Review Reports

Round 1

Reviewer 1 Report

In this interesting article, the differences in the composition of the leaf and the kernel of walnuts are studied. The objective of the work has been to find markers of the tree's water status through the study of some components of the leaf and to determine the effect that different irrigation regimes can have on some kernel metabolites. In general, the experimental strategy is adequate, the results are clearly presented and the conclusions seem coherent with them.

From my point of view, the most important weakness of the article is the use of non-specific methods for the determination of phenols, flavonoids and sugars. The three methods used are colorimetric and offer very little and very non-specific information on the composition of the leaf. Probably a more detailed study by high resolution chromatographic methods could have offered better markers.

Author Response

Dear reviewer, we sincerely appreciate your valuable comments and recommendations for improving the manuscript.

The decision to use spectrophotometric techniques to measure the concentration of total phenols, total flavonoids and total soluble sugars in leaves was made even though it was not very specific, because they offer some advantages such as: speed of execution, easy sample preparation, high repeatability and low demand for analytical equipment. These are desirable characteristics in a water deficit biomarker complementary to the traditional crop water status indicators. Unfortunately, we did not observe a significant association between these potential biomarkers and water regime.

On the other hand, we are currently working on a new manuscript about changes in walnut leaf metabolome under different water regimes. For the metabolite profiling in this work we will use the GC-MS technique presented here and a new technique that we are adjusting by HPLC-QqQ-MS/MS.

Some comments have been included in the revised ms.

 

 

 

Reviewer 2 Report

Forte:

The article submitted has great potential. The information is presented clearly and it is easy to follow. The methods are clearly described.

The article presents differences that occur when water deficit appears in walnut trees. The authors selected 3 separate water irrigation type in order to see the differences in walnuts. They also studied specific aspects that are correlated to water deficit seeing how walnuts become more and more needed on the market and producers must use land with few water resources that can be overused.

Issues:

There are only a few modifications necessary but nothing scientific, such as name of the species with italic or the chemical formulas. I also suggested a graphic representation so that some information can be more easily followed. Besides that, the article is well done and interesting.

The article is interesting and has valuable information.

There are a few changes necessary:

Page 4- carefully check how the chemical formulas are written

Page 6, Page 10, Page 11, Page 14- species name in italic (Throughout the paper)

Please add a graphic representation (column) for table 1 where it would be easy to follow the data.

Kind regards,

Suzana Biris

Comments for author File: Comments.pdf

Author Response

Dear reviewer, thank you very much for your comments and suggestions for improving the manuscript. We are very pleased that you find our work interesting. All changes made to the ms are highlighted.

A2: There are a few changes necessary:

Page 4- carefully check how the chemical formulas are written

Q2: Ok, accepted and adjusted. We have revised the subscripts and superscripts throughout the manuscript.

A3: Page 6, Page 10, Page 11, Page 14- species name in italic (Throughout the paper)

Q3: Ok, accepted and adjusted. We have revised the scientific names and changed their format to italic.

A4: Please add a graphic representation (column) for table 1 where it would be easy to follow the data.

Q4: Thank you very much for warning us about the complications associated with the presentation of the information in (old) Table 1. We have replaced it with two figures. The new Figure 1 presents the means and standard deviations of MDA, PRO, SST, FEN and FLA classified by collection date and irrigation regime, while the new Figure 2 presents the same information but only classified by collection date.

Based on their recommendation, we have also changed the order of appearance of the leaf biomarkers: 1-MDA, 2-PRO, 3-SST, 4-FEN, 5-FLA. In the same way, we order the appearance of the irrigation regimes as follows: 1-T100, 2-T75 and 3-T50. We believe that this slight modification will help the reader's better understanding.

Finally, we made minor changes in the graphic section of the article, such as the unification of the font types in the figures, change colors in the Pearson correlation scale of Figure 5, and improved the format of Table 1 (old Table 2).

Author Response File: Author Response.docx