Saffron Cultivation in Marginal Alpine Environments: How AMF Inoculation Modulates Yield and Bioactive Compounds

Round 1

Reviewer 1 Report

The manuscript is very well written except for the presentation of the results. The work is interesting. The study was well conceived and designed. The statistical analysis seems appropriate. Conclusions are consistent with the results. The only issue I found is the presentation of the results. The authors should present the main effect and interaction for all the parameters measured on the field experiments. The first question, did the location had any significant effect? There is no mention of how the data of the two locations were treated in the Table title or note. Was the location considered as a factor in the analysis? It is not clear. In general, the information that accompanies each table is not enough because there is no mention of the locations, or there is no mention of if and how data were averaged across location, year or across AMF inoculation. In table 3 and 5, for example, is presented the effect of the year, while in table 4 and 6 is presented the effect of the AMF inoculum. These effects should be presented in the same table, along with the significance of the main effects and interaction. Then when the interaction is significant it will be good to present the interaction either in a table or in a graph, at least for the most important parameters, because the interaction is more important than the single effect. In my opinion, the paper will improve if the data are presented as suggested.

Check the spacing throughout the manuscript as some spaces are missing between reference number for example, or at line 295. Also, the symbol × should be used instead of the x.

Author Response

Dear Reviewer 1,

we thank you for your valuable and useful comments and suggestions that help us to improve the quality and clarity of the paper.

Here we listed the main changes to the text.

- We modified the text and we divided the Section 3. Results and Discussion in 3. Results and 4. Discussion, accordingly to the Editor suggestions.

- Tables 3 and 4 were merged as well as for Tables 5 and 6.

- In the new Tables 3 and 4, we added all the requested statistical effects and interactions. Results were batter explained in the two new separated Sections 3. Results and 4. Discussion.

- Since fields and blocks were analysed as random factors, the effects of the two sites were not indicated in the text because they represent replicates in the applied mixed model. In the paragraph 'Statistical analysis' (lines 228-230) were indicated all the information.

- The whole text was checked for editing mistakes and we modified according to Reviewer suggestions.

Author Response File: Author Response.doc

Reviewer 2 Report

In the present study, the effect of arbuscular mycorrhizal fungi (AMF) on Crocus sativus was studied. For this purpose, the efficiency of root colonization with two AMFs and their effect on yield and quality of saffron plants cultivated open-field in two sites for two growing periods were evaluated, while a pot experiment in agreenhouse was also conducted.

Specific comments (refer also to the attached pdf file)

Line 41: The second keyword has to be shortened.

Line 45: correct to "is reproduced"

Lines 128, 183: correct to "sown"

Line 180: correct to "was"

Line 267: replace with "detected"

Line 372: cite Table 5 ata the end of the sentence (Line 376).

Line 385: correct to "(Table 5)"

In Tale 5, the single effect of AMF should be also presented.

Correct the highlighted references.

Overall, the manuscript is well written and the results are sufficiently discussed. The authors have to check statistics inTable 5 by presenting the single effect of AMFs.

Comments for author File: Comments.pdf

Author Response

Dear Reviewer 2,

we thank you for your valuable and useful suggestions and comments that help us to improve the quality of the paper.

Here we listed the main changes to the text.

- We modified the text and we divided the Section 4. Results and Discussion in the new Sections 3. Results and 4. Discussion, accordingly to Editor suggestions.

- We modified the text according to all the specific comments indicated in the attached pdf file.

- We modified the Tables according to suggestions and we added the single effects of AMF.

Author Response File: Author Response.doc

Reviewer 3 Report

Dear Editor, the manuscript “Saffron cultivation in marginal alpine environments: How AMF inoculation modulates yield and bioactive compounds” from Matteo Casser et al. deals with the evaluation productivity and accumulation of bioactive compounds of saffron after Arbuscular mycorrhizal fungi (AMF) inoculation. Overall, the text is well written and expose the results and the conclusions clearly.

However, I have some comment to the authors that should be addressed before its publication.

L41 Keywords. I would suggest to authors to change “biofertilization with arbuscular mycorrhizal fungi” by two keywords: “biofertilization” and “arbuscular mycorrhizal fungi”.

L126 Materials and Methods. It is necessary to include  “Chemicals and reagents” point where specify the brand of all analytical standards used in L252, as well as all reagents used during all the tests.

L139 I would be interesting to include the meters above sea level as in the same way that open field conditions. This could lead to a comparison between the glasshouse and open field environments.

L171 Supplementary Figure 2. From my point of view, instead of a continuous graphic, this figure should be a bar graph to state the number of picked flowers and the flowers per m2 on a determinate date that is a discrete value.

L192 Please, remove the space between 1 and % to be consistent throughout the text.

L196 Please, check the size of letters in formula (idem in L201, L217).

L237 Check the size of letters, please.

L266 Results and Discussion. Why are not data for a second year in trials at pot scale? A second year would strength the conclusions achieved.

L266 It would be interesting the analysis of quality parameters of saffron obtained at pot scale to compare with the results obtained in open field trials. Why the authors did not analyse them?

L295 Please, add a space between “year while”.

L302 Supplementary Figure 1. Please, correct P Olsen mg kg^-1 (superscript) and Electrical conductivity µS cm^-1 (superscript).

L342 Table 3. At the end of table 3, there is a missed space between “Carotenoids” and their units.

L375 Please, remove the “m” after 23.

L387 There seems to be an extra space between “values” and “ranges”.

L402 Table 5. I would ask authors if they have any explanation for the fact that in the first year, higher amounts of isoquercitrin, quercitrin, ellagic acid, safranal, and total vitamin C were obtained. They have commented in the manuscript the higher ISO3632 traits in the second year but, the majority of single bioactive compounds have higher accumulations in the first year. Can you discuss it?

L425 In my opinion, the word “antioxidants” should be replaced by “antioxidant activity”.

Author Response

Dear Reviewer 3,

we thank you for your valuable and useful suggestions and comments that help us to improve the quality of the paper.

Here we listed the main chenges to the text.

- We modified the text and we divided the section 3. Results and discussion in the new Sections 3. Results and 4. Discussion, accordingly to Editor suggestions.

- We modified the keywords accordingly.

- We included the paragraph "Chemicals and reagents" in Material and Methods Section.

- We added the sea above meters level of the glasshouse location.

- We checked for editing mistakes on the whole text and we modified accordingly. 

- The Glasshouse trial was performed only to evaluate the AMF interactions with saffron roots. As indicated in Material and Methods Section (lines 130-131), in pot cultivation no flowers were obtained due to the small caliber of the used corms.

- We added more specific and detailed information about the obtained results in the new Sections 3. Results and 4. Discussion.

Author Response File: Author Response.doc