
Reviewer 2 Report
This manuscript reports the comparative metabolic analysis of Opisthopappus longilobus wild and transplant flowers, and O. taihangensis flowers. The authors found that the aromatic related compounds were significantly correlated with the lowest temperatures in sampled sites and some aromatic metabolites played important roles in response to environmental changes. The results reveal the potential response mechanisms of O. longilobus to different habitats via the synthesis and regulation of aromatic substances. In general, the present study is of value and importance to be published in Plants. However, there are several issues need to be addressed before publication.
Major comments.
Materials and Methods. “In 2016, we established a reciprocal transplant garden (the O. longilobus individuals from Xiangtang Mountains was transplanted to garden) in the foot of Xiangtang Mountains, where O. longilobus flowers were collected from July to October of 2020. Concurrently, O. taihangensis wild flowers were collected as comparative samples from the Shennong Mountains (TableA3).”
From the description above, the reciprocal transplant garden is at the foot of Xiangtang Mountains, and the O. longilobus individuals from Xiangtang Mountains was transplanted to the garden. Then the authors compared the O. longilobus from Xiangtang Mountains, O. longilobus from the garden, and O. taihangensis from Shennong Mountains.
What is the different climates between the Xiangtang Mountain and its foot? Since significant different climates between Xiangtang and Shennong, why did not transplant the O. longilobus to Shennong? Or transplant the O. taihangensis to the garden?From the aspect of different environmental climates (different habitats), it seems that the transplant between the two long-distance places (Xiangtang and Shennong) is called reciprocal transplant, rather than the mountain and its foot.
“O. taihangensis wild flowers were collected as comparative samples from the Shennong Mountains.” Here, O. taihangensis is different from O. longilobus at the species level. To compare the environmental factor effects, why did not use the same species O. longilobus under same genetic background to perform the reciprocal transplant between the two Mountains? Why used the two different species in two different places?
Why O. longilobus wild flower was named CLW, O. longilobus reciprocal transplant flower was named CLT, and O. taihangensis wild flower was named TH ?
Since the manuscript is about the flower fragrances and aromatic compounds, some recent reviews on flower fragrances should be referenced, such as Front Plant Sci. 2022; 13: 860157.
Author Response
Dear Editor and reviewers，
We deeply appreciate the editor and reviewers taking your valuable time to review our manuscript entitled ‘Responses of aroma related metabolic attributes of Opisthopappus longilobus flowers to environmental changes (plants-2261211)’. Those comments are valuable and very helpful. We have read through comments carefully and have made corrections. Based on the instructions provided in your letter, we uploaded the file of the revised manuscript. The responses to the reviewer's comments are presented following:
Response to the reviewer 2’s comments
This manuscript reports the comparative metabolic analysis of Opisthopappus longilobus wild and transplant flowers, and O. taihangensis flowers. The authors found that the aromatic related compounds were significantly correlated with the lowest temperatures in sampled sites and some aromatic metabolites played important roles in response to environmental changes. The results reveal the potential response mechanisms of O. longilobus to different habitats via the synthesis and regulation of aromatic substances. In general, the present study is of value and importance to be published in Plants. However, there are several issues need to be addressed before publication.
Answer: Thank you for your valuable comments. According your comments, we carefully revised our manuscript.
- Materials and Methods. “In 2016, we established a reciprocal transplant garden (the O. longilobus individuals from Xiangtang Mountains was transplanted to garden) in the foot of Xiangtang Mountains, where O. longilobus flowers were collected from July to October of 2020. Concurrently, O. taihangensis wild flowers were collected as comparative samples from the Shennong Mountains (TableA3).”
From the description above, the reciprocal transplant garden is at the foot of Xiangtang Mountains, and the O. longilobus individuals from Xiangtang Mountains was transplanted to the garden. Then the authors compared the O. longilobus from Xiangtang Mountains, O. longilobus from the garden, and O. taihangensis from Shennong Mountains.
What is the different climates between the Xiangtang Mountain and its foot? Since significant different climates between Xiangtang and Shennong, why did not transplant the O. longilobus to Shennong? Or transplant the O. taihangensis to the garden? From the aspect of different environmental climates (different habitats), it seems that the transplant between the two long-distance places (Xiangtang and Shennong) is called reciprocal transplant, rather than the mountain and its foot.
Answer: Thank you much for your comments. Generally, the environmental conditions are different between lands and mountains. O. longilobus grows on the cliffs of Taihang Mountains. Our transplant site accurately was in the Xiangtangshan National Park of China. This park locates in Fengfeng Mining District, with an annual dryness of 1.4 degrees. Maximum wind speed in the calendar year is 14m/s. Spring precipitation accounts for 12.7% of the region's annual precipitation (Li, 2021). The environmental conditions of the park, such as annual average temperature, extreme maximum temperature, and annual average rainfall, are different with the Xiangtang Mountains through both belonging to the same climatic zone (Ye et., 2021; Liu et al., 2023). Thus, when O. longilobus was transplanted into the Xiangtangshan National Park, it also faces to the changed surroundings. The detailed descriptions have been displayed in the section 4.1.
References:
① Li, W.Q. Monitoring of vegetation change in mining area based on multi-source remote sensing data: a case study of Fengfeng mining area. Hebei University of Engineering 2021.
② Ye, H., et al. Localized environmental heterogeneity drives the population differentiation of two endangered and endemic Opisthopappus Shih species. BMC Ecology and Evolution 2021, 21, 56.
③ Liu, L., et al. Geographic distribution pattern and ecological niche differentiation of endangered Opisthopappus in Taihang Mountains. Brazilian Journal of Botany 2023, 46, 217-226.
- “O. taihangensis wild flowers were collected as comparative samples from the Shennong Mountains.” Here, O. taihangensis is different from O. longilobus at the species level. To compare the environmental factor effects, why did not use the same species O. longilobus under same genetic background to perform the reciprocal transplant between the two Mountains? Why used the two different species in two different places?
Answer: Dear, we collected O. taihangensis wild flower as comparative samples for the following reasons:
① The wild flowers of O. longilobus were sampled in the same growing site based on such a consideration of the same background.
② The phylogenetic relationship among these two species, O. taihangensis was a descendant of O. longilobus.
③ In terms of geographical distribution, Xiangtang Mountains was a growing area only for O. longilobus, while Shennong Mountains only for O. taihangensis distributed. However, these two sties both belong to the center of Taihang Mountains, the environment of them is similar but different. Under this background, the differential metabolites more reflect in response to the environmental heterogeneity.
3. Why longilobus wild flower was named CLW, O. longilobus reciprocal transplant flower was named CLT, and O. taihangensis wild flower was named TH?
Answer: In this manuscript, CL and TH was from the name of Chinese phonetic transcriptions of O. longilobus and O. taihangensis respectively. Wild was abbreviated for W, while Transplant for T. For O. taihangensis, the flowers are only from wild that remarked with TH.
4.Since the manuscript is about the flower fragrances and aromatic compounds, some recent reviews on flower fragrances should be referenced, such as Front Plant Sci. 2022; 13: 860157
Answer: The literature has been cited in the paper, thank you much.
Sincerely,
Zhixia Liu and Yiling Wang
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
Revision #2
We deeply appreciate the editor and reviewers taking your valuable time to review our manuscript entitled ‘Responses of aroma related metabolic attributes of Opisthopappus longilobus flowers to environmental changes (plants-2261211)’. Those comments are valuable and very helpful. We have read through comments carefully and have made corrections. Based on the instructions provided in your letter, we uploaded the file of the revised manuscript. The responses to the reviewer's comments are presented following:
Response to the reviewer 1’s comments
The manuscript titled “Responses of aroma related metabolic attributes of Opisthopappus longilobus flowers to environmental changes” reports data regarding the difference in metabolites extracted from flowers of two Opisthopappus species: O. longilobus wild and transplanted flowers, and O. taihangensis wild flowers. The work is interesting and result are convincing, however, they have to be better described. English should be carefully revised.
Answer: Thank you much. We carefully revised our manuscript based on your comments.
Please, explain abbreviation the first time they appear (for example CLW, CLT, TH).
Answer: The abbreviation of CLW, CLT, TH was explained in the manuscript when first time presented. Thank you much.
“KEGG enrichment analysis revealed that the differential metabolites were enriched
into 47 pathways in CLW/CLT, 59 metabolic pathways in TH/CLW, and 61 pathways in
TH/CLT. Meanwhile, the significantly different metabolic pathways were all involved in
secondary plant metabolite biosynthesis, phenylpropanoid biosynthesis, and arginine
biosynthesis between TH/CLT, TH/CLW, and CLW/CLT.”: the results with statistical parameters have to be shown.
Answer: Dear, according your comments, we added an affiliated table A1. In this table, the detailed information about the pathways and statistical parameters were displayed.
OK
Aromatic compound expression: which is the unit utilized?
Answer: Based on the analyzed methods of metabonomics, the expression of aromatic compounds is only a relative value which obtained by calculating the peak area of the characteristic ion for each substance. Consequently, there are no units for these metabolites’ expression.
OK, where possible write “relative units”
Is the Variation coefficient expressed as %?
Answer: The relative sentences were reviewed, thank you.
OK
Data reported in the tables should not be repeated in the text. Therefore, sentences could result more readable. I.e.:
For CLW, the most highly expressed metabolite was phenyl acetate with a content of 148542.13, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate, with relative contents of 120830.08, 56960.99, 39622.58, and 26437.30, respectively. The least expressed metabolite was p-Cresol with a relative content of 514.12.
Should became:
For CLW, the most highly expressed metabolite was phenyl acetate, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate. The least expressed metabolite was p-Cresol. Etc.
Answer: According your suggests, the sentences have been rewritten.
NOT DONE
In the Network diagram, authors write: Most of the metabolites were relatively dispersed. The plot shows that most of the metabolites were well connected. Otherwise, please explain better.
Answer: The sentences were carefully reviewed, thank you much.
DONE
Methods
Authors should better explain what reciprocal transplantation is. We expect that O. longilobus individuals from Xiangtang Mountains were transplanted to Shennong Mountains and vice versa. Otherwise, the transplantation is not reciprocal. Differently, the reader understand that plants were transplanted to a garden that is not the environment of the other species.
Moreover, table A3 is not informative. It seems that CLW and CLT are from the same location. What does CL stand for? Why CLW1-3 are separated from CLW4? Please, also add a column with species name.
Answer: Thank your comments much. Yes, our transplantation was not reciprocal only transplanting. The accurate transplant site should be Xiangtangshan National Park of China. Generally, the environmental conditions are different between lands and mountains. As know, O. longilobus grows on the cliffs of Taihang Mountains. When transplanted into the Xiangtangshan National Park, it also faces to the changed surroundings, such as average temperature, maximum temperature, and average rainfall. And the detailed descriptions have been displayed in the section 4.1. In addition, the contents of table A3 was also reviewed.
References:
① Li, W.Q. Monitoring of vegetation change in mining area based on multi-source remote sensing data: a case study of Fengfeng mining area. Hebei University of Engineering 2021.
② Ye, H., et al. Localized environmental heterogeneity drives the population differentiation of two endangered and endemic Opisthopappus Shih species. BMC Ecology and Evolution 2021, 21, 56.
③ Liu, L., et al. Geographic distribution pattern and ecological niche differentiation of endangered Opisthopappus in Taihang Mountains. Brazilian Journal of Botany 2023, 46, 217-226.
Since your transplantation was not reciprocal, please remove the word reciprocal throughout the text (10 occurrences)
7.The Extraction of total metabolite method is not clear. It seems that collected samples were liquid. Please explain better.
Answer: This section was rewrote, thank you.
Ok
Results and/or Table A2: Could authors report at least the other most important environmental factors considered?
Answer: We reviewed the contents of the table and added the results of PCA.
OK
Table 2. What does asterisk mean?
Answer: The note of asterisk was added, thank you.
OK
Figure 2 and Table 1: add the measure unit
Answer: The relative parts were reviewed.
Please, add “(relative units)” in table 2 title.
Figure 3: substance names are not readable.
Answer: Thank you, this figure was reviewed.
OK
Figure A1: PCA diagram is not readable
Answer: Figure A1 was reedited.
Unfortunately, the figure is still confused.
Figure 4 is too small.
Answer: Figure 4 was reviewed, thank you.
OK
Author Response
Dear Editor and reviewers，
We appreciate the editor and reviewers valuable comments. Based on these comments, we have carefully reviewed our manuscript. The responses to the reviewer's comments piont- to -point as following:
Response to the reviewer 1’s comments
Data reported in the tables should not be repeated in the text. Therefore, sentences could result more readable. I.e.:
For CLW, the most highly expressed metabolite was phenyl acetate with a content of 148542.13, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate, with relative contents of 120830.08, 56960.99, 39622.58, and 26437.30, respectively. The least expressed metabolite was p-Cresol with a relative content of 514.12.
Should became:
For CLW, the most highly expressed metabolite was phenyl acetate, followed by chrysoeriol, hesperetin, dioctyl phthalate, caffeate. The least expressed metabolite was p-Cresol. Etc.
Answer: According your suggests, the sentences have been rewritten.
NOT DONE
Answer: Dear, according your comments. The sentences have been rewritten in manuscript.
Since your transplantation was not reciprocal, please remove the word reciprocal throughout the text (10 occurrences).
Answer: The word reciprocal was removed, thank you.
Figure 2 and Table 1: add the measure unit
Answer: The relative parts were reviewed.
Please, add “(relative units)” in table 1 title.
Answer: Thank your comments much. “(relative units)” was added in the table 1 title.
Figure A1: PCA diagram is not readable
Answer: Figure A1 was reedited.
Unfortunately, the figure is still confused.
Answer: Dear, we added the detailed notation about the contents of this figure. Here, we presented a scatter plot with PC1 and PC2 as axes in PCA analysis. The black, red, and green dots represent samples from the CLT, CLW, and TH groups, respectively. The arrows in the figure indicated the feature vectors of the aromatic metabolites in three groups. The projection of the arrows to the coordinate axis represented the contribution of the original feature to each principal component in the new space. The longer the projection, the greater the contribution of this variable to this principal component. Based on the feature vectors of metabolites, it can be clearly seen that the samples from different groups were far apart from each other, while samples from the same group were gathered together.
Reference:
ZHENG Sunyuan, ZHU Hong, JIN Siyu, et al. Environment-dependent phenotypic variation of Osmanthus fragrans [J]. Journal of Nanjing Forestry University: natural Science Edition, 2019, 43 (2): 9.
Sincerely,
Zhixia Liu and Yiling Wang
Author Response File: Author Response.docx
Reviewer 2 Report
Since CL and TH was from the name of Chinese phonetic transcriptions of O. longilobus and O. taihangensis respectively, the authors should make a clarification on this in Materials and Methods.
In addition, all deletions should not appear in the manuscript.
Author Response
Dear Editor and reviewers，
We appreciate the editor and reviewers valuable comments. Based on these comments, we have carefully reviewed our manuscript. The responses to the reviewer's comments piont- to -point as following:
Response to the reviewer 2’s comments
Since CL and TH was from the name of Chinese phonetic transcriptions of O. longilobus and O. taihangensis respectively, the authors should make a clarification on this in Materials and Methods.
In addition, all deletions should not appear in the manuscript.
Answer: Thank you. The description of abbreviation of CL and TH presented in section 4.1 of Materials and Methods. And the deleted contents were not appeared in our paper.
真诚地
刘志霞、王一玲
Author Response File: Author Response.docx
