Global Methylation Analysis Using MSAP Reveals Differences in Chilling-Associated DNA Methylation Changes during Dormancy Release in Contrasting Sweet Cherry Varieties
, Claudio Meneses 4,5 and Andrea Miyasaka Almeida 6,7,*Round 1
Reviewer 1 Report
This manuscript aims to reveal the difference in DNA methylation associated with dormancy transition in three cherry varieties. The authors firstly compared the chilling requirements of three cherry varieties and then analyzed the methylation changes using MSAP. Here are a few comments before the consideration of acceptance.
1. The introduction is too breif. Please include some information about the biological understanding of ENDO and ECO in trees.
2. Why not use young plants following the dormancy process and chilling treatment? Cuttings were directly used for the chilling requirement test by placing in 4 °C. Thus there will be additional environmental stresses beside Chilling, such as wounding and dehydration, which may affect the methylation profiles.
3. In Table 3. There are different bands detected in three cherry varieties, 6 in RD, 7 OM GR and 3 in K. Why there are only 8 genes identified in Table 5?
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Dear Authors
Undoubtedly, DNA-methylation and de-methylation plays important role in plants’ life cycle. I appreciate the effort and great idea but unfortunately the work has no experimental replicate. The bud samples were collected during June 2015. What about the next years? Can we trust one year experiment? My suggestion is to repeat experiment at least for 2 years. Moreover, it is not clear from how many trees per each genotype you did sampling and who many time you performed MSAP.
Best wishes
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 3 Report
It's a very interesting report about epigenetic memory on dormancy release of buds taking into account cherry as a biological model. The paper is very well written and, therefore, I recommend accepting it.
Author Response
Reviewer 3. Comments and Suggestions for Authors
It's a very interesting report about epigenetic memory on dormancy release of buds taking into account cherry as a biological model. The paper is very well written and, therefore, I recommend accepting it.
Answer: Many thanks for the comment.
Round 2
Reviewer 1 Report
I appreciate the authors' responses basically addressed my concerns.
Reviewer 2 Report
Dear Authors
Having low chill genotypes from different region and high chill genotype from other region is not really accurate way to study the dormancy even if the genotypes are well characterized. There is only 3 genotypes 2 low and 1 high chill. I don’t think only one or two genotype is enough. The candidate genes and their methylation statue in Table 5 make me to hesitate about the reliability of the results. More genotypes and at least 2 years of experiment are needed.
Best wishes
