Callogenesis and Plant Regeneration in Peony (Paeonia × suffruticosa) Using Flower Petal Explants
and Xiangtao Zhu 1,*Round 1
Reviewer 1 Report
Dear Authors,
I have conducted a detailed reading and review of the manuscript entitled ‘Callogenesis and plant regeneration in peony (Paeonia suffruticosa) using flower petal explants’. Compliments on the very extensive and very detailed investigation presented in the paper. I have some suggestions that should be solved prior to paper acceptance and publication.
In the title and throughout the entire manuscript it would be more appropriate to use Paeonia × suffruticosa, due to their hybrid origin.
Abstract and the keywords are appropriately written.
The Introduction section is very well written, covering all important aspects and clear research aims.
Line 71: correct ‘laying a foundation' by ‘providing a basis’ or ‘providing a foundation’.
Line 82: please be specific about which petals? ‘After the sterilization, the petals were cut from the unfolded buds and used for tissue culture inoculation.’ Did you discard the outer layers that might have been damaged during the sterilization process?
Otherwise, the Materials and Methods section is very detailed and accurate.
Results section
Lines 182 – 187 should precede table 3, and thus be placed in line 173.
Figure 1 should be placed after the corresponding text in lines 202 – 213.
The discussion and conclusion sections are appropriately written.
Please explain why you opted for SI:
"Omics Technologies and Their Applications in Vegetable Plant Research"
I do not see a connection between the presented work with this call for papers and future papers assembly.
Author Response
Changes in the revised manuscript
Herewith, the below are the points that highlights any changes to the manuscript mainly based on the reviewer’s comments.
Please note: the reviewer’s comments are in black, the authors’ responses are in blue. We showed these points one by one, thus, you can easily find and review all the changes in the revised manuscript attached.
Moreover, we also submitted a final version in which any changes to the text in red.
Finally, we expect that you would be satisfied with these revisions. Also, if you have any questions, please fell free to contact with us.
(1) Reviewer 1’s comments and response:
Comment 1: In the title and throughout the entire manuscript it would be more appropriate to use Paeonia × suffruticosa, due to their hybrid origin.
Response: Thanks to the reviewer for the valuable comments. Now I have replaced " Paeonia suffruticosa " with" Paeonia × suffruticosa "in the title and the entire manuscript.
Comment 2: Line 71: correct ‘laying a foundation' by ‘providing a basis’ or ‘providing a foundation’.
Response: Now I have use " providing a foundation " instead of" laying a foundation "in the revised manuscript. (Line 72)
Comment 3: Line 82: please be specific about which petals? ‘After the sterilization, the petals were cut from the unfolded buds and used for tissue culture inoculation.’ Did you discard the outer layers that might have been damaged during the sterilization process?
Response: Yes, here we cut off all the petals except those damaged during the sterilization process for inoculation. I have re-described it more appropriately in the revised manuscript. (Line 83)
Comment 4: Lines 182 – 187 should precede table 3, and thus be placed in line 173.
Response: Now this part has been placed in the front of Table 3 according to the comments of reviewers in the revised manuscript.
Comment 5: Figure 1 should be placed after the corresponding text in lines 202 – 213.
Response: Thanks to the reviewer for the comments. Since Figure 1 also contains images from the resulting sections 3.1 and 3.2, we place the figure in the middle of sections 3.1,3.2, and 3.3,3.4.
Comment 6: Please explain why you opted for SI: "Omics Technologies and Their Applications in Vegetable Plant Research"
Response: Peony is not only an excellent garden plant, but also developed into oil and medicine in recent years, and is a new economic crop. With peony seed oil by process can produce high quality cooking oil, medicinal peony root bark has clear heat cool blood, promoting blood circulation to remove blood stasis, the peony flowers can also be processed into the peony flower tea, peony pollen contains a variety of nutrients, so peony pollen can be used to make peony pollen health noodles and vegetables juice beer, pollen yam yogurt, not only unique flavor, and has the health care function. It can be seen that planting peony with vegetable edible value can not only improve the ecological environment, but also increase farmers' income and promote regional economic development, so it is a new cash crop with a good development prospect. Tissue culture technology is the basis of modern agriculture, and its application is very wide, especially in the field of plant transgenic technology. Along with the development of multiple omics technology, for mining, verify omics data as a result, the need to establish a more mature, suitable genetic transformation system combining with the study, the peony current haven't perfect genetic system can be used both at home and abroad, this study aims to provide theoretical basis for peony build a mature system of genetic based, and lay a foundation for multiple omics technology combined with use.
Reviewer 2 Report
The manuscript describes interesting results on plant regeneration from petals. The present from needs improvements including english editing. All detailed comments are placed in pdf version attached to this review.
Comments for author File: 
Comments.pdf
Author Response
Changes in the revised manuscript
Herewith, the below are the points that highlights any changes to the manuscript mainly based on the reviewer’s comments.
Please note: the reviewer’s comments are in black, the authors’ responses are in blue. We showed these points one by one, thus, you can easily find and review all the changes in the revised manuscript attached.
Moreover, we also submitted a final version in which any changes to the text in red.
Finally, we expect that you would be satisfied with these revisions. Also, if you have any questions, please fell free to contact with us.
(2) Reviewer 2’s comments and response:
Comment 1: Please explain in more details what value it is. (medicinal value)
Response: Thanks to the reviewer for the valuable comments. Now I have explained and added the medicinal value of peony in my revised manuscript. (Line 37-40)
Comment 2: Please be more specific, as not all peonias are sterile (asexual plants)
Response: Yes, the original description does create ambiguity, I have re-edited the sentence in my revised manuscript (Line 46-47).
Comment 3: what it means? (plant detoxification)
Response: The "plant detoxification" means the elimination of viruses that have infected by plants.
Comment 4: lines 46-54 Please be more specific in this paragraph -Plant tissue cultures does not equals with micropropagation. It is not known to which of the mentioned above Authors refers to.
Response: Thanks to the reviewers for the reminder, now I have corrected all incorrect descriptions in the revised manuscript. (Line 47-55)
Comment 5: Young flower petals were clipped from the unfolded peony buds as explants. “clipped” not clear, please replace with other word.
Response: Now we use "cut" instead of " clipped " in the revised manuscript. (Line 78)
Comment 6: please replace “callus increment” by other wording.
it is not clear how the parameters were calculated. What means transferred callus? It is not known at what period that measurements were taken. (Line 98-101)
Please enter here a clear and complete description what and when it has been done
Response: After the callus induced by the petals as an explant is transferred to the proliferation medium, the number and weight of the callus will continue to increase with the continuous division of the cells. Therefore, in order to screen the most suitable callus proliferation medium, we counted the callus increment and proliferation rate, and now I have re-described this part in the revised manuscript, adding when and how to calculate. (Line 98-103)
Comment 7: please insert the temperature of culturing. (Line 106)
Response: I have supplemented the temperature of culturing in the revised manuscript. (Line 108)
Comment 8: please specify concentrations (Line 123)
Response: I have specified the concentrations of sucrose, NAA and IBA in the revised manuscript. (Line 125-126)
Comment 9: This sentence is not clear, please rephrase. (Line 125-126)
Response: I have rephrased this sentence to "After about 30 days of aseptic tissue culture seedling root growth, the plants with ro-bust growth were selected for the transplanting test." (Line 128-129)
Comment 10: please specify how hardening was performed (Line 126-127)
Response: We directly opened the lid of the culture bottle to allow the tissue culture seedlings to adapt. I have added the content of how to domesticate the tissue culture seedlings in the revised manuscript. (Line 129-131)
Comment 11: the sentence is not clear, please rewrite. (Line 128-129)
Response: I have rephrased the sentence in the revised manuscript. (Line 131-132)
Comment 12: what it means - it was not mentioned in M&M (Line 147)
Response: It means that the explants under treatment 5 can form callus in 98.52% of the cases. I have made changes to the inappropriate description in the original manuscript. (Line 150-151)
Comment 13: profiled? please use proper word (Line 158)
Response: I have changed " were profiled " to " can proliferate " in the revised manuscript. (Line 161)
Comment 14: Tables 1-3. It is not known at what period of the culture the results were collected. Please Insert such information in the table title
Response: Thanks to the reviewer for the valuable comments. I have inserted such information in the table title in the revised manuscript. (Table 1-3)
Comment 15: The 'CK' either explain abbreviation or replace by 'control'
Response: I have changed " CK " to " control " in the revised manuscript. (Table 1-3)
Comment 16: The whole description should be more precise, as not all pictures refers to scaning microscopy. Please add here an informations given in the methods on callus description (white, compact green etc). The description of picturesJ-K is sparse, please add infor on the prolideration rate, and what arrow means. Lack of description of picture L (Figure 1)
Response: I have redescribed the annotations in Figure 1. (Line 188-193)
Comment 17: Statistics in table 4 shows it is not significant with the combinations 3-7, so combination 4 cannot be presented as separate group.
Response: Thanks for the reviewer's reminding, I have deleted the inappropriate description in the revised manuscript.
Comment 18: ' There were relatively few protrusions, the roots of the protrusions were cracked...' nolt clear, please change (Line 208)
Response: Now the “the roots of the protrusions were cracked” have been changed to “the bottom of the protrusions was cracked” in the revised manuscript. (Line 212)
Comment 19: apical buds are expected to produce shoots instead of callus, so that is not a good example (Line 227)
Response: Yes, we agree with your comment. Now we have given additional examples in the revised manuscript. (Line 230-232; 362)
Comment 20: please insert % (Line 240)
Response: I have added the percentage of differentiation in the revision manuscript. (Line 244-245)
Comment 21: Is such selection possible macroscopically as Authors observed it under scaning microscope?
Response: Yes, macroscopically, we select by the morphological characteristics of the callus, and we further observe the microstructure of the selected callus by scanning electron microscopy to analyze the reasons for the different differentiation rates under different morphologies, which also provides a basis for further selection.
