Identification of the Brassica Campestris ssp. Chinensis BcHY5 Gene Important for Hypocotyl Length

Round 1

Reviewer 1 Report

1)      Graphical representation of abstract is missing

2)      Have not compare any experiment with wild type Brassica campestris or with other sub species of Brassica campestris.

3)       As from the Fig-1b, Arabidopsis and Brassica species share more recent common ancestor than that of tobacco but experiments have done in tobacco plant while it can be done in more closely related species of NHCC or Brassica campestris.

4)      In fig 1b, even though the phylogenetic tree shows higher similarity with BnHY5 and BrHY5, but AtHY5 has been chosen as orthologue. Kindly explain.

5)      In Fig-1a, c result of motif analysis have shown that the identical motif exist in BcHY5, BrHY5, BnHY5, and RsHY5 , which was similar to that in AtHY5 but have not shown similarity with Nicotiana tabacum (NtHY5) and how much percentage of similarity in structure of motifs between different species also have not discussed.

6)       Provide possible reasons for increase then decrease in relative expression of HY5 during salt and dark treatment (Fig-2b).

7)      If BcHY5 expression occur during light condition then why there is less expression of BcHY5 under minimum time of exposure to dark period (Fig-2b).

8)      A control graph showing BcHY5 expression during light condition at different time points should be given to compare other graphs with treatment (Fig-2b).

9)      Define clearly how expression of HY5 is related to AtHY2 and AtYUC8 expression in Fig-3d.

10)  Whether BcBBX24 form some protein complex to repress HY5 expression.

11)  Repetitions of sentences in Experiment 2.4 of Materials and methods.

12)  In fig-1a, b, c there is incorrect abbreviation used for tobacco i.e. NaHY5 instead of NtHY5.

13)  Provide a good resolution images of Fig. 4a (Yeast two hybrid) and Fig.6 a,b (Yeast one hybrid).

14)  As there can be more than one GC box present in the promotor of BcHY5 (as shown in Fig. 6a), there is a need to specify the particular GC Box Where BcBBX24 Bind.

15)  In the fig 6a, the estimated size and position of the sequence used for yeast one hybrid is not mark.

16)  Additionally, the interaction between the BcHY5 and BcBBX24 can be supported by in-vivo studies, like Chip-qPCR.

17)  In the figure 6, you perform only in vitro experiment to check interaction between BcHY5 and BcBBX24. You should also check In vivo interaction between BcHY5 and BcBBX24, like Chip q-PCR.

18)  Check all the reference thoroughly.

Author Response

Please see the attachment. Thanks！

Author Response File: Author Response.pdf

Reviewer 2 Report

Summary:

In this manuscript, Li et al. found the BcHY5 gene in Brassica campestris ssp. Chinensis, which is a homolog of the AtHY5 gene in Arabidopsis thaliana. In Arabidopsis thaliana, overexpressing BcHY5 can shorten the plant's hypocotyl. Additionally, the BcBBX24 protein was found to interact with BcHY5 and reduce BcHY5 expression by binding to the promoter region of BcHY5. In conclusion, The manuscript is well-written and this work will improve our understanding of BcHY5 function and how BcBBX24 modifies BcHY5 expression level.

Some minor suggestions:

1. Fig. 1 and Fig. 4 need to be adjusted because of some improper stretching.

2. The histograms are all of low quality. Please try to create your histograms using Prism or any softwares that can produce high-quality histograms in PDF files.

3. In the ABA area of Fig. 2b, please set the Y axis to two segments so that the histogram displays both values below 0.5 and above 5.0.

4. In Fig. 4a, pGBTK7 + pGADT7-BcBBX24 should be added as a negative control for the Y2H experiment.

5. The caption "BcHY5-OX*BcBBX24-OX" in the x axis of Fig. 5b-5d is too wide. Please change it.

Author Response

Please see the attachment. Thanks.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

May be accepted in the current format