Establishment, Multiplication, and Biochemical Analysis of Embryogenic Lines of the Amazonian Palm Euterpe precatoria Mart. under Suspension Culture
, Joane dos Santos Neves 2, Anderson Marcos de Souza 1, André LuÃs Xavier de Souza 3 and Jonny Everson Scherwinski-Pereira 3,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsPlease do correction according to manuscripts. Abstract need to be clear and divide the scope accordingly (Intro, Problem statement, Methods/ Missing bar scale and the title need to be concise, Some title in Results need to change, Conclusion). Methods need to be corrected and Conclusion need to be rewrite accordingly. See the comments accordingly. A lot of grammatical error in the manuscript. Suggestion to get proof read if possible.
Comments for author File: 
Comments.pdf
Need to prof read accordingly if possible.
Author Response
All reviewers' comments have been carefully evaluated and addressed. The detailed response to the reviewers can be found in the file "v1_AuthorResponses_Forests_.docx".
Author Response File: Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThis manuscript on "embryogenic" callus propagation touches an important aspect of clonal palm production in general. The authors describe a comparison in methodology based on medium type and provenance of starting material to propagate this type of callus. They tried to fit Gompertz curves to the acquired growth data and did a principal component analysis of growth, as well as a biochemical analysis of sugar and protein content.
The introduction was very nice and to the point, although some recent economic numbers at line 46-47 could have been provided to set the actual importance of açai palm.
In the methods section, at various occasions it is mentioned that medium is "similar to the MS medium described above" (line 103, 114) it is not clear whether these media still contain charcoal, and all the hormones. It would be best to write the additions in full for clarity.
The study of the growth kinetics is warranted, but lacks early time points and therefore very valuable information (to the authors admittance at the beginning of 4.2) to properly fit a model to the measurements. With the current data points, it is probably equally possible to fit a linear growth model instead of the Gompertz. I would therefore suggest to repeat the effort with the "best" end result, using additional early time points. It would also be good to have error bars on the data points of the growth curves.
In the PC analysis graph the label L7 is missing. And a confusing figure reference is given in line 557.
Although the PCA seemingly yields separation of components, I urge the authors to condense the analysis and stick to the relevant points, that can be interpreted afterwards in biological terms.
For the Biochemical analysis (3.4), it would be good to know what the incentive is of doing such analysis, and why it is important to understand the growth of the callus. What is the biological significance of the findings?
I noticed that TSS in the results becomes AST in the discussion, and TSP becomes PTS. Please uniformize.
A major point that is missing in the study is to know whether all callus lines are indeed embryogenic, and led to the production of plants in the end. Such data would increase the value of the study enormously. At least what the authors conclude to be the "best" treatments, should be proven to still be leading to plant production.
The discussion is rather lengthy and can be shortened, while the conclusions are concise. A framing of the work in a broader context, what this work contributes to clonal production of açai, what is the take home message for producers and academics is warranted.
Comments on the Quality of English Language
Everything is understandable. line 16 has some superfluous words.
Author Response
All reviewers' comments have been carefully evaluated and addressed. The detailed response to the reviewers can be found in the file "v1_AuthorResponses_Forests_.docx".
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsDear authors, to improve the article it is necessary to clarify certain points.
Line 93: The colon at the end of the sentence must be removed.
Lines 94-136: For each stage of induction and multiplication, carefully indicate which and in what quantity of gelling agent was used or not.
Abbreviations must be deciphered the first time they are mentioned (MS, Y3. etc.).
Figure 3 is of poor quality and uninformative.
Dear authors, why do you call callus cultures embryogenic? What new structures are you writing about (lines 306-307)? Provide evidence of the formation of embryoids in callus cultures. Accordingly, the text needs to be revised.
The statement «Analyzing the curves, it is noticeable that the lag phase was imperceptible in the 454 growth of all analyzed lineages» is incorrect (lines 454-455). Intervals for analyzing growth characteristics are too long.
Author Response
All reviewers' comments have been carefully evaluated and addressed. The detailed response to the reviewers can be found in the file "v1_AuthorResponses_Forests_.docx".
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
Comments and Suggestions for AuthorsThis is the second time I read this manuscript. The authors have addressed many of the comments I made, and I am particularly pleased to see they did an analysis of callus lines to test for embryogenic character.
However, the authors also ignored my comment on the lack of data in the growth analysis to perform a decent modelling.
"The study of the growth kinetics is warranted, but lacks early time points and therefore very valuable information (to the authors admittance at the beginning of 4.2) to properly fit a model to the measurements. With the current data points, it is probably equally possible to fit a linear growth model instead of the Gompertz. I would therefore suggest to repeat the effort with the "best" end result, using additional early time points. It would also be good to have error bars on the data points of the growth curves."
Trying to fit the Gompertz model at this point seems somewhat superfluous, and would need earlier data points in the graphs. If the authors choose to keep the Gompertz modelling, they should add early data time points. The story by itself I believe is also worthwhile publishing without the Gompertz modelling.
Comments on the Quality of English LanguageHave an ultimate check up by a native speaker.
Author Response
Dear Reviewer,
We would like to thank you for your suggestions for improving our manuscript.
All responses to suggestions can be found in the file "v2_AuthorResponses Forests.docx" sent as an attachment.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsDear authors, you have significantly improved the manuscript.
Please add an explanation of the abbreviation for medium Y3. It is Eeuwens and Blake medium (Eeuwens CJ, and Blake J. Mineral requirement for growth and callus initiation of tissue explants excised from mature coconut palms (Cocos nucifera L.) and culture in vitro. Physiol. Plant. 1976; 36:23-28.)
Correct the abbreviation 'ss' to 'se' (somatic embryo) in Figure 12. In Figure 12 B, correct 'es' to 'se'.
Author Response
Dear Reviewer,
We would like to thank you for your suggestions for improving our manuscript.
All responses to suggestions can be found in the file "v2_AuthorResponses Forests.docx" sent as an attachment
Author Response File: Author Response.pdf
