The Effects of Altitude on Fruit Characteristics, Nutrient Chemicals, and Biochemical Properties of Walnut Fruits (Juglans regia L.)

Round 1

Reviewer 1 Report

The manuscript entitled "The Effects of Altitude on Fruit Characteristics, Chemical, Biochemical, Sugar, Organic Acid and Fatty Acid Contents of Walnut Fruits (Juglans regia L.)"  by Muhammet Ali Gündeşli et al., reported the changes in nutrient chemicals, such as total phenolic content, total organic acids, sugars, protein, fatty acids contents, as well as biochemical properties, such as total antioxidant capacity, DPPH inhibition, and DPPH  radicals,  as a function of varing altitudes from 500 m to 1200 m under Mediterranean 13 climatic conditions.   The study provided meaningful rationale and experimental designs, the investigations and experiments were well designed and performed, data provided unique information for our understanding of the effects of altitudes on the nutrient quality of Walnut Fruits, which may have directional roles in agricultural practices.    Therefore, the manuscript is basically valuable for the community.   However, English writing and manuscript preparation needs a lot of improvement.  1. The title needs to be revised to correctly reflect the contents and the authors' purpose.  maybe "The Effects of Altitude on Fruit Characteristics, Nutrient Chemicals, and Biochemical Properties of Walnut Fruits (Juglans regia L.)"?   2. The Abstract should be precisely condensed and revised, the current Abstract contains much information only should be shown in Materials and Methods,   3. Many mistakes in writing and spelling need to be corrected, such as listed below, but not limited to them:    line 14, 10 genotypes and 3 walnut cultivars (Maraş-18, Chandler, and Franquette) were used as research material, please rephrase the sentence.   Line 24, total fat content varied between 53.44% and... should be total fatty acid?  Line 131,  2.1.2 Preparation of plant material and extractions, should be "materials"? Line 144, 2.2. Metod,  should be "Method"? ....

minor revision on English writing 

Author Response

Dear Reviewer, Thank you for spending your valuable time to review the article.

1) The title needs to be revised to correctly reflect the contents and the authors' purpose.  maybe "The Effects of Altitude on Fruit Characteristics, Nutrient Chemicals, and Biochemical Properties of Walnut Fruits (Juglans regia L.)"?  

Necessary corrections were made according to your suggestion. The title of the article has been changed to “The Effects of Altitude on Fruit Characteristics, Nutrient Chemicals, and Biochemical Properties of Walnut Fruits (Juglans regia L.)”

2) The Abstract should be precisely condensed and revised, the current Abstract contains much information only should be shown in Materials and Methods.

Necessary corrections were made according to your suggestion. The abstract of the article has been changed to “This study aimed to investigate changes in fruit characteristics, total phenolics, total antioxidant capacity, organic acids, sugars, and fatty acid compositions of 10 walnut genotypes and 3 cultivars grown at different altitudes under Mediterranean climate conditions. At both 500 m and 1200 m altitudes, total phenolics ranged between 237.51 and 412.96 mg GAE/100 g dw and 234.94 and 392.31 mg GAE/100 g dw, respectively. Total antioxidant capacity ranged from 54.65% to 71.99% DPPH inhibition and 47.65% to 64.99% DPPH radical at 500 m altitude, and 64.76% to 75.92% DPPH inhibition and 57.76% to 68.92% DPPH radical at 1200 m altitude. Oil rate varied between 53.44% and 76.17% at 500 m altitude and 43.11% and 67.03% at 1200 m altitude. Protein rate ranged from 13.71% to 21.32% at 500 m altitude and 12.07% to 17.79% at 1200 m altitude. At both 500 m and 1200 m altitudes, linoleic acid was major fatty acid. It was followed by linolenic and oleic acids. Linoleic acid ranged from 58.82% to 62.44% at 500 m altitude 57.33 and 59.38 at 1200 m altitude. Linolenic acid varied between 7.93 and 9.72 % at 500 m altitude and 8.80 and 10.79 at 1200 m altitude. Malic acid was the major organic acid, ranging from 1.35% to 7.33% at both altitudes. Sucrose content in the cultivars ranged from 1.34% to 3.13%. In conclusion, this study showed that walnut seeds are rich in oil, protein, total phenolics, antioxidant activity, and fatty acids, with variations depending on the cultivar/genotype, especially affected by altitude”

3) Many mistakes in writing and spelling need to be corrected, such as listed below, but not limited to them:    line 14, 10 genotypes and 3 walnut cultivars (Maraş-18, Chandler, and Franquette) were used as research material, please rephrase the sentence.   Line 24, total fat content varied between 53.44% and... should be total fatty acid?  Line 131,  2.1.2 Preparation of plant material and extractions, should be "materials"? Line 144, 2.2. Metod,  should be "Method"?

Necessary corrections were made according to your suggestion. The abstract of the article has been revised. Line 131 has been changed to “2.1.2 Plant materials” and line 144 has been changed to “2.2 Method”

 

Moreover,
The publication has been extensively language edited by an expert researcher whose native language is English.The editing document is attached.

Author Response File: Author Response.pdf

Reviewer 2 Report

This is an interesting and straight forward type of study; it has practical interest, especially for breeders. 

Samples - 10 genotypes and three cultivars / varieties. Authors should indicate what is the difference among them, and their basic characteristics, as well as where these materials come from, and its specific source.

Abstract - Very long description; aceptable, but very long. However, art the end of Abstract description is rather poor. Authors need to indicate the basic and important messages of this long report.

 

Objectives - They should be better described; authors mention a "biochemical" item, but they should indicate what they want to express by this so wide term.

Section 2.2.8. Evaluation of results; it should be termed statistical assessments. 

Total protein - Authors only quantified nitrogen by Kjeldhal; it would be interesting to have determined protein by one of the methods for protein determination, in addition to nitrogen.

Biochemical content - Authors need to describe the meaning of this statement.

Results and conclusions - Results are about the same of other similar and previous studies. Authors need to state very clearly what is the advantage and importance of results obtained in this experimentation versus previous works done by other colleagues; otherwise, this work will appear to have nearly cero originality, and no any importance for breeders and other people interested in scientific characteristics of these genetic materials affected by altitude.  

Author Response

Dear Reviewer, Thank you for spending your valuable time to review the article.

1) Samples - 10 genotypes and three cultivars / varieties. Authors should indicate what is the difference among them, and their basic characteristics, as well as where these materials come from, and its specific source.

Necessary corrections were made according to your suggestion. Information about 10 genotypes has been added to the material section of the article.

2) Abstract - Very long description; aceptable, but very long. However, art the end of Abstract description is rather poor. Authors need to indicate the basic and important messages of this long report.

Necessary corrections were made according to your suggestion. The abstract of the article has been changed to “This study aimed to investigate changes in fruit characteristics, total phenolics, total antioxidant capacity, organic acids, sugars, and fatty acid compositions of 10 walnut genotypes and 3 cultivars grown at different altitudes under Mediterranean climate conditions. At both 500 m and 1200 m altitudes, total phenolics ranged between 237.51 and 412.96 mg GAE/100 g dw and 234.94 and 392.31 mg GAE/100 g dw, respectively. Total antioxidant capacity ranged from 54.65% to 71.99% DPPH inhibition and 47.65% to 64.99% DPPH radical at 500 m altitude, and 64.76% to 75.92% DPPH inhibition and 57.76% to 68.92% DPPH radical at 1200 m altitude. Oil rate varied between 53.44% and 76.17% at 500 m altitude and 43.11% and 67.03% at 1200 m altitude. Protein rate ranged from 13.71% to 21.32% at 500 m altitude and 12.07% to 17.79% at 1200 m altitude. At both 500 m and 1200 m altitudes, linoleic acid was major fatty acid. It was followed by linolenic and oleic acids. Linoleic acid ranged from 58.82% to 62.44% at 500 m altitude 57.33 and 59.38 at 1200 m altitude. Linolenic acid varied between 7.93 and 9.72 % at 500 m altitude and 8.80 and 10.79 at 1200 m altitude. Malic acid was the major organic acid, ranging from 1.35% to 7.33% at both altitudes. Sucrose content in the cultivars ranged from 1.34% to 3.13%. In conclusion, this study showed that walnut seeds are rich in oil, protein, total phenolics, antioxidant activity, and fatty acids, with variations depending on the cultivar/genotype, especially affected by altitude”

3) Objectives - They should be better described; authors mention a "biochemical" item, but they should indicate what they want to express by this so wide term.

Necessary corrections were made according to your suggestion. The biochemical characteristics in the study are stated in the abstract and introduction part.

4) Section 2.2.8. Evaluation of results; it should be termed statistical assessments.

Necessary corrections were made according to your suggestion. The section 2.2.8. "Evaluation of results" has been changed to "2.2.8 Statistical Assessments”

5) Total protein - Authors only quantified nitrogen by Kjeldhal; it would be interesting to have determined protein by one of the methods for protein determination, in addition to nitrogen.

The laboratory conditions where the study was conducted were suitable for the Kjeldhal method. Therefore, the Kjeldhal method was preferred.

 

6) Biochemical content - Authors need to describe the meaning of this statement.

Necessary corrections were made according to your suggestion . The biochemical contents are stated in the introduction part of the article. It is stated as “……biochemical content (total phenolics, total antioxidant capacity, organic acids, sugars, and fatty acid compositions)” on lines 173 and 174.

7) Results and conclusions - Results are about the same of other similar and previous studies. Authors need to state very clearly what is the advantage and importance of results obtained in this experimentation versus previous works done by other colleagues; otherwise, this work will appear to have nearly cero originality, and no any importance for breeders and other people interested in scientific characteristics of these genetic materials affected by altitude.

Necessary corrections were made according to your suggestion. The conclusion of the article has been changed to “In this study, the effects of altitude on the quality characteristics and biochemical properties (total fat, total protein, total phenolic, total antioxidant, sugars and organic acid content) of walnut fruit in subtropical climate conditions were investigated. In our research, the differences between varieties and genotypes were effective in de-ter-mining the biochemical content of walnut seeds and when the results obtained for bi-ochemical content were analyzed, it was determined that these differ-ences were sta-tistically significant (p<0.05). Among the biochemical contents of the analyzed walnut genotypes and varieties, it was found that fat had the highest value and linole-ic acid was the most abundant unsaturated fatty acid, followed by oleic and linolenic acids, respectively. In our study, the most dominant and major organic acid among all geno-types and cultivars at 500 m and 1200 m altitude was malic acid, followed by cit-ric, succinic, and oxalic acid. Sucrose and glu-cose were found to be the most common major sugars in the genotypes and cultivars examined. In our study, it was determined that altitude had significant effects on the bio-chemical contents of walnut genotypes and cultivars. It was determined that some contents increased and some contents de-creased due to the increase in altitude. While our study results are similar to the stud-ies in the literature, there are differences in terms of some characteristics. The differ-ences between the studies may be due to genetic factors, the effect of environ-ment and differences in methods.

In conclusion, the effect of altitude on walnut fruits was found to be significant. It was observed that there are limited number of studies indicating the effect of altitude on walnut fruits.On the other hand, there are not enough studies on organic acid and sugar content of walnut in the literature. With this study, the results of the effects of altitude on fruit quality and biochemical properties of walnuts, which are missing in the literature, are seen. Temperature is the most important factor affecting fruit quali-ty in walnuts. While high temperatures in summer cause inner darkening in walnuts low temperatures and cold winds negatively affect vegetative development and the physical and chemical structure of the fruit. As altitude increases, climate change changes the yield and quality of walnuts.

We believe that these genotypes will serve as a source for fu-ture studies, can be used as parents in walnut breeding studies, and will help interna-tional walnut breed-ers and researchers interested in the development of varieties with superior and better qualities. It is recommended that this study be carried out in a new collection in a dif-ferent season (dry/rainy).”.

Reviewer 3 Report

The article is of interest in the scientific community. The information in the manuscript is clear, organized and detailed. However, the results are not solid because there are factors that may be involved in the quality of the fruits, such as climate, frequency of irrigation, age of the crop, crop management, fertilization, incidence of pests and diseases during the growth and development of the trees.

 

In the summary the author indicates that he harvests in two different districts, however, the methodology does not describe this information in detail.

 

For this reason, it is suggested that the author provide information on the climate, irrigation frequency, crop management, fertilization, incidence of pests and diseases during the growth and development of the trees in the plantations where the fruits were harvested.

 

The conclusion should indicate the most outstanding results of the study and not describe the work of other authors or indicate perspectives.

 

The author must review the references in the text, in some cases he does not include the reference number.

 

The author must correctly cite the following paragraphs:

 

Line 255-257: Bayazıt et al (2020) investigated the performance of Chandler walnut variety at 100 m, 400 m, 800 m and 1100 m altitude. The researchers reported that fruit weight decreased after 400 m, and the weight of the kernel and the ratio of the kernel decreased after 800 m altitude.

 

 

Line 305- 315: In the study conducted by Muradoglu et al. (2010), the average fat content of walnut genotypes was found as 58.2%. Kafkas et al. (2017) determined that the fat content was between 49.4% and 70.70%. In another study, Kafkas et al. (2020) determined that the protein content was between 13.57% and 25.72%. In another study, Okatan et al. (2021) reported that total fat was between 50.88-64.28% in different walnut genotypes. It is observed that the studies conducted in previous years are in accordance with the findings obtained from our study. In addition, in our results, it was determined that with the in- crease in altitude, total protein and total fat contents decreased on average. However, in studies on Chandler walnut variety, Bayazıt et al. (2020) reported that total protein and total fat content increased with altitude

 

Line 335-337: Doğan and Akgül (2005), in their study of some walnut (Juglans regia L.) varieties from Eastern Anatolia, palmitic acid content ranged between 5.61% and 5.82%, while trace amounts of myristic acid

 

Line 342-344: In another study, Bayazıt and Sümbül (2012) found that palmitic acid values ranged 342 between 6.98% and 8.77%, stearic acid values ranged between 3.22% and 4.99% according 343 to the results obtained from the genotypes they selected from the Eastern Mediterranean 344 region of Türkiye.

 

Line 347-355: In another study, Kafkas et al. (2017) determined that palmitic acid content ranged between 5.61% and 5.82% and myristic acid was trace amount (<0.1%) in different walnut varieties. In another study 349 conducted by Kafkas et al. (2020), palmitic acid values ranged between 5.74% and 9.79% 350 and stearic acid values ranged between 2.04% and 3.00% in different walnut varieties. In 351 another study conducted by Okatan et al. (2021), palmitic acid content ranged between 352 6.72% and 9.06%, stearic acid between 0.07% and 0.66% and myristic acid between 0.12% 353 and 0.74% depending on the variety. When the results of our study are compared with 354 the findings of other studies conducted in previous years, it is seen that our results are 355 consistent and compatible>0.1%) in different walnut varieties. In another study conducted by Kafkas et al. (2020), palmitic acid values ranged between 5.74% and 9.79% and stearic acid values ranged between 2.04% and 3.00% in different walnut varieties. In another study conducted by Okatan et al. (2021), palmitic acid content ranged between 6.72% and 9.06%, stearic acid between 0.07% and 0.66% and myristic acid between 0.12% and 0.74% depending on the variety. When the results of our study are compared with the findings of other studies conducted in previous years, it is seen that our results are consistent and compatible.

 

Line 377-379: Kafkas et al. (2017) determined that the values of monounsaturated fatty acids 377 (palmitoleic and oleic acid) ranged between 0.11% and 14.36%, 0.13% and 27.57%, respec- 378 tively.

 

Line 381-382: Kafkas et al. (2020), oleic acid ranged between 10.85% and 20.65%, pal mitoleic acid between 0.06% and 0.46%

 

Line 395-399: Beyhan et al. (2017) found that linoleic acid varied between 53.23% and 63.62%, linolenic acid between 10.75% and 15.24% and oleic acid between 14.73% and 24.17%. Kafkas et al. (2017) determined that the values of polyunsaturated fatty acids (linoleic acid and linolenic acid) varied between 7.83% and 13.16%, 53.24% and 62.92%, respectively. In an- other study conducted in 2020, Kafkas et al. (2020) determined that linoleic acid varied between 58.96% and 66.07%, linolenic acid between 8.41% and 16.14%.

 

Linea407-409: Büyüksolak et al. (2020) reported that oleic and linolenic acid contents in walnuts increased with altitude. Our results were consistent and in accordance with the previously reported studies.

 

Line 480-482: In our study, it was found that malic acid, succinic acid and citric acid were found intensively in walnuts. This result supports the results of the study conducted by Koç et al. (2019).

Author Response

Dear Reviewer. Thank you for valuable comments.

• The article is of interest in the scientific community. The information in the manuscript is clear, organized and detailed. However, the results are not solid because there are factors that may be involved in the quality of the fruits,such as climate, frequency of irrigation, age of the crop, crop management, fertilization, incidence of pests and diseases during the growth and development of the trees.

We added the desired correction to both the method section and the conlusion section.

• In the summary the author indicates that he harvests in two different districts, however, the methodology does not describe this information in detail.

In both study regions, fruits were harvested at the same maturity levels. Subsequently, these collected samples underwent prompt transportation to the Department of Horticulture Laboratory within the Faculty of Agriculture at Çukurova University. This transportation was meticulously executed within a cold chain transportation system to preserve the samples' integrity. Cultural practises such as irrigation, fertilization and pruning are routinely carried out in the orchards.

• For this reason, it is suggested that the author provide information on the climate, irrigation frequency, crop management, fertilization, incidence of pests and diseases during the growth and development of the trees in the plantations where the fruits were harvested.

The following paragraph has been added to the method section.

Irrigation and fertilization of the plants were carried out regularly. A fertilization program was applied to the plants with approximately 800 g net Phosphorus and Potassium and 1500 g net Nitrogen per plant. Phosphorus and potassium were applied in a single dose in February, and Nitrogen was applied 1/3 in three months, with one-month intervals, starting from March. Microelement application was applied to the leaves as of the end of April. Irrigation was carried out using the drip irrigation method at approximately 15-day intervals from the end of May to the end of September.

• The conclusion should indicate the most outstanding results of the study and not describe the work of other authors or indicate perspectives.

The conclusion section of the study has been rewritten.

• The author must review the references in the text, in some cases he does not include the reference number.

 

In the study, relevant references and all other references were reviewed and necessary edits were made.

Reviewer 4 Report

The manuscript "The Effects of Altitude on Fruit Characteristics, Chemical, Biochemical, Sugar, Organic Acid and Fatty Acid Contents of Walnut Fruits (Juglans regia L.) makes an important contribution by evaluating different genotypes of walnut fruits, determining different characteristics of the fruits grown in regions with a Mediterranean climate, and comparing them with studies in the literature.

The results are very well presented, and my comments are below:

- In the abstract I suggest closing the text, emphasizing the relevance of the contributions of the results found, the way it was presented the abstract has a greater focus on the results found in the analyzes.

- Wouldn't it be interesting in a future study to carry out a new collection in a different season (dry/rainy) to check that there are no significant variations in the results found? Discuss in the text possible influences of the time of collection on the constitution of the fruit.

- The introduction mentions that the walnut has a wide range of uses and cites works showing these applications. In addition, it is mentioned that there are more than 22 species, the results found are valid only for the type studied, mention.

- In figure 1, determine what the values on the axes of the graph would be. Evaluate whether it would be necessary to keep the figure or only put the most important data in the text. The samples were collected in the years 2021-2022 but it is not clear whether in the rainy and/or dry season.

- Lines 285-289: "Studies related to the effect of elevation on biochemical properties in walnuts are insufficient. It has been reported that besides genetic structure, tree age, cultural practices, diseases and pests, ecological conditions and the number of fruits on the plant are also effective on biochemical contentes in fruits". Explain how the factors mentioned interfere with the biochemical content of the fruit and how to determine this.

Author Response

Dear Reviewer, thank you for spending your valuable time to review the article.

1) In the abstract I suggest closing the text, emphasizing the relevance of the contributions of the results found, the way it was presented the abstract has a greater focus on the results found in the analyzes.

Necessary corrections were made according to your suggestion. The abstract of the article has been changed to “This study aimed to investigate changes in fruit characteristics, total phenolics, total antioxidant capacity, organic acids, sugars, and fatty acid compositions of 10 walnut genotypes and 3 cultivars grown at different altitudes under Mediterranean climate conditions. At both 500 m and 1200 m altitudes, total phenolics ranged between 237.51 and 412.96 mg GAE/100 g dw and 234.94 and 392.31 mg GAE/100 g dw, respectively. Total antioxidant capacity ranged from 54.65% to 71.99% DPPH inhibition and 47.65% to 64.99% DPPH radical at 500 m altitude, and 64.76% to 75.92% DPPH inhibition and 57.76% to 68.92% DPPH radical at 1200 m altitude. Oil rate varied between 53.44% and 76.17% at 500 m altitude and 43.11% and 67.03% at 1200 m altitude. Protein rate ranged from 13.71% to 21.32% at 500 m altitude and 12.07% to 17.79% at 1200 m altitude. At both 500 m and 1200 m altitudes, linoleic acid was major fatty acid. It was followed by linolenic and oleic acids. Linoleic acid ranged from 58.82% to 62.44% at 500 m altitude 57.33 and 59.38 at 1200 m altitude. Linolenic acid varied between 7.93 and 9.72 % at 500 m altitude and 8.80 and 10.79 at 1200 m altitude. Malic acid was the major organic acid, ranging from 1.35% to 7.33% at both altitudes. Sucrose content in the cultivars ranged from 1.34% to 3.13%. In conclusion, this study showed that walnut seeds are rich in oil, protein, total phenolics, antioxidant activity, and fatty acids, with variations depending on the cultivar/genotype, especially affected by altitude”

2) Wouldn't it be interesting in a future study to carry out a new collection in a different season (dry/rainy) to check that there are no significant variations in the results found? Discuss in the text possible influences of the time of collection on the constitution of the fruit.

Necessary corrections were made according to your suggestion. It is explained as "“It is recommended that this study be carried out in a new collection in a different season (dry/rainy). Temperature is the most important factor affecting fruit quality in walnuts. While high temperatures in summer cause inner darkening in walnuts low temperatures and cold winds negatively affect vegetative development and the physical and chemical structure of the fruit. As altitude increases, climate change changes the yield and quality of walnuts" in the conclusion section of the article.

The article aims to explain the effect of altitude, not the difference between time of collection of the fruit. For this reason, collection times are not mentioned.

3) - The introduction mentions that the walnut has a wide range of uses and cites works showing these applications. In addition, it is mentioned that there are more than 22 species, the results found are valid only for the type studied, mention.

Juglans regia is the most produced species in the world. Therefore, studies of this type are mentioned.

4) In figure 1, determine what the values on the axes of the graph would be. Evaluate whether it would be necessary to keep the figure or only put the most important data in the text. The samples were collected in the years 2021-2022 but it is not clear whether in the rainy and/or dry season.

We believe that Figure 1 will be more visually striking. For this reason, climate data is given as a figure.

There is a sentence "Kahramanmaraş with Mediterranean climate characteristics has warm and rainy winters and dry and hot summers." regarding the climate structure of the region.

5) Lines 285-289: "Studies related to the effect of elevation on biochemical properties in walnuts are insufficient. It has been reported that besides genetic structure, tree age, cultural practices, diseases and pests, ecological conditions and the number of fruits on the plant are also effective on biochemical contentes in fruits". Explain how the factors mentioned interfere with the biochemical content of the fruit and how to determine this.

The sentence "In fact, these factors affect nutritional physiology. Additionally, it has been stated by many researchers that temperature has an effect on the contents of fruits. Thefore, many factors are effective on the biochemical contents of fruits" has been added under the heading 3.2.Total Phenolics and Total Antioxidant Contents.

Round 2

Reviewer 1 Report

I only see the changed paper title according to my suggestion, other parts were basically not revised, particularly, the abstract contains too much wordy descriptions of experiment methods or data, which should be included in the Methods and Results sections. 

Also many unnecessary words in the Abstract, for example,

since the authors accepted my suggestions regarding title changes, they should also not put "fruit characteristics and chemical, biochemical, sugar, organic acid and fatty acid contents of walnuts" together in parallel, because these concepts in different scopes. 

 also " Among genotypes and cultivars, myristic, palmitic and stearic acid were determined as saturated fatty acids; linoleic, linolenic, oleic and palmitoleic acid as unsaturated fatty acids for both altitudes."  "genotypes and cultivars" can not be mentioned in parallel. 

myristic, palmitic and stearic acid have been regarded as saturated fatty acids for a long time as common sense.  So do linoleic, linolenic, oleic and palmitoleic acid as unsaturated fatty acids. You do not need to define them, you only need to determine their contents and changes in different proportions of walnut fruits harvested in different varieties grown under different altitudes.

I do not want to mention many other writing problems. The manuscript needs to be revised, according to what I mentioned in the last round of review.  Particularly, the Abstract needs to be concisely rewritten, and other parts also need to be precisely revised.  

 

 

 

 authors should do that

Author Response

Dear Reviewer 1;

First of all, thank you for your detailed evaluation of our article. We have partially made the revisions you requested. However, since the publication went to 3 other reviewers other than you, and they gave a “publishable” opinion, we cannot make some of your revisions in order not to compromise the integrity of the study. To summarize these;

1) I only see the changed paper title according to my suggestion, other parts were basically not revised, particularly, the abstract contains too much wordy descriptions of experiment methods or data, which should be included in the Methods and Results sections.

We changed the article title in the 1st revision in line with your suggestions. We have updated the Abstract section once again in line with your suggestion. We have shortened the summary section considerably. Additionally, the method section has been revised in line with your suggestions.

2. Since the authors accepted my suggestions regarding title changes, they should also not put "fruit characteristics and chemical, biochemical, sugar, organic acid and fatty acid contents of walnuts" together in parallel, because these concepts in different scopes.

We strongly agree with your opinion that fruit characteristics and other examined characteristics should be evaluated separately. However, our aim was to examine whether altitude has an effect on fruit characteristics as well as these characteristics. You will also appreciate that we have worked hard on these features, and in the literature review, there are studies that combine various features such as biochemical properties, sugar, and fatty acid content. That's why we don't want to exclude fruit characteristics from the study. I share some of these studies with you.

10.3390/foods11050761

10.5586/aa.7333

https://scholar.google.com/scholar?hl=tr&as_sdt=0%2C5&q=Effect+of+altitude+on+fruit+and+biochemical+properties+of+nut+fruits&btnG=#d=gs_qabs&t=1695566507450&u=% 23p%3D_O1h5mu6LmcJ

3. Also "Among genotypes and cultivars, myristic, palmitic and stearic acid were determined as saturated fatty acids; linoleic, linolenic, oleic and palmitoleic acid as unsaturated fatty acids for both altitudes." "genotypes and cultivars" cannot be mentioned in parallel.

We absolutely agree with your opinion. However, the genotypes used in the study are at the "variety registration" stage. Therefore, it is an important step in breeding to compare genotypes with varieties registered at different altitudes and conditions such as soil properties. In this way, the advantages and disadvantages of the varieties to be registered compared to the existing varieties are important for the breeder. Therefore, we regretfully could not make this revision.

 

4) Myristic, palmitic and stearic acid have been regarded as saturated fatty acids for a long time as common sense. So do linoleic, linolenic, oleic and palmitoleic acid as unsaturated fatty acids. You do not need to define them, you only need to determine their contents and changes in different proportions of walnut fruits harvested in different varieties grown under different altitudes.

The revision you requested has been made. All parts of the article have been revised.

5. I do not want to mention many other writing problems. The manuscript needs to be revised, according to what I mentioned in the last round of review. Particularly, the Abstract needs to be concisely rewritten, and other parts also need to be precisely revised.

The article has been extensively revised, especially the abstract section. All changes made to the article uploaded to the system have been made in line with the revisions you requested. We hope the article is suitable for acceptance.

We genuinely appreciate your feedback and your willingness to review our work again after the major revisions have been completed. Your valuable insights and input are crucial in ensuring the quality and rigor of our research. We look forward to addressing your comments and making the necessary improvements to meet your expectations if need be. Thank you for your time and commitment to the review process.

Author Response File: Author Response.pdf

Reviewer 2 Report

It has been substantially improved. It is ready for its acceptation. 

Author Response

Thank u for efforts