Evaluation of the Effect of Water Stress on Clonal Variations of Cv. Monastrell (Vitis vinifera L.) in South-Eastern Spain: Physiology, Nutrition, Yield, Berry, and Wine-Quality Responses
, Francisco M. del Amor 3 and Josefa MarÃa Navarro 1Round 1
Reviewer 1 Report
The manuscript contained a collection of data and results that contribute to the development of this field of study. However, this manuscript may require some clarifications before it is ready for publication.
· There were some references that did not follow with the style of the journal, such as lines 137 and 771.
· I am unable to distinguish between the control and stress in figures 3a, 4 and 6.
· Generally, despite the positive results, the discussion of these results was inadequate. There is a repetition of the results without discussion, as well as a substantial amount of text that does not emphasize the results. In my opinion, it needs to be reconsidered and made more focused. In addition, I would like to inquire about the following comments.
· “The values of WUEyield reported in field grown deficit irrigated grapevines vary widely depending on the rootstock, edaphocli matic and irrigation conditions [42], ranging between 9–18 kg m-3 in Monastrell vines grafted on different rootstocks in southeast Spain [24]. However, the clone effect on WUEy ield in Monastrell vines observed in this study under similar semiarid conditions was stronger, maximizing WUEyield until reaching an averaged of 28-30 kg m-3 during the period 2018-2020 (in 94 and 188, the most productive clones, Table 5), similar to the highest WUEyield values found in cv. Gaglioppo in Italy (9–29 kg m-3) [43].” Despite the fact that there was a significant difference between your results and the previous results in this section, you failed to explain what factors may have contributed to this difference.
· What does the reference in this sentence refer to? “Future reseach combining high-WUE drought tolerant rootstocks [24] and high-WUE drought tolerant Monastrell clones under semiarid-DI/rainfed conditions would be necessary, because could reduce reliance on supplemental irrigation and making vineyards more sustainable in semiarid areas, even for dry farming.”
- In the paragraphs between lines 624 and 636 and between 701 and 710; although you cited numerous sources, you simply repeated your results without discussing these sources.
- The authors mentioned “Significant relationships between long-term A/gs and WUEyield have been also found in Monastrell and other grapevine varieties, both positive and negative, reflecting the complexity of this relationship [48, 49, 50, 11]. In our study, improvements in WUEyield were not related to an increase in A/gs (Figures 2A, 2C)” What do you believe the reason is for the absence of this positive or negative relationship in your results?
Author Response
x
ACTIONS MADE TAKING INTO ACCOUNT THE COMMENTS OF THE REVIEWERS AND EDITOR
Reviewer 1
Reviewer 1: There were some references that did not follow with the style of the journal, such as lines 137 and 771.
Answer: Please, can you specify better where is the style of the reference is not correct?. We don't really know what it means. In lines 137 is “an electrical conductivity of 0.68-0.81 dS m−1. The climate is Mediterranean semiarid.”In line 771 is “gs and A/gs (negative), indicate that differences in A among clones, come mainly from…..”
Reviewer 1: I am unable to distinguish between the control and stress in figures 3a, 4 and 6.
Answer: We have revised again Figures 3A, 4 and 6 and black bars represent control and white bars stress treatment. We think that it is clear in the figures. In figure 6, we do not show control and stress treatments, only early morning and midday for each clone.
Reviewer 1: Generally, despite the positive results, the discussion of these results was inadequate. There is a repetition of the results without discussion, as well as a substantial amount of text that does not emphasize the results. In my opinion, it needs to be reconsidered and made more focused. In addition, I would like to inquire about the following comments.
Answer: In accordance to the reviewer1, we have made an effort to reduce discussion section and to discuss better the results, avoiding repetition of results without discussion and focussing more important topics.
We have removed the following paragraphs in the discussion section because are repetitive and confussing and they don´t add anything new to the discussion:
We have removed the next paragraphs in the discussion section:
“The combination of DI and more resilient, efficient and drought adapted Monastrell genotypes (e.g. clones) can be an useful adaptation measure to climate change (CC) in order to maintain the sustainability of vineyards [37, 38, 39] - especially in the semiarid, warm, and more vulnerable winegrowing regions of Southern Europe (such as our study area, with very limited water availability).
“The decreasing water potential status is an initial step that plants recognize dehydration stress conditions [59”
These results suggest the diversification of rootstocks and clones (exploring possible interactions) as a way of increasing genetic variability, avoiding the cultivation of a single rootstock and clone [7].
Reviewer 1: “The values of WUEyield reported in field grown deficit irrigated grapevines vary widely depending on the rootstock, edaphocli matic and irrigation conditions [42], ranging between 9–18 kg m-3 in Monastrell vines grafted on different rootstocks in southeast Spain [24]. However, the clone effect on WUEy ield in Monastrell vines observed in this study under similar semiarid conditions was stronger, maximizing WUEyield until reaching an averaged of 28-30 kg m-3 during the period 2018-2020 (in 94 and 188, the most productive clones, Table 5), similar to the highest WUEyield values found in cv. Gaglioppo in Italy (9–29 kg m-3) [43].” Despite the fact that there was a significant difference between your results and the previous results in this section, you failed to explain what factors may have contributed to this difference.
Answer: In accordance to the reviewer 1, we have made some modifications in this paragraph in order to clarify this part of the discussion.
Reviewer 1: What does the reference in this sentence refer to? “Future reseach combining high-WUE drought tolerant rootstocks [24] and high-WUE drought tolerant Monastrell clones under semiarid-DI/rainfed conditions would be necessary, because could reduce reliance on supplemental irrigation and making vineyards more sustainable in semiarid areas, even for dry farming.”
Answer: In accordance to the reviewer 1, we have modified slightly the sentence to clarify better the meaning. In earlier experiment we identifid very high WUE drought tolerant rootstocks (such as 140Ru) and in this experiment we have identified very high WUE drought tolerant clones, such as clones 4 and 188. The idea of the next experiment is combine both, high WUE clones and rootstocks under DI and rainfed conditions.
- Reviewer 1: In the paragraphs between lines 624 and 636 and between 701 and 710; although you cited numerous sources, you simply repeated your results without discussing these sources.
Answer: This paragraph (624-636) has been modified.
- Reviewer 1: The authors mentioned “Significant relationships between long-term A/gs and WUEyield have been also found in Monastrell and other grapevine varieties, both positive and negative, reflecting the complexity of this relationship [48, 49, 50, 11]. In our study, improvements in WUEyield were not related to an increase in A/gs (Figures 2A, 2C)” What do you believe the reason is for the absence of this positive or negative relationship in your results?
Answer: We have modified this paragraph in order to avoid confussion and clarify the meaning
The correct sentence is “Significant relationships between long-term A/gs and WUEyield have been also found in Monastrell and other grapevine varieties [48, 49, 50, 11]”.
Reviewer 2 Report
Thanks for providing me the opportunity to evaluate the article entitled “Evaluating intracultivar genetic diversity in Cv. Monastrell (Vitis vinífera L.) in Souteastern of Spain: Physiology, nutrition, yield and berry and wine quality response”. The study will be great interest about the intrageneric diversity of the grape’s cultivars under drought condition. I have some concerns about the article. The introduction part needs attention and rephrasing.
The second most important thing is data analysis. Can authors explain how they manage the experiment without replications and analyze the data without replication. It means that each experimental unit data is considered as mean. Or each block considered as replication while neglecting soil nutrient gradient shift and environmental effect. How we fulfilled the assumptions of ANOVA and two-way anova. Mean comparisons were managed through DMRT. DMRT is good for 8 treatments, while in your case 7 cultivars x 2 stress level =14 so LSD is good option for means separation.
Table 1: Total/annual average in the same table ETo showed total value however, other showed average value in the same table. Correct it
Table 2: check the formatting of the table
Some specific comments are presented below.
In the abstract there were some confusing results presented 19-28. Need rephrasing for intended and clear picture of the results.
Line 17: Leaf gas exchange, vegetative growth, leaf mineral nutrition, yield, water use efficiency and grape and wine quality.
The authors said that the drought tolerant clone or sensitive: Line 19-28 need revision
and high VPD: used full form at first sight
Line 61-64 rephrase the sentence for intended meaning.
Line 65-69. rephrase the sentence for intended meaning.
Line 74-75: and indeed, respects the cultural and social components of wine grape cultivation and the wine typicity associated with local terroir. rephrase the sentence for intended meaning.
Be consistent in standardized wording, In the abstract authors used word under water stress (stress) while line # 106 under sever water restriction strategy (stress). Or DI.
There is a need to modify the last paragraph for objective for intended meaning.
Line # 122 (38-36% silt) or 36-38%.
P asimilable is a Spanish word, try to use English word, as whole article written in English
Line # 127: dry summers or summer.
Line 153: 50-100L. ha-1 month-1) why it is varying “clones received the same annual amount of organic fertilizer”.
Line 154-155: There is a need to add the proper punctuation for readability sentence, “In the stressed vines fertilizer was applied directly into the soil when irrigation was cut”.
Due to its small size and soil homogeneity of the vineyard, the experimental design was elaborated without repetitions of complete blocks of each clone.
I am wondering, the authors used complete block design while they are not using the replications.
Line 158-159: Need rephrasing for intended meaning.
Line 143: What is RDI???
Even though the conditions were homogenized the repetitions of the treatments should be necessary???? Due to its small size and soil homogeneity of the vineyard,
Line # 200: and K, Mg, Ca, Na, P, Fe, Mn, Zn, and B?????
Line 222-224? Need citation
Line 227: 23–24° Brix measuring device????
Line 886-888: Need revision
Conclusion should be under separate headings.
Comments for author File: 
Comments.docx
Author Response
REVIEWER 2
Reviewer 2 Thanks for providing me the opportunity to evaluate the article entitled “Evaluating intracultivar genetic diversity in Cv. Monastrell (Vitis vinífera L.) in Souteastern of Spain: Physiology, nutrition, yield and berry and wine quality response”. The study will be great interest about the intrageneric diversity of the grape’s cultivars under drought condition. I have some concerns about the article. The introduction part needs attention and rephrasing.
Answer: Thanks for the interest in ths study. The introduction section and the whole text has been corrected by an english editor.
Reviewer 2: The second most important thing is data analysis. Can authors explain how they manage the experiment without replications and analyze the data without replication. It means that each experimental unit data is considered as mean. Or each block considered as replication while neglecting soil nutrient gradient shift and environmental effect. How we fulfilled the assumptions of ANOVA and two-way anova. Mean comparisons were managed through DMRT. DMRT is good for 8 treatments, while in your case 7 cultivars x 2 stress level =14 so LSD is good option for means separation.
Answer:
A completely randomized design was used with two factors: Clone and irrigation treatment, with 15 repetitions per combination. With 7 different clones and 2 irrigation conditions (Deficit irrigation and rainfed conditions). We don´t have blocks, because the orchard is very small and there were not significant differences in soil characteristics, such as texture, organic matter or pH, or cation exchange capacity. In accordance to the reviewer 2, we have modified slightly the paragraph of the experimental design in material and methods section in order to clarify it better.
For the statistical analysis of the data, a bifactorial ANOVA (two-way anova) was used, and for the comparison of means, Duncan's Multiple Range test. This test avoids errors inherent to the indiscriminate use of the DSM test (minimum significant difference). It requires progressively higher values for the significance between means, to the extent that they are more widely separated. And it is used more appropriately when various unrelated treatments are included in the experiment (Little et al., 1989), as is in our case, with clones, which we are interested to compare all the posible combinations between them in each irrigation treatment and also the number of pairwise comparisons is very large, as is the case (14). They offer a compromise solution between the desired global error rate and an individual error rate that is too small and therefore unacceptable. These methods are preferred in the sense of producing narrower confidence intervals.
Little, T. M. 1. 8., Hills, F. J. 1. 7., Paula Crespo, A. d. t., Silveira de Jasa, M. i. r. t., & Flores Alcántara, R. A. r. t. (1989). Métodos estadísticos para la investigación en la agricultura. México, DF : Trillas
Reviewer 2. Table 1: Total/annual average in the same table ETo showed total value however, other showed average value in the same table. Correct it
Answer: We think Table 1 is clear because Total refers to ETo and rainfall and anual average to the rest of the climatic parameters.
Reviewer 2: Table 2: check the formatting of the table.
Answer: Done
Reviewer 2: In the abstract there were some confusing results presented 19-28. Need rephrasing for intended and clear picture of the results.
Answer: Done
Reviewer 2: Line 17: Leaf gas exchange, vegetative growth, leaf mineral nutrition, yield, water use efficiency and grape and wine quality.The authors said that the drought tolerant clone or sensitive: Line 19-28 need revision
Answer: Done
Reviewer 2: and high VPD: used full form at first sight
Answer: Done.
Reviewer 2: Line 61-64 rephrase the sentence for intended meaning. Line 65-69. rephrase the sentence for intended meaning.
Answer: Done. The whole text has been revised by a native english editor.
Reviewer 2: Line 74-75: and indeed, respects the cultural and social components of wine grape cultivation and the wine typicity associated with local terroir. rephrase the sentence for intended meaning.
Answer: We have removed this sentence.
Reviewer 2: Be consistent in standardized wording, In the abstract authors used word under water stress (stress) while line # 106 under sever water restriction strategy (stress). Or DI. There is a need to modify the last paragraph for objective for intended meaning.
Answer: In accordance top the reviewer 2, we have revised the whole text to be more consistent with the words. We have removed the word DI. We have modified the last paragraph for objective.
Reviewer 2: Line # 122 (38-36% silt) or 36-38%.
Answer: Done
Reviewer 2: P asimilable is a Spanish word, try to use English word, as whole article written in English.
Answer: Done
Reviewer 2: Line # 127: dry summers or summer.
Answer: We think that it is correct, dry summer….Because it is usually very dry…
Reviewer 2: Line 153: 50-100L. ha-1 month-1) why it is varying “clones received the same annual amount of organic fertilizer”.
Answer: The first year of the experiment we applied 100 L /ha /month, but the rest of the years we reduced the dose to 50 L/ha/month. All clones received the same fertilizer amount. We have modified the text in order to clarify and avoid confusion
Reviewer 2:Line 154-155: There is a need to add the proper punctuation for readability sentence, “In the stressed vines fertilizer was applied directly into the soil when irrigation was cut”.
Answer: Done
Reviewer 2:Due to its small size and soil homogeneity of the vineyard, the experimental design was elaborated without repetitions of complete blocks of each clone.
I am wondering, the authors used complete block design while they are not using the replications.
Answer: We have explained better the experimental design in Material and Methods and in this document, above.
Reviewer 2: Line 158-159: Need rephrasing for intended meaning.
Answer: Done
Reviewer 2: Line 143: What is RDI???. Even though the conditions were homogenized the repetitions of the treatments should be necessary???? Due to its small size and soil homogeneity of the vineyard.
Answer: Done (RDI) and explained in Material and Methods
Reviewer 2: Line # 200: and K, Mg, Ca, Na, P, Fe, Mn, Zn, and B?????
Answer: Yes
Reviewer 2: Line 222-224? Need citation
Answer: Done
Reviewer 2: Line 227: 23–24° Brix measuring device????
Answer: Done
Reviewer 2: Line 886-888: Need revision
Answer: Done
Reviewer 2: Conclusion should be under separate headings.
Answer: In accordance to the reviewer, We have modified and shortened the last section of the discussion and the conclusions to clarify it.
Reviewer 3 Report
The manuscript titled “Evaluating intracultivar genetic diversity in Cv. Monastrell (Vitis vinífera L.) in Southeastern of Spain: Physiology, nutrition, yield and berry and wine quality response” has successfully reported significant phenotyping results on a range of physiological and agronomical traits including leaf gas exchange, vegetative growth, leaf mineral nutrition, yield, water use efficiency (WUE), grape and wine quality in Southeastern Spain. All the phenotyping measurements of the seven clones of Cv. Monastrell were also tested with some significantly different results, indicating substantial clonal variations do exist for the varied clones of Cv. Monastrell in respond to water stress treatment. Since this research only covered water stress responses of the seven clones of Cv. Monastrell rather than genotyping of the genetic diversity of the cultivar, a more appropriate title for this research could be: Evaluation of clonal variations within Cv. Monastrell (Vitis vinifera L.) in Southeastern Spain: physiology, nutrition, yield, berry and wine quality responses.
Generally, the overall experimental design, material and methods, and data analysis in this manuscript are well organized. Phenotyping profiles of physiological and agronomical traits described in this research are standard and meet current international viticulture practices. Characteristics of plant water relations were successfully determined on vine water status, leaf gas exchange, leaf hydraulic conductivity, stem water potential, isotope carbon composition, leaf mineral analysis, main shoot length and total leaf area, yield response, water use efficiency (WUE) and sink/source ratios, berry composition and berry quality index, as well as certain standards for wine quality on microvinification, wine color intensity, wine quality index, and quantification of volatile aromatic compounds in the wine. Findings from this research especially the range of specific phenotyping methods could be applicable for similar viticulture research practices, as well as for the Spanish and/or international wine production industry.
In conclusion, this paper is suitable for publication once their English language expression is reviewed by a professional academic editor to make their communication more succinct.
Some specific editing suggestions:
1. All the titles of the Tables and Figures need to be made more concise, so as to directly represent the information contained therein.
2. L115: change ‘The trial was’ into ‘All the trials and experiments were’.
3. The following abbreviations should initially be spelt in full: L143 (RDI); L179 (VPD); L182 (PPFD); L29 (TSS).
4. L205: The text states that “TLA per plant was measured once a year at the end of July” – does “once a year at the end of July" mean in July of every year from 2018 to 2021?
5. L215: Does “each year at harvest” mean every year from 2018 to 2021?
6. L240-255: The specific control treatment for the microvinification tests need to be added to the Method here. Similarly, all the control treatments used overall need to be described in detail in the Material and Methods section.
Author Response
REVIEWER 3
Reviewer 3: Since this research only covered water stress responses of the seven clones of Cv. Monastrell rather than genotyping of the genetic diversity of the cultivar, a more appropriate title for this research could be: Evaluation of clonal variations within Cv. Monastrell (Vitis vinifera L.) in Southeastern Spain: physiology, nutrition, yield, berry and wine quality responses.
Answer: In accordance to the Reviewer 3, we have changed the title of the article.
Reviewer 3: In conclusion, this paper is suitable for publication once their English language expression is reviewed by a professional academic editor to make their communication more succinct.
Answer: Done. A professional native English academic editor has revised the manuscript.
Reviewer 3: All the titles of the Tables and Figures need to be made more concise, so as to directly represent the information contained therein.
Answer: In accordance to the reviewer 3, we have revised the legends of the tables and figures and we have shortened slightly. We consider that the legends explain well the figures and tables. Sometimes are long because we have to explain a lot of parameters or graphics in the same figure or table.
Reviewer 3: L115: change ‘The trial was’ into ‘All the trials and experiments were’.
Answer: Done
Reviewer 3. The following abbreviations should initially be spelt in full: L143 (RDI); L179 (VPD); L182 (PPFD); L29 (TSS).
Answer: Done
Reviewer 3: 4. L205: The text states that “TLA per plant was measured once a year at the end of July” – does “once a year at the end of July" mean in July of every year from 2018 to 2021?
Answer: Yes. We have modified slightly in order to clarify the meaning.
Reviewer 3: 5. L215: Does “each year at harvest” mean every year from 2018 to 2021?
Answer: Yes. We have modified slightly in order to clarify the meaning.
Reviewer 3: 6. L240-255: The specific control treatment for the microvinification tests need to be added to the Method here. Similarly, all the control treatments used overall need to be described in detail in the Material and Methods section.
Answer: Done.
Round 2
Reviewer 1 Report
No comments
Author Response
ACTIONS MADE TAKING INTO ACCOUNT THE COMMENTS OF THE REVIEWERS AND EDITOR
Reviewer 1
Thank you very much for your comments.
Reviewer 2 Report
Line: 166
It is difficult to find homogeneous experimental units in all respects and hence CRD is seldom suitable for field experiments.
Line 248: remove the parenthesis (total soluble solids
Line 810: use journal format for the (Zufferey et al., 2020)
Line 921 and 924: why —.
Line 949: 41 and 97%, respectively
Used % singed for 41.
Table 1. Total/anual or annual
Table 2 check the formatting
Table 5: -stress why – with word stress
Comments for author File: Comments.docx
Author Response
ACTIONS MADE TAKING INTO ACCOUNT THE COMMENTS OF THE REVIEWERS AND EDITOR
Reviewer 2:
It is difficult to find homogeneous experimental units in all respects and hence CRD is seldom suitable for field experiments.
Answer: Yes, we agree with that. But in these experimental conditions and very small orchard we think that can be valid.
Line 248: remove the parenthesis (total soluble solids
Answer: Done
Line 810: use journal format for the (Zufferey et al., 2020)
Answer: Done
Line 921 and 924: why —
Line 949: 41 and 97%, respectively. Used % singed for 41.
Answer: Done
Table 1. Total/anual or annual
Answer: Done: Annual
Table 2 check the formatting
Answer: Done
Table 5: -stress why – with word stress
I do not understand this. I´m sorry.
Author Response File: Author Response.docx
Reviewer 3 Report
Overall, I am happy with the revised manuscript. I have just thought of one last suggestion about the title. It should include reference to water stress. My suggested title is: "Evaluation of clonal variations due to water stress on cv. Monastrell (Vitis vinifera L.) in south-eastern Spain: physiology, nutrition, yield, berry, and wine-quality responses".
Author Response
Reviewer 3
Overall, I am happy with the revised manuscript. I have just thought of one last suggestion about the title. It should include reference to water stress. My suggested title is: "Evaluation of clonal variations due to water stress on cv. Monastrell (Vitis vinifera L.) in south-eastern Spain: physiology, nutrition, yield, berry, and wine-quality responses".
Answer: Thank you for the suggestion. In accordance to the reviewer 3, We have added water stress in the title and has been revised by an english native editor.
Evaluation of the effect of water stress on clonal variations of cv. Monastrell (Vitis vinifera L.) in south-eastern Spain: physiology, nutrition, yield, berry, and wine-quality responses.
Author Response File: Author Response.docx
