Genetic Stability, Phenolic, Flavonoid, Ferulic Acid Contents, and Antioxidant Activity of Micropropagated Lycium schweinfurthii Plants

Round 1

Reviewer 1 Report

The authors presented a manuscript entitled “Genetic Stability, Phenolic, Flavonoid, Ferulic Acid Contents and Antioxidant Activity of Micropropagated Lycium schweinfurthii Plants.” This article presents is extremely interesting. 

 The results are very important to food science and for the understanding of in vitro micropropagation of L. schweinfurthii and assess the genetic stability of micropropagated plants (MiPs) as well as to estimate phenolic, flavonoid, ferulic acid contents, and antioxidant activity in the leaves of micropropagated plants.

 I think it is necessary for the authors to improve the resolution of the subtitles in Figure 3.

In addition, it is not necessary to present the standard curve graphs for gallic acid and catechin (Figure 6). This information can be added to the text.

I suggest the authors put in a table the results of total phenolics, flavonoids, DPPH, ABTS, and ferulic acid, it will be more informative for the authors, as it is necessary information.

Author Response

Dear reviewer 1,

Thank you very much for your valuable comments that should improve our article. Please find the attached file of the revised manuscript.

Point 1: I think it is necessary for the authors to improve the resolution of the subtitles in Figure 3.

Response 1: We agree with you about that. We removed the borders between different gels photos to save more space but it was still too much. Maybe, putting only two subfigures (one for RAPD and one for ISSR) in the paper and the rest on the supplementary material solve this problem.

Point 2: In addition, it is not necessary to present the standard curve graphs for gallic acid and catechin (Figure 6). This information can be added to the text.

Response 2:We removed Figure 6 from the manuscript and added some details in the text instead. They are highlighted with green color.

Point 3: I suggest the authors put in a table the results of total phenolics, flavonoids, DPPH, ABTS, and ferulic acid, it will be more informative for the authors, as it is necessary information.

Response 3: We added Table 4 that summarizes their results. The table caption is highlighted with green in the manuscript.

Best regards,

Authors

Author Response File: Author Response.docx

Reviewer 2 Report

The paper described an in vitro micropropagation of Lycium schweinfurthii. Authors described the motivation of studying this plant based on its richness in secondary metabolites. To reach their targets, authors have carried out multiple culture assays with changing the composition of the fortified media. Then, authors controlled the genetic stability of obtained plantlets by carrying out biochemical tests. Finally, authors have estimate the total antioxidative compounds in micro propagated plants.

The manuscript is very interesting and the experiments are well described. Nevertheless, there is lack of details repeatedly especially in Figures. In addition, the quality of some figures is poor and should be considered if this research paper will be published. In the following section, you will find some remarks and comments to improve your paper.

 

Line 26-27 : The sentence should be more simple to express the finding in the dried leaves.

Line 59 -60 : Correct the name of species and put it in italic form in all over the manuscript.

Adding picture of described plant would be appreciated in the supplementary data for seeds, and plants.

Line 168 : please correct twenty microliter to 20µL

Please define MS media line 139.

Figure 1 : more details are required in the legend of the photo.

It is difficult to understand the figure, what represent the “x” axis? How do you interpret this enormous error bars? For the media ½ MS+IBA 0.4, the error bar is approximatively equal to the half of your result.

Figure 2 is not well described in the text.

Figure 4: more details should be added on the figure to clarified the differences. You can add for example circle to show the difference. You can also add clearer picture of your results.

Figure 6: please improve your results presentation, if this curve is the result of duplicates, please plot the average value and then add the error bar for each value. You can use (a) and (b) to distinguish both graphs. And add more details in the legend namely the solvent used in your preparation and the studied concentration range.

Figure 7: add rack number on your HPTLC plates.

Author Response

Dear reviewer 2,

Thank you very much for your valuable comments that should improve our article. Please find the attached file of the revised manuscript

All Changes in the manuscript related to your comments are highlighted in yellow color.

Point 1: Line 26-27 : The sentence should be more simple to express the finding in the dried leaves.

Response 1: We tried to simplify the sentence.

Point 2: Line 59 -60 : Correct the name of species and put it in italic form in all over the manuscript.

Response 2: This was related to problems that occurred during submission and changing the manuscript to the journal format. We double-checked the scientific names in the whole manuscript. We removed also the extra dash (-) that appeared within some words.

Point 3: Adding picture of described plant would be appreciated in the supplementary data for seeds, and plants.

Response 3: They will be added as supplementary materials.

Point 4: Line 168 : please correct twenty microliter to 20µL

Response 4: Corrected.

Point 5: Please define MS media line 139.

Response 5: we defined it in the first mention time on the context lines 67, 68.

Point 6: Figure 1 : more details are required in the legend of the photo.

It is difficult to understand the figure, what represent the “x” axis? How do you interpret this enormous error bars? For the media ½ MS+IBA 0.4, the error bar is approximatively equal to the half of your result.

Response 6: The “x” axis in Figure 1 (It is now named Figure 2) represents both root length in cm and the number of roots that emerged per plant. The number of replicates used in the rooting stage was 8. Not all the 8 replicates responded to the plant growth regulators from the point of root formation (some of them didn’t form roots, with 0 roots and 0 cm length) while others formed high number of long roots. During the calculation of standard error, the error should be high. If we select only the responded replicates (3 or 4 replicates), the means will be higher and the standard error will be lower than now. We didn’t prefer to do so to avoid bias in selection.

There is also a suggestion that to calculate the percentage of rooting response (in the 8 replicates) and then analyze the responded replicates (3 to 4 replicates) from the point of root length and number of roots formed.

Point 7: Figure 2 is not well described in the text.

Response 7: You are right. It was mentioned only by the end of the rooting paragraph. It is mentioned now three times (one for each sub-figure). We changed the Figure number to Figure 1 due to the first citation.

Point 8: Figure 4: more details should be added on the figure to clarified the differences. You can add for example circle to show the difference. You can also add clearer picture of your results.

Response 8: The figure with relatively better resolution is added with some modifications to show examples of similarities.

Point 9: Figure 6: please improve your results presentation, if this curve is the result of duplicates, please plot the average value and then add the error bar for each value. You can use (a) and (b) to distinguish both graphs. And add more details in the legend namely the solvent used in your preparation and the studied concentration range.

Response 9: According to the opinion of Reviewer 1 “In addition, it is not necessary to present the standard curve graphs for gallic acid and catechin (Figure 6). This information can be added to the text.”, we removed the standard curves and wrote the necessary information within context (highlighted with green) showing the number of replicates and concentrations used to obtain the equation. We preferred to do so to avoid the confusion of the reader with these results as they are not of that importance. It is just routine work to generate the equation to be used to quantifying phenolics and flavonoids in relation to gallic acid and catechin equivalence. Furthermore, the replication and using different concentrations, in this case, is already reflected in the generated equation and its accuracy in calculation.

Point 10: Figure 7: add rack number on your HPTLC plates.

Response 10: Rack number is added. The figure changed to Figure 6 as a result of gallic acid and catechin standard curves figure removal.

Best regards,

Authors

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

All the comments were considered.

Author Response

Dear Reviewer 2,

We would like to thank you very much one more time for your comments that helped in improving our article.

Best regards,

Authors