Novel Seed Size: A Novel Seed-Developing Gene in Glycine max

Round 1

Reviewer 1 Report

The manuscript by Mingxia Zhang et al. “Novel Seed Size, a novel seed developing gene in Glycine max“ describes the identification of a new gene involved in the soyabean seed development. The novel gene titled Novel Seed Size gene (NSS) was found using analysis of a T-DNA insertion ramdom mutant (S006 mutant). This mutant of a transgenic soybean line has seed phenotypes with small size and brown seed coat. It is known, that seed size not only determines yield but also influences plant fitness and adaption. Therefore, the identification of genes that control seed size and the study of the related mechanisms is of particular interest. It has been shown that NSS gene encodes a functional unknown-protein, that contains a peptidase domain as a potential DNA helicase subunit and provides small seed size of the S006 mutant. Metabolite profiles of S006 mutant seeds and wild type seeds were compared. A new pathway has been discovered that controls the development of soybean seeds. It has been shown that suppression of NSS expression results in small seed size. The genes responsible for brown pigmentation of the seed coat also have been characterized. Transcriptome analysis confirmed that seed coat pigmentation of the S006 mutant may be due to the change in the expression level of genes related to flavonoid synthesis pathway. The article contains new important data on the mechanisms of regulation of soyabeen seeds development and can be published in its current form.

Page 2, line 55. Misprint: Phynotypes. Should be: phenotypes.

Author Response

Point 1: Page 2, line 55. Misprint: Phynotypes. Should be: phenotypes.

Response 1: Thank you for your kind and critical comments, and the misprint of "phynotypes" has been corrected.

Reviewer 2 Report

Within this work, soybean seed development is controlled in multiple aspects as many known regulating genes. The Authors identified a novel gene, Novel Seed Size (NSS), involving in seed development, by analyzing a T-DNA mutant (S006). S006 mutant is a random mutant of GmFTL4pro:GUS transgenic line with phenotypes of small and brown seed coat.

This is an extensive research, with a lot of experimental and numerical analysis. Thematically the work is interesting for the researchers and professionals and the proposed manuscript is relevant to the scope of the journal.

I found it appropriate for publication in the International Journal of Molecular Sciences, but only after some modifications and clarification from the Authors.
The abstract reflects realistically the substance of the work. It covers the research context and background, motivation, hypothesis, method, main results, and conclusion, underlining the implications of the main findings.

The overall organization and structure of the manuscript are appropriate. The paper is well written and the topic is appropriate for the journal.
The aim of the paper is well described and the discussion was well approached, its results and discussion are correlated to the cited literature data.

The literature review could be more properly done, by introducing a few newer references?

The novelty of the work must be more clearly demonstrated. This could be arranged by correlate the results of the study to newer literature references?
The significance of the Work: Given the large number of analyzed data, this is an interesting study with a possible significant impact in this area.
Statistical interpretation of the analytical data was well approached, but perhaps the graphics could be larger? Consider the changing the graphics' font type to Palantino?.

Other Specific Comments: The work is properly presented in terms of the language. The work presented here is very interesting and well done, it is presented in a compact manner.
In general, there are no doubtful or controversial arguments in the manuscript. The methodology applied in the research is presented in clear manner, so that it is repeatable by other authors.
The results are presented in a logical sequence and the discussion and analysis of the results are properly elaborated. The claims in the section "Conclusion" are reasonable and supported by the presented results.
 In my opinion, the authors should put additional effort to demonstrate that the present work gives a substantial contribution in the research area.

The manuscript should be improved from technical/graphical viewpoint.

Author Response

Please see the attachment

Author Response File: Author Response.docx