Exogenous Application of Aloe vera Leaf Extract Improves Silybin Content in Silybum marianum L. by Up-Regulating Chalcone Synthase Gene

Round 1

Reviewer 1 Report

The manuscript number: Agriculture-1893959 entitled “The Alteration of the Silybin Content of Silybum marianum L. by Aloe vera Leaf Extract via the Modification of Chalcone Synthase Gene Expression” authored by Alkuwayti et al., is crude phyto-extract application for plant growth attributes and silybin content improvement based study. The author tried to find out an optimum crude extract concentration by analyzing plant growth, biochemical and gene expression parameters. The reviewer gone through the manuscript and found that the manuscript is presented as per journal guidelines, the contents of manuscript is in define format, table and figures are presented appropriately. Followings are the query and suggestion for improving the quality of the manuscript-

Title

The title “The Alteration of the Silybin Content of Silybum marianum L.  by Aloe vera Leaf Extract via the Modification of Chalcone Synthase Gene Expression” can be modified as “Exogenous Application of Aloe vera Leaf Extract Improves Silybin Content in Silybum marianum L. by up-regulating Chalcone Synthase Gene”

Abstract:

1.      Rewrite “Biotic elicitors have been shown to stimulate growth and modify the bioactive composi tion of various plant species” for better clarity line no. 9-10

2.      Italicize “chalcone synthase” line no. 12

3.      Modify “Further, the activation of  chalcone synthase by ALE has been demonstrated by gene expression research.” As “Further, the activation of  chalcone synthase by ALE has been analyzed  by gene expression analysis.”

4.      Rewrite “while 40 mL/L ALE resulted in enhancement effects on silybin content and the expression” as “while 40 mL/L ALE resulted an enhancement in silybin content and the expression” line no. 17-18

5.      Italicize “chalcone synthase” line no. 18

6.      Replace “towards” with “against” line no. 20

Keywords:

1.      Add “anti-microbial activity and  Bio-elicitors” in  keywords

 Introduction:

1.      Rewrite “There are currently challenges related to crop production (e.g., the reduction in the 25 area and quality of arable lands; the irrigation of water due to climate change, pollution, 26 and non-sustainable farming practices)” as “There are currently challenges related to crop production due to erosion of agri-resources, climate change, pollution, and non-sustainable farming practices.” Line no. 25-26

2.      Give suitable citation for the sentence “ In view of this, the ……over the past few decades.” Line no. 30-31

3.      What is mean by “nutritional efficiency” line no.33

4.      Delete “examples” line no. 35

5.      Replace “environmentally friendly” with “environment friendly” line no. 36

6.      Give a global scenario of Aloe vera and Silybum marianum cultivation and economic value.

7.      Rewrite “In medical and …..seeds are critical [8,9]”. For better clarity line no. 46-47

8.      Replace “The goal of this paper” with “The goal of this study” line no. 62

9.      Replace “vegetative growth” with “growth attributes” line no. 63

10.  Replace “molecular events” with “ molecular mechanisms” line  no. 66

 Method and materials:

1.      Mention the geographical location of Al Sahab farm  in Al Ahsa, Saudi Arabia.

2.      Mention the geographical location of Agriculture and Veterinary Research and Training Centre, King Faisal University, Saudi Arabia

3.      What was the intensity of light during the experiment mention in line no. 84

4.      What was the age of seedlings at the time of transplantation mention in line no. 87

5.      Mention the days for the vegetative growth stage of  S. marianum.

6.      Merge 2.4 and 2.5 to reduce subheading

7.      Rewrite “At 270 days after planting, plant leaf samples were collected and dried for 48 h at 60 °C.” for better clarity

8.      Why author selected “third bottom” leaf

9.      Delete heading 2.9

10.  Merge 2.9.1 and 2.9.2 subheading and make heading 2.9 as “Total RNA Isolation, cDNA Preparation and Gene Expression Analysis”

11.  Mention the name of reagent company and city with country do not mention cat. No. however incase of instrument, model with city and country may be mentioned

12.  What was the concentration of cDNA  mention in line no. 159

13.  Replace “An amount” with “A total volume” in line no. 190-191

 Results:

1.      Results are written and presented in a nice way but there are use of many unscientific words like uncovered (214), diverse (215), toward (290) etc.

2.      It is better to put only up to three digits after decimal for standard deviations, so the tables can be modified

3.      Figure 1 is ok, but mention only letter of significant not value.

4.      Italicize “S. aureus and E. coli in table 9

5.      Rewrite “Indeed, a decrease in anti-E. coli activity was observed in extracts from plants treated  with 40 mL/L ALE compared to that from control plants.” For better clarity” line no. 297-298

Discussion

1.      Replace “cultivated by” with “in” line no. 315

2.      Replace “biggest” with “highest” line no. 316

3.      Shift the sentence “Silybin was present in various amounts in the S. marianum plant, and has been shown to have great potential in the treatment of Alzheimer’s disease [33] and cancer [34]. Indeed, the majority of the hepatoprotective effects of silymarin are due to silybin [17].” In introduction section

4.      The discussion section is written nicely and the interpretation of results in discussion is good but there is use of irrelevant and unscientific words.

 Conclusion

1.      The conclusion section is ok, but can be improved for better soundness

Overall, the manuscript presented in a nice manner and there are very few reports available on the present study. The present study will also open a new avenue for bio-stimulators and will help in sustainable agriculture. 

Author Response

Dear reviewer,

Thank you for your time and valuable suggestions and comments. Attached is the list of responses. 

Author Response File: Author Response.docx

Reviewer 2 Report

There is need to improve statements for English Language as there are some of statements which are not easy to understandable.

What was the pH of growing media of plants and how it was maintained through irrigation water??

What was the correlation of using??? Foliar spray of ALE was applied around 8 a.m. Approximately 50 mL of 93 ALE solution (20, 40, and 60 mL/L) or distilled water (control) was sprayed throughout the individual plant.

There must be correlation analysis of ALE applications with S. marianum and interrelation of recorded traits.

The data were analyzed using ANOVA/MANOVA but there is no any table to show the significance of results.

Zone of inhibition (mm) plate photos should also be given

The accession number of both bacteria should be mentioned or remove form Escherichia coli ATCC8739 (E. coli),

Author Response

Dear reviewer,

Thank you for your time and valuable suggestion and comments. Attached is the list of responses. 

Author Response File: Author Response.docx