Saffron Stigmas Apocarotenoid Contents from Saffron Latent Virus (SaLV)-Infected Plants with Different Origins and Dehydration Temperatures
, Natalia Moratalla-López 1, Habibullah Bahlolzada 3, Rosario Sánchez-Gómez 1, Akbar Dizadji 2,* and Gonzalo L. Alonso 1,*
Round 1
Reviewer 1 Report
Followings are the suggestions for the improvement.
Page 3 of 12, line 11-24
The explanation of the biosynthetic pathway of apocarotenoids is very complicated and difficult to understand. I strongly recommend that the authors should illustrate the diagram of saffron’s apocarotenoids biosynthesis pathway as figure.
Figure 2
Since the title includes the virus name, the authors should describe the possible effect of SaLV infection on biosynthesis of apocartenoids in the discussion, although all the samples tested were SaLV-positive, making analysis difficult. The authors have been keeping healthy saffron plants as a negative control, so I think that the effects of SaLV infection on spice components could be methodologically investigated.
Figure 3a
Because the principal component loading data, which represent the degree of correlation with each principal component, are not shown, it is not clear what kind of component has a strong influence on PC1 and PC2. If possible, it is better to name the axes based on the principal component loadings. For example, PC1 is ‘the tendency to contain more cosins’, and PC2 is ‘the tendency to contain more HTCCs’, something like that.
Author Response
We thank the reviewer for his suggestion, we consider that the manuscript has greatly improved.
Page 3 of 12, line 11-24
La explicación de la vía biosintética de los apocarotenoides es muy complicada y difícil de entender. Recomiendo encarecidamente que los autores ilustren el diagrama de la vía de biosíntesis de apocarotenoides del azafrán como figura.
Agradecemos al revisor por su sugerencia, hemos agregado la Figura 2 que describe la biosíntesis de apocarotenoides del azafrán y aclara su relación estequiométrica para seguir mejor el razonamiento de su discusión.
Figura 2
Dado que el título incluye el nombre del virus, los autores deben describir el posible efecto de la infección por SaLV en la biosíntesis de apocarotenoides en la discusión, aunque todas las muestras analizadas fueron positivas para SaLV, lo que dificulta el análisis. Los autores han mantenido plantas de azafrán sanas como un control negativo, por lo que creo que los efectos de la infección por SaLV en los componentes de las especias podrían investigarse metodológicamente.
A sugerencia del revisor, hemos insertado el siguiente comentario en la página 7:
La comparación de diferentes metabolitos secundarios del estigma del azafrán liofilizado entre muestras infectadas y no infectadas por SaLV de las mismas provincias de Irán mostró que el contenido de metabolitos secundarios fue mayor en las muestras no infectadas por SaLV que en las infectadas por SaLV. Parece que el contenido de varios metabolitos secundarios del azafrán podría ser modificado por la presencia de SaLV en muestras de azafrán de la misma provincia. Además, algunas características ambientales (altitud, temperatura y precipitación/precipitación) de diferentes orígenes geográficos podrían afectar el contenido de varios compuestos de los estigmas de C. sativus y la presencia de SaLV podría modificar estos efectos [22].
Figura 3a
Debido a que no se muestran los datos de carga del componente principal, que representan el grado de correlación con cada componente principal, no está claro qué tipo de componente tiene una fuerte influencia en PC1 y PC2. Si es posible, es mejor nombrar los ejes en función de las cargas de los componentes principales. Por ejemplo, PC1 es "la tendencia a contener más crocinas", y PC2 es "la tendencia a contener más HTCC", algo así.
Siguiendo la sugerencia del revisor, se muestran los datos para PC1 y PC2, para lo cual se ha agregado la tabla correspondiente en la Figura 4 y comentarios en el texto.
Author Response File: Author Response.odt
Reviewer 2 Report
The aim of this article was to study the concentration of the main saffron metabolites derived from corms planted under the same soil, climatic conditions, and cultural practices and infected with the same virus in order to determine whether the origin of the first planted corm significantly influences the metabolic composition of the spice obtained from them and to examine whether there is a difference between dehydrating saffron at 40 ºC or 50 ºC. This article compares the content of saffron metabolites from six production areas in Iran and neighboring countries infected with the saffron latent virus (SaLV) and dehydrated at two temperatures. The results revealed that after two years of remaining in the same soil and climatic conditions, subjected to consistent cultural practices, and infected with the same virus, they did not completely retain the memory of their original origin. Overall, this manuscript is of great significance for the research on the virulence of SaLV. It is suggested to be published after major revision.
Questions:
1. The introduction provides insufficient background and does not include all relevant references. In addition, some new research results need to be added.
2. “A secondary objective was to examine whether there is a difference between dehydrating saffron at 40 ºC or 50 ºC, temperatures at which protein denaturation typically occurs.” Why are the dehydration temperatures in Table 2 40 °C and 55 °C?
3. There is no title in Table 1.
4. “Saffron aqueous extracts were prepared following the standard procedures outlined by the International Organization for Standardization (ISO:3632-2, [26] with a minor modification.” “)” is missing from this sentence.
5. The authors describe that “an initial denaturing step as 94°C for 3 min, followed by 35 cycles of 94°C for 1 min, 57°C for 30 s and 72°C for the 70s; and a final extension time step at 72°C for 5 min..” There should be a space between the number and the unit. The use of periods is repeated in this sentence.
6. In Table 2 and Table 4, "Kg" should be replaced by "kg".
7. “Table 3 shows the Multivariate Analysis of Variance (MANOVA) of different saffron metabolites depending on the zone and dehydration temperature.” The font size of this sentence was used incorrectly.
8. In Figure 3b, “componente” should be replaced by “component”.
9. The literature cited in the main text uses hyperlinks. Please check and modify with the requirements of the submitting journal.
10. The "2" in the correlation coefficient (R2) should be superscripted. Please correct similar errors.
11. In the heading “2.5. Nomenclature for crocetin esters”, the part "cis" is not italicized.
12. The reference application format is not standardized, such as references 2, 3, and 6, etc.
Author Response
We thank the reviewer for his suggestions and corrections that have made the manuscript improve.
- The introduction provides insufficient background and does not include all relevant references. In addition, some new research results need to be added.
We have added in the Introduction comments and references from this last year, they are marked in red in the manuscript. We hope these will satisfy the reviewer.
- “A secondary objective was to examine whether there is a difference between dehydrating saffron at 40 ºC or 50 ºC, temperatures at which protein denaturation typically occurs.”Why are the dehydration temperatures in Table 2 40 °C and 55 °C?
We have dehydrated at 40º C, which is considered the temperature limit where proteins are not generally denatured, and their tertiary structure is not broken. At a temperature of 55ºC it is considered that the tertiary structure of proteins is destroyed.
- There is no title in Table 1.
The title of Table 1 has been corrected.
- “Saffron aqueous extracts were prepared following the standard procedures outlined by the International Organization for Standardization (ISO:3632-2, [26] with a minor modification.” “)” is missing from this sentence.
The “)” has been written.
- The authors describe that “an initial denaturing step as 94°C for 3 min, followed by 35 cycles of 94°C for 1 min, 57°C for 30 s and 72°C for the 70s; and a final extension time step at 72°C for 5 min..”There should be a space between the number and the unit.The use of periods is repeated in this sentence.
This has been corrected in the text.
- In Table 2 and Table 4, "Kg" should be replaced by "kg".
This has been corrected in the text.
- “Table 3 shows the Multivariate Analysis of Variance (MANOVA) of different saffron metabolites depending on the zone and dehydration temperature.” The font size of this sentence was used incorrectly.
This has been corrected in the text.
- In Figure 3b,“componente” should be replaced by “component”.
This has been corrected in Figure 3b.
- The literature cited in the main text uses hyperlinks. Please check and modify with the requirements of the submitting journal.
Literature is verified against journal requirements.
- The "2" in the correlation coefficient (R2) should be superscripted.Please correct similar errors.
Those errors in the text have been corrected.
- In the heading “2.5. Nomenclature for crocetin esters”, the part "cis" is not italicized.
It has been corrected in the text.
- The reference application format is not standardized, such as references 2, 3, and 6, etc.
The format is in accordance with the model of the Horticulturae editorial.
Author Response File: Author Response.odt
Reviewer 3 Report
The main results of this ms. are effect of two treatments, 40 °C for 120 minutes and 55 °C for 45 minutes, on metabolites. Because all the samples are infected with SaLV, this aspect should not be emphasized. I suggested: First, only indicated two temperatures was not the reality. Second, effect of different zones can be re-analyzed and compared. And, several other concerns are below:
1. abstract should provide some data;
2. the title of table 1 is not good;
3. 2.1 Plant materials, more information such as soil and field culture are encouraged to add.
4. table 2, comparison in the same column has no meaning, should in the same line.
5. the conclusions should be re-write.
Author Response
We thank the reviewer for his suggestions and corrections that have made the manuscript improve.
- abstract should provide some data;
The following sentence related to the synthesis of apocarotenoids has been added.
In general, the ratio of the sum of mmol/kg of HTCC derivatives to the sum of the mmol of crocins is greater than 2. This implies that the biosynthesis of saffron apocarotenoids due to the degradation of β-carotene towards HTCC is more important than that of zeaxanthin formation, which later gives HTCC and crocetindialdehyde
- the title of table 1 is not good;
The title of Table 1 has been changed.
- 2.1 Plant materials, more information such as soil and field culture are encouraged to add.
The information requested in section 2.1 Plant materials has been completed.
- table 2, comparison in the same column has no meaning, should in the same line.
The reviewer is right, there was an error in the comment.
- the conclusions should be re-write.
The conclusion has been rewritten.
Author Response File: Author Response.odt
Round 2
Reviewer 2 Report
The authors made necessary improvements and provided the answers to the concerns raised in a satisfactory manner. I recommend the paper for publishing in its current form.
Reviewer 3 Report
I suggest to accept this revised ms.
