PbZAT12, Independent of PbMYB10, Activates Structural Genes to Promote Anthocyanin Biosynthesis in ‘Red Zaosu’ Pear Fruit (Pyrus bretschneideri Rehd.)

Round 1

Reviewer 1 Report

23.6.2023

 

The revised manuscript entitled "PbZAT12, rather than PbMYB10, activates structural genes to promote anthocyanin biosynthesis in 'Red Zaosu' pear fruit (Pyrus bretschneideri Rehd.)" was reviewed.

The manuscript delivers novel insights in anthocyanin biosynthesis in pear fruit peel. A very nice manuscript, that I enjoyed reading. However, it was very biased to PbZAT12 compared to MYB10. In addition, RNAseq data should be presented and deposited to the Genbank and discussion needs more details. Therefore, I do not recommend the publication of this manuscript in its current form in "Horticulturae" unless the following comments are considered in a revised version.

 

1. General:

- The title is very aggressive. The data delivered in this work sports the importance of PbZAT12 in anthocyanin biosynthesis. Nonetheless, previous reports support a similar role for MYB10, therefore, modifying the title is necessary!!!

For example: PbZAT10, a novel player that activates …….

 

- The scientific name should be italicized in the title (Punica granatum L.).

- The manuscript language is good but it needs some corrections, please check the entire manuscript for weak phrases and common structural mistakes. Few corrections were mentioned in the "Abstract", please see below.

- Do not leave space between degree and symbol. Change “80 °C” to “80°C”.

- On the other hand, keep a space for all other units, e.g. change “800bp” to “800 bp”, and “200uM” to 200 uM”.

 

2. Abstract:

- Line 18: replace “determined to localize” to “localized”.

- Line 18: replace "favorable” to “positive”.

- Line 20: remove “gene”.

- Line 21: replace “the direct” to “a direct”.

- Line 21: replace “to the” with “to”.

- Line 22: replace “as” with “which was”.

- Line 22: replace “the dual” with “dual”.

- Line 25: add “biosynthesis” after “anthocyanin”.

 

3. Introduction:

- Lines 69-75: These are brief results replicated from “Abstract”. Please remove!

- Lines 76-70: these are brief results replicated from “Conclusions”. Please remove!

- Line 79: Add a explicit sentence indicating the study objective!

 

3. Materials and Methods:

- Lines 95-99: important details are missing including RNA isolation procedure or kit, DNA digestion, Library type, read length and nature, total reads, assembly, annotation, data analysis, … etc. !!!

- Line 97: replace “(CS)” with “(RS)” and vice versa!!!

 

4. Results:

- Lines 171-181: You did RNAseq for three tissues. The data is missing! You need to characterize them for example using a volcano plot” and indicate quality control features (coverage, annotations, … etc.).

- Lines 173-174: according to “Fig. 1a” “PbZAT12” IS NOT “the most different RS vs GS expression”!!! but it is rather “PbMYB114” according to exhibited heat map color scale!!!

Please you need to correct that and indicate the importance of both genes in anthocyanin biosynthesis!!!  

- Line 174: what is “FPKM”? Many readers would not know what does it stand for!

- Line 202: scale bars are needed for panels a and b.

- Line 232: scale bars are needed for all.

 

5. Discussion:

- The first paragraph (lines 255-274) has nothing to do with discussion! It covers a part of “introduction” and some details about “the problem statement”. Please re-write!

- This keeps us with only two paragraphs, which are very good. I really enjoy reading the second paragraph (lines 275-297), however, more details are needed to compare between expected and proved actions of both PbZAT12 and PbMYB10b!

- Therefore, the discussion is relatively SHORT and it is recommended to tackle other aspects of anthocyanin biosynthesis with more details!

- A representative activation model based on this work and your previous studies would deliver a clearer picture and would elevate the value of this manuscript.

 

6. Data availability:

- Line 328: RNAseq data should be deposited in the Genbank as SRA. This is the norm to benefit the scientific community!!!

 

7. References:

- It is recommended to incorporate more RECENT articles as only one fifth (11 out from 57) of cited articles were published in the last five years. 

- The manuscript language is good but it needs some corrections, please check the entire manuscript for weak phrases and common structural mistakes. Few corrections were mentioned in the "Abstract" comments.

Author Response

Thank you for your valuable feedback on our research. Your insightful suggestions have greatly contributed to our further studies. I have provided a detailed response to each of your comments in the attached supplementary document.

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript by Liu et al. explored a novel transcription factor ZAT12, which plays a key role in anthocyanin accumulation during pear fruit development. The authors gave reliable evidence that ZAT12 can directly bind to the promoters of anthocyanin synthesis-related genes and positively regulate their transcription. And this regulation mode does not depend on MYB10. In addition, the author verified the role of ZAT12 in anthocyanin synthesis by transient expression method. The article is well written and the results support the conclusions.

Minor comments：

1. The title should be changed to “PbZAT12, independent of PbMYB10, activates structural gene to promote anthocyanin biosynthesis in XXXX”.

2. Line 24, MYB10 should be PbMYB10

3. Line 82-90, the authors should give a specific time of injection, such as days after full bloom.

4. Line 95-99, how is the transcriptome library constructed and how is the transcriptome data analyzed? Authors should cite references.

5. Line 103-106, the authors only describe the extraction method of anthocyanins, but the measurement method is missing.

6. Line 108-110, the authors should describe how the QRT-PCR was set up.

7. Line 253, Here the author needs to explain the meaning of * and **.

Author Response

Thank you for your valuable feedback on our research. Your insightful suggestions have greatly contributed to our further studies. I have provided a detailed response to each of your comments in the attached supplementary document.

Author Response File: Author Response.docx