Optimizing Transplanting Practices for Potted Tree Peony Based on Non-Structural Carbohydrates Accumulation
and Guoan Shi 1,*
O. Roger Anderson
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript addresses a relevant topic, and the authors have demonstrated considerable effort in conducting the study and presenting the results. The research has a well-founded experimental design, and the conclusions are supported by the data presented. However, some areas require revision to enhance clarity, scientific consistency, and the overall presentation of the work. The quality of the language, although comprehensible, can be improved to meet the standards of an international journal.
Below, I present the specific comments, focused on scientific aspects, that I consider essential for the improvement of the work:
Comment 1: page 1, lines 27-29: I suggest improving the phrasing of: "The optimal protocol identified involved 25% root pruning followed by irrigation with a solution containing 750 mg·L-1 rooting agent and 20 million units·mL-1 of Metarhizium anisopliae by range analysis." to: "The optimal protocol identified through range analysis involved 25% root pruning, followed by irrigation with a solution containing 750 mg·L-1 rooting agent and 20 million units·mL-1 of Metarhizium anisopliae."
Comment 2: page 1, lines 13 and 35: The term "engineering seedlings" is used and is not universally understood. Please provide a definition or clarification of what this term implies in the context of peony cultivation (e.g., does it refer to high-quality or standardized seedlings for specific commercial purposes?).
Comment 3: page 3, line 130 (Table 1): The unit "million U·mL-1" for Metarhizium anisopliae could be more precise. For greater specificity, I suggest using "million CFU·mL-1" or "million spores·mL-1".
Comment 4: page 4 (Figure 1 caption), Page 7 (Table 2 note), Page 8 (Table 3 note), Page 8 (Table 4 note), Page 9 (Table 5 note), Page 9 (Figure 2 caption), the same applies to the figure captions: The note "Different lowercase letters represent the obvious differences of various indicators in different soil types (P < 0.05)" is incorrect, as you did not work with different soil types. Please correct this note in all occurrences to something like "Different lowercase letters represent significant differences (P < 0.05) among the various indicators".
Comment 5: page 4, lines 139 and 141: I was confused by the description of the number of replicates. For photosynthetic performance, "each combination had three replicates" is mentioned, while for morphological indicators, it's "Selecting three representative plants from each combination". I would like to clarify: does "each combination had three replicates" refer to three plants per combination, or three independent biological replicates, each with 40 plants? Please adjust this section for greater clarity and to avoid ambiguity.
Comment 6: pages 6, 9, and 11: Please improve the quality of the figures.
Comment 7: page 10, line 260 (Table 6): There is a typo in "Metarhizium ansopliae". Please adjust to "Metarhizium anisopliae".
Comment 8: page 11, lines 296-298: The statement "Although the FRNSC has a negative direct impact on TNSCA, it benefits the latter through indirect effects with other indicators..." caused confusion. If the direct impact is negative, the "beneficial" nature of the indirect effects needs to be explained for better understanding, perhaps by discussing the physiological pathways involved.
Comment 9: page 13, lines 333-334: The mention that "there was no scarab in all the potting substrates" is information that needs to be better integrated and emphasized in the discussion. The observed benefits of Metarhizium anisopliae application in the present study are likely attributable to its plant growth-promoting capacity (as your own references [19,20] indicate endophytic colonization of roots and enhanced growth), and not primarily to its function as a biopesticide in pest control. I suggest that this distinction be more clearly emphasized in the discussion to highlight an important and perhaps underestimated aspect of Metarhizium anisopliae is role in this context, thereby strengthening the study's contribution.
Comments on the Quality of English LanguageGrammatical errors, typos, and awkward phrasing in the manuscript hinder clarity.
Author Response
Response letter
Dear Editor and Reviewers
Thank you for offering us an opportunity to improve the quality of our submitted manuscript (horticulturae-3785717). We appreciated very much the reviewers’ constructive and insightful comments. In this revision, we have addressed all of these comments. We hope the revised manuscript has now met the publication standard of Horticulturae.
We highlighted all the revisions in blue.
Next, our point-to-point responses to the queries raised by the reviewers are listed.
Comment 1: page 1, lines 27-29: I suggest improving the phrasing of: "The optimal protocol identified involved 25% root pruning followed by irrigation with a solution containing 750 mg·L-1 rooting agent and 20 million units·mL-1 of Metarhizium anisopliae by range analysis." to: "The optimal protocol identified through range analysis involved 25% root pruning, followed by irrigation with a solution containing 750 mg·L-1 rooting agent and 20 million units·mL-1 of Metarhizium anisopliae."
Response 1: We have adopted this suggestion and incorporated the corresponding modifications in the manuscript (Line 29).
Comment 2: page 1, lines 13 and 35: The term "engineering seedlings" is used and is not universally understood. Please provide a definition or clarification of what this term implies in the context of tree peony cultivation (e.g., does it refer to high-quality or standardized seedlings for specific commercial purposes?).
Response 2: Engineering seedlings refers to standardized seedlings produced through controlled facility cultivation, enabling transplantation unrestricted by growing seasons or geographical limitations. This approach serves as a critical methodology for rapid large-scale afforestation and landscape construction in non-native regions. We have explained the engineering seedlings mentioned in the manuscript and revised the sentence to “Potted cultivation serves as a vital strategy for industrialized production of standardized tree peony engineering seedlings capable of year-round and off-site transplantation” (Lines 13-14).
Comment 3: page 3, line 130 (Table 1): The unit "million U·mL-1" for Metarhizium anisopliae could be more precise. For greater specificity, I suggest using "million CFU·mL-1" or "million spores·mL-1".
Response 3: We have adopted this suggestion and replaced U with spores. (Lines 30, 124, 132, 263, 266, 349, 394)
Comment 4: page 4 (Figure 1 caption), Page 7 (Table 2 note), Page 8 (Table 3 note), Page 8 (Table 4 note), Page 9 (Table 5 note), Page 9 (Figure 2 caption), the same applies to the figure captions: The note "Different lowercase letters represent the obvious differences of various indicators in different soil types (P < 0.05)" is incorrect, as you did not work with different soil types. Please correct this note in all occurrences to something like "Different lowercase letters represent significant differences (P < 0.05) among the various indicators".
Response 4: Thank you for pointing out these obvious mistakes. I have revised these sentences as “Different lowercase letters represent statistically significant differences among the various indicators (P < 0.05).” (In the notes of figure 1 and 2, and table 2, 3, 4 and 5)
Comment 5: page 4, lines 139 and 141: I was confused by the description of the number of replicates. For photosynthetic performance, "each combination had three replicates" is mentioned, while for morphological indicators, it's "Selecting three representative plants from each combination". I would like to clarify: does "each combination had three replicates" refer to three plants per combination, or three independent biological replicates, each with 40 plants? Please adjust this section for greater clarity and to avoid ambiguity.
Response 5: We selected three representative plants in per combination, each with 40 plants. We revised the sentences as” Three representative plants were selected for each combination, with a total of 40 plants per combination, and three leaves were measured per plant.” (Lines 142-143).
Comment 6: pages 6, 9, and 11: Please improve the quality of the figures.
Response 6: The image resolution has been enhanced to improve graphical clarity.
Comment 7: page 10, line 260 (Table 6): There is a typo in "Metarhizium ansopliae". Please adjust to "Metarhizium anisopliae".
Response 7: We have replaced “Metarhizium ansopliae” with “Metarhizium anisopliae” (Table 6)
Comment 8: page 11, lines 296-298: The statement "Although the FRNSC has a negative direct impact on TNSCA, it benefits the latter through indirect effects with other indicators..." caused confusion. If the direct impact is negative, the "beneficial" nature of the indirect effects needs to be explained for better understanding, perhaps by discussing the physiological pathways involved.
Response 8: The growth of fibrous roots requires the consumption of NSCs. Furthermore, there is a significant positive correlation between FRNSC and fibrous root biomass, indicating that a high input of NSCs benefits fibrous root growth. This is consistent with the negative direct path coefficient between FRNSC and TNSCA. However, increased fibrous root biomass helps enhance the plant's capacity to absorb water and nutrients, which are essential material conditions for photosynthesis. By enhancing the plant's absorption capacity, this indirectly promotes NSCs accumulation, offsetting the NSCs consumed during root growth and thus exerting an indirect positive effect on TNSCA. We revised the sentences as follows: “Although the direct path coefficient between FRNSC and TNSCA is negative, the indirect path coefficient of FRNSC on TNSCA is positive. This may be related to the fact that during fibrous roots growth, plants increase their input of NSCs, which promotes the growth of fibrous roots and enhances their capacity to absorb water and nutrients, thereby facilitating the accumulation of NSCs. This indicates that transporting NSCs to fibrous roots and promoting their growth represents an important survival strategy for potted tree peonies.” (Lines 305-310)
Comment 9: page 13, lines 333-334: The mention that "there was no scarab in all the potting substrates" is information that needs to be better integrated and emphasized in the discussion. The observed benefits of Metarhizium anisopliae application in the present study are likely attributable to its plant growth-promoting capacity (as your own references [19,20] indicate endophytic colonization of roots and enhanced growth), and not primarily to its function as a biopesticide in pest control. I suggest that this distinction be more clearly emphasized in the discussion to highlight an important and perhaps underestimated aspect of Metarhizium anisopliae is role in this context, thereby strengthening the study's contribution.
Response 9: We fully accept this suggestion and have revised the manuscript to emphasize Metarhizium anisopliae's role as a plant growth promoter in tree peonies rather than its insecticidal effects. We revised the sentences of “In addition, there was no scarab in all the potting substrates, so the effectiveness of Metarhizium anisopliae in pest control is worthy of affirmation.” to “Therefore, the effect of Metarhizium anisopliae on root signaling in tree peonies and their growth deserves further investigation.” (line 344-345)
Comment 10: Grammatical errors, typos, and awkward phrasing in the manuscript hinder clarity.
Response 10: The manuscript has been polished by Prof. M. Shaaban to improve its English quality
Best regards,
Guoan Shi
Aug 17, 2025
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThis is an interesting and generally clearly written manuscript that provides scientific and practical experimental horticultural evidence for improving the culture of potted tree peonies (Paeonia suffruticosa Andr.). I have only some general comments for the authors to consider to clarify a few aspects of the text.
Line Comment or recommended rewording
40 It usually has been used as a field ornamental since ancient times.
45 ----- requirements, potted cultivation has gained increasing-----
55 ------ leading to uneven growth of the stem [8,9].
56 ------ pruning to adapt the roots to the relatively restricted conditions of pot culture.
69–71 While the optimal concentration of root agent is influenced by its composition and crop species, the optimal concentration of its application in conjunction with other treatments has yet to be verified by experiments. [removed the word ‘and’].
82 ----- but can also endophytically colonize plant roots, enhancing ----
110 It would be helpful if the authors add a clarifying sentence explaining what the ‘rooting agents’ were that were added.
141 Plant morphological indicators were assessed by selecting three representative plants from ----
146 Leaf area was analyzed using Digimizer image analysis software (MedCalc Software Ltd., Ostend, Belgium).
153 recommend rewording of this sentence.
----- of each plant; total non-structural carbohydrate accumulation (TNSCA) in each plant was the sum of NSCs accumulated in each organ of the plant.
163–165 Under slight and medium root pruning conditions, the number of stems tended to increase with the enhancement of Metarhizium anisopliae; and severe root pruning significantly reduced the stem length of potted tree peonies.
165–169 Comparing the effects of different treatments on biomass, revealed that root pruning had a negative effect on tree peony’s biomass, in which the biomass of leaves, --------
171 The analysis of different plant organ’s biomass revealed that the-------
198 ---- of the soluble sugar content in different organs of the tree peony----
In the English language, it is not necessary to use the plural form of sugars content. Please check that you correct this throughout the paragraph to be soluble sugar content, not soluble sugars content.
207 ---- increase of root pruning. Slight root pruning was beneficial ----
211 Table 3. Soluble sugar content in different organs --------
193–194 Recommend rewording of the Note at bottom of table.
Note: The data are shown as means ± SE (n = 3). Different lowercase letters represent statistically significant differences of various indicators in different soil types (P < 0.05).
Please consider using the above formatting for the note inserted in the caption for each of your tables with statistical data, and also in your figure captions (such as Figure 2).
319–320 ---- root morphology, so it is widely used in seedling ----
326–327 --- which bi-directionally promotes above ground and below ground growth of seedlings [12].
A more general comment on terminology.
The authors are advised that the abbreviation SPAD is not appropriate given the method of measuring the chlorophyll content that the authors used. SPAD is specifically a proprietary acronym used for chlorophyll measurements [The SPAD-502 Chl meter (Soil Plant Analysis Development)] made with the Konica Minolta chlorophyll meter (URL: Konica Minolta SPAD-502Plus Chlorophyll Meter).
The authors used a CCM200 chlorophyll meter (Opti-Sciences, Hudson, NH, U.S.A.) not a Konica Minolta SPAD meter.
To be more accurate, I recommend that the authors use terminology such as leaf chlorophyll content (LCC). And instead of SPAD substitute LCC, or some equivalent abbreviation, other than SPAD.
Thus, in line 137, I recommend the authors consider the composition of the text as follows.
Leaf chlorophyll content (LCC) was measured in the top leaves using a CCM200 chlorophyll meter (Opti-Sciences, Hudson, NH, U.S.A.). Three leaves were measured per plant, and each combination had three replicates.
[Or if the authors purchased the CCM200 from a different vendor than Opti-Sciences, please insert the name and address of the vendor that was used]
Then throughout the manuscript replace SPAD with the abbreviation LCC. OR, if you prefer a different abbreviation than LCC, please consider your preferred abbreviation to replace SPAD.
Author Response
Response letter
Dear Editor and Reviewers
Thank you for offering us an opportunity to improve the quality of our submitted manuscript (horticulturae-3785717). We appreciated very much the reviewers’ constructive and insightful comments. In this revision, we have addressed all of these comments. We hope the revised manuscript has now met the publication standard of Horticulturae.
We highlighted all the revisions in red.
Next, our point-to-point responses to the queries raised by the reviewers are listed.
Comment 1: Line 40. It usually has been used as a field ornamental since ancient times.
Response 1: We accept this suggestion and have made revisions (line 41) in the manuscript.
Comment 2: Line 45. ----- requirements, potted cultivation has gained increasing-----
Response 2: We accept this suggestion and have made revisions (line 46) in the manuscript.
Comment 3: Line 55. ------ leading to uneven growth of the stem [8,9].
Response 3: We accept this suggestion and have made revisions (line 56) in the manuscript.
Comment 4: Line 56. ------ pruning to adapt the roots to the relatively restricted conditions of pot culture.
Response 4: We accept this suggestion and have made revisions (line 57) in the manuscript.
Comment 5: Line69–71. While the optimal concentration of root agent is influenced by its composition and crop species, the optimal concentration of its application in conjunction with other treatments has yet to be verified by experiments. [removed the word ‘and’].
Response 5: We accept this suggestion and have made revisions (line 72) in the manuscript.
Comment 6: Line 82. ----- but can also endophytically colonize plant roots, enhancing
Response 6: We accept this suggestion and have made revisions (line 84) in the manuscript.
Comment 7: Line 110. It would be helpful if the authors add a clarifying sentence explaining what the ‘rooting agents’ were that were added.
Response 7: We added an explanation that root agent was “Our laboratory's patented technology [27]” (line 120)
Comment 8: Line 141. Plant morphological indicators were assessed by selecting three representative plants from ----
Response 8: We accept this suggestion and have made revisions (line 145) in the manuscript.
Comment 9: Line 146. Leaf area was analyzed using Digimizer image analysis software (MedCalc Software Ltd., Ostend, Belgium).
Response 9: We accept this suggestion and have made revisions (line 151) in the manuscript.
Comment 10: Line 153. Recommend rewording of this sentence----- of each plant; total non-structural carbohydrate accumulation (TNSCA) in each plant was the sum of NSCs accumulated in each organ of the plant.
Response 10: We accept this suggestion and revised the sentences as follows: “NSC accumulation (NSCA) in each organ was obtained by multiplying of the organ biomass by its NSCs content, while the total non-structural carbohydrate accumulation (TNSCA) of each plant was calculated as the sum of NSCs across all organs.” (lines 157-160).
Comment 11: Line 163–165. Under slight and medium root pruning conditions, the number of stems tended to increase with the enhancement of Metarhizium anisopliae, and severe root pruning significantly reduced the stem length of potted tree peonies.
Response 11: We accept this suggestion and have made revisions (line 170) in the manuscript.
Comment 12: Line 165–169. Comparing the effects of different treatments on biomass, revealed that root pruning had a negative effect on tree peony’s biomass, in which the biomass of leaves, --------
Response 12: We accept this suggestion and have made revisions (line 174) in the manuscript.
Comment 13: Line 171. The analysis of different plant organ’s biomass revealed that the-------
Response 13: We accept this suggestion and revised the sentences as follows: The analysis of biomass distribution among different organs revealed that (line 178)
Comment 14: Line 198. ---- of the soluble sugar content in different organs of the tree peony----In the English language, it is not necessary to use the plural form of sugars content. Please check that you correct this throughout the paragraph to be soluble sugar content, not soluble sugars content.
Response 14: We accept this suggestion and have made revisions (lines 206, 208, 209, 212, 214) in the manuscript and replaced “soluble sugars” with “soluble sugar”.
Comment 15: Line 207. ---- increase of root pruning. Slight root pruning was beneficial ----
Response 15: We accept this suggestion and have made revisions (lines 214-215) in the manuscript.
Comment 16: Line 211. Table 3. Soluble sugar content in different organs --------
Response 16: We accept this suggestion and have made revisions (line 218) in the manuscript.
Comment 17: 193–194. Recommend rewording of the Note at bottom of table.
Note: The data are shown as means ± SE (n = 3). Different lowercase letters represent statistically significant differences of various indicators in different soil types (P < 0.05). Please consider using the above formatting for the note inserted in the caption for each of your tables with statistical data, and also in your figure captions (such as Figure 2).
Response 17: We accept this suggestion and have made revisions in the notes of figures 1, 2. and tables 2, 3, 4, 5 and revised the sentences as follows: Different lowercase letters represent statistically significant differences among the various indicators.
Comment 18: 319–320. ---- root morphology, so it is widely used in seedling ----
Response 18: We accept this suggestion and have made revisions (line 331) in the manuscript.
Comment 19: 326–327. - which bi-directionally promotes above ground and below ground growth of seedlings.
Response 19: We revised the sentences as follows: thereby simultaneously enhancing above-and below-ground growth of seedlings (lines 337-338).
Comment 20: A more general comment on terminology. The authors are advised that the abbreviation SPAD is not appropriate given the method of measuring the chlorophyll content that the authors used. SPAD is specifically a proprietary acronym used for chlorophyll measurements [The SPAD-502 Chl meter (Soil Plant Analysis Development)] made with the Konica Minolta chlorophyll meter (URL: Konica Minolta SPAD-502Plus Chlorophyll Meter). The authors used a CCM200 chlorophyll meter (Opti-Sciences, Hudson, NH, U.S.A.) not a Konica Minolta SPAD meter. To be more accurate, I recommend that the authors use terminology such as leaf chlorophyll content (LCC). And instead of SPAD substitute LCC, or some equivalent abbreviation, other than SPAD. Thus, in line 137, I recommend the authors consider the composition of the text as follows. Leaf chlorophyll content (LCC) was measured in the top leaves using a CCM200 chlorophyll meter (Opti-Sciences, Hudson, NH, U.S.A.). Three leaves were measured per plant, and each combination had three replicates. [Or if the authors purchased the CCM200 from a different vendor than Opti-Sciences, please insert the name and address of the vendor that was used] Then throughout the manuscript replace SPAD with the abbreviation LCC. Or, if you prefer a different abbreviation than LCC, please consider your preferred abbreviation to replace SPAD.
Response 20: I apologize for the mistake in the instrument name in the manuscript. After further confirmation, it turns out that the instrument I used for the measurements is the SPAD-502Plus. Below is a picture of the instrument I used. I have revised the instrument name to SPAD-502 Plus Chlorophyll Meter (Konica Minolta Sensing, Osaka, Japan) in line 141.
Best regards,
Guoan Shi
Aug 17, 2025
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe authors satisfactorily carried out the necessary corrections, addressing the points raised in the previous review. The manuscript was improved in its clarity, scientific consistency, and presentation.
