GIS Bioclimatic Profile and Seed Germination of the Endangered and Protected Cretan Endemic Plant Campanula cretica (A. DC.) D. Dietr. for Conservation and Sustainable Utilization
, Ioannis Anestis 1, Elias Pipinis 2, Stefanos Kostas 1, Georgios Tsoktouridis 3, Stefanos Hatzilazarou 1,* and Nikos Krigas 3,4,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsXVC
General comments.
- There is a serious problem with this manuscript. Unless, an author investigates the embryos in seeds, he/she cannot tell if the seeds have physiological dormancy or morphophysiological dormancy. That is, based on the requirements to break dormancy and promote germination, embryo work is required to tell if seeds have morphophysiological dormancy or only physiological dormancy. Thus, all the references and discussion related to kind of dormancy in this manuscript need to be deleted.
Please note that Frattaroli only speculated about the kind of dormancy in seeds of Campanula, but she did not determine if the embryo in the seeds grew prior to germination. Thus, the reference to the work of Frattaroli and others needs to be deleted.
You have a good paper without mentioning the kind of dormancy. However, I hope that in the future you will do some embryo work. See methods found in Amer. J. Bot. 96 (6): 1086-1095. 2009 for an idea of how you might be able to do this.
- It seems that you did not test fresh seeds. That is, seeds were stored from August to December, which is about 4 months. You need to talk about this is the Discussion. The high germination you present in Figure 2, may in fact reflect the fact that much breaking of physiological dormancy occurred during the storage period. Loss of dormancy during dry storage is called ‘after-ripening’. In the Discussion you need to tell the reader that some after-ripening may have occurred. You need to warn readers who may be interested in propagating this species that they may need to give the seeds a period of after-ripening.
Specific comments
Line 22. Delete “different light conditions (“ and delete the “)” after “darkness
Lines 27-28. Delete The seeds exhibited non-deep physiological dormancy type 1, as” Begin sentence with “Pre-treatments with GA3 and cold …”
Line3 30-32. I suggest you rewrite this sentence and tell the reader what you have learned that will facilitate conservation of C. cretica. What you have now is very vague.
Line 58. Not clear what you mean by “troubled geological history” Need to make this clear. Do you mean mountain building?
Lines 64-92. Delete. You have not studied embryo growth, and it seems that some of the studies you cite did not investigate embryo growth either. It is better not to even mention the kind of dormancy in seeds of C. cretica until you have done embryo work and know for sure what kind of dormancy the seeds have.
Line 182. You need to tell the reader how the seeds were stored until they were tested for germination. In the future, someone may want to repeat your work and get seedlings of C. cretica. They need to know the storage temperatures and the relative humidity. Without information on seed storage conditions, your work cannot be repeated.
Line 190. I am curious as to why you only gave the seeds 25 days of cold stratification, when your environmental data clearly show (Fig. 4) that seeds may received several months of cold stratification in the field. Did 25 days of cold stratification promote germination because much of the dormancy had been broken during the after-ripening period?
Line 207. For the second experiment, how old were the seeds (how long had they been stored) when you started the experiment? Need to add this information to the methods.
Line 307. Delete “incubation temperature of”
Line 314. Delete “treatment” , delete “percentage” and delate “incubation temperature of”
Line 355. Change “untreated” to “nontreated”
Line 356. Change “declined” to “decreased”
Line 359. Delete “the genus”
Line 367. Change wording to “monthly temperatures decreased enough to be within the range of those required for germination, as herein…”
Lines 431-463. Delete. You do not know fi seeds of morphophysiological dormancy of physiological dormancy. It is best not to speculate about all this. It is best to have another paper in which you deal with the kind of dormancy in the seeds – but first you need to do some embryo work.
Line 465. Change wording to “Species of Campanulaceae typically produce….”
References – several references need to be deleted.
Author Response
Reviewer 1
General comments.
- There is a serious problem with this manuscript. Unless, an author investigates the embryos in seeds, he/she cannot tell if the seeds have physiological dormancy or morphophysiological dormancy. That is, based on the requirements to break dormancy and promote germination, embryo work is required to tell if seeds have morphophysiological dormancy or only physiological dormancy. Thus, all the references and discussion related to kind of dormancy in this manuscript need to be deleted.
Please note that Frattaroli only speculated about the kind of dormancy in seeds of Campanula, but she did not determine if the embryo in the seeds grew prior to germination. Thus, the reference to the work of Frattaroli and others needs to be deleted.
You have a good paper without mentioning the kind of dormancy. However, I hope that in the future you will do some embryo work. See methods found in Amer. J. Bot. 96 (6): 1086-1095. 2009 for an idea of how you might be able to do this.
Authors’ response: We would like to thank the reviewer for his/her kind words and mainly for the suggestions given. Consequently, we have rephrased a considerable part of the introduction and discussion in the revised version of the manuscript to accommodate the suggestions provided by the reviewer (see track changes).
- It seems that you did not test fresh seeds. That is, seeds were stored from August to December, which is about 4 months. You need to talk about this is the Discussion. The high germination you present in Figure 2, may in fact reflect the fact that much breaking of physiological dormancy occurred during the storage period. Loss of dormancy during dry storage is called ‘after-ripening’. In the Discussion you need to tell the reader that some after-ripening may have occurred. You need to warn readers who may be interested in propagating this species that they may need to give the seeds a period of after-ripening.
Authors’ response: We would like to thank the reviewer for his/her suggestions. The information that seeds stored for 4 months were used in the germination experiments and that this may have affected to some extent the dormancy of the seeds has been mentioned in the revised manuscript. (see track changes).
Specific comments
Line 22. Delete “different light conditions (“ and delete the “)” after “darkness
Authors’ response: Corrected (see track changes)
Lines 27-28. Delete The seeds exhibited non-deep physiological dormancy type 1, as” Begin sentence with “Pre-treatments with GA3 and cold …”
Authors’ response:
Corrected (see track changes)
Line3 30-32. I suggest you rewrite this sentence and tell the reader what you have learned that will facilitate conservation of C. cretica. What you have now is very vague.
Authors’ response: We would like to thank the reviewer for this suggestion; in the revised version of the manuscript, we have added in the abstract the conclusions in parenthesis as new pieces of knowledge gained (see track changes).
Line 58. Not clear what you mean by “troubled geological history” Need to make this clear. Do you mean mountain building?
Authors’ response: The reviewer is right; in the revised version of the manuscript, we have clarified that we refer to mountain building and tectonic movements (see track changes).
Lines 64-92. Delete. You have not studied embryo growth, and it seems that some of the studies you cite did not investigate embryo growth either. It is better not to even mention the kind of dormancy in seeds of C. cretica until you have done embryo work and know for sure what kind of dormancy the seeds have.
Authors’ response:
We have rephrased a considerable part of the introduction in the revised version of the manuscript to accommodate the suggestion given by the reviewer (see track changes).
Line 182. You need to tell the reader how the seeds were stored until they were tested for germination. In the future, someone may want to repeat your work and get seedlings of C. cretica. They need to know the storage temperatures and the relative humidity. Without information on seed storage conditions, your work cannot be repeated.
Authors’ response: In sub section “Seed Collection and Storage”, information about the storage conditions of seeds is provided, Specifically, it has been stated that “Upon collection, the capsules were placed on filter paper and were maintained for a week under laboratory conditions” and “After thorough cleaning, the seeds were stored in glass containers at a temperature of 3–5 °C in a low-humidity environment (<15%) until the initiation of the experiments.”. To be more accurate, the duration of the natural drying of the capsules before the seed cleaning has been added to the revised manuscript (see track changes).
Line 190. I am curious as to why you only gave the seeds 25 days of cold stratification, when your environmental data clearly show (Fig. 4) that seeds may received several months of cold stratification in the field. Did 25 days of cold stratification promote germination because much of the dormancy had been broken during the after-ripening period?
Authors’ response: In the submitted manuscript, we mentioned that “In the present study, the seeds of C. cretica were not stratified for more than 25 days, due to a pre-experiment conducted showing that seed germination was observed by the end of the fourth week of cold stratification (data not shown).” (see lines 427-430).
However, in order to be more accurate, the “In the present study, the seeds of C. cretica were not stratified for more than 25 days, due to a pre-experiment conducted showing that germination of seeds stored for about four months was observed by the end of the fourth week of cold stratification (data not shown).” has been added to subsection “Seed germination experiments” in the revised manuscript (see track changes).
Unfortunately, since fresh seed germination was not studied in the present study, there is no possibility to compare the effect of cold stratification on germination between fresh and four months stored seeds. Therefore, we do not know if seed storage reduced the required duration of cold stratification.
Line 207. For the second experiment, how old were the seeds (how long had they been stored) when you started the experiment? Need to add this information to the methods.
Authors’ response: In the revised manuscript, we have reported that the second experiment started immediately after the completion of the germination test of control seeds of the first experiment which lasted for 36 days (see track changes). Thus, it can be understood that the seeds of the second experiment were stored at 3-5°C for 36 days longer than in the first experiment.
Furthermore, it has been stated that the seeds from the seedlot used in the first experiment were used for the second experiment (see track changes).
Line 307. Delete “incubation temperature of”
Authors’ response: Corrected (see tracked changes)
Line 314. Delete “treatment” , delete “percentage” and delate “incubation temperature of”
Authors’ response: Corrected (see tracked changes)
Line 355. Change “untreated” to “nontreated”
Authors’ response: Corrected (see tracked changes)
Line 356. Change “declined” to “decreased”
Authors’ response: Corrected (see tracked changes)
Line 359. Delete “the genus”
Authors’ response: Corrected (see tracked changes)
Line 367. Change wording to “monthly temperatures decreased enough to be within the range of those required for germination, as herein…”
Authors’ response: Corrected (see tracked changes)
Lines 431-463. Delete. You do not know fi seeds of morphophysiological dormancy of physiological dormancy. It is best not to speculate about all this. It is best to have another paper in which you deal with the kind of dormancy in the seeds – but first you need to do some embryo work.
Authors’ response: Deleted (see tracked changes)
Line 465. Change wording to “Species of Campanulaceae typically produce….”
Authors’ response: Corrected (see tracked changes)
References – several references need to be deleted.
Authors’ response: All references have been re-enumerated (see tracked changes)
Submission Date
28 April 2025
Date of this review
08 May 2025 14:37:09
Reviewer 2 Report
Comments and Suggestions for AuthorsThis study systematically evaluated the ecological requirements and seed germination mechanisms of the Cretan endemic endangered plant Campanula cretica through GIS bioclimatic data and seed germination experiments, providing critical insights for its conservation and sustainable utilization. The work holds significant scientific merit and practical value, though certain details require refinement to enhance completeness.
- I recommend adding the species name “Campanula cretica”to the Keywords.
- Line 157: The authors collected climate data from 1970–2000 for the geographical locations where Campanula cretica grows. These data seem outdated. Why not use recent data (e.g., 1990–2020) for analysis?
- Is there a connection between Sections 3.1 and 3.2 of the Results? Did the temperature ranges identified in Section 3.1 inform the temperature settings for the indoor germination experiments in Section 3.2?
- Line 295: The figure is labeled "ns²", but the caption uses "ns" .Please standardize the notation.
- Lines 177–180: To protect genetic diversity and ensure seed representativeness, the authors collected seeds from multiple locations in Crete between 2022–2025. Please clarify the geographical distribution, ecological environments of these collection sites, and their potential impacts on experimental results. Are these sites geographically distant? Do seeds from different locations exhibit variations in germination traits?
Author Response
REVIEWER 2
This study systematically evaluated the ecological requirements and seed germination mechanisms of the Cretan endemic endangered plant Campanula cretica through GIS bioclimatic data and seed germination experiments, providing critical insights for its conservation and sustainable utilization. The work holds significant scientific merit and practical value, though certain details require refinement to enhance completeness.
Authors’ response: We would like to thank the reviewer for his/fer kind words.
- I recommend adding the species name “Campanula cretica”to the Keywords.
Authors’ response: As this scientific name is already mentioned in the manuscript’s title, there is no need to add it as keyword. However, profiting from the reviewer’s comment, we have added Campanulaceae as new keyword in the revised version of the manuscript (see tracked changes).
- Line 157: The authors collected climate data from 1970–2000 for the geographical locations where Campanula cretica grows. These data seem outdated. Why not use recent data (e.g., 1990–2020) for analysis?
Authors’ response: This is the kind of data provided by the open-access WorldClim database. For comparability with previous own research articles, we have preferred to use the same dataset.
- Is there a connection between Sections 3.1 and 3.2 of the Results? Did the temperature ranges identified in Section 3.1 inform the temperature settings for the indoor germination experiments in Section 3.2?
Authors’ response: This issue is discussed in the second and the third paragraph of the discussion which has been improved in the revised version of the manuscript (see track changes).
- Line 295: The figure is labeled "ns²", but the caption uses "ns" .Please standardize the notation.
Authors’ response: Corrected (see track changes)
- Lines 177–180: To protect genetic diversity and ensure seed representativeness, the authors collected seeds from multiple locations in Crete between 2022–2025. Please clarify the geographical distribution, ecological environments of these collection sites, and their potential impacts on experimental results. Are these sites geographically distant? Do seeds from different locations exhibit variations in germination traits?
Authors’ response: We would like to thank the reviewer for this comment; we have added at the beginning of the discussion section of the revised manuscript new text to accommodate the reviewers’ suggestion (see track changes).
Round 2
Reviewer 1 Report
Comments and Suggestions for Authorsno additional comments
