Comparative Transcriptome Analysis Revealed Two Alternative Splicing bHLHs Account for Flower Color Alteration in Chrysanthemum

Round 1

Reviewer 1 Report

Manuscript: ijms-1472269 Comparative transcriptome analysis revealed two alternative splicing bHLHs account for flower color alteration in Chrysanthemum

In the manuscript, the authors to investigate the underlying mechanism of this process a comparative transcriptome was analyzed between white and turning red ‘Jimba’ chrysanthemum. Authors found the structural and regulatory genes of anthocyanin pathway were significantly up-regulated in turning red ‘Jimba’. Among them, two alternative splicings CmbHLH2 and CmbHLH2.1 showed the most significantly up-regulated in turning red ‘Jimba’ chrysanthemum. Both CmbHLH2 and CmbHLH2.1 could interacted with CmMYB6 to activate CmDFR promoter, which involved in regulation flower color alteration. The aim of the paper is comparative analysis of transcriptome and transcription factors between white and turning red ‘Jimba’. It is important for fundamental science.
The uniqueness of the text is about 90% by AntyPlagiarism.net.
The manuscript is written well. English is proper, well understandable. But I found some mistakes.

Reviewer has some comments:
Line 17 - remove dot between - of this process. A comparative. - It should be one sentence. Combine them.
Line 36 - flower - should be - flowers.
Line 37 - turning - should be - turn.
Line 40 - were - should be - was.
Line 50 - are belong to IIIf subfamily which recognized - should be - belong to IIIf subfamily which is recognized.
Line 51 - were - should be - was.
Line 85 - were - should be - was.
Line 91 - database - should be - databases.
Line 100 - libraries - should be - library.
Line 150 - tree - should be - trees.
Line 152 - were - should be - was.
Line 152 - shown the close - should be - showed a close.
Line 159 - also analyzed - should be - was also analyzed.
Line 172 - regulator were - should be - regulators were.
Line 175 - tree were - should be - trees were.
Line 175 - constucted - should be -  constructed.
Line 183 - 614 amino acid - should be  - 614 amino acids.
Line 188 - gene structural analysis - should be - gene structure analysis.
Line 203 - including - should be - includes.
Line 213 - were conducted - should be - was conducted.
Line 216 - could be grow - should be - could be growing.
Line 231 - at 4th day - should be - on the 4th day.
Line 233 - anthcocyanin - should be - anthocyanin.
Line 233 - at 8th day - should be - on the 8th day.
Line 234 - paches - should be - patches.
Line 237 - were analyzed - should be - was analyzed.
Line 266 - assay were - should be - assays were.
Line 289 - which regulated - should be - which is regulated.
Line 302 - chrysanthmum - should be - chrysanthemum.
Line 302 - who also involved - should be - which also is involved.
Line 303 - could induced - should be - could induce.
Line 342 - were cultivated - should be - was cultivated.
Line 336 - It worth to further investigate - should be - It is worth further investigating.
Line 332 - fails to regulate - should be - failing to regulate.
Line 330 - as far as we known - should be - as far as we know.
Line 322 - were further confirmed - should be - was further confirmed.
Line 324 - the regulatory effects is far - should be - the regulatory effects are far.
Line 323 - could formed a complex - should be - could form a complex.
Line 318 - region binds DNA - should be - region that binds DNA.
Line 317 - which been constituted - should be - which has been constituted.
Line 314 - ensures a strong - should be - ensure a strong.
Line 348 -  used for total RNA extraction was described previously - should be -  is used for total RNA extraction as described previously.
Line 431 - each experiment were carried out more - should be  - Each experiment was carried out with more
Line 417 - were transformed - should be was transformed.
Line 420 - were tested - should be - was tested.
Line 416 - were into pAbAi - should be - was into pAbAi.
Line 405 - according the - should be - according to the.
Line 443 - assay were - should be - assays were.
Please add to the Diss\cussion part some discussion about Figure 6. Protein-protein and protein-DNA interaction in yeast. Please explain these interactions in detail.

Please check English one more time.

No other comments.

Minor revisions are required.

Comments for author File: Comments.pdf

Author Response

Point 1：Mistakes in English grammar

Response 1：Mistakes in English grammar have been modified throughout.

Point 2：Please add to the Diss\cussion part some discussion about Figure 6. Protein-protein and protein-DNA interaction in yeast. Please explain these interactions in detail.

Response 2: It has been add into the Discussion section (Line347-355).

Point3: Please check English one more time.

Response 3: It has been checked.

Reviewer 2 Report

The presented article is devoted to the investigation of mechanisms of
Chrysanthemum flower color alteration. Presented material is an interesting
model for the evaluation of the genes expression mechanisms, which may be
changed and lead to an alternative phenotype expression. Also it was shown
that alternative splicing plays the main role in flowers coloring in mutant
form of Chrysanthemum.
Unfortunately, there are many mistakes in English grammar. The article can be
published after correction of these mistakes.

Author Response

Point 1: There are many mistakes in English grammar. The article can be
published after correction of these mistakes.

Response 1: Mistakes in English grammar have been modified throughout.