FTIR Screening to Elucidate Compositional Differences in Maize Recombinant Inbred Lines with Contrasting Saccharification Efficiency Yields
, Rogelio Santiago 1, Rosa Ana Malvar 2, Daniel Martín 3, Inês Pereira dos Santos 3, Luís A. E. Batista de Carvalho 3, Laura Faas 4, Leonardo D. Gómez 4 and Ricardo M. F. da Costa 3,5Round 1
Reviewer 1 Report
The authors report on the characterization of the maize MAGIC population that showed variability for saccharification efficiency.
The obtained results were expected to deepen the available knowledge on the relationship between cell wall composition and biorefining potential.
A FTIR-ATR spectroscopy were applied to elucidate the compositional differences between high and low saccharification yielders.
It was found that high and low saccharification efficiency groups differed in cell wall compositional features: high saccharification RILs displayed higher proportions of S subunits, aromatic compound, and hemicellulose in contrast to low saccharification efficiency groups for which FTIR analysis showed higher proportions of lignin richer in subunits G, greater proportions of crystalline cellulose and acetyl methyl esters.
The aim of the study is interesting. The authors compiled a large amount of characterization data.
Yet, the authors should the precise indicate the novelty of presented paper and should be more specific in the Experimental part - how they ensured spectral comparability.
Author Response
Dear Reviewer,
Please find enclosed the revised version of the manuscript ID: agronomy-1214120 Title: " FTIR Screening to Elucidate Compositional Differences in Maize Recombinant Inbred Lines with Contrasting Saccharification Efficiency Yields" submitted to Agronomy
We have addressed responses to all your main suggestions and made changes accordingly
Comments and Suggestions for Authors
The authors report on the characterization of the maize MAGIC population that showed variability for saccharification efficiency.
The obtained results were expected to deepen the available knowledge on the relationship between cell wall composition and biorefining potential.
A FTIR-ATR spectroscopy were applied to elucidate the compositional differences between high and low saccharification yielders.
It was found that high and low saccharification efficiency groups differed in cell wall compositional features: high saccharification RILs displayed higher proportions of S subunits, aromatic compound, and hemicellulose in contrast to low saccharification efficiency groups for which FTIR analysis showed higher proportions of lignin richer in subunits G, greater proportions of crystalline cellulose and acetyl methyl esters.
The aim of the study is interesting. The authors compiled a large amount of characterization data.
Yet, the authors should the precise indicate the novelty of presented paper and should be more specific in the Experimental part - how they ensured spectral comparability.
A conclusion section has been included in order to highlight the main take home messages and sum the main findings of the study.
We would like to thank your comments; I hope all your inquiries are properly addressed.
We consider the manuscript has improved substantially with your input.
Sincerely,
Dr. Ana López Malvar
Author Response File: 
Author Response.docx
Reviewer 2 Report
The research investigates maize recombinant inbred lines for their saccharification efficiency yields using FTIR spectroscopy. The background information in introduction and the Materials and Methods section is well presented. The research results are clearly written and discussed. Minor spell check might be needed.
Overall, the reader can comprehend the research and its findings, but I believe that the manuscript structure in unnecessarily complicated. Why do you prefer to include first your Results & Discussion section followed by Materials and Methods? It might be easier for someone to read first your research design and methods and afterwards to comprehend your findings. It is the logical sequence unless there is a reason for this preference.
Furthermore, since your Results are combined with the Discussion section maybe you should consider a conclusions section, ending your paper more smoothly. Nevertheless, the last paragraph of your manuscript summarizes your main conclusions.
Author Response
Dear Reviewer,
Please find enclosed the revised version of the manuscript ID: agronomy-1214120 Title: " FTIR Screening to Elucidate Compositional Differences in Maize Recombinant Inbred Lines with Contrasting Saccharification Efficiency Yields" submitted to Agronomy
We have addressed responses to all your main suggestions and made changes accordingly
Comments and Suggestions for Authors
The research investigates maize recombinant inbred lines for their saccharification efficiency yields using FTIR spectroscopy. The background information in introduction and the Materials and Methods section is well presented. The research results are clearly written and discussed. Minor spell check might be needed.
Minor spell mistakes have been corrected in the manuscript
Overall, the reader can comprehend the research and its findings, but I believe that the manuscript structure in unnecessarily complicated. Why do you prefer to include first your Results & Discussion section followed by Materials and Methods? It might be easier for someone to read first your research design and methods and afterwards to comprehend your findings. It is the logical sequence unless there is a reason for this preference.
In agreement with the reviewer comments, we have move the Material and Methods section before the Results and Discussion in order to gain comprehension.
Furthermore, since your Results are combined with the Discussion section maybe you should consider a conclusions section, ending your paper more smoothly. Nevertheless, the last paragraph of your manuscript summarizes your main conclusions.
A conclusion section has been included in order to highlight the main take home messages and sum the main findings of the study.
We would like to thank your comments; I hope all your inquiries are properly addressed.
We consider the manuscript has improved substantially with your input.
Sincerely,
Dr. Ana López Malvar
Author Response File: Author Response.docx
Reviewer 3 Report
Very interesting work and authors have tried to present their research findings succinctly; however, major revision is needed to be considered further. Please see the attached review comments
Comments for author File: 
Comments.pdf
Author Response
Dear Reviewer,
Please find enclosed the revised version of the manuscript ID: agronomy-1214120 Title: " FTIR Screening to Elucidate Compositional Differences in Maize Recombinant Inbred Lines with Contrasting Saccharification Efficiency Yields" submitted to Agronomy
We have addressed responses to all your main suggestions and made changes accordingly
Comments and Suggestions for Authors
Very interesting work and authors have tried to present their research findings succinctly; however, major revision is needed to be considered further. Please see the attached review comments
Review Comments:
It is very well designed study and authors have taken significant efforts to present their research findings concisely. However, authors should have provided the lines to review the manuscript since it gets very difficult to keep track of it. Please provide the lines numbers in the revised draft. Also this manuscript still needs some more work to be done to accept this manuscript. Please see the below comments.
Line numbers have been included in order to facilitate further revisions.
Abstract: Currently there are 192 words in the abstract which is lower than the allowed word count (200 words). Abstract is very well written and authors have succinctly presented the background, method, results, and usefulness of this research in future.
In order not to exceed the limit of allowed words, we consider that the length of the abstract is sufficient to include the necessary information to encourage the reading of the article.
Introduction:
Very well written authors have provided sufficient information on the background and supplemented with relevant literature; however, objectives and hypotheses is missing.
The objectives and the novelty of the study has been included in the last paragraph of the introduction
Paragraph 3: “The conversion…..to ethanol [9]” should be merged with paragraph 4 “The main obstacle…..be fermented”.
Paragraphs have been merged
Paragraph 7: Seems more of a result interpretation and conclusions. Please revise this paragraph by incorporating the objectives and hypotheses of this study. Current paragraph should not be in the introduction.
The objective has been included at the end of the introduction. As now included in the text: The objective has been included. The aim of this study is to take advantage of this spectroscopy technique applied to contrasting saccharification efficiency groups, in order to elucidate differences in the cell wall composition and architecture of high and low sugar-yielding RILs. In addition, as far as we now, this is first time that FTIR-PCA was used on maize RILs to correlate cell wall traits with saccharification. The results obtained in this research would show the relationship between the composition of the wall and the cellulose bioconversion process in stover samples from highly variable RILs, that could reveal key aspects to optimize the conversion of lignocellulosic biomass bio-refinery, but also other promising bio-products such as biogas via fermentation with anaerobic bacteria or non-biofuel related products derived from fermentation of lignocellulose derived sugars.
Results and Discussion: Authors have tried to present the results in a succinct manner but seems that more results should be supported to conclude. I also wonder why authors not considered assessing the kernel composition and cross-check the correlation between starch, ethanol, oil, and protein so that more clear trends could have been seen. For reference, please refer to the recently publish work of Nankar and Pratt (2020) on NIR calibration development and kernel compositional analysis. https://doi.org/10.3390/plants9121775.
In our study, the kernels did not contribute in saccharification efficiency estimations. We focused on maize stover, the debris that remains after the grain has been harvested. Bioethanol derived from maize stover presents the added advantage of not diverting basic goods to industrial uses making possible a double exploitation of the crop as source of food or feed and energy. Ethanol produced from non-grain plant material is considered as second generation biofuel.
Since there are not many figures and tables in the current draft, why authors didn’t provided results for 2016, 2017, and across years ANOVA in the text since it may add the value to quality of the manuscript.
As suggested by the reviewer, saccharification efficiency values (2016, 2017) for every RIL included in the study has been incorporated as a Table in the main manuscript: “Table 1: Saccharification efficiency data, means and range, through two years (2016, 1017) in selected RILs from a MAGIC population classified in high and low saccharification yielders. “
Materials and methods: Overall experiment is designed well, only component seems to be missing is assessment of other kernel compositional contents.
See comment above explaining why kernels didn’t contribute on saccharification efficiency estimations.
Conclusion: There is no conclusion provided so please add a paragraph (maybe revised the last paragraph from the introduction to highlight main take home messages). Missing take home messages (THMs), please revise and provide refined THMs
A conclusion section has been included in order to highlight the main take home messages and sum the main findings of the study.
We would like to thank your comments; I hope all your inquiries are properly addressed.
We consider the manuscript has improved substantially with your input.
Sincerely,
Dr. Ana López Malvar
Author Response File: Author Response.docx
Round 2
Reviewer 3 Report
Authors have made substantial revision and manuscript has improved significantly. I do not have any more comments and decision is in favor to be accepted if Editor feels the manuscript appropriate fit for the journal format.
