Non-Invasive Single-Grain Screening of Proteins and Other Features by Combination of Near-Infrared Spectroscopy and Nuclear Magnetic Resonance

Round 1

Reviewer 1 Report

Dear authors, 

The paper is well presented and describe an interesting analytical approach to study seeds individually. This work has a great potential to influence high-quality studies in the future, regarding not only agronomy, but also spectroscopy field. 

In addition, I would like just to recommend a detailed discussion about the results achieved using time-domain NMR. For example, in the page  6, lines 182-184, the authors described that TD-NMR has an almost a perfect prediction about the seeds. However, it is not clear the reasons for this achievement....

Therefore, I suggest in the discussion section, a more detailed explanation comparing the reasons about the differences for the results achieved using IR and TD-NMR. In this ways, the paper will be clear and highly suitable, especially for the audience that it is not used with these spectroscopies. 

Finally, I highly recommend this work for publication. The innovation and the quality of this paper is suitable for the Agronomy readers. 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Dear authors,

Thank you for an interesting and well formulated paper.

From the abstract it was not very clear why oats are chosen to do this research on. If you intend to create an in-line sorting procedure to enhance varietal identity and purity of a seed lot, then the application would be interesting for high-value seed types; for oats it would very likely to be too costly. Screening mutant populations (line 211) appears to me as a breeder a more viable option.

It could indeed also be an interesting research tool to test large numbers of samples in genebanks (as you do). In addition, I wonder whether it would be valuable in agronomic research to identify effects of soil/water etc gradients in a trial field on the seed characteristics. 

Related this I wonder in fig 6. to what extent is the kernel composition determined by the genetics of the variety; and to what extent by the growing conditions/age/storage conditions of the seed? It could thus be an interesting research tool for genebank seed research as well.

Finally: line 235. it would be interesting to expand a little in this discussion to what extent your findings could inform work on other seeds, i.e. is it realistic to assume that your findings have a much wider value than just for oats? If that is the case, then this deserves a sentence in the abstract, in my view.

Some minor questions/suggestions: 

31. reference instead of reverence

223. breeding’s ??

110. what is the meaning of the word ‘biological’  in this sentence?

Author Response

Please see the attachment.

Author Response File: Author Response.pdf