Wheat, Barley, and Triticale Response to Nitrogen Fertilization in Pannonian Environment

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Nitrogen is essential for plant growth and development. Nowadays, a large of nitrogen fertilizer is applied, resulting in low nitrogen utilization efficiency, waste of resources, and harm to the environment. It is urgent to explore the rational crop type and nitrogen fertilizer application. This study explores the response of small grain producers to nitrogen gradients in the Pannonian environment and can help small grain producers improve nitrogen uptake and utilization in rain-fed systems. Some insights on the utilization of nitrogen fertilizer in Pannonia were provided. However, there are still many deficiencies in this study, and it is recommended to accept it after modification.

Major comments

 

1. Abstract: L10-L14 ‘Small grains producers in the southern Pannonian plain prefer winter barley production on poor soils and drought-prone areas’------------ No clear explanation as to why these crops were chosen.

 

2. Lines 10-11 “Small grains producers in the southern Pannonian plain prefer winter barley production on poor soils and drought-prone areas, assuming higher resource use efficiency in barley than in wheat.” -------------Whether the soil properties is the same as that of the test site.

 

3. L15-L16: ‘Therefore, this study aimed to compare the performance of wheat, triticale, six-rowed, and two-rowed barley under different fertilization levels in the southern Pannonian plain’------------- Lack of specific experimental design and expected objectives.

 

4. Lines 15-20 “Nitrogen fertilization had a positive effect on the grain yield and nitrogen uptake in all studied crops. Grain yield was closely related to nitrogen uptake. Nitrogen use efficiency showed a positive relationship with nitrogen uptake efficiency, while the relationship with nitrogen utilization efficiency was insignificant. There was a notable difference between crops in terms of grain yield and nitrogen use efficiency traits.” --------------------That seems to be a fact. In the abstract, a lot of exposition of facts is a bad performance, the author should pay more attention to the in-depth exploration of research results.

 

5. Since this study takes small grain crops as the research object, its concept and importance should be fully explained in introduction.

 

6. Lines 61-91 There are some repetitions, and the manuscript needs to straighten out his thinking.

 

7. The manuscript lacks hypotheses.

 

8. Experimental Design and Methods: L113 ‘Table 1. Basic description of experimental locations in the southern Pannonian plain’------------- More basic information about soil properties should to be displayed, such as PH and organic matter

 

9. Lines 126-127 “Although there is a significant genotypic variation within each species, to efficiently manage different locations and N fertilization combinations during the crops growth cycle the two most representative cultivar for each species were selected.” By what means does the manuscript identify the most representative cultivar. Supplement it.

 

10. L131-L132------------- Lack of specific soil measurement methods

 

11. L155-L156 ‘For each treatment the mean difference was analyzed using the Tukey test at a 0.05 probability level’------------- The wording is not clear enough to be misunderstood as ‘a Tukey test is performed for the mean difference of each treatment’, in fact the Tukey test is used to compare the mean difference between multiple treatment groups.

 

12. Result: “The results analysis section appears to be somewhat complex and includes a considerable amount of detailed information. While these details may be informative, they may also make it challenging for readers to quickly grasp the key findings. Such as L161-L162 ‘(4818 - 9418 kg ha-1 in wheat, 6317 - 10016 kg ha-1 in triticale, 5001 – 9900 kg ha-1 two-rowed barley and 4673 - 10655 kg ha-1 in six-rowed barley; Fig 1).’, L186-L188-------------- I suggest the authors highlight the most critical results and explanations.

 

13. I think the authors should add the yield of each crop, nitrogen fertilizer use efficiency and other relevant data in the supplementary materials.

 

14. The use of a crop is closely related to its benefit. This manuscript considers its use, but does not mention its benefit. I believe that benefit is the main influence factor for producers to consider the type of crop. I think you should mention the economic benefits.

 

15. The manuscript reports trials conducted during the 2019-20 and 2020-21 seasons. However, the results section lacks a comprehensive comparison of data across years.

 

16. L175-L176, ‘In approximately 2/3 combination of treatments,’----------- The manuscript would benefit from the use of more precise and rigorous scientific terminology.

 

17. L196, ‘3.2. Differences in nitrogen use efficiency’-----------delate ‘Differences’

 

 

18. L197-L198’ There were significant differences between winter wheat, triticale, two-rowed and six-rowed barley in the NUE across different locations and fertilization levels (Fig 3).’------------ Lack of significance tests (e.g., p-values). Additionally, reporting the coefficient of determination (R²) would provide valuable information about the goodness of fit of the models.

 

19. Discuss: 1) The influence of region × rainfall on yield and nitrogen fertilizer is not discussed. In Table1, there is a large difference in rainfall between years. In addition, there is a large difference in rainfall between regions in the same year, such as Novi Sad and Sombor in 2020. 2) The possible physiological mechanisms (such as root nitrogen absorption efficiency) of triticale for high yield under low nitrogen conditions need to be further explored, rather than citing previous studies. 3) L296-L300---- The discussion of a restricted triticale market is weakly associated with scientific conclusions

 

Minor comments

 

20. Fig1-------The symbols (such as □, ▲) may be difficult to distinguish when printing, it is recommended to use solid/hollow shapes or add legend instructions.

 

21. Fig2 ------- The regression line should be marked with significance level to enhance the reliability of the results.

 

22. Fig units (e.g. "kgcrain kg available N⁻¹" and "kggrain kg Nin biomass⁻¹") should be unified into international standard abbreviations (e.g. "kgkg⁻¹").

 

23. Table1----- Please ensure that all chemical formulas and mathematical expressions are accurately formatted and consistent throughout the manuscript, such as Ca2Co3 subscripts.

 

24. What is “2R” “6R” “W” “T”? Supplement in the manuscript.

 

25. The formula and its composition should be described and explained in detail in the manuscript.

 

26. Standard chart format.

 

27. References should be rearranged according to the journal's format.

Comments on the Quality of English Language

The writing of the paper needs improvement with the assistance of English editing

Author Response

Nitrogen is essential for plant growth and development. Nowadays, a large of nitrogen fertilizer is applied, resulting in low nitrogen utilization efficiency, waste of resources, and harm to the environment. It is urgent to explore the rational crop type and nitrogen fertilizer application. This study explores the response of small grain producers to nitrogen gradients in the Pannonian environment and can help small grain producers improve nitrogen uptake and utilization in rain-fed systems. Some insights on the utilization of nitrogen fertilizer in Pannonia were provided. However, there are still many deficiencies in this study, and it is recommended to accept it after modification.

Major comments

 

1. Abstract: L10-L14 ‘Small grains producers in the southern Pannonian plain prefer winter barley production on poor soils and drought-prone areas’------------ No clear explanation as to why these crops were chosen.

Authors: We thank the reviewer for their observation regarding the explanation of crop selection in the abstract. Due to the strict character limit for the abstract, we were unable to include a detailed rationale for the choice of crops. However, a comprehensive explanation of why winter barley and other small grains are preferred in the southern Pannonian plain—particularly for poor soils and drought-prone areas—is provided in the Introduction section.

 

1. Lines 10-11 “Small grains producers in the southern Pannonian plain prefer winter barley production on poor soils and drought-prone areas, assuming higher resource use efficiency in barley than in wheat.” -------------Whether the soil properties is the same as that of the test site.

Authors: This sentence aimed to briefly describe the differences between the crops and was not directly related to the experiment. Regarding soil properties, the experiment was conducted at three representative locations, and soil conditions were uniform due to randomization.

 

1. L15-L16: ‘Therefore, this study aimed to compare the performance of wheat, triticale, six-rowed, and two-rowed barley under different fertilization levels in the southern Pannonian plain’------------- Lack of specific experimental design and expected objectives.

Authors: Two sentences describing the experimental design have been added to the abstract, along with a clearer statement of the study’s objective.

 

1. Lines 15-20 “Nitrogen fertilization had a positive effect on the grain yield and nitrogen uptake in all studied crops. Grain yield was closely related to nitrogen uptake. Nitrogen use efficiency showed a positive relationship with nitrogen uptake efficiency, while the relationship with nitrogen utilization efficiency was insignificant. There was a notable difference between crops in terms of grain yield and nitrogen use efficiency traits.” --------------------That seems to be a fact. In the abstract, a lot of exposition of facts is a bad performance, the author should pay more attention to the in-depth exploration of research results.

Authors: Specific numerical results have been added to the abstract to better highlight key findings.

 

1. Since this study takes small grain crops as the research object, its concept and importance should be fully explained in introduction.

Authors: A several sentences elaborating on the significance of small grains, including wheat, barley, and triticale, have been incorporated into the Introduction.

 

1. Lines 61-91 There are some repetitions, and the manuscript needs to straighten out his thinking.

Authors: The sentence “Although the information on N management and NUE is widely available for the different European regions, as far as we are aware the data about the comparisons of the winter bread wheat, triticale, two-rowed, and six-rowed barley within the same trials are unavailable for the Pannonian environment.” was similar to another statement and has been removed to improve clarity.

 

1. The manuscript lacks hypotheses.

Authors: A hypothesis has been added before the Objectives section.

 

1. Experimental Design and Methods: L113 ‘Table 1. Basic description of experimental locations in the southern Pannonian plain’------------- More basic information about soil properties should to be displayed, such as PH and organic matter

Authors: We thank the reviewer for their valuable suggestion. In response to the comment, we have added detailed information regarding soil properties, including pH and organic matter content, to Table 1.

 

1. Lines 126-127 “Although there is a significant genotypic variation within each species, to efficiently manage different locations and N fertilization combinations during the crops growth cycle the two most representative cultivar for each species were selected.” By what means does the manuscript identify the most representative cultivar. Supplement it.

Authors: We appreciate the reviewer's comment regarding the selection of cultivars. To clarify, the manuscript does not aim to identify or define the most representative cultivars. Instead, the selection of the two cultivars for each species was based on their widespread use, adaptability, and relevance to the study's objectives, as explained in the preceding text. These cultivars were chosen to ensure the results are applicable to common agricultural practices in the region.

 

1. L131-L132------------- Lack of specific soil measurement methods

Authors: In response to this comment, we have added the specific methodology used for soil analysis in the revised manuscript.

 

1. L155-L156 ‘For each treatment the mean difference was analyzed using the Tukey test at a 0.05 probability level’------------- The wording is not clear enough to be misunderstood as ‘a Tukey test is performed for the mean difference of each treatment’, in fact the Tukey test is used to compare the mean difference between multiple treatment groups.

Authors: We thank the reviewer for their insightful comment regarding the clarity of the statistical analysis description. We acknowledge that the original wording could have been misinterpreted. To address this, we have revised the text

 

1. Result: “The results analysis section appears to be somewhat complex and includes a considerable amount of detailed information. While these details may be informative, they may also make it challenging for readers to quickly grasp the key findings. Such as L161-L162 ‘(4818 - 9418 kg ha-1 in wheat, 6317 - 10016 kg ha-1 in triticale, 5001 – 9900 kg ha-1 two-rowed barley and 4673 - 10655 kg ha-1 in six-rowed barley; Fig 1).’, L186-L188-------------- I suggest the authors highlight the most critical results and explanations.

Authors: We thank the reviewer for their valuable suggestion to the Results section. In response, we have revised the text to highlight key results and their significance, reducing unnecessary complexity while retaining essential information. Additionally, as suggested, we have incorporated percentage changes to provide a clearer and more intuitive understanding of the observed variations.

 

1. I think the authors should add the yield of each crop, nitrogen fertilizer use efficiency and other relevant data in the supplementary materials.

Authors: In response, we have created a supplementary file containing the average values for each crop, fertilization treatment, year, and location, along with other relevant traits. However, due to the complexity of the experimental design, which involves multiple factors, we have not included interaction results in the supplementary materials. Presenting these interactions would further complicate the data analysis and potentially overwhelm readers. Instead, we have focused on providing clear and concise summaries of the key findings in the main text

 

14. The use of a crop is closely related to its benefit. This manuscript considers its use, but does not mention its benefit. I believe that benefit is the main influence factor for producers to consider the type of crop. I think you should mention the economic benefits.

Authors: In response, we have added a section to the Discussion that addresses the economic benefits of cultivating wheat, triticale, and barley under different nitrogen fertilization levels.

 

1. The manuscript reports trials conducted during the 2019-20 and 2020-21 seasons. However, the results section lacks a comprehensive comparison of data across years.

Authors: In response, we have added a supplementary table that provides a detailed comparison of the results between the 2019-20 and 2020-21 seasons. Additionally, we have included a discussion in the Results and Discussion sections to highlight the differences observed between the two seasons. However, as the primary focus of this study is to compare the performance of different crops (wheat, triticale, and barley) under varying nitrogen fertilization levels, the analysis is predominantly centered on crop-related differences rather than year-to-year or location-specific variations.

 

1. L175-L176, ‘In approximately 2/3 combination of treatments,’----------- The manuscript would benefit from the use of more precise and rigorous scientific terminology.

Authors: We thank the reviewer for their suggestion to improve the precision of our scientific terminology. In response, we have replaced the phrase "In approximately 2/3 combination of treatments" with the more precise and rigorous term "In the majority of treatment combinations." This change ensures greater clarity and aligns with standard scientific writing practices

 

17. L196, ‘3.2. Differences in nitrogen use efficiency’-----------delate ‘Differences’

Authors: n response, we have removed the word "Differences" from the heading

 

 

1. L197-L198’ There were significant differences between winter wheat, triticale, two-rowed and six-rowed barley in the NUE across different locations and fertilization levels (Fig 3).’------------ Lack of significance tests (e.g., p-values). Additionally, reporting the coefficient of determination (R²) would provide valuable information about the goodness of fit of the models.

Authors: To clarify, Figure 3 was designed to provide a visual comparison of nitrogen use efficiency (NUE) across different crops, locations, and fertilization levels, rather than to present regression analysis or statistical significance. The graph uses a straightforward approach, where the position of each symbol relative to the diagonal line (from the lower left to the upper right corner) indicates the relative NUE performance of each crop. Symbols in the lower right quadrant represent higher NUE values for the crop on the x-axis, while those in the upper left quadrant indicate higher NUE values for the crop on the y-axis.

 

1. Discuss: 

1) The influence of region × rainfall on yield and nitrogen fertilizer is not discussed. In Table1, there is a large difference in rainfall between years. In addition, there is a large difference in rainfall between regions in the same year, such as Novi Sad and Sombor in 2020.

Authors: In response, we have added a discussion on the variability in yield between seasons and locations, including the impact of rainfall differences, as highlighted in Table 1. However, the primary focus of this study remains on comparing the performance of different crops (wheat, triticale, and barley) under varying nitrogen fertilization levels, rather than on year-to-year or location-specific variations

 

2) The possible physiological mechanisms (such as root nitrogen absorption efficiency) of triticale for high yield under low nitrogen conditions need to be further explored, rather than citing previous studies.

Authors: We agree with the reviewer that exploring the physiological mechanisms, such as root nitrogen absorption efficiency, could provide valuable insights into triticale's high yield under low nitrogen conditions. However, our study did not include an analysis of the root system, which limits our ability to directly address this aspect. Nevertheless, in the Discussion section, we have incorporated additional text supported by existing literature to highlight the potential role of triticale's root system and its nitrogen uptake efficiency in contributing to its performance under low nitrogen conditions.

3) L296-L300---- The discussion of a restricted triticale market is weakly associated with scientific conclusions

Authors: We thank the reviewer for their valuable comment regarding the discussion of triticale's market limitations. In response, we have revised the text to better align with the scientific conclusions of the study.

 

Minor comments

 

1. Fig1-------The symbols (such as □, ▲) may be difficult to distinguish when printing, it is recommended to use solid/hollow shapes or add legend instructions.

Authors: In response, we have added further clarification to the figure caption to ensure the symbols are easily distinguishable.

 

1. Fig2 ------- The regression line should be marked with significance level to enhance the reliability of the results.

Authors: We acknowledge that this issue also occurred in Figure 1, likely due to an error during the figure preparation process. To address this, we have corrected both figures by adding the significance levels to the regression lines. Additionally, we have updated the captions and text below the figures to provide clearer explanations of the statistical significance, ensuring the reliability and transparency of the results.

 

1. Fig units (e.g. "kgcrain kg available N⁻¹" and "kggrain kg Nin biomass⁻¹") should be unified into international standard abbreviations (e.g. "kgkg⁻¹").

Authors: We thank the reviewer for their comment regarding the units used in the figures. The units "kg grain kg available N⁻¹" and "kg grain kg N in biomass⁻¹" were adopted from the study by Cossani et al. (2012) (Cossani, C.M., Slafer, G.A. and Savin, R., 2012. Nitrogen and water use efficiencies of wheat and barley under a Mediterranean environment in Catalonia. Field Crops Research, 128, pp.109-118; https://doi.org/10.1016/j.fcr.2012.01.001), as they are widely recognized and used in the field of agronomy and crop science. We believe these units are appropriate and align with established scientific conventions.

 

1. Table1----- Please ensure that all chemical formulas and mathematical expressions are accurately formatted and consistent throughout the manuscript, such as Ca2Co3 subscripts.

 

1. What is “2R” “6R” “W” “T”? Supplement in the manuscript.

Authors: We thank the reviewer for pointing out the need to clarify the abbreviations "2R," "6R," "W," and "T" in the manuscript. In response, we have added explanations for these abbreviations in the figure captions and relevant sections of the text.

 

1. The formula and its composition should be described and explained in detail in the manuscript.

Authors: We have included the formula for calculating nitrogen use efficiency (NUE) in the manuscript, along with a clear explanation of its components. Additionally, we have referenced the study from which the methodology for calculating NUE was adopted.

 

1. Standard chart format.

Authors: If this refers to Chart 1, we have revised it to align with standard formatting guidelines, ensuring clarity and consistency in the presentation of data

 

1. References should be rearranged according to the journal's format.

Authors: we have carefully rearranged and formatted all references according to the journal's guidelines. Each reference now follows the required structure, including proper abbreviations, punctuation, and order of elements

 

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

The authors determined the effect of different nitrogen doses (50 and 100 kg/ha) on selected parameters in the cultivation of different varieties of wheat, triticale, two-row and six-row barley in different locations and growing seasons. The concept and scope of the conducted research seem appropriate, but extremely uninnovative. The work does not contribute much new to the existing state of knowledge. A large part of the observations can be predicted based on the existing state of knowledge, without conducting these experiments. Many of the presented discoveries are truisms, widely known and used for a long time in agricultural practice.

In addition, the manuscript gives the impression of being generally well-written. The exception is the chapter "Materials and Methods", which should be corrected. The authors should describe the NPK fertilization schedule in detail. A dose of 100 kg N/ha is not a high dose.

I have serious doubts about the method of calculating the NUE index. The differential method is commonly used, and it indicates the part of nitrogen taken up by the plant in relation to the amount of nitrogen introduced into the soil. The authors proposed a completely different indicator hidden under the name NUE (nitrogen use efficiency). This misleads the reader. Please provide a reference to the literature or describe in detail the meaning of calculating this index. The same applies to the NUpE indicator.

Authors should consider calculating NUE using traditional formulas.

All indicators should be provided with clear mathematical formulas and their explanation.

Examples from the "Discussion" chapter:

Lines 256-258: "These results challenge the generally accepted belief that barley is more resilient to low-fertility soil and stress than other cereal crops, favoring barley production in low input and yielding conditions." - this is absolutely not true

Lines 304-306: "Our findings demonstrate a positive response of various small-grain cereals to nitrogen fertilization treatments, with the yield response predominantly higher under increased nitrogen levels." - a truism

Lines 341-343: "The greater NUE of triticale compared to other cereals was mostly the result of increased nitrogen (N) uptake, indicating higher N soil recovery by triticale." The NUE index calculated as described in the M&M chapter does not indicate anything like that.

Best regards,

Reviewer

Author Response

Dear Authors,

The authors determined the effect of different nitrogen doses (50 and 100 kg/ha) on selected parameters in the cultivation of different varieties of wheat, triticale, two-row and six-row barley in different locations and growing seasons. The concept and scope of the conducted research seem appropriate, but extremely uninnovative. The work does not contribute much new to the existing state of knowledge. A large part of the observations can be predicted based on the existing state of knowledge, without conducting these experiments. Many of the presented discoveries are truisms, widely known and used for a long time in agricultural practice.

 

In addition, the manuscript gives the impression of being generally well-written. The exception is the chapter "Materials and Methods", which should be corrected. The authors should describe the NPK fertilization schedule in detail. A dose of 100 kg N/ha is not a high dose.

Authors: We added aplied doses of PK fertilazers in Table 1. Concerning the dose of 100 kg N/ha, we acknowledge the reviewer's observation that this may not be considered a high dose in some contexts. However, in the conditions of the Panonian Plain, where there has been a recent trend of reduced fertilizer application due to regulatory and market factors, we referred to this dose as "high" to reflect the current agricultural practices and constraints in the region.

 

I have serious doubts about the method of calculating the NUE index. The differential method is commonly used, and it indicates the part of nitrogen taken up by the plant in relation to the amount of nitrogen introduced into the soil. The authors proposed a completely different indicator hidden under the name NUE (nitrogen use efficiency). This misleads the reader. Please provide a reference to the literature or describe in detail the meaning of calculating this index. The same applies to the NUpE indicator.

Authors should consider calculating NUE using traditional formulas.

All indicators should be provided with clear mathematical formulas and their explanation.

Authors: We sincerely thank the reviewer for their thorough and insightful comments regarding the methodology used for calculating the Nitrogen Use Efficiency (NUE) and Nitrogen Uptake Efficiency (NUpE) indices. We acknowledge the concerns raised and would like to clarify the approach taken in our study.The methodology used for calculating NUE and NUpE in our work is based on well-established definitions and calculations from the literature. Specifically, we followed the approach described by Moll et al. (1982) in their seminal paper, where NUE is defined as grain production per unit of N available in the soil (Moll RH, Kamprath EJ, Jackson WA. Analysis and interpretation of factors which contribute to efficiency of nitrogen utilization. Agronomy Journal. 1982;74(3):562-564. https://doi.org/10.2134/agronj1982.00021962007400030037x). In this work, NUE is calculated as the ratio of grain yield to the total N available to the crop from both soil and fertilizer sources. This definition aligns with our calculation, where NUE was determined by dividing the grain yield by the amount of N available to the crop from the soil and applied fertilizer.

Furthermore, Moll et al. (1982) also describe two primary components of NUE: (1) the efficiency of N uptake (NUpE), calculated as the ratio of total N in the plant at maturity (Nt) to the N available in the soil (Ns), and (2) the efficiency of N utilization (NUtE), calculated as the grain yield (Gw) per unit of N taken up by the plant (Nt). Our approach to calculating NUpE and NUtE follows this framework, which has been widely adopted in subsequent studies.

For instance, Hawkesford and Riche (2020) provide a similar definition of NUE as the yield of grain produced per unit of N available to the crop, expressed as kg yield per kg of available N. They also emphasize that NUE is the product of NUpE and NUtE, where NUpE is the ratio of N taken up by the crop to the N available from the soil and fertilizer, and NUtE is the grain yield produced per unit of N taken up by the plant (Hawkesford MJ, Riche AB. Impacts of G x E x M on nitrogen use efficiency in wheat and future prospects. Frontiers in Plant Science. 2020;11:1157. https://doi.org/10.3389/fpls.2020.01157).

Additionally, similar methodologies have been employed in more recent studies, such as Biradar et al. (2024), who define NUE as the yield produced per unit of available N to the crop, expressed in terms of kg yield per kg of available N. They also highlight that NUE is the product of NUpE and NUtE, consistent with our approach (Biradar SS, Patil MK, Desai SA, et al. Nitrogen use efficiency in bread wheat: Genetic variation and prospects for improvement. PLoS ONE. 2024;19(4):e0294755 ).

We recognize that there may be differing definitions of NUE in the literature, ranging from agronomic to physiological perspectives. However, our methodology was not intended to mislead readers but rather to align with established definitions and calculations from foundational and contemporary studies. We have now included additional references and a more detailed explanation of the calculations in the revised manuscript to ensure clarity and transparency.

 

 

Examples from the "Discussion" chapter:

Lines 256-258: "These results challenge the generally accepted belief that barley is more resilient to low-fertility soil and stress than other cereal crops, favoring barley production in low input and yielding conditions." - this is absolutely not true

Authors: We agree with the reviewer's comment regarding the statement in lines 256-258. Upon reflection, we recognize that the claim about barley's resilience to low-fertility soil and stress compared to other cereal crops may have been overstated and not fully supported by our data. We have revised this sentence to better reflect the findings of our study and to acknowledge that barley's performance is highly dependent on the specific genotype.

In the revised manuscript, we have added that the performance of barley in our study varied significantly depending on the genotype, with notable differences in yield between six-rowed and two-rowed barley types. This clarification aligns with our results and provides a more nuanced interpretation of the data.

Thank you for pointing this out, as it has allowed us to improve the accuracy and clarity of our discussion.

 

Lines 304-306: "Our findings demonstrate a positive response of various small-grain cereals to nitrogen fertilization treatments, with the yield response predominantly higher under increased nitrogen levels." - a truism

Authors: We thank the reviewer for their observation regarding the statement in lines 304-306. We agree that the original sentence may have come across as a truism, and we have revised it to provide more specific and meaningful insights. The sentence has been expanded to highlight the differences in NUE (Nitrogen Use Efficiency) and NUE-related traits (such as N uptake efficiency and N utilization efficiency) among the crops, which adds depth to the discussion and avoids the issue of stating the obvious. We believe the revised version better reflects the scientific contribution of our findings.

 

Lines 341-343: "The greater NUE of triticale compared to other cereals was mostly the result of increased nitrogen (N) uptake, indicating higher N soil recovery by triticale." The NUE index calculated as described in the M&M chapter does not indicate anything like that.

Authors: We thank the reviewer for their comment regarding the interpretation of the NUE results. We hope that with the additional clarification of the methodology provided in the revised manuscript, it is now clearer how the NUE index was calculated. Based on the established methodology, which defines NUE as grain yield per unit of N available to the crop, and considering the components of NUE (N uptake efficiency and N utilization efficiency), we believe that the conclusion regarding triticale's higher NUE being linked to increased nitrogen uptake and soil recovery is acceptable

 

 

 

Reviewer 3 Report

Comments and Suggestions for Authors

Comments and suggestions for authors attached.

Comments for author File: Comments.pdf

Author Response

Reviewer: After objective add experimental site, experimental design and treatments.

Authors: As suggested, we have included detailed information about the experimental site, experimental design, and treatments in the revised manuscript. This

 

Reviewer: Add the percentage increase.

Authors: We have incorporated the percentage changes in the relevant sections of the manuscript, as recommended by the reviewer.

 

Reviewer: Increase in-text citations.

Authors: We thank the reviewer for their suggestion to enhance the discussion and we incorporated new in-text citations to support our findings and strengthen the scientific context of our work

Reviewer 4 Report

Comments and Suggestions for Authors

The title is basically adequate, but it would be better to emphasize in the Mediterranean climate than the Pannonian lowland.
The order of keywords should be changed slightly - first list all the species studied and then grain yield and NUE.
The weather conditions should be described in a slightly broader way. It would be necessary to add information on whether the conditions in the years studied were similar to the multi-year average and whether the distribution of precipitation was relatively even.This is important because the research was conducted in only two growing seasons.
The conclusions should be changed. In this chapter, reference should be made primarily to the inter-species differences in NUE, because this was the main goal of the work.
There is no doubt that in practice, the choice of species for cultivation is determined by economic considerations, so it is good that the authors drew attention to this in the discussion.
In general, the work is understandable and clear and can be an interesting reference for other studies on the efficiency of nitrogen use by different species (varieties) in different conditions, e.g. related to environmental stresses.

Author Response

Reviewer: "The title is basically adequate, but it would be better to emphasize the Mediterranean climate rather than the Pannonian lowland."

Authors: We appreciate the reviewer’s comment. However, there are notable climatic differences between the Mediterranean region and the Pannonian lowland. In particular, winters in the Pannonian lowland are colder, precipitation is more evenly distributed throughout the year, and the climate is more continental with less maritime influence. That being said, in recent years, these differences have been decreasing. If the reviewer agrees, we would prefer to retain "Pannonian environement" in the title.

 

Reviewer: "The order of keywords should be changed slightly – first list all the species studied and then grain yield and NUE."

Authors: The keywords have been rearranged according to the reviewer’s suggestion.

 

Reviewer: "The weather conditions should be described in a slightly broader way. It would be necessary to add information on whether the conditions in the years studied were similar to the multi-year average and whether the distribution of precipitation was relatively even. This is important because the research was conducted in only two growing seasons."

Authors: We have added columns with multi-year averages and commented on the weather conditions in accordance with the reviewer’s suggestions.

 

Reviewer: "The conclusions should be changed. In this chapter, reference should be made primarily to the inter-species differences in NUE, because this was the main goal of the work."

Authors: The conclusions have been revised to emphasize inter-species differences in NUE, in line with the reviewer’s comments.

 

Reviewer: "There is no doubt that in practice, the choice of species for cultivation is determined by economic considerations, so it is good that the authors drew attention to this in the discussion."

Authors: We have added a sentence highlighting the importance of wheat and other crops in the production systems of the studied region.

 

In general, the work is understandable and clear and can be an interesting reference for other studies on the efficiency of nitrogen use by different species (varieties) in different conditions, e.g. related to environmental stresses.

 

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

1. L136’ In order to minimize possible N losses, N was applied in two doses after emergence (GS11-12) and prior to the start of the stem elongation period (GS30)’---------------Supplement the specific amount of nitrogen applied each time

 

2. These are many factors (such as cultivars, N levels, locations), ------------However, the interactions between these factors are not adequately explored in the results and discussion sections.

 

3. Some conclusions drawn in this study differ from previous research such as L316-318 ‘barley is more resilient to low-fertility soil and stress’ -------------A more in-depth discussion is needed here, preferably with comparisons to existing studies, to better understand the discrepancies and the implications of the new findings.

 

4. The economic analysis presented is somewhat weak -------------Could benefit from additional supporting literature. Alternatively, the authors may consider supplementing the analysis with cost-effectiveness data or other relevant economic indicators to provide a more complete picture.

 

5. L415 Reference does not correctly cite '[341]'

 

6. Table 1, the data for "Rainfall mm" in Novi Sad 2020 shows a discrepancy between "72.7" and the previous value of "42,7"-------------- The author has replaced the data without providing an explanation. A clarification regarding this data change is necessary to maintain the integrity and transparency of the dataset.

 

Author Response

1. L136’ In order to minimize possible N losses, N was applied in two doses after emergence (GS11-12) and prior to the start of the stem elongation period (GS30)’---------------Supplement the specific amount of nitrogen applied each time

Authors: We have supplemented the text with the exact amounts of nitrogen applied during each application. Nitrogen was applied in two equal doses: the first after emergence (GS11-12) and the second prior to the start of the stem elongation period (GS30). For the moderate N level (50 kg N ha⁻¹), 25 kg N ha⁻¹ was applied at each timing (25 + 25 kg ha⁻¹), and for the high N level (100 kg N ha⁻¹), 50 kg N ha⁻¹ was applied at each timing (50 + 50 kg ha⁻¹).

 

1. These are many factors (such as cultivars, N levels, locations), ------------However, the interactions between these factors are not adequately explored in the results and discussion sections.  

Authors: We agree with the reviewer that the data from this study did not fully explore the aspects of cultivars, locations, years, and their interactions. The primary idea and main goal of this study were to highlight the differences between plant species in response to nitrogen application. To ensure the study was representative and comprehensive, we included two cultivars of each plant species and conducted the research across three locations over two years.However, we acknowledge that a more detailed analysis of the effects of location, cultivar × location interactions, or location × year interactions could provide additional insights. That said, we believe that delving deeper into these aspects might divert attention from the core objective of the study. We suggest that future research could explore these factors in greater detail, particularly through climate-related analyses, to further investigate these interactions.

 

1. Some conclusions drawn in this study differ from previous research such as L316-318 ‘barley is more resilient to low-fertility soil and stress’ -------------A more in-depth discussion is needed here, preferably with comparisons to existing studies, to better understand the discrepancies and the implications of the new findings.

Authors: We acknowledge the reviewer’s observation that some conclusions in this study differ from previous research, particularly regarding the statement that ‘barley is more resilient to low-fertility soil and stress’ (L309-339). To address this, we have expanded the discussion to provide a more in-depth analysis, including comparisons with existing studies, to better understand the discrepancies and implications of our findings. We also identified an error in the discussion, where wheat was incorrectly compared to two-rowed barley instead of six-rowed barley. This has been corrected, and we have added new references to support the discussion, particularly regarding the previously claimed advantages of barley. These adjustments aim to strengthen the discussion and provide a clearer context for the differences observed in our study compared to earlier research. 

 

1. The economic analysis presented is somewhat weak -------------Could benefit from additional supporting literature. Alternatively, the authors may consider supplementing the analysis with cost-effectiveness data or other relevant economic indicators to provide a more complete picture.

Authors: We appreciate the reviewer’s suggestion regarding the economic analysis. However, the primary focus of this study was not to evaluate economic profitability but rather to examine the differences in Nitrogen Use Efficiency (NUE) among various plant species grown together. None of the authors specialize in agroeconomics, and we do not feel competent to conduct detailed economic analyses, which we believe are not a priority for this study. Including a detailed economic analysis in the results and discussion would shift the focus away from the core objective of the paper. We suggest that a separate study, potentially involving agroeconomists, could be conducted to explore the economic aspects in greater depth. For the current manuscript, we kindly ask the reviewer to accept this response and acknowledge that the economic analysis is beyond the scope of this work.

 

1. L415 Reference does not correctly cite '[341]'

Authors: We thank the reviewer for pointing out the error in the citation. During the track changes process, the number “3” was inadvertently left in the text. The correct citation should be “[41].” This has now been corrected in the manuscript.

 

1. Table 1, the data for "Rainfall mm" in Novi Sad 2020 shows a discrepancy between "72.7" and the previous value of "42,7"-------------- The author has replaced the data without providing an explanation. A clarification regarding this data change is necessary to maintain the integrity and transparency of the dataset.  

Authors: We apologize for the oversight and any confusion caused by the discrepancy in the rainfall data for Novi Sad 2020. Upon reviewing the table, we identified a typographical error where “72.7” was mistakenly entered as “42.7” in the initial version. While correcting this error as per the reviewer’s request, we inadvertently failed to provide an explanation for the change. The correct value for rainfall in Novi Sad 2020 is indeed 72.7 mm. We sincerely apologize for this oversight and appreciate the reviewer’s attention to detail, which has helped us maintain the integrity and transparency of the dataset.

Reviewer 2 Report

Comments and Suggestions for Authors

The "References" section needs checking.

Author Response

We have reviewed the references and made the necessary corrections.