Continuous Flooding or Alternate Wetting and Drying Differently Affect the Accumulation of Health-Promoting Phytochemicals and Minerals in Rice Brown Grain

Round 1

Reviewer 1 Report

The manuscript is generally well written and the authors answered an important question whether AWD practices will influence rice grain quality, compared to continuous flooding, by measuring a variety of grain traits after harvest. The reviewer suggests the following revisions before it could be published: 1) explain explicitly why all analyses were performed on whole grains while brown rice and/or polished rice are more common commercial/consumable products; 2) provide all the quality control data in the supplementary information. Specially, listed all the recovery values of the certified reference materials used in this study; 3) improve the language by shortening some long and hard-to-read sentences, e.g., line 33-35.

Author Response

Dear Sir,

thanks for your kind and deep revision work. We have revised the MS (ID: agronomy-589563) “Continuous Flooding or Alternate Wetting and Drying Differently Affect the Accumulation of Health-Promoting Phytochemicals and Minerals in Rice Brown Grain” by Orasen et al. accomplishing all of your suggestions.

Every change is highlighted in the text having used the “Track Change” function.

Since the Discussion and Conclusion Sections (and, to a lesser extent, the Introduction) have been extensively changed and shortened according to the suggestions of referee 2, a few references have been deleted and their numbering changed accordingly. Discussion and Conclusions have been completely substituted, leaving track in the text of the former version.

In detail concerning your comments:

Point 1- Explain explicitly why all analyses were performed on whole grains while brown rice and/or polished rice are more common commercial/consumable products.  

Answer - Indeed, all analyses and measurements were performed on brown rice, i.e., the commercial/consumable product. The term “whole” rice has been substituted with “brown” rice all throughout the text.

Point 2 - provide all the quality control data in the supplementary information. Specially, listed all the recovery values of the certified reference materials used in this study.

Answer- Concerning the request of providing the quality control data, we wish to kindly point out that rationale of the MS was to verify at the physiological level the potential effects of specific water managements, in two different seasons, on the concentrations of different grain components in the three cultivars considered, rather than to assess, at a purely analytical level, absolute concentrations. All the methods for the assay of the different compounds considered are based upon widely validated protocols concerning specifically, in most cases, rice grain. Repeatability data are expressed in the SDs reported in the Supplemental Tables S3-S8. We have answered to your request concerning recovery values only for the ionomic data since in this case we had at our disposal certified material. We have added a Supplementary Table (S9) where the recovery values of the certified reference materials used for ionome contents are listed. In this Table, the Ni data are not present since the concentration of this element was not considered in the certified material used. We have also better detailed the ionomic procedure in the M&M section concerning the calibration for As and P. Concerning the measurements of Apparent Amylose content, we did not perform experiments evaluating the amylose recovery. For better clarity, we have added in the M&M section the trade names of the amylose standards used. For all compounds and analytes all the R² of the calibration curves were higher than 0.98.

Point 3- improve the language by shortening some long and hard-to-read sentences, e.g., line 33-35.

Answer - Lines 33-35: we have shortened some long and hard to read sentences where explicitly suggested and all throughout the text, according also to the suggestions of Reviewer 2.

Reviewer 2 Report

I have no major concerns with the present study but I nevertheless have a number of minor suggestions that I think the authors should consider in a revised version.

Lines 38-39 - do not repeat words that already appear in the title; these are automatically indexed.

Lines 55-58 – very convoluted sentence; please consider rephrasing.

Line 61 – there is something wrong with this sentence.

Line 72 – there is something wrong with this sentence.

Lines 79-83 – very convoluted sentence; please consider rephrasing.

Line 89 – there is something wrong with this sentence.

Lines 76-91 – I strongly suggest to formulate one or two scientific hypotheses that this study is aiming at testing. The aim is clearly stated but the Introduction would greatly benefit from hypotheses. These could also be presented in the Abstract and should be referred to in the Discussion later on.

Line 109 – why ”therefore”; please help the reader.

Line 110 – text is missing here; see ”About.”

Lines 121-122 – this information is perhaps unnecessary since the reader would expect such a procedure.

Line 125 – initiation

Line 147 – I assume that the authors have treated these samples as three independent replicates and not 6 cf. line 202 that explains the procedure?

Line 204-207 – please provide details on how you tested if the ANOVA assumptions were met for each data set tested.

Figure 1 – it would be helpful for the reader if the year were plotted on top of the figure; it makes it unnecessary difficult if the reader has to consult the caption. For the water potential, I suggest to add a horizontal dotted line at -20 to indicate the threshold that the authors referred to in the Introduction.

Lines 226-227 – please define “dry periods”?

Lines 237-246 – I would find it interesting if you could highlight some of the differences that you refer to.

Table 1 – indicate that Baldo, Cladio and Loto are cultivar names.

Line 276-277 – provide info on the statistical tests used.

Table 2 – indicate that Baldo, Cladio and Loto are cultivar names.

Line 287-288 – provide info on the statistical tests used.

Table 3 – indicate that Baldo, Cladio and Loto are cultivar names.

Line 308-309 – provide info on the statistical tests used.

Table 4 – indicate that Baldo, Cladio and Loto are cultivar names.

Line 332-333 – provide info on the statistical tests used.

Table 5 – indicate that Baldo, Cladio and Loto are cultivar names.

Line 362-364 – provide info on the statistical tests used.

Table 6 – indicate that Baldo, Cladio and Loto are cultivar names.

Line 396-397 – provide info on the statistical tests used.

Lines 399-420 – this text is not really a Discussion but rather a literature review that belongs in a condensed form to the Introduction.

Line 398-588 – I have severe difficulties following a logical line through the Discussion and it becomes particularly problematic since it is so long. I suggest to condense the Discussion and further consider if the current order of topics is the best.

Lines 590-618 – this is a very long Conclusion and I strongly suggest to substantially condense it.

Author Response

Dear Sir,

thanks for your kind and deep revision work. We have revised the MS (ID: agronomy-589563) “Continuous Flooding or Alternate Wetting and Drying Differently Affect the Accumulation of Health-Promoting Phytochemicals and Minerals in Rice Brown Grain” by Orasen et al. accomplishing all of your suggestions.

Every change is highlighted in the text having used the “Track Change” function.

Since the Discussion and Conclusion Sections (and, to a lesser extent, the Introduction) have been extensively changed and shortened according to your suggestions, a few references have been deleted and their numbering changed accordingly. Discussion and Conclusions have been completely substituted, leaving track in the text of the former version.

In detail concerning your comments:

Lines 38-39 - do not repeat words that already appear in the title; these are automatically indexed.

We have cancelled the words already appearing in the title, i.e., alternate wetting and drying and continuous flooding, grain health related compounds and added japonica ssp.

Lines 55-58 – very convoluted sentence; please consider rephrasing.

We have rewritten the sentence

Line 61 – there is something wrong with this sentence.

We have rewritten the sentence making it clearer.

Line 72 – there is something wrong with this sentence.

We have rewritten the sentence.

Lines 79-83 – very convoluted sentence; please consider rephrasing.

We have rewritten the sentence, that has also been split after introducing, as suggested, a few concepts from the initial paragraph of the Discussion section

Line 89 – there is something wrong with this sentence.

The whole paragraph which the sentence belongs to has been rephrased.

Lines 76-91 – I strongly suggest to formulate one or two scientific hypotheses that this study is aiming at testing. The aim is clearly stated but the Introduction would greatly benefit from hypotheses. These could also be presented in the Abstract and should be referred to in the Discussion later on.

We have formulated the working hypothesis, and introduced this concept also in the Abstract and in the Discussion

Line 109 – why ”therefore”; please help the reader.

As requested, we have deleted the word “therefore”.

Line 110 – text is missing here; see ”About.”

As requested, we have deleted “About”.

Lines 121-122 – this information is perhaps unnecessary since the reader would expect such a procedure.

As requested, we have deleted the unnecessary information.

Line 125 – initiation

We have corrected “Initiation” to “initiation” as suggested.

Line 147 – I assume that the authors have treated these samples as three independent replicates and not 6 cf. line 202 that explains the procedure?

Yes, the assumption is correct, but, in order to avoid any misunderstanding on the number of analytical replicates, we deleted this sentence refer, like we did for all other analyses, to the paragraph in the M&M section 2.5 “Statistical analyses”.

Line 204-207 – please provide details on how you tested if the ANOVA assumptions were met for each data set tested.

We have added a sentence concerning the check of the homogeneity of the variances (Levene’s test) for traits subjected to ANOVA.

Figure 1 – it would be helpful for the reader if the year were plotted on top of the figure; it makes it unnecessary difficult if the reader has to consult the caption. For the water potential, I suggest to add a horizontal dotted line at -20 to indicate the threshold that the authors referred to in the Introduction.

We have modified the Figure as suggested

Lines 226-227 – please define “dry periods”?

We have better defined “dry periods” as requested

Lines 237-246 – I would find it interesting if you could highlight some of the differences that you refer to.

We have highlighted, as requested, and better detailed some remarkable differences that we referred to.

Table 1– indicate that Baldo, Cladio and Loto are cultivar names.

We have indicated in all tables that Baldo, Gladio and Loto are cultivars, by both adding single quotes before and after the names and writing “cv.” in the Tables.

Line 276-277 – provide info on the statistical tests used.

For better clarity, we have provided here and elsewhere, as requested, information on the statistical tests used also in the legends of all Tables.

Lines 399-420 – this text is not really a Discussion but rather a literature review that belongs in a condensed form to the Introduction.

After shortening, the indicated text has been moved to the Introduction section.

Line 398-588 – I have severe difficulties following a logical line through the Discussion and it becomes particularly problematic since it is so long. I suggest to condense the Discussion and further consider if the current order of topics is the best.

We agree with you that the Discussion was exceedingly long and of poorly readable; therefore this Section has been extensively rewritten and shortened (by almost 30%) and simplified.

Lines 590-618 – this is a very long Conclusion and I strongly suggest to substantially condense it.

The Conclusion section has been substantially condensed and reduced by almost 50%.