The Isolation and Identification of Anthocyanin-Related GSTs in Chrysanthemum
Round 1
Reviewer 1 Report
Here are few issues needed to be addressed.
Authors can use amino acid sequences for the preparation of phylogenetic trees.
include all the 94 accession numbers used for 94 CmGSTs genes in the phylogenetic tree.
use italics in the tree for genes name
Authors can revise the 2C qRT-PCR result and convert it to fold change data.
Provide accession numbers used in Figure 3.
What was the query sequence taken for the BLAST study?
Authors can show the BLAST result as an additional figure including all the scores ( queries coverage, percent identity, values, etc.)
The writing of the results section can be improved. Analyze the finding in each result section.
Some gene name is not in italics in the abstract section. please correct them.
Author Response
Point 1: Authors can use amino acid sequences for the preparation of phylogenetic trees.
Response 1: Amino acid sequences were used to construct phylogenetic tree in revised manuscript (Figure 1).
Point 2: include all the 94 accession numbers used for 94 CmGSTs genes in the phylogenetic tree.
Response 2: 94 accession numbers have been presented in phylogenetic tree in revised manuscript (Figure 1), which download from chrysanthemum genomic database (http://mum-garden.kazusa.or.jp).
Point 3: use italics in the tree for genes name
Response 3: Amino acid sequences were used in present phylogenetic tree and no gene names.
Point 4: Authors can revise the 2C qRT-PCR result and convert it to fold change data.
Response 4: Fold change data has been presented. To keep consistent with phylogenetic tree, gene name were replaced by accession numbers in revised manuscript (Figure 2).
Point 5: Provide accession numbers used in Figure 3.
Response 5: Genbank accession numbers have been added in the legend of Figure 3 (Line 233-238 ) in revised manuscript.
Point 6: What was the query sequence taken for the BLAST study? Authors can show the BLAST result as an additional figure including all the scores ( queries coverage, percent identity, values, etc.)
Response 6: We used CmGST protein sequence from chrysanthemum genome to construct phylogenetic tree (Figure 1), the sentence ‘....local-blast was used to search 94 GSTs in Chrysanthemum genomic database’ has been changed as ‘Protein sequence of 94 CmGSTs were screened from Chrysanthemum genome (http://mum-garden.kazusa.or.jp) based on annotation’ (Line 173-174) as well.
Point 7: The writing of the results section can be improved. Analyze the finding in each result section.
Response 7: Result sections 3.1, 3.3, 3.4, 3.5 have been improved, and the finding has been analyzed in each result section.
Point 8: Some gene name is not in italics in the abstract section. please correct them.
Response 8: It has been corrected.
Reviewer 2 Report
Well designed experiments. It was important to show the role of the transcription factors which would be required to boost anthocyanin levels sufficiently to allow for visualization of sequestration. You may wish to cite Scientia Hoticulturae 189:132.
Author Response
Point 1: It was important to show the role of the transcription factors which would be required to boost anthocyanin levels sufficiently to allow for visualization of sequestration.
Response 1: Re-writing of result section 3.6 (transient overexpressed assay) and 3.7 (dual-luciferase assays), suggested that CmMYB6 and CmbHLH2 could sufficiently to activate CmGST1 and boost anthocyanin levels.
Point 2: You may wish to cite Scientia Hoticulturae 189:132.
Response 2: It has been cited in revised manuscript (Line 343-344).
Reviewer 3 Report
This paper is presented very well and well written. Only a few minor errors. The research and conclusion looks to be very strong.
I congratulate the authors
Comments for author File: 
Comments.pdf
Author Response
Point 1: This paper is presented very well and well written. Only a few minor errors. The research and conclusion looks to be very strong.
Response 1: It has been revised.
Round 2
Reviewer 2 Report
Suitable revisions have been made. You may want to change the introductory line to "Chrysanthemum production is one of the four largest cut flower markets in the world". It is not the fourth largest cut flower as there are many cut flowers that are larger in physical size.
Author Response
Point1: Suitable revisions have been made. You may want to change the introductory line to "Chrysanthemum production is one of the four largest cut flower markets in the world". It is not the fourth largest cut flower as there are many cut flowers that are larger in physical size.
Response1: The introductory line "Chrysanthemum is one of the four largest cut flower in the world" has been changed as ''Chrysanthemum (Chrysanthemum morifolium Ramat.) is one of the most popular ornamental plant in the world'' (Line 29-30)
Reviewer 3 Report
The paper corrections are excellent, but one missed
line 51 could't should be couldn't
Author Response
Point1: The paper corrections are excellent, but one missed
line 51 could't should be couldn't
Response1: It has been revised
