Brassinolide Alleviates Chilling Injury of Sweet Cherry (Prunus avium L. cv. Tieton) during Cold Storage

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The authors proposed a new treatment to reduce chilling injury in cherries using a product obtained from Brassinolide.

-          The objective of the study should be included in the abstract.

-          The preparation of the BR treatment should be explained.

-          In the introduction, a review of other methodologies to reduce chilling injury in cherries could be added. Besides, a review of the main parameters related to cherry quality could also be added (and the methodology used to meassure them).

-          Lines 78-79: How many fruits were used (for the measurements) in each set of 2 kg? Were the tested fruits selected from the ones in the set of 2 kg according to any criteria?

-          Which was the base to decide the concentrations of BR used in the study?

-          The number of fruits that were tested in each measurement (destructively and non-destructively) should be indicated.

-          Line 113-116: ten fruits were tested from each group (firmness test); how many repetitions were developed per fruit? Considering the small size of the fruit (the difficulties in carrying out repetitions per fruits) and the variability of fruit firmness in each group (Figure 1 shows high visually variability), more than ten fruits would have been necessary.

-          Figure 1: it is crucial to show a photo with a higher number of cherries. There are 5 fruits per group and day except in day 28. Were some fruits seriously deteriorated after 28 days storage?

-          Figure 1: the variability in each group/day combination is high. A photo with the amount of fruit tested per day treatment would be very representative.

-          In Figures 2,3,4,5, and 6 the number of repetitions per day and treatment should be indicated.

-          A two-way Anova analysis could be developed to assess the significant effect of the factors BR treatment level and day.

-          Figure 3: The colour parameters shown in Figure 3 do not agree with the images shown in Figure 1. For example, in day 28 the treatments CK and CL2 look very similar compared to the treatment CL. However, in the colour parameters curves shown this result is completely different.

-          In the Discussion section a clearer reference to the results obtained with the study could be added.

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors

Thanks for your invitation. Cherry is among the most studied garden plant species. Therefore, its preservation is also important. This study is an original study in this respect. I am of the opinion that it will make significant contributions to the literature. The article is generally well prepared, written in a professional language, and modern techniques are used. It is well discussed with other studies in the literature. However, significant revisions are needed at some points;

Minor revisions

The article abstract is poorly written. It is recommended to add some numerical data from the study results. Which type was used should be written in the abstract.

A paragraph can be added to the introduction section that includes topics such as the world production value of cherries, their importance for human health, etc.

Please write the full name where the abbreviations are first used.

Check the use of uppercase letters in chemical symbols.

Check the punctuation throughout the article.

 

Major recommendations

Image 1, and other graphics presented in the text, have poor image quality. Enlarge Picture 1. Adjust the size of other images so that they can be clearly understood. Additionally, graphically generated data can be exported as a supplementary file.

 

material method,

Conditions such as climate, rootstock, cultural processes, etc. have an effect on storage. There is no data in the article regarding these parameters

The variety, rootstock, soil and climate values used in the study should be given.

Presenting correlation analysis on the parameters examined will make significant contributions to the scientific nature of the study.

The conclusion of the study is poorly written. Please elaborate here.

To summarize;

I think the article will be in a better position after the suggestions are made. The article contains scientifically important results for the cherry species and will contribute to science. Congratulations to the authors.

Author Response

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Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors have detailly answered most of the questions asked by the reviewer. However, there are some information that could be added:

Comment 6: The number of fruits that were tested in each measurement (destructively and non-destructively) should be indicated.

Response 6: Thanks to your suggestion, we have added the following to the manuscript (lines 107-111). Fifteen fruits were randomly selected from each treatment group every 7 d for determination of chilling injury index, stem freshness index, weight loss, color, firmness of 10 fruits, and sugar-acid ratio, vitamin C , total phenolics and flavonoids, and antioxidant status of 15 fruits, with three replicates in each group.

Does it mean that 15 fruits were tested every 7 days from each treatment for all the magnitudes measured (except firmness that 10 fruits were tested)?

Why were not the non-destructive measurements carried out with a higher number of fruits? Only fifteen fruits for weight loss, chilling injury index, stem freshness index seems to be a reduced number and colour. For example, the high variability of the colour parameters could be seemed in Figure 1 since day 0.

Were the 15 fruits non-destructively tested always the same fruits?

Which criteria was used to select the 10 fruits of the firmness measurement from each treatment and day?

Were the same 10 fruits of the firmness measurement from each treatment and day used for the other destructive tests?

 

Comment 11: A two-way Anova analysis could be developed to assess the significant effect of the factors BR treatment level and day.

Response 11: Thank you for your insightful suggestion to conduct a two-way ANOVA analysis. After careful consideration, we have decided to maintain the one-way ANOVA analysis for the following reasons: Our study was primarily designed to assess the impact of different concentrations of brassinolide (BR) on the quality of sweet cherries during cold storage, rather than to explore the interaction effects with time. The one-way ANOVA has provided clear evidence of the effect of BR treatment on the quality indicators of interest, which aligns with our research objectives. We have ensured that the assumptions of one-way ANOVA, including homogeneity of variances and normality of residuals, are adequately met by our data. Introducing a two-way ANOVA may complicate the analysis without significantly enhancing our understanding of the effects of BR treatment. Our findings, based on one-way ANOVA, are robust and consistent across all measured quality indicators, we believe that the one-way ANOVA is the most appropriate statistical method for our study design and objectives, and it has allowed us to present a clear and concise analysis of the impact of BR treatment on sweet cherry quality during cold storage. We appreciate the opportunity to clarify our approach and hope this explanation addresses your concerns.

Thank you for the response. However, considering that the experimental design has two important factors (both of them clearly studied in the experiment) the interaction of both (treatment and storage time) it would be very interesting and could be used to summarize the results.

Comment 12: Figure 3: The colour parameters shown in Figure 3 do not agree with the images shown in Figure 1. For example, in day 28 the treatments CK and CL2 look very similar compared to the treatment CL. However, in the colour parameters curves shown this result is completely different.

Response 12: Thanks for your correction. We apologize for the low quality of the images we provided causing the CK and CL2 groups to look similar in color. We have re-processed and resubmitted these images to improve the clarity and you can see that on day 28, the cherries 3 in the CK group were so badly decayed that the surface color had turned reddish-brown or even greyish-brown. In the CL2 group, the color of the fruit was bright red.

Thank you for improving the images. However, the information provided in Figure 3 (related to colour) still does not agree with the colour of the cherries shown in Figure 1 (specially day 28 CL1, colour parameters in Figure 3 and visual colour in Figure 1). This fact should be checked. Maybe colour parameter “a” could be shown.

-         -->  The authors have provided the data file but only with the average values. It would be important to show the measured magnitude values from each fruit tested (the completely data file).

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The author made the revisions I suggested.

Author Response

Thank you for your valuable time and professional review.

Round 3

Reviewer 1 Report

Comments and Suggestions for Authors

-          The authors indicated in their responses that the number of fruits were “…45 fruits sampled at one time for each treatment group…” and for firmness 30 fruits. The y also replied that “Additionally, we uploaded the measurements for each tested fruit in an additional attachment “. However, in the provided data file the repetitions (per day and treatment) are different (3 repetitions per day and treatment for all, the variables except firmness and L that there are 10 repetitions per day and treatment). It is crucial to clarify this fact.

Author Response

Comments: The authors indicated in their responses that the number of fruits were “…45 fruits sampled at one time for each treatment group…” and for firmness 30 fruits. The y also replied that “Additionally, we uploaded the measurements for each tested fruit in an additional attachment “. However, in the provided data file the repetitions (per day and treatment) are different (3 repetitions per day and treatment for all, the variables except firmness and L that there are 10 repetitions per day and treatment). It is crucial to clarify this fact.
Response: Thank you for your review. We reviewed the raw data and processing methods. We found an error in our description and have now made changes in lines 111 and 144-146 of the manuscript.
The three indicators （chilling injury index, stem fresh-ness index,  and weight loss） do not suit the representation of data from individual fruits, so we provide the data in the form of three parallel data for each group.
For the determination of the data other than firmness and color, the amount of cherries needed was small and the unit of sampling needed was g. To reduce the experimental error due to the large individual differences in cherries, so we took the mass of the required sample after cutting or grinding and mixing the fruits of each group and made three parallels. Therefore the data presented represent each parallel.
The firmness was measured twice in the equatorial plane of each fruit and averaged for each fruit to make the data more accurate, and the minimum and maximum values were removed. For L* and a*, data from 15 fruits were measured, the minimum and maximum values were removed and 10 data were retained for consistency with the number of firmness data.