Identification and Introgression of a Novel HMW-GS Gene from Aegilops tauschii
Maria Itria Ibba
Round 1
Reviewer 1 Report
I enjoyed reading the manuscript which is well written. The study was able to show the value of introgressing directly genes from Aegilops tauschii into bread wheat. This has allowed, to identify novel gene for HMW-GS1, to clone it, to identify the substitution of cysteine by arginine and to assess the impacts of derived lines on quality of bread wheat. The three lines showed improved CSB quality attributes and improvements in several yield components.
No errors are found in the manuscript except for the Latin names of species in the references which should be in Italics and few mistakes highlighted in yellow in the attached version. In the discussion, it will be good to add a bit on the way forwards for using the lines.
Comments for author File: 
Comments.pdf
Author Response
We have revised these mistakes in the manuscript. Thanks so much for your comments.
Reviewer 2 Report
In this article, the authors cloned a novel HMW-GS gene from Aegilops tauschii, and successfully transferred it into an elite winter wheat through direct hybridization approach. The dough quality and agronomic performance of three introgression lines were investigated. The topic is interesting. Some minor corrections are needed before publication.
1. Authors need to perform a significance analysis for Table 1 and Figure 4, respectively.
2. For Figure 3, what are the “*”, “**”, and “***” ? If they represent the significant difference, please labeled them on the column chart.
3. Grammar checks should be carried out for proper English.
4. Some formats are wrong. For example, the Journal Name in the reference should be abbreviated. Please check the structure of format presentation for Agronomy.
Author Response
Point 1: Authors need to perform a significance analysis for Table 1 and Figure 4, respectively.
Response 1: We have revised Table 1 and Figure 4 in the manuscript. Thanks.
Point 2: For Figure 3, what are the “*”, “**”, and “***” ? If they represent the significant difference, please labeled them on the column chart.
Response 2: We have labeled them on the column chart in Figure 3. Thanks.
Point 3: Grammar checks should be carried out for proper English.
Response 3: We have conducted the grammar checks in the manuscript. Thanks so much for your suggestions.
Point 4: Some formats are wrong. For example, the Journal Name in the reference should be abbreviated. Please check the structure of format presentation for Agronomy.
Response 4: We have revised the formats mistakes in the reference. Thanks.
Reviewer 3 Report
In the present study, the authors report the characterization of a novel HOMW-GS allele from Ae. tauschii and its introgression into a bread wheat variety. Overall, the study is very interesting and included a significant number of experiments that supported the conclusions of the authors. For these reasons, I believe that the manuscript is worthy to be published and would only need a few minor revisions which could improve its overall quality.
1) It would be good if the authors could report the HMW-GS profile of the bread wheat variety (Jimai22) used to introgress the Glu-Dt1 allele. Does this variety have the Glu-D1a (2+12) allele? Or the Glu-D1d (5+10) allele? Which are the alleles at the Glu-A1 and Glu-B1 loci? This information would help better understand the quality profile of this variety and the variation in the quality profile observed after introgressing the Glu-Dt1 locus.
2) Based on the reported results, the introgression of the Glu-Dt1 allele was associated with an overall lower gluten strength but also with a higher steam bread quality. Since different products require flour with different quality profiles, it would be nice to briefly report either in the introduction or in the discussion, a short summary of the quality profile required to obtain high-quality steamed bread.
Apart from these, there are a few other modifications that I would suggest to improve the quality of the manuscript.
1) Line 210. Maybe you want to start this paragraph with: "as reported in table 1". If you keep it like this it would seem like this part is actually the caption of the table whereas it is part of the results.
2) Line 35. Better “HMW-GS are” and not “were”
3) Line 37. “In common wheat” is repeated
4) Line 41. Eliminate the dot after HMW-GSs
5) Line 67. Replace “has” with “have”
6) Line 105. “To minimize error” is repeated
7) Line 109. Change “analysis” to “analyze”
8) Table 1. Change “Stable time” into “mixograph stability time”
Author Response
Point 1: It would be good if the authors could report the HMW-GS profile of the bread wheat variety (Jimai22) used to introgress the Glu-Dt1 allele. Does this variety have the Glu-D1a (2+12) allele? Or the Glu-D1d (5+10) allele? Which are the alleles at the Glu-A1 and Glu-B1 loci? This information would help better understand the quality profile of this variety and the variation in the quality profile observed after introgressing the Glu-Dt1 locus.
Response 1: We have revised it in the manuscript. Thanks so much for your suggestions.
Point 2: Based on the reported results, the introgression of the Glu-Dt1 allele was associated with an overall lower gluten strength but also with a higher steam bread quality. Since different products require flour with different quality profiles, it would be nice to briefly report either in the introduction or in the discussion, a short summary of the quality profile required to obtain high-quality steamed bread.
Response 2: We have briefly reported it in the discussion. Thanks for your suggestion.
Point 3: Apart from these, there are a few other modifications that I would suggest to improve the quality of the manuscript.
1) Line 210. Maybe you want to start this paragraph with: "as reported in table 1". If you keep it like this it would seem like this part is actually the caption of the table whereas it is part of the results.
2) Line 35. Better “HMW-GS are” and not “were”
3) Line 37. “In common wheat” is repeated
4) Line 41. Eliminate the dot after HMW-GSs
5) Line 67. Replace “has” with “have”
6) Line 105. “To minimize error” is repeated
7) Line 109. Change “analysis” to “analyze”
8) Table 1. Change “Stable time” into “mixograph stability time”
Response 3: We have revised these mistakes in the manuscript. Thanks so much for your suggestions.
Reviewer 4 Report
I am grateful for the opportunity to give my considerations to this manuscript.
I suggest that study authors look again at the publication guidelines as well as a scientific English-language review.
All my observations and suggestions are highlighted or written in the text.
Comments for author File: Comments.pdf
Author Response
Point 1: All my observations and suggestions are highlighted or written in the text. I suggest that study authors look again at the publication guidelines as well as a scientific English-language review.
Response 1: We have revised the suggestions highlighted in the manuscript. Thanks so much for your suggestions.
