Review Reports - Morphological and Molecular Characterization of <i>Micromeria croatica</i> (Lamiaceae), an Endemic and Potentially Valuable Horticultural Species of the Dinaric Alps

Round 1

Reviewer 1 Report

1. The article is devoted to the topical topic of the study of the characteristics of species living in conditions of high biodiversity, taking into account molecular and morphological characteristics.,

2. Conclusion "It is known that the fragmentation of the natural habitat may have contributed to the differentiation of populations. Hilpold et al. 2011 [40] found that sandy beaches and river mouths probably acted as barriers for representatives of the Sicilian Cornflower group cineraria group (family of Compound flowers), which are limited to rocky outcrops near the sea. It is also possible that partial isolation of populations of M. Croatia by river valleys and mountain ranges has led to genetic differentiation among populations over a long period of time.", requires additions for other representatives of the flora. I believe that in addition to the factor of geographical isolation, other types of factors determining taxonomic diversity are possible: height of growth above sea level, exposure, type and illumination of the slope, biological features of the species. The universal sets of adaptive markers include lipid composition (Shavikin et al., 2022), which was previously shown (Kremer et al., 2022).

3. It is recommended to formulate conclusions more clearly and fully after the "Discussions" section.

Kremer, D.; Dunkich, V.; Radosavljevic, I.; Bogunich, F.; Ivanova, D.; Ballian, D.; Steshevich, D.; Matevski, V.; Rangelovich, V.; Eleftheriadou, E.; Shatovich, Z.; Lieber, Z. Phytochemicals and their correlation with molecular data in Micromeria and Clinopodium (Lamiaceae) Taxa. Plants 2022, 11, 3407. https://doi.org/10.3390/plants11233407

Shavikin K.P.; Menkovich N.R.; Zdunich G.M.; Tasich S.R.; Ristich M.S.; Stevich T.R.; Daich-Stevanovich Z.P. Chemical composition and antimicrobial activity of essential oils of Micromeria thymifolia (Scop.) Fritsch., M. dalmatica Benth. and Satureja cuneifolia Ten. and its secretory elements. J. Essent. Oil, 2010, 22, 91-96.

Author Response

Corresponding author:

Dario Kremer

University of Zagreb

Faculty of Pharmacy and Biochemistry

A. Kovačića 1,

10000 Zagreb

Croatia

e-mail: kremer@pharma.hr

Tel.: + 38-51-4619-422

Zagreb, March, 08, 2023

Dear Reviewer,

Thank you very much for your comments. Please find enclosed answers on your comments on manuscript "Morphological and molecular characterization of Micromeria croatica (Lamiaceae), an endemic and potentially valuable horticultural species of the Dinaric Alps".

List of comments author responses and are given below.

Reviewer’s comments

The article is devoted to the topical topic of the study of the characteristics of species living in conditions of high biodiversity, taking into account molecular and morphological characteristics.

Comment: Conclusion "It is known that the fragmentation of the natural habitat may have contributed to the differentiation of populations. Hilpold et al. 2011 [40] found that sandy beaches and river mouths probably acted as barriers for representatives of the Sicilian Cornflower group cineraria group (family of Compound flowers), which are limited to rocky outcrops near the sea. It is also possible that partial isolation of populations of M. Croatia by river valleys and mountain ranges has led to genetic differentiation among populations over a long period of time.", requires additions for other representatives of the flora. I believe that in addition to the factor of geographical isolation, other types of factors determining taxonomic diversity are possible: height of growth above sea level, exposure, type and illumination of the slope, biological features of the species. The universal sets of adaptive markers include lipid composition (Shavikin et al., 2022), which was previously shown (Kremer et al., 2022).

Author response: The reviewer is right. We added other representatives of the flora from literature in two paragraph.

Comment: It is recommended to formulate conclusions more clearly and fully after the "Discussions" section.

Author response: The comment is accepted and the section Conclusions was modified.

Sincerely, Dario Kremer

Author Response File: Author Response.pdf

Reviewer 2 Report

The submitted manuscript “Morphological and molecular characterization of Micromeria croatica (Lamiaceae), an endemic and potentially valuable horticultural species of the Dinaric Alps” by Liber et al. is a well planned and executed work. However, I addressed some concerned issues:

Comments:

1. Abstract: The authors must clarify the study's objectives and findings in the abstract. The authors must correct any spelling errors and grammatical errors. Lines 33 and 198; change laves to leaves.

2. Introduction: Please delete figure1 from introduction part.

3. Materials and Methods: Line 191; Please change 2.3. Morphological Analysis with 2.4. Morphological Analysis.

4. Results:

4.1 Line 309; Please change 3.1. Morphological variability with 3.2. Morphological variability.

4.2 Resolution of figure 6 should be improved.

5. Discussion: Please add some sub-headers in the discussion section to help the reader.

6. The authors should be checked carefully for references.

Comments for author File: Comments.docx

Reviewer 3 Report

The manuscript submitted by Liber et al. describes a study on the characteristics of 10 populations of M. croatica in the Balkans. The authors concluded that STRUCTURE and BAPS analysis allowed them to divide these 10 populations into two or three genetic groups, respectively. Furthermore, a combined analysis of morphological and genetic studies by CANDISC suggested that there is a distinction between western and eastern populations. The topic addressed is interesting. However, there are several concerns as follows that need to be addressed and corrected.

The authors collected detailed data for the growth conditions, such as altitude, for the different 10 populations. However, other factors such as day length, humid and nutrition in the soil, and temperature should give more direct impact for the plant growth. If it is available, these data should be shown.

In this manuscript, the number of flowers per shoot was analyzed. Seed counts are important for maintaining and propagating populations under natural growing conditions. The number of seeds per shoot and their germination rate should be analyzed.

Fig. 2; The shape and color of symbols should be the same as Fig. 4 and other figures. Personally, I don't think the symbol color of Mn should be the same as that of the BH population. Further, a scale bar should be added.

Fig. 7 and 8; The authors should clearly define what the length of arrows indicates.

Minor comments

Line 309; 3.1. should be 3.2.

Line 474; CR2 should be Cr2.

Author Response

Corresponding author:

Dario Kremer

University of Zagreb

Faculty of Pharmacy and Biochemistry

A. Kovačića 1,

10000 Zagreb

Croatia

e-mail: kremer@pharma.hr

Tel.: + 38-51-4619-422

Zagreb, March, 08, 2023

Dear Reviewer,

List of comments author responses and are given below.

Reviewer’s commentsThe manuscript submitted by Liber et al. describes a study on the characteristics of 10 populations of M. croatica in the Balkans. The authors concluded that STRUCTURE and BAPS analysis allowed them to divide these 10 populations into two or three genetic groups, respectively. Furthermore, a combined analysis of morphological and genetic studies by CANDISC suggested that there is a distinction between western and eastern populations. The topic addressed is interesting. However, there are several concerns as follows that need to be addressed and corrected.

Comment: The authors collected detailed data for the growth conditions, such as altitude, for the different 10 populations. However, other factors such as day length, humid and nutrition in the soil, and temperature should give more direct impact for the plant growth. If it is available, these data should be shown.

Authors’ response: Bioclimatic variables are derived from the monthly temperature and rainfall values in order to generate more biologically meaningful variables. These are often used in species distribution modelling and related ecological modelling techniques. Concerning to the day length, the distance between the most northern (44°45') and the most southern (42°52') population is below the two degree of geographical latitude. Therefore, small differences in the day length among populations are to be expected. The soil were not collected and analysed in this investigation. Consequently, authors have no information about soil humid and nutrition. But, humid in soil depend of precipitation which is enclosed (precipitation of wettest and driest month, precipitation seasonality, precipitation of wettest and driest quarter, precipitation of warmest and coldest quarter) in Table S1.

Comment: In this manuscript, the number of flowers per shoot was analyzed. Seed counts are important for maintaining and propagating populations under natural growing conditions. The number of seeds per shoot and their germination rate should be analyzed.

Authors’ response: The seeds were not collected and analysed in this investigation. This is why the research proposed by Reviewer is not possible.

Comment: Fig. 2; The shape and color of symbols should be the same as Fig. 4 and other figures. Personally, I don't think the symbol color of Mn should be the same as that of the BH population. Further, a scale bar should be added.

Authors’ response: The comment is accepted.

Comment: Fig. 7 and 8; The authors should clearly define what the length of arrows indicates.

Authors’ response: The reviewer is right. Thank you. A sentence indicating the length of the arrows has been added to the figure captions:

Figure 7. The length of the vector is proportional to the eigenvector of the variable, scaled by the square root of the eigenvalue.

Figure 8. The length of the vector is proportional to the discriminatory power of the variable.

Minor comments

Comment: Line 309; 3.1. should be 3.2.

Authors’ response: The comment is accepted. Thank you for the carefully reading of the manuscript.