Shade-Induced Effects on Essential Oil Yield, Chemical Profiling, and Biological Activity in Some Lamiaceae Plants Cultivated in Serbia

Round 1

Reviewer 1 Report

The manuscript entitled “Influence of shading on content, chemical composition and biological activities of essential oils from Lamiaceae plants cultivated in Serbia” presents a significant work. But manuscript should be revised in the following points:

In lines 23,104 should be Escherichia

In line 33 please write latin names of studied species

In lines 35, 45 please remove subscript

Please add information about known biological activities for each studied species

The methodology lacks information from which parts of the plant the essential oil was obtained

In lines 183-189 there is no reference to literature and no examples

In line 218, please write the names of compounds in lower case

Please improve the quality of the drawn compound structures in figures 2, 3, 4

In lines 243-250 and 284-289 , please write the names of compounds in lower case

Please, correct reference 51

 

Author Response

Answer to Reviewer 1.

 

In lines 23,104 should be Escherichia

You are right, we make changes

In line 33 please write latin names of studied species

We add full Latin names for all plants

In lines 35, 45 please remove subscript

We remove subscript

Please add information about known biological activities for each studied species

As main compounds from thyme essential oils (TEOs), thymol and carvacrol inhibit lipid peroxidation and manifest strong antimicrobial activity against different kinds of microorganisms. Polyphenolic phytochemicals in thyme act as powerful antioxidants, which aroused an increasing interest in their application in functional food. There is a possibility of future use of TEO in extending the shelf life of bakery products and it could find application in the storage of root vegetables Based on the antimicrobial activity of the vapor phase, they found that the essential oil has the potential to be used to protect packaged foods [4 ].

 

Mint essential oils (MEOs) rich in monoterpenes and sesquiterpenes having as major constituents menthone and menthol exerted potent to treat muscle pain and neuralgic pain, and the ones correlated with gastrointestinal disorders (relief of abdominal pain, flatulence, repletion, obstipation, and diarrhea). Inhalation use is effective for relieving symptoms like nausea, vomiting and anorexia form patients receiving chemotherapy. Other biological effects of MEO are insecticidal, antibacterial, antiviral, antiallergenic, antioxidant, and cytotoxic activities [5].

 

According to the biological studies, Melissa officinalis L possesses high amount of antioxidant activity through its chemical compounds including high amount of flavonoids, rosmaric acid, gallic acid, phenolic contents. Geranial, neral, citronellal and geraniol are the main components of lemon balm essential oils (LEOs). The pharmacological effects of the extracts are mainly assigned to the presence of large amounts of polyphenolic compounds, such as antioxidant, antimicrobial, antiproliferative, and cytotoxic effects, and so on [6].

 

[4] Galovičová, L.; Borotová, P.; Valková, V.; Vukovic, N.L.; Vukic, M.; Štefániková, J.; Ďúranová, H.; Kowalczewski, P.Ł.; Čmiková, N.; Kačániová, M. Thymus vulgaris essential oil and its biological activity. Plants (Basel). 2021, 19, 10(9), 1959.     doi: 10.3390/plants10091959.

[5] Efe Ertürk, N.; Ta¸scı, S. The Effects of peppermint oil on nausea, vomiting and retching in cancer patients undergoing chemotherapy: An open label quasi-randomized controlled pilot study. Complement. Ther. Med. 2021, 56, 102587.

[6] Petrisor, G.; Motelica, L.; Craciun, L.N.; Oprea, O.C.; Ficai, D.; Ficai, A. Melissa officinalis: composition, pharmacological effects and derived release systems-A review. Int J Mol Sci. 2022, 23(7), 3591.

The methodology lacks information from which parts of the plant the essential oil was obtained

We add…..        Uniform shoots with leaves without any injuries or defects were selected and dried without the presence of light and ventilation at room temperature (about 25–30 ◦C) as air-dry herbs for analysis.

In lines 183-189 there is no reference to literature and no examples

We add reference …  [7]

In line 218, please write the names of compounds in lower case

Names of compounds are in lower case

Please improve the quality of the drawn compound structures in figures 2, 3, 4

We improve figures 2,3,4 with higher resolution

In lines 243-250 and 284-289 , please write the names of compounds in lower case

Yes…..all name of compounds are in lower case

Please, correct reference 51

We correct reference 51

 

 

Author Response File: Author Response.docx

Reviewer 2 Report

 Dear Authors, 

I have completed my review of the submission entitled "Influence of shading on content, chemical composition and biological activities of essential oils from Lamiaceae plants cultivated in Serbia". This contains important data that can be recommended for publication in "Horticulturae". However, I see that the manuscript must be revised carefully to improve its presentation. Please find below my comments and suggestions to improve the submission: 

1-The text must be revised carefully for typo and language mistakes, 

2-The Title has to be rephrased, what do you think about this suggestion?: Shade Induced Effects on Essential Oil Yield, Chemical Profiling, and Biological Activity in Some Serbian Lamiaceae Plants",

3-Lines 13-15, authors must compare shaded/not shaded instead of providing different values without comparison,

4- For plant species, please use full scientic names at first mention and then the corresponding abbreviations, e.g, Thymus vulgaris L. (T. vulgaris),

5-Some recent references can be added to fortify the state of the-art-of research in the Introduction section but also to enrich discussion on biological acitivity: https://doi.org/10.1016/j.bcab.2022.102569,

6- The Authors must highlight the gap of knowledge before stating the objectives,

7-Regarding the abbreviations of different essential oil from different plant species, please see the following suggestion: T. vulgaris essential oil : TVEO, if you agree, please revise accordingly throughout the manuscript, 

 8- For plant material, I think it would be better if you use a table, in which you summarize, plant species with common and scientiifc name, family, wild or cultivated, plant part used, voucher (and identification), etc.,

9-Climatic and edapahic data of sampling site must be provided, 

10- line 119, please remove "multivariate" as it is obvious, 

11-Table 3, different values must be compared statistically. The same comment for the subsequent tables (mean values comaprison),

12- Figures 8 and 9 must be improved,

13- I see that other multivariate statistical analyses can be used to better exploit the obtained data: correlatiosn, regressions, cluster analysis, etc. 

Regards.

Author Response

Answer to Reviewer 2.

I have completed my review of the submission entitled "Influence of shading on content, chemical composition and biological activities of essential oils from Lamiaceae plants cultivated in Serbia". This contains important data that can be recommended for publication in "Horticulturae". However, I see that the manuscript must be revised carefully to improve its presentation. Please find below my comments and suggestions to improve the submission: 

1-The text must be revised carefully for typo and language mistakes, 

We make extensive English revisions by lector and improve manuscript body

 

2-The Title has to be rephrased, what do you think about this suggestion?: Shade Induced Effects on Essential Oil Yield, Chemical Profiling, and Biological Activity in Some Serbian Lamiaceae Plants",

 

We accept your suggestion  and create new Title

Shade induced effects on essential oil yield, chemical profiling, and biological activity in some Lamiaceae plants cultivated in Serbia

 

3-Lines 13-15, authors must compare shaded/not shaded instead of providing different values without comparison,

Yes we accept your suggestion………Abstract: Thyme, mint and lemon balm were used to determine whether shading conditions could improve yield, composition, antioxidant and antimicrobial activity in plant essential oils (EOs) in comparison with non-shaded plants from open field.  ……….

 

 

4- For plant species, please use full scientic names at first mention and then the corresponding abbreviations, e.g, Thymus vulgaris L. (T. vulgaris),

Yes we respect your comment and use full scientic names at first mention and then the corresponding abbreviations, e.g, Thymus vulgaris L. (T. vulgaris) in all Latin name during the text of manyscript

 

5-Some recent references can be added to fortify the state of the-art-of research in the Introduction section but also to enrich discussion on biological acitivity: https://doi.org/10.1016/j.bcab.2022.102569,

 

We add one sentence from these article in part of Introduction……….These plants are rich sources of minerals (especially K, Ca, and Mg), proteins, and other bioactive compounds. In addition, these plants could contribute to the production of EOs and other aromatic extracts as a source of natural antioxidant compounds.

[3] Ait Bouzid,A.; Oubannin, S.; Ibourki, M.; Bijla, L.; Hamdouch, A.; Sakar E.H.; Harhar, H.; Majourhat, K.; Koubachi, J.; Gharby,  S. Comparative evaluation of chemical composition, antioxidant capacity, and some contaminants in six Moroccan medicinal and aromatic plants. Biocat Agric Biotech, 2023. 47, 102569doi.org/10.1016/j.bcab.2022.102569                                                                     

 

6- The Authors must highlight the gap of knowledge before stating the objectives,

 

Due to the insufficient of experience and literature data on the influence of shading and plant density on the biological and chemical activity of cultivated medicinal plants, experiments were carried out, the aim of study was to investigate the influence of shading on the EOs content and composition of thyme, mint and lemon balm and their antioxidant and antimicrobial activity.

 

7-Regarding the abbreviations of different essential oil from different plant species, please see the following suggestion: T. vulgaris essential oil : TVEO, if you agree, please revise accordingly throughout the manuscript, 

We accept your suggestion and revise during the all text thyme essential oils as TEOs; mint essential oils as MEOs;   Lemon balm essential oils as LEOs

 

 8- For plant material, I think it would be better if you use a table, in which you summarize, plant species with common and scientiifc name, family, wild or cultivated, plant part used, voucher (and identification), etc.,

We have scientific (Latin ) and common name in the text , we use herb (leaf and flowers), and all plant was cultivated (with shade or with nononshading-open field condition)…its same for all species……

 

9-Climatic and edapahic data of sampling site must be provided, 

We added soil data and climacteric data from experimental garden

 

Table 1. Basic agrochemical properties of the soil

CaCO₃ (%)	рH in KCl	Humus (%)	N - Total	P₂O₅	К₂O
			%	mg/100g	mg/100g
2.36	6.94	3.69	0.18	16.8	30.8

 

 

 

 

 

 

 

 

Table 2. Temperature, solar radiation and precipitation

 

Month	TS 2020.      2021.	TOD 2020.   2021.	TX 2020.     2021.	TM 2020.   2021.	MSR 2020.     2021.	MSRA	RR 2020.   2021.	RO
Мay	16,4        17,2	-0,2         0,6	22,9       24,5	11,0       11,0	173,2     226,8	219,0	67,0      29,4	66,7
Јune	19,8        21,5	0,3         2,0	25,7       28,9	14,9       14,6	183,7     245,7	237,2	186,7    30,3	69,7
Јuly	22,3        25,5	1,0         4,2	29,1       32,8	15,9       18,6	290,9     293,2	289,0	78,6      39,8	43,6
Аugust	22,6        23,5	1,5         2,4	30,0       31,8	16,5       16,1	265,5     302,1	276,0	78,6      39,6	43,3
Sept.	20,3        18,0	3,1         0,8	28,0       25,8	13,6       14,1	224,8     200,9	210,0	14,9      21,0	43,6

 

TS-mean monthly air temperature ( ◦C); TOD- temperature deviation for 30-year average ( ◦C); TX- mean daily temperature maximum for month ( ◦C); TM- mean daily temperature minimum for month ( ◦C); MSR-monthly solar radiation (h ).MSRA- monthly solar radiation (h) for 30-year average; RR-total  monthly precipitation (mm); RO  monthly precipitation (mm) for 30-yearaverage

 

 

10- line 119, please remove "multivariate" as it is obvious, 

We are remove "multivariate"

 

11-Table 3, different values must be compared statistically. The same comment for the subsequent tables (mean values comaprison),

In case of differences between shaded and non shaded, Student's t-test was used, while for effect of

essential oils on antimicrobial activity ANOVA was used to analyze the significance with Duncan’s

multiple range tests (with a level of 0.05). For explanatory data analysis, principal component

analysis was used. All statistic calculation was performed with STATISTICA software (TIBCO Software

Inc. Palo Alto, CA, USA. 2020, version 14.0.015.)

 

 

12- Figures 8 and 9 must be improved,

Yes we are improve Fig 8 and 9

 

13- I see that other multivariate statistical analyses can be used to better exploit the obtained data: correlatiosn, regressions, cluster analysis, etc. 

The main goal of manuscript is to investigate shading effects on difference in EO composition in

plant species (thyme, mint and lemon balm) from Laminaceae family. Main results are difference in

EO composition and those differences are now statistically processed, as You suggested in in

comment 11. Use of multivariate statistics, as you suggested, will draw attention from differences,

since for analysis only essential oils which are find in all three species in both shaded and control

plants should be used. In addition, even if such analysis is performed, due to analysis of different

plant species any analysis which will group presented results will only bring confusion in manuscript.

Use of PCA in antimicrobial activity serves, beside difference between shaded and non shaded EO to

further differentiate plants according to microbiological activity. So authors opinion is that, from

stand point of manuscript goal, presented statistical analysis is adequate to give answer to

manuscript goals.

Author Response File: Author Response.docx

Reviewer 3 Report

Comments to the Authors

The manuscript entitled “Influence of shading on content, chemical composition and biological activities of essential oils from Lamiaceae plants cultivated in Serbia” has been reviewed. The manuscript is interesting and complete, I suggest to accept the MS with very minor revisions.

A minor revision of the language is suggested.

1 - Introduction

Lines 35 – Please, check the word “used”.

Lines 45 – Please. check the word “ontogenesis”.

 

1 - Results

Line 222 – Perhaps it is better to lower the text of line 222 to the page 7.

Line 251 – Perhaps it is better to lower the text of line 251 to the page 9.

 

3 - Conclusion

Line 408 – I suggest to increase the conclusions adding new possible trend concerning the influence of shading on essential oil composition and biological activities.

 

Author Response

Answer to Reviewer 3.

 

A minor revision of the language is suggested.

We make extensive English revisions and improve manuscript body

1 - Introduction

Lines 35 – Please, check the word “used”.

Yes we add have been ,,used,,

Lines 45 – Please. check the word “ontogenesis”.

Yes we replace otogenesis with new word ,,development stage,,

 

1 - Results

Line 222 – Perhaps it is better to lower the text of line 222 to the page 7.

Line 251 – Perhaps it is better to lower the text of line 251 to the page 9.

 Yes it’s a technical procedure

3 - Conclusion

Line 408 – I suggest to increase the conclusions adding new possible trend concerning the influence of shading on essential oil composition and biological activities.

We add sentences on effect of shading on essential oil composition and biological activities.

………………It is evident that the modification of light intensity can act as a physiological tool via shade nets to improve the yield of essential oils and their constituents, phytochemical quality and antioxidant activity. These plant species tolerate shading well, so it is recommended to grow them under shading nets. The EOs have antimicrobial activity against a wide range of microorganisms. The TEOs are the most active in terms of antimicrobial and antifungal activity…………………….

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Dear authors,

The manuscript was significantly improved and therefore I can be accepted for publication in Horticulturae. Typo mistakes must be corrected at proof stage.

Regards. 

Author Response

Thank you so much for your comments.

Author Response File: Author Response.docx