Evaluation of Greek Tobacco Varieties (Nicotiana tabacum L.) Grown in Different Regions οf Greece

Round 1

Reviewer 1 Report

I have suggested some changes in the attached PDF, which are self-explanatory.

Abstract, Introduction, M & M, Results, Discussion, and Conclusion must be improved. Most of the sentences are jumpy and break the sequence of information. All sections must be improved. Although it is stated in the M & M that the statistical design was three-factorial, however, the ANOVA tables did not match it.  The morphological parameters studies are also limited.

Comments for author File: Comments.pdf

Extensive editing of the English language is required.

Author Response

Thank you very much for having considered our manuscript. All your comments  were considered, and English language were checked by a native speaker.

More specifically, regarding the quality of the figures, the reviewer is right to argue that it was poor in the original version of the manuscript. In the revised version of the manuscript, the figures are improved in quality to address the concerns of the reviewer.

Regarding the comment on the statistical design, and the three-factorial ANOVA, we have already stated in the M. & M. in the original version of the manuscript that the three-factor analysis of variance (ANOVA) was employed for each dependent variable, to assess these three factors as main effects (year, area/location, and variety), along with their pairwise interactions (the three-way interaction was not assessed). The reason that the three-way interaction was not assessed is that we have performed three-factor ANOVA without replication (based on the available dataset), thus there is no interaction of the three factors, but there are pairwise interactions. In the revised version of the manuscript, we have now modified the relevant text in M. & M., and clearly state that we have performed three-factor analysis of variance (ANOVA) without replication.

Regarding the addition of the hybrids NC 297 and NC7L in the Table 1, we would like to inform you that these hybrids were not genetic material of the Hellenic Agricultural Organization DIMITRA, whose characteristics are registered and may be published, but bred by North Carolina State University and were used only as references.

Finally, regarding the use of the term “location” instead of “area”, we keep the term “area”, which is more general, because the tobacco in Greece is cultivated in big areas of the country, and we would like to emphasize this fact.

Reviewer 2 Report

1. What is the main question addressed by the research?

The main issue considered in this paper is the ecological variety testing of tobacco breeding material of different geographical and ecological origin and the selection of the best varieties and breeding samples in the conditions of the regions of Greece, which are important for the cultivation of this valuable industrial crop.

 2. Do you consider the topic original or relevant in the field? Does it
address a specific gap in the field?

Ecological variety testing is a key moment in breeding work and evaluation of genetic material. This work is devoted to the solution of this issue.

Works in this area are important and interesting for specialists all over the world who deal with various agricultural crops. Therefore, I consider this work original and in demand.

 3. What does it add to the subject area compared with other published material?

Each culture is different, but the approaches that scientists in different regions of the world use are expanding our understanding of breeding science.

To present the results, the authors used an approach that makes it possible to single out the share of the influence of both organized factors (genetic characteristics of varieties, influence of habitats) and random factors (for example, weather conditions). Studies of tobacco culture in the Balkans are not enough. Many questions require clarification. The work of the authors to a certain extent fills this gap.

 4. What specific improvements should the authors consider regarding the
methodology? What further controls should be considered?

The authors used UPOV methods to evaluate the breeding material, which makes it possible to obtain comparable scientific data that are of interest to the general world scientific community. Using the methods used by the authors in their work makes it possible to apply them not only in Greece, but also in other regions of the world where tobacco culture is practiced. The studies were carried out in five regions of Greece for three years, which allows drawing reasonable conclusions based on the results obtained. Experimental plots were laid out in triplicate. This allows us to say that the results obtained are valid.

The authors presented the results in additional tables, and in the main text they presented the results of mathematical analysis, which confirm significant differences in individual indicators.

 5. Are the conclusions consistent with the evidence and arguments presented
and do they address the main question posed?

In connection with the questions that have arisen in the article, I think that the authors need to expand the conclusions, strengthen them with digital material in order to convincingly show the novelty and originality of the studies, as well as their importance for the world breeding science and, especially, practice.

 6. Are the references appropriate?

The authors used to prepare a review of the literature a sufficient number of scientific sources that characterize the essence of the problem under study. All references to the literature are justified and correct. There is no excessive self-citation - the authors referred to 2 of their works correctly and reasonably.

 7. Please include any additional comments on the tables and figures.

The authors presented a large number of tables relating to research and mathematical processing, trying to show the scientific community that they did everything right: they organized the experiment, conducted chemical analyzes, and extensive mathematical processing of the data obtained.

  In my opinion, even an excessive amount of additional data.

The material that the authors provide in tables S1-S34 confirms the graphs in the text of the article. But this can be waived. The graphs in the article are convincing enough on their own.

The quality of the graphs needs to be improved. But I am sure that the editors of a respected journal will help the authors to solve this small technical problem.

 I confirm my opinion that this article, after making minor corrections and eliminating technical comments, can be considered in a journal.

Author Response

We would like to thank the reviewer for finding our work origina land in demand, and for stating that to a certain extent fills this gap concerning studies of tobacco culture in the Balkans.

Regarding point 5, the conclusion session has been rewritten in order to accommodate the comment of the reviewer.

Regarding point 7 raised by the Reviewer. We agree with the Reviewer that we have provided a large amount of additional data, in particular, Supplementary Tables S1-S34. However, these tables are necessary to support the results in the main text, since they provide all relevant information for the post hoc analysis, enabling to better understand the statistically significant results originally obtained based on the three-factor ANOVA. However, to address the concerns of the Reviewer, we have now reduced to a certain extent the amount of data presented in the Results Section in the main text.

Regarding the quality of the graphs, the reviewer is right to argue that it was poor in the original version of the manuscript. In the revised version of the manuscript, the figures are improved in quality to address the concerns of the reviewer.

Reviewer 3 Report

This manuscript proves to be new innovation regarding tobacco production in Greece, however,, minor changes are required in order to make it acceptable for publication.

Title: Add the botanical name of tobacco in the title.

Line 21: what type of interaction was detected between area and variety? Either it was negative or positive. Incorporate it in the abstract for a clear understanding of the results.

Line 32-33: was some variety from flue and air cured types perform better in their respective studied areas?

Keywords: Arrange keywords in alphabetical order.

Introduction:

Line 37: Italicize the botanical name of tobacco.

Paragraph 38-47: Incorporate the literature regarding tobacco growing conditions and how Greece's environmental conditions favour tobacco growth in Greece.

Methodology:

Line 75-77: Either use the word oriental or sun-cured, avoid using both word in the manuscript to prevent readers from confusion.

Why only 3 locations were used for B type group instead of 5 (as in A type).

Table 2: Were both types (A, B) planted at the same time in different locations?

Discussion: What factors play an important role in the adaptability of these varieties with respect to their locations? Was it climate, soil conditions or cultural practices etc.

Paragraph 423-432: What factor apart from genotype influences nicotine and sugar content in varieties? 

Conclusion: there was no mentioned of location and season's affect on variety quality parameters. 

 

 

The English language needs to be modified by native speaker. Minor mistakes related to grammar are detected in manuscript

Author Response

We appreciate the reviewers' suggestions and we attach the file with the additions and the corrections.

This manuscript proves to be new innovation regarding tobacco production in Greece, however, minor changes are required in order to make it acceptable for publication.

Title: Add the botanical name of tobacco in the title.

Done

Line 21: what type of interaction was detected between area and variety? Either it was negative or positive. Incorporate it in the abstract for a clear understanding of the results.

The term interaction (between factors, particularly area and variety in line 21, but also in general throughout the text) in this context is used to represent that the effect of one factor (in the dependent variable) is dependent on the conditions in the other factor. Thus, if there is no interaction, then the two factors are acting independently to each other. That said, the interaction cannot be characterized as negative or positive, but only as present or absent. In the particular example of the interaction that was detected between area and variety related to yield, this is mainly based on the fact that variety K53 exhibited an evidently different behavior (related to yield) in comparison to the remaining four varieties (Figure 2(A)). In particular, variety K53 exhibited a much larger mean value in Katerini (1979.633 kg/ha), compared to the remaining four varieties in Katerini (overall mean value of 1309.075 kg/ha), while in Aitoloakarnania, and in Xanthi, it exhibited similar and even lower mean values compared to the remaining varieties. This shows that when the yield is assessed based on both area and variety, their specific categories should be taken into account, since they interact, and this interaction is significant, as shown in Table 3 (p=0.023).

Line 32-33: was some variety from flue and air cured types perform better in their respective studied areas?

According to Supplementary Table 24 A and B only the variety Burley 21E had lower yields compared to other varieties of the same group B.

Keywords: Arrange keywords in alphabetical order.

Done.

Introduction:

Line 37: Italicize the botanical name of tobacco.

Done.

Paragraph 38-47: Incorporate the literature regarding tobacco growing conditions and how Greece's environmental conditions favour tobacco growth in Greece.

Done.

Methodology:

Line 75-77: Either use the word oriental or sun-cured, avoid using both word in the manuscript to prevent readers from confusion.

Thank you for your comment, we are going ahead with this change.

Why only 3 locations were used for B type group instead of 5 (as in A type).

The B types of varieties were not used to be cultivated in all the tobacco growing areas of Greece and for this reason we focused the experimentation only in the areas of Aitoloakarnania, Karditsa and Xanthi.

Table 2: Were both types (A, B) planted at the same time in different locations?

Yes, both types of varieties were transplanted at the same time, but the type A varieties were cultivated in Xanthi, Serres, Katerini, Karditsa and Aitoloakarnania while B type varieties only in three regions as previously stated.

Discussion: What factors play an important role in the adaptability of these varieties with respect to their locations? Was it climate, soil conditions or cultural practices etc.

A relevant paragraph is added to the discussion section.

Paragraph 423-432: What factor apart from genotype influences nicotine and sugar content in varieties? 

A relevant paragraph is added to the discussion section.

Conclusion: there was no mentioned of location and season's affect on variety quality parameters. 

The conclusion section was rewritten.

Overall, we would like to thank the reviewer for providing valuable comments/corrections that aided us to improve the quality and clarity of the manuscript.

Reviewer 4 Report

The authors did good and quality research and wrote a correct scientific paper. In this study, five oriental and six flue and air cured type tobacco varieties were cultivated in five regions of Greece during three seasons. The authors evaluated the effect of variety to the yield as well as to nicotine, sugar and nitrate content in tobacco leaves. The results obtained in this study are helpful for tobacco producers and tobacco breeders to know the yield and the chemical contents in studied tobacco varieties and its interaction with different environments. There is no need for corrections in the work, except to supplement the list of flue and air cured type tobacco varieties (lines 78-79) with tobacco variety Burley 21E. Thanks to the authors for the interesting contribution.

Author Response

We would like to thank the reviewer for finding our work interesting and helpful for tobacco producers and breeders. The required addition for variety Burley 21E was done.

Round 2

Reviewer 1 Report

Paper is modified and is OK for publication.

English language is improved.