Evaluation of Magonia pubescens A. St.-Hill. Roots Extract against Phytopathogens: Searching for Eco-Friendly Crop Protection Products
, Guacimara Espinel 3, Grasiely F. Sousa 2, Lucienir P. Duarte 2, Ignacio A. Jiménez 1, Raimundo Cabrera 3 and Isabel L. Bazzocchi 1,*Round 1
Reviewer 1 Report
Please see the PDF for comments. Lots of issues are raised in the paper, need to answer all the mentioned issues.
Comments for author File: 
Comments.pdf
The English is acceptable.
Author Response
Response to Reviewer 1
Remark 1.its insecticidal potential
ANWER: This point has been corrected in the revised version
Remark 2. Take information form this paper and cite the references.
- Development and evaluation of emulsifiable concentrate formulation containing Sophora alopecuroides L. extract for the novel management of Asian citrus psyllid
ANWER: This point has been corrected in the revised version following the reviewer’s suggestion, and references 2 and 18 have been included.
Remark 3. This paragraph is the concise information of the decade. So need to add more literature evidence
Need to cite
- Efficacy of some selected synthetic chemical insecticides and bio-pesticides against cotton mealybug, Phenacoccus solenopsis Tinsley (Sternorrhyncha: Pseudococcidae)
- Application of essential oils as natural biopesticides; recent advances. Critical Reviews in Food Science and Nutrition.
ANWER: This point has been corrected in the revised version following the reviewer’s suggestion, and the two cited references (22 and 23) have been added.
Remark 4. I am sure there will be a literature available regarding the photochemistry of this plant. Need to add literature regarding the bio-active compounds in this plant. At least cite literature what are the chemical groups or compounds present in this plant.
ANWER: In the Introduction section, an extensive description on the biological evaluation and chemical composition of different plant parts of Magonia pusbences are already included, together with several references (lines 80-96, references 29-36).
“The fruits and seeds of the plant are used to prepare a soap for the treatment of dermatitis, lice infestations and as an insecticide and larvicide [29]. Larvicidal activity of ethanolic extracts from the M. pubescens stem have been reported against different species of the Aedes genera, the main vector of the dengue disease [30]. There is also a report on the leishmanicidal activity of the stem bark ethanolic extract [31]. The volatile and flavonoid profile in flowers and leaves from M. pubescens have also been investigated, suggesting it could be effective against free radicals [32]. In addition, essential oils from M. pubescens inflorescences and their cytotoxic activity has been studied [33]. Infusions of the roots are also used as a tranquilizer, and its ethanolic extracts are used to treat wounds and pain [34]. This extract shows inhibitory activity against insects, fungi, and bacteria. In addition, the ethanolic extract of M. pubescens roots displays strong cytotoxic activity against MDA-MB-435 breast cancer cell lines (IC50 7.9 μg/mL) [29]. Furthermore, a study on the insecticide irritability effect of the ethanol extract from the roots of M. pubescens demonstrated that this induced significantly greater insecticide-irritability behavior than the control, permethrin, against Sitophilus zeamais in stored maize [35]. Analysis of the external coat mucilage from M. pubescens seeds has also been reported [36].”
In addition, in Results and Discussion, section 3.2 (lines 313-319), compounds present in the plant are cited, and extended in the revised manuscript.
“Furthermore, several chemical components have been identified in different parts of the plant, including tannins, isolated from the steam bark [47], and 2-O-methylinositol and proanthocyanidin from the peel of the fruits [48]. Moreover, flavonoids and volatile oils constituents of its flowers and leaves [32], and essential oils from inflorescences [33] have been reported. In addition, a preliminary phytochemical survey allowed the identification of steroids, tannins, alkaloids, saponins and flavonoids in the leaves, and flavonoids and alkaloids in the bark of the tree trunks [49].”
Remark 5. Need to add picture of Zone of inhibition and leaf disk bio assay picture to make paper more inters-ting and realistic.
ANWER: As suggested by the Reviewer, additional pictures for both assays, in vitro test-assay on mycelium and leaf-disk bioassay, have been added to the Supporting Information (Figures S1 and S2) in the revised manuscript.
Remark 6. Statistical lettering are missing thought the difference are clear. But need to add letterings or sign of significance.
ANWER: This point has been corrected in the revised version following the reviewer’s suggestion
Remark 7. How you calculate the disk consumed. Either by weighing the leaf disk before and after feeding?
ANWER: As describe in Material and Methods, section 2.6, the methodology used for the leaf-disk bioassay, indicates that: “In the choice assay, leaf disks (5 cm diameter) were alternatively treated with the tested sample solution in ethanol (5 µL per disk–0.2 mg/cm2), or solvent as control, and to calculate the FR in each dish, the leaf area consumed was measured with the ImageJ 1.53e program, and used the formula: FR = [1 - (% Treated disks consumed / % Control disks consumed)] x 100.”
Moreover, a statistical analysis Section (2.7) has been included in the revised manuscript.
Remark 8. You already explain these in results. While discussing the results n need to compare your results with literature.
ANWER: Regarding this point, the only study on the potential insecticide activity of M. pubescens is already cited (ref. 35). Furthermore, comparison with other results using different experimental conditions and/or experimental procedures does not give too much information regarding the comparative potency of plant extracts. Because that, our results were compared with a commercial insecticide, and this comparison suggests the insecticidal potential of M. pubescens.
Lines 309-312: “The only study regarding the potential insecticide activity of M. pubescens is by Silva et al [35], who reported the insecticide irritability effect (avoidance after contact) similar to permethrin in its stem ethanolic extract against Sitophilus zeamais (maize weevil), a serious pest of stored maize.”
Lines 304-308: “Notably, the crude ethanolic extract showed similar effects as Steward 30WG [42], the commercial insecticide used as a positive control, exhibiting a FR of 92.14% and 95.16% in the choice and non-choice feeding assays, respectively. This fact suggest that the plant could be used, after a simple and accessible procedure as a biopesticide against this phytopathogenic insect.”
Remark 9. The readers are totally blind by reading your paper. What this plant contain? what are the possible bio-active chemicals groups? Need to make a table of previous e literature of this plant about its activities.
ANWER: Regarding this point, in Results and Discussion, section 3.2 (lines 313-319), compounds present in the plant are cited, and extended in the revised manuscript. There is not much literature about Magonia pubescens phytochemical analysis, that is our first motivation to study this plant. Specially, as mentioned in the paper, phytochemical analysis of the roots has not been reported.
Author Response File: Author Response.pdf
Reviewer 2 Report
After reviewing the manuscript "Evaluation of Magonia pubescens extract against phytopathogens: searching for eco-friendly crop protection products" I have some suggestions.
I suggest writing a full botanical name when introducing species (also in the title). I recommend specifying in the title that it is a root extract.
The introduction section is too long. Specifically, write the hypothesis of this research.
Line 138 - ...in October 2019 by Maria Olívia Mercadante-Simões, PhD (UNIMONTES). Remove the name from the core text. There is an acknowledgment section for that.
Line 143 - "Kg" to"kg"
Line 179 - I suggest avoiding links in the text.
Add statistical analysis to the results. Currently, results convey nothing, and this is more than necessary to do.
The results and discussion section is short and needs more comparison to similar research. Compare your results with other research on this topic, e.g. root of another plant and its potential. Add value to why M. pubescens deserves more attention with respect to the other natural source of biopesticides.
There are many small errors in the text. Also, botanical names need to be checked and italicized. The full botanical name should be introduced when at the first mention.
Author Response
Response to Reviewer 2
Remark 1. I suggest writing a full botanical name when introducing species (also in the title). I recommend specifying in the title that it is a root extract.
ANSWER: This point has been corrected in the revised version following the reviewer’s suggestion (Title and Abstract).
Remark 2. The introduction section is too long. Specifically, write the hypothesis of this research.
ANSWER: This point has been corrected in the revised version, and the introduction length has been substantially reduced.
Remark 3. Line 138 - ...in October 2019 by Maria Olívia Mercadante-Simões, PhD (UNIMONTES). Remove the name from the core text. There is an acknowledgment section for that.
ANSWER: This point has been corrected in the revised version.
Remark 4. Line 143 - "Kg" to"kg"
ANSWER: This point has been corrected in the revised version
Remark 5. Line 179 - I suggest avoiding links in the text.
ANSWER: This point has been corrected in the revised version, and the link has been removed from the text.
Remark 6. Add statistical analysis to the results. Currently, results convey nothing, and this is more than necessary to do.
ANSWER: As suggested by the Reviewer, a statistical analysis Section (2.7) has been included in the revised manuscript.
“2.7. Statistical Analysis
All data were shown as mean ± standard deviation (SD). In the test-assay on mycelium, the percentage of inhibition was analyzed using one-way analysis of variance (ANOVA). Tukey HSD multiple comparisons of means was used to compare concentrations and treatments. Differences of p < 0.05 were considered statistically significant. All the analyses were performed using Social Science Statistics, 2018.
Leaf-disk bioassay, using three fifth-instar larvae of C. chalcites, were conducted in sextuple for each experiment. Results were expressed in terms of refusal rate (FR), and the leaf area consumed was measured with the ImageJ 1.53e program. Differences of p < 0.05 were considered statistically significant. Analysis of variance was determined by one-way ANOVA [47].”
Remark 7. The results and discussion section is short and needs more comparison to similar research. Compare your results with other research on this topic, e.g. root of another plant and its potential. Add value to why M. pubescens deserves more attention with respect to the other natural source of biopesticides.
ANSWER: Regarding this point, the only study on the potential insecticide activity of M. pubescens is already cited (ref. 35). Furthermore, comparison with other results using different experimental conditions and/or experimental procedures does not give too much information regarding the potency of the plant extract. Because that, results were compared with a commercial insecticide, and this give us information on the insecticidal potential of M. pubescens.
Remark 8. Comments on the Quality of English Language
There are many small errors in the text. Also, botanical names need to be checked and italicized. The full botanical name should be introduced when at the first mention.
ANSWER: The full botanical name has been included in Title, Abstract and when at the first mention.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments:
To the Editor Prof.
The manuscript “Evaluation of Magonia pubescens extract against phytopathogens: searching for eco-friendly crop protection products, investigate the suppression effect of the ethanolic extract of the Magonia pubescens roots against the phytopathogenic fungi, Botritis cinerea, Fusarium oxysporum, and Alternaria alternata. The study is interesting and of great practical importance. The experimental studies are mostly carried out professionally. The article satisfies the criteria of the Applied Science. As detailed data and interpretation, I recommend it for an international audience in this journal, however some points have to be précised and a minor revision is requested.
1. Materials and methods, Line 165, mention the source of phytopathogenic strains i.e, host, disease caused, identification, pathogenicity ….etc.
2. Materials and methods, Where Statistical Analyses section? Authors mentioned that under the figures, Data are presented as meas ± SD (Standard Deviation, n = 8). Not satisfy.
3. Results and Discussion, Line 212-215. Transfer this paragraph to Abstract Section.
4. Results and Discussion, Improve discussion in this scection.
With best regards
Author Response
Response to Review 3
Remark 1. Materials and methods, Line 165, mention the source of phytopathogenic strains i.e, host, disease caused, identification, pathogenicity ….etc.
ANSWER: Currently, in Materials and Methods, Sections 2.5. and 2.6, the sources of phytopathogenic strains are included.
Lines 149-152: “..Strains of B. cinerea (B05.10) and A. alternata (Aa 100) were isolated from Vitis vinifera and Lycopersicon esculentum, respectively, both supplied by the Universidad de La Laguna, Tenerife. F. oxysporum f. sp. lycopersici (2715) strain, isolated from Lycopersicon esculentum, was provided by the Colección Española de Cultivos Tipo (CECT) from Valencia, Spain.”
Lines 168-169: A laboratory colony of Chrysodeixis chalcites was initiated using larvae collected from banana (Musa acuminata, var. Cavendish) crops in Tenerife.
Moreover, diseases and pathogenicity caused by the phytopathogens are already included in the Introduction section (lines 44-52 for the fungi under study, and lines 54-60 for Chrysodeixis chalcites)
Remark 2. Materials and methods, Where Statistical Analyses section? Authors mentioned that under the figures, Data are presented as meas ± SD (Standard Deviation, n = 8). Not satisfy.
ANSWER: As suggested by the Reviewer, a statistical analysis Section (2.7) has been included in the revised manuscript. Moreover, some additional pictures for both assays, in vitro test-assay on mycelium and leaf-disk bioassay, have been added to the Supporting Information (Figures S1 and S2).
“2.7. Statistical Analysis
All data were shown as mean ± standard deviation (SD). In the test-assay on mycelium, the percentage of inhibition was analyzed using one-way analysis of variance (ANOVA). Tukey HSD multiple comparisons of means was used to compare concentrations and treatments. Differences of p < 0.05 were considered statistically significant. All the analyses were performed using Social Science Statistics, 2018.
Leaf-disk bioassay, using three fifth-instar larvae of C. chalcites, were conducted in sextuple for each experiment. Results were expressed in terms of refusal rate (FR), and the leaf area consumed was measured with the ImageJ 1.53e program. Differences of p < 0.05 were considered statistically significant. Analysis of variance was determined by one-way ANOVA [47].”
Remark 3. Results and Discussion, Line 212-215. Transfer this paragraph to Abstract Section.
ANSWER: This point has been corrected in the revised version.
Remark 4. Results and Discussion, Improve discussion in this section.
ANSWER: Discussion has been improved in the revised manuscript.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
The revised versions is quite good. However, the title still need improvement. Regarding the pictures need to add in the main text not in supplimentary file. See the PDF.
Comments for author File: Comments.pdf
English Language is acceptable for publicatiopn.
Author Response
Response to Reviewer 1
Remark 1. The manuscript’s title.
ANWER: Regarding this point, and although the Reviewer suggets chancing the manuscript’s Title as: “Evaluation of antifungal and insecticidal activity of Magonia pubescens A. St.-Hill. roots extract as a potential biopesticide”, I think the current title “Evaluation of Magonia pubescens A. St.-Hill. roots extract against phytopathogens: searching for eco-friendly crop protection products” sounds better and could catch the attention of more readers. Even though if the Reviewer insists it could be changed.
Remark 2. Regarding the pictures need to add in the main text not in supplementary file.
ANWER: As suggested by the Reviewer, additional pictures for both assays, in vitro test-assay on mycelium and leaf-disk bioassay, have been added to the text (Figures 4 and 5) in the revised manuscript.
Author Response File: Author Response.pdf
Reviewer 2 Report
The authors have addressed all of the issues. I can recommend this manuscript for publication.
Minor editing of the English language is required.
Author Response
Response to Reviewer 2
Remark 1. Minor editing of the English language is required.
ANWER: This point has been corrected in the revised version following the reviewer’s suggestion, and English language has been checked by a native English speaker.
Author Response File: Author Response.pdf
