The Influences of Genotype and Year on Some Biologically Active Compounds in Honeysuckle Berries
, Ivona Cristina Enescu (Mazilu) 2,*, Georgiana Badea 3 and Loredana Elena Vijan 4,*
Round 1
Reviewer 1 Report
Introduction. The introduction should be more related to the purpose of the experience. Some of the important articles are missing, f.e. : Gołba, M.; Sokół-Łętowska, A.; Kucharska, A.Z. Health Properties and Composition of Honeysuckle Berry Lonicera caerulea L. An Update on Recent Studies. Molecules 2020, 25, 749. https://doi.org/10.3390/ecue omol07les25039 4
Sampling. Lack of important information; 200 g from one bush or one cultivar or what? How many bushes were cultivated in one replicate? How many replicates? It is not clear.
Table 1.
A sum of rainfall for each season out of 6 months should be presented. The same for sunshine hours. This information is needed for discussion and explaining the results obtained. Moreover, the weather conditions differ each year, hence the year can not be regarded as the factor of the experiment. Instead of that, the experiment should be continued for more years, and based on the obtained results a significant conclusion can be drawn.
Statistical analysis.
Some analyses were carried out only in one year, i.e. one for each year of study. Bearing in mind the weather diversity in the years of study, the such performing analysis seems to be unbelievable.
Discussion.
The discussion is incorrectly conducted. It is not enough to give the results of other authors, without citing specific honeysuckle cultivars, but you have to justify your own results and physiological relationships based on the knowledge available in the literature.
Conclusions.
Lack of any specific conclusions obtained from results. The precise recommendation for each investigated cultivar should be presented. The influence of weather conditions on biochemical compound content changes in honeysuckle berries in particular years should be shown and explained. For example: vitamin C content in relation to high rainfall in May and June in 2020 before fruit harvest (look table 1 and table 4).
L. 411 - not clear
Author Response
Dear Editor,
The authors of this manuscript express their sincere thanks to the reviewers for their excellent and constructive suggestions. The authors have acted upon the recommendations of the reviewers which have resulted in a significant enhancement of the quality of this paper. A point by point response to the reviewers’ comments is outlined below.
Your consideration of the manuscript would be greatly appreciated.
Sincerely yours,
Loredana Vijan
Introduction. The introduction should be more related to the purpose of the experience. Some of the important articles are missing, f.e. : Gołba, M.; Sokół-Łętowska, A.; Kucharska, A.Z. Health Properties and Composition of Honeysuckle Berry Lonicera caerulea L. An Update on Recent Studies. Molecules 2020, 25, 749. https://doi.org/10.3390/ecue omol07les25039 4
Answer: thanks for the suggestion, the author has been cited.
Sampling. Lack of important information; 200 g from one bush or one cultivar or what? How many bushes were cultivated in one replicate? How many replicates? It is not clear.
Answer: additional information was added regarding the composition of the sample (... 200 g per cultivar and harvest) were harvested at full maturity, visually assessed by the appearance of intense violet-blue coloring and the easy detachment of the berries from the plant. Three harvests were performed for each genotype, between the last week of May and the first week of June, and the samples were kept at -18°C until the time of extract preparation (about 14 days).
Table 1.
A sum of rainfall for each season out of 6 months should be presented. The same for sunshine hours. This information is needed for discussion and explaining the results obtained. Moreover, the weather conditions differ each year, hence the year can not be regarded as the factor of the experiment. Instead of that, the experiment should be continued for more years, and based on the obtained results a significant conclusion can be drawn.
Answer: Table 1 shows a climatic characteristic of the area during the research years. The new manuscript followed the influence of each climatic factor on the composition characteristics, the variation of the chemical composition according to the climatic year, with all its characteristics, was highlighted. The experiment will continue, and based on the results obtained, a significant conclusion will be drawn regarding the influence of environmental factors on the composition.
Statistical analysis.
Some analyses were carried out only in one year, i.e. one for each year of study. Bearing in mind the weather diversity in the years of study, the such performing analysis seems to be unbelievable.
Answer: analyzes were performed every year except for the content of lycopene, β-carotene, chlorogenic acid (CA), neochlorogenic acid (NCA), crypto-chlorogenic acid (CCA), catechin (C), rutin (R), and isoquercetin. All analyses were performed in three replicates.
Discussion.
The discussion is incorrectly conducted. It is not enough to give the results of other authors, without citing specific honeysuckle cultivars, but you have to justify your own results and physiological relationships based on the knowledge available in the literature.
Answer: the chapter has been improved.
Conclusions.
Lack of any specific conclusions obtained from results. The precise recommendation for each investigated cultivar should be presented. The influence of weather conditions on biochemical compound content changes in honeysuckle berries in particular years should be shown and explained. For example: vitamin C content in relation to high rainfall in May and June in 2020 before fruit harvest (look table 1 and table 4).
Answer: additional information has been added.
L. 411 - not clear
Answer: the phrase has been reformulated.
Author Response File: 
Author Response.docx
Reviewer 2 Report
The manuscript is well-written, well-structured and clear, and the language is fluent and precise. There isn’t anything novel, and the results are not very interesting but the work is solid. The scientific methods are valid and clearly outlined.
Author Response
Dear Editor,
The authors of this manuscript express their sincere thanks to the reviewers for their excellent and constructive suggestions. The authors have acted upon the recommendations of the reviewers which have resulted in a significant enhancement of the quality of this paper. A point by point response to the reviewers’ comments is outlined below.
Your consideration of the manuscript would be greatly appreciated.
Sincerely yours,
Loredana Vijan
Reviewer #2:
The manuscript is well-written, well-structured and clear, and the language is fluent and precise. There isn’t anything novel, and the results are not very interesting but the work is solid. The scientific methods are valid and clearly outlined.
Answer: We thank you for your patience to read our work, and for your appreciation!
Reviewer 3 Report
Dear Authors,
I am glad that I had the opportunity to review the manuscript entitled: "The influence of genotype and year on some biologically active compounds in honeysuckle berries" by Sina Cosmulescu, Ivona Cristina Enescu (Mazilu), Georgiana Badea and Loredana Elena Vijan.
The purpose of the research presented in this manuscript was to investigate the chemical composition of the berries of three Romanian Lonicera caerulea cultivars.
The research is interesting, although very niche. Nevertheless, they give a good picture of the chemical composition of different cultivars of honeysuckle berries. Undoubtedly, the results of this research should encourage more frequent planting of this species in orchards and gardens.
I believe that in terms of methodology, the research was carried out correctly and the results were well presented. Discussion is interesting and I really like the Conclusions section. However, I have a few comments that should contribute to the improvement of this manuscript:
1) Abstract - the results are discussed in too much detail here, in my opinion it is not necessary to give the numerical values of the tested chemical compounds. In addition, a short research hypothesis and one sentence summarizing the practical significance of the research should be added.
2) Keywords - there are only 3, I suggest expanding this list (you need to add, among other things, the scientific name of the species), and also please arrange these words alphabetically.
3) The biggest disadvantage of this manuscript is the lack of general characteristics of the tested cultivars, because the Authors want to encourage more frequent planting of this plant also in gardens, where it can additionally play the role of an ornamental plant. This description can be found in the Materials and Methods chapter or in the Introduction section (Lines 67-70).
4) Lines 71-75 - please make a clear hypothesis here.
5) Line 78 - please add the scientific name of the species.
6) Overall, the entire manuscript does not fit well with the template required by the journal Horticulturae. Please pay particular attention to the tables. In addition, please check the notation of units if it is in accordance with the SI system.
In conclusion, I believe that the journal Horticulturae should consider publishing this manuscript.
Author Response
Dear Editor,
The authors of this manuscript express their sincere thanks to the reviewers for their excellent and constructive suggestions. The authors have acted upon the recommendations of the reviewers which have resulted in a significant enhancement of the quality of this paper. A point by point response to the reviewers’ comments is outlined below.
Your consideration of the manuscript would be greatly appreciated.
Sincerely yours,
Loredana Vijan
1) Abstract - the results are discussed in too much detail here, in my opinion it is not necessary to give the numerical values of the tested chemical compounds. In addition, a short research hypothesis and one sentence summarizing the practical significance of the research should be added.
Answer: The abstract was reformulated. A short research hypothesis and the practical significance of our research were also added.
2) Keywords - there are only 3, I suggest expanding this list (you need to add, among other things, the scientific name of the species), and also please arrange these words alphabetically.
Answer: We reformulated the Keywords section as suggested.
3) The biggest disadvantage of this manuscript is the lack of general characteristics of the tested cultivars, because the Authors want to encourage more frequent planting of this plant also in gardens, where it can additionally play the role of an ornamental plant. This description can be found in the Materials and Methods chapter or in the Introduction section (Lines 67-70).
Answer: Thank you for this advice! We added a short presentation of the three cultivars in the Introduction section.
4) Lines 71-75 - please make a clear hypothesis here.
Answer: We reformulated the hypothesis.
5) Line 78 - please add the scientific name of the species.
Answer: Thank you again! We added the scientific name of the species.
6) Overall, the entire manuscript does not fit well with the template required by the journal Horticulturae. Please pay particular attention to the tables. In addition, please check the notation of units if it is in accordance with the SI system.
Answer: We checked the manuscript and corrected all found mistakes.
Author Response File: Author Response.docx
Reviewer 4 Report
This research maybe helps to highlight the more valuable honeysuckle cultivar to be planted and exploited. However, some of the presentations are not clear or inaccurate, revisions are needed, questions and suggestions are as following.
1. Please specify the abbreviations for the first time in the abstract, such as GAE, CE, etc. The key words should be reconsidered.
2. Please provide information about the functional compounds and their potential health benefits previously reported.
3. Please check throughout the manuscript about mistakes, for example, ml to mL.
4. For extraction of carotenoid, did you refer to someone’s method? Why so long time (72h) was used in the extraction?
5. Line 394, what’s this mean? And please revise 0.192-0.852% GAE.
6. Line 402, EQ/100?
7. Line 443, mg/g SU? Please specify the abbreviations.
8. Discussion section could be reorganized.
9. Please give a summary of your research in the conclusion section. You did not summarize your results.
Author Response
Dear Editor,
The authors of this manuscript express their sincere thanks to the reviewers for their excellent and constructive suggestions. The authors have acted upon the recommendations of the reviewers which have resulted in a significant enhancement of the quality of this paper. A point by point response to the reviewers’ comments is outlined below.
Your consideration of the manuscript would be greatly appreciated.
Sincerely yours,
Loredana Vijan
- Please specify the abbreviations for the first time in the abstract, such as GAE, CE, etc. The key words should be reconsidered.
Answer: Initially, we made these changes in the abstract. The reason why we used abbreviations was to fit within the word limit. However, following the suggestions of the second reviewer, we reformulated the text. We hope you will agree with the new version!
- Please provide information about the functional compounds and their potential health benefits previously reported.
Answer: I added this information. Thanks for the suggestion!
- Please check throughout the manuscript about mistakes, for example, ml to mL.
Answer: Thank you! Indeed, we had many such mistakes!
- For extraction of carotenoid, did you refer to someone’s method? Why so long time (72h) was used in the extraction?
Answer: Carotenoid extraction was performed with a solvent mixture frequently used in laboratory practice. The ratio between the components of the extractant mixture and the long-time procedure was chosen, based on a previous experiment conducted by the author L. Vijan (data have not yet been published).
- Line 394, what’s this mean?
Answer: Regarding line 394, we are not sure what you are referring to. However, we noticed that we have a different line numbering. For this reason, we hope that the reformulation made is the correct one. Therefore, we reformulated as follows: “Tannins are responsible for food astringent taste, felt following the interaction between salivary proteins and (hydrolyzable) tannins [27]. Large quantities of tannins are found in unripe fruits discouraging their consumption, until the moment their seeds become mature.”. We consider that fruits’ main purpose is to perpetuate species. Therefore, for this to happen, it is necessary for the seeds to reach the appropriate substrate at the optimal time, not before. As frugivores represent one of the plant species spreading vectors, the high tannin content (unpleasant fruit taste) could be a strategy developed by the species to prevent unnecessary seed wastage.
And please revise 0.192-0.852% GAE.
Answer: Thank you! We made the correction.
- Line 402, EQ/100?
Answer: We corrected it and added the explanation in the text: quercetin equivalents (QE).
- Line 443, mg/g SU? Please specify the abbreviations.
We corrected it and added the explanation in the text: dry weight (DW).
- Discussion section could be reorganized.
Answer: The section has been completed.
- Please give a summary of your research in the conclusion section. You did not summarize your results.
Answer: We added our research summary in the conclusion section.
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
The authors did not follow the recommendations to improve statistical calculations. According to statistical knowledge the year cannot be treated as a research factor.
L. 443 by 25 ??? not clear
Author Response
Dear Editor,
The authors of this manuscript express their sincere thanks to the reviewers for their excellent and constructive suggestions. The authors have acted upon the recommendations of the reviewers which have resulted in a significant enhancement of the quality of this paper. A point-by-point response to the reviewers’ comments is outlined below.
Your consideration of the manuscript would be greatly appreciated.
Sincerely yours,
Sina Cosmulescu
Reviewer #1:
The authors did not follow the recommendations to improve statistical calculations. According to statistical knowledge, the year cannot be treated as a research factor.
Answer: This study was carried out in the period 2020–2022 to highlight the content of bioactive compounds in the berries of the three varieties of Lonicera caerulea grown in Romania, i.e. "Cera", "Kami" and "Loni". Two-way ANOVA analysis followed by the Duncan Multiple Range tests (P<.05) was used to study the effect of cultivar and experimental year on total phenolics, tannins, flavonoids, monomeric anthocyanins, and vitamin C. The composition was different from one year to another, within the same variety, and, as a result, statistical analysis was applied to see if the fluctuations recorded from one to another had a greater or lesser significance. The analysis of the influence of each climatic factor (temperature, light, etc.) on the composition was not followed. The climatic characteristics of the research area are presented in the material and method chapter.
- 443 by 25 ???
Answer: We have corrected it.
