Biological Activity of Four Trichoderma Species Confers Protection against Rhizoctonia solani, the Causal Agent of Cucumber Damping-Off and Root Rot Diseases
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors
Manuscript ID: sustainability-2348656
The paper entitled “Biological activity of four Trichoderma species confer protection against Rhizoctonia solani, the causal agent of cucumber damping-off and root rot diseases” was carefully reviewed. This paper provides valuable information on the use of Trichoderma species as a biological control agent against the plant pathogen R. solani in cucumber crops. The authors have done extensive research on the topic and have presented an interesting work. However, the manuscript needs to be revised.
Detailed comments:
- Line 31: Replace “The four Trichoderma species that increased defense-enzymes activities, T. asperellum KSATC and KSAT1E, were the greatest in increasing peroxidase, catalse, and chitinase enzymes activities in cucumber plants” by “T. asperellum KSATC and KSAT1E, were the greatest in increasing catalase, peroxidase, and chitinase enzymes activities in cucumber plants”.
- Line 31: Correct “catalse” by “catalase”.
- Line 34: Add quantitative data to this statement: “Conversely, a significant (P<0.05) elevation in polyphenoloxidase enzyme and total phenol content was recorded in cucumber plants treated with T. harzianum KSATR89 and KSATR10”.
- Line 95: replace “…against R. solani throughout greenhouse and laboratory trials” by “… against R. solani throughout laboratory and greenhouse trials”.
- The authors refer to some methods from previous studies (e.g., Sneh et al., [12], Barnett and Hunter [39], Misawa et al. [42], MEGA [43]) without providing sufficient details about these methods. It would be helpful to include relevant information for these methods to allow readers to access the full protocols for better understanding.
- The authors indicate that disease assessment was done according to Hassan et al. [44], but it does not provide any details about the specific methods used for assessing pre-emergence damping-off, post-emergence damping-off, survival seedlings, and disease incidence. Please provide a brief description of the methods used for these assessments.
- Move Table 1 (Rhizoctonia solani and Trichoderma species of the current study and those retrieved from GenBank used in phylogenetic analysis) to the Materials & Methods section.
- The authors mention that their findings are in agreement with previous studies, but do not provide sufficient context or comparison with previous research (e.g. lines 603-604, lines 661-663, lines 672-673). It would be helpful to provide more information on the previous studies that reported similar results and discuss how the current work contributes to the existing knowledge in the field. This would provide readers with a better understanding of the novelty and significance of the findings. THerfore, I recommend providing more context regarding this comment.
- The discussion section is not well enough written. I recommend rewriting it to better understand and express the relevance of the results to prior knowledge. In addition, many "key" and recent references have been omitted from the discussion section:
1. Behiry, S.; Soliman, S.A.; Massoud, M.A.; Abdelbary, M.; Kordy, A.M.; Abdelkhalek, A.; Heflish, A. Trichoderma pubescens Elicit Induced Systemic Resistance in Tomato Challenged by Rhizoctonia solani. J. Fungi 2023, 9, 167. https://doi.org/10.3390/jof9020167
2. Shalaby, T.A.; Taha, N.; El-Beltagi, H.S.; El-Ramady, H. Combined Application of Trichoderma harzianum and Paclobutrazol to Control Root Rot Disease Caused by Rhizoctonia solani of Tomato Seedlings. Agronomy 2022, 12, 3186. https://doi.org/10.3390/agronomy12123186
3. Abbas, A., Mubeen, M., Zheng, H., Sohail, M. A., Shakeel, Q., Solanki, M. K., Iftikhar, Y., Sharma, S., Kashyap, B. K., Hussain, S., Zuñiga Romano, M. D. C., Moya-Elizondo, E. A., & Zhou, L. (2022). Trichoderma spp. genes involved in the biocontrol activity against Rhizoctonia solani. Frontiers in Microbiology, 13. https://doi.org/10.3389/fmicb.2022.884469
4. Anugya Bhattarai, Amrit Sharma, Ritesh Kumar Yadav, Pradeep Wagle, 2022. Interacting effects of botanicals, biocontrol agents, and potting media on Rhizoctonia solani led damping-off of okra seedlings, Journal of Agriculture and Food Research, Volume 10, 100410, https://doi.org/10.1016/j.jafr.2022.100410
5. Hlokwe, Mapula T.R, Kena, Mapotso A., & Mamphiswana, David N. (2020). Application of plant extracts and Trichoderma harzianum for the management of tomato seedling damping-off caused by Rhizoctonia solani. South African Journal of Science, 116(11-12), 1-5. https://dx.doi.org/10.17159/sajs.2020/7860
6. hao WANG, Wen-ying ZHUANG, 2019. Evaluating effective Trichoderma isolates for biocontrol of Rhizoctonia solani causing root rot of Vigna unguiculata, Journal of Integrative Agriculture, Volume 18, Issue 9, Pages 2072-2079. https://doi.org/10.1016/S2095-3119(19)62593-1
7. Nawrocka, J., Małolepsza, U., Szymczak, K. et al. Involvement of metabolic components, volatile compounds, PR proteins, and mechanical strengthening in multilayer protection of cucumber plants against Rhizoctonia solani activated by Trichoderma atroviride TRS25. Protoplasma 255, 359–373 (2018). https://doi.org/10.1007/s00709-017-1157-1
Numerous oversights in terms of spelling and grammar, as well as areas where phrasing may benefit from refinement, can be observed.
Author Response
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Author Response.pdf
Reviewer 2 Report
The present work deals with the isolation and characterization of Rhizoctonia solani pathogens and Trichoderma species. The authors also showed i) pathogenicity of Rhizoctonia and ii) biocontrol activity of Trichoderma. In general, the work is good and fits well within the scope of the Journal. Some comments are given below.
1. Further describe the isolation part. What was the frequency of Rhizoctonia and Trichoderma isolation?
2. For Rhizoctonia, why didn't the authors show the data of the remaining isolates (especially for the identification part)? Besides, there seemed to be a confusion between the strains used (see PPDURS10 and PPDURS5, for example).
3. Briefly describe the results from the BLAST search of both Rhizoctonia and Trichoderma.
4. For Trichoderma, show their colony morphology to support the molecular data used for their identification.
5. Clearly explain why the PPDURS2 was selected for further study. The data obtained from pathogenicity seemed to be varied among the strains tested. Also, recheck the data of PPDURS2 for % survival (0) and % DI (0).
6. How about PPDURS10? Was this strain not a pathogen?
7. For Fig. 4. 4a was not important; 4b, control can be removed.
8. In Table 3, further explain why some strains of antagonistic Trichoderma showed an increase in percentage of post damping off, compared to the infected control. Besides, what was % efficacy? Also, why the infected control did not show any DI?
9. Revise the explanation of the enzyme activity studied. See related work regarding this. Then correct the y-axis of Fig. 6. Also, correct the y-axis of Fig. 7.
10. In the Abstract, cfu was used inappropriately.
This can be rechecked.
Author Response
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Reviewer 3 Report
In this research, the author studied the association of three AGs of R.solani with the root rot of cucumber. It was found that there is no obvious difference between T. asperellum KSATC and Rizolex-T fungicide in the treatment of root rot and damping-off. However, the concentrations of three defense-related enzymes (PO, CAT, and CHT) increase in the treated cucumber plants.
This paper was carefully prepared with solid experimental proof. The reviewer believes it can be published with the current version. If the following detailed issues are well-addressed, this reviewer believes that the essential contribution of this paper is important for the against of damping-off and root rot diseases of cucumber.
- There are at least two grammatical and spelling errors in the manuscript, such as, in line 109, “0.3 g L-1“ should be “0.3 g.L-1“; in line 159, should have a space between number and unit, like “10 g“.
- Authors need to unify the format of all references, such as whether to add the DOI, like Refs 6, 8; whether to use both the starting and ending page numbers, like Refs 3, 7, and 9.
Author Response
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Reviewer 4 Report
Reviewer report
The paper “Biological Activity of Four Trichoderma Species Confer Protection against Rhizoctonia solani, the Causal Agent of Cucumber Damping-off and Root Rot diseases” is well written, and English writing is clear but editing is required to correct errors or improve readability. Authors investigated the antifungal activity of four Trichoderma species, providing a safe and effective way to combat damping-off and root rot of cucumber. From my point of view, the topic is interesting and meaningful in the field of sustainable agriculture, but the manuscript needs to be improved with minor revisions to be suitable for publication in Sustainability journal. Some questions need to be addressed and some suggestions are presented below:
Abstract:
Line 16- Please put the scientific name after cucumber and remove family name.
Line 19-20: use in vitro and in vivo instead of laboratory and greenhouse conditions.
Line 21: describe the virulence of different AGs on cumber plant.
Line 28: replace CFU with conidia.
Introduction:
Line 49: “the essential root rot causal factors in a wide range of plants worldwide” please use scientific word i. e. Common instead of essential, and agents instead of factors.
Line 50: replace influences with infects.
Line 51: “Damping-off accounts for the most prevalent symptom of Rhizoctonia” replace accounts for with is.
Line 95: replace native with indigenous.
Materials and Methods
Line 104: replace expressing with exhibiting.
Line 105: replace specimens with samples.
Line 112: replace acquired with obtained.
Line 213: Influence with effect.
Results
Line 296- 306: The authors presented directly molecular characterization of Rhizoctonia solani while they must describe the results of morphological characteristics at first then results about sequence based identification.
Please remove Table 1 to supplementary file
In the Table 2, disease incidences for isolates PPDURS2, PPDURS9 are 0%. While survival plants were 0 and 93.33% respectively, could you please clarify this?
Discussion
Start the discussion with morphological characterization of R. solani isolates and number of nuclei, which group was more pathogenic multi or binuclear strain.
Line 591- 592: Do not start sentences with quoting a Ref like this “Yıldırım and Erper [60] stated” or Erper et al. [62] ascribed, please discus the results then mention the ref at the end of sentences. Correct all next citation as I mentioned:-
Line 617: Zhang et al. [68] stated that T. longibrachiatum caused
Line 633 : Huang et al. [70] point out that T. harzianum strain SQR
Line 640- 643: Inbar et al. [71] have reported similar, Melo and Faull [72] demonstrated , El-Kazzaz et al. [73] observed the lysis.
Author Response
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Round 2
Reviewer 2 Report
1. Line 178, use 'Pre-'.
2. Line 222, use superscript to denote the spore numbers.
3. Describe your response to the Comments no. 5 and 8 in the text where appropriate.
4. For Comment 7, Fig. 4a and 4b are the same result. You can choose one in which the appropriate is 4b. Besides, no need to show the control data. If the authors want to show the data of the control, you can just show the Fig. of the Rhizoctonia mycelial growth without Trichoderma.
5. No revision for Comment 9 yet. Check the related paper and revise how to define the enzyme activity. Also, for the Fig. showing the data of enzyme activity, recheck the Unit used for this enzyme activity.
Recheck the English used.
Author Response
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