Turning Waste into Wealth: Utilizing Trichoderma’s Solid-State Fermentation to Recycle Tea Residue for Tea Cutting Production

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The paper explores the utilization of Trichoderma guizhouense NJAU 4742 (NJAU 4742) in promoting the growth of tea cuttings through solid-state fermentation (SSF) using tea residue.

n the optimized solid-state fermentation (SSF) conditions, using tea residue as a substrate, NJAU 4742 demonstrated significant potential for spore production. The optimal conditions included a 7:3 (w/w) ratio of tea residue to rice bran, a material thickness of 3 cm, an inoculum concentration of 15% (v/w), and an incubation time of 4 days, resulting in a spore count of 1.83×10^9 CFU/g.

A subsequent pot experiment assessed the influence of nursery substrates with varying concentrations of NJAU 4742 spores on tea cutting growth and rhizosphere fungal communities. The results indicated that the application of NJAU 4742 spore products significantly increased the biomass of tea cuttings. Higher concentrations of NJAU 4742 spores yielded better growth performance, with the most economically viable option identified as the application of a nursery substrate with 1×10^7 CFU/mL spores.

Notably, the study revealed a positive correlation between the presence of Trichoderma and the fresh weight of tea cuttings. Among the top ten fungal genera, Trichoderma stood out for its positive association, while others exhibited a negative correlation.

In conclusion, utilizing tea residue for SSF to produce NJAU 4742 spores proved to be a feasible approach, demonstrating its potential to enhance the growth of tea cuttings. The findings contribute valuable insights into the economic production of Trichoderma spores and their application in sustainable agriculture.

 

 

While the study presents valuable insights, there are areas for improvement:

-The figures quality needs to be improved.

-The introduction could benefit from providing more context on the significance of Trichoderma in agriculture and the specific challenges or gaps in existing research that the study aims to address.

-While the study outlines the optimized conditions for spore yield, more detailed information on the experimental setup, parameters, and statistical analyses would enhance the transparency and reproducibility of the study.

-The economic viability analysis is mentioned, but there could be a more detailed discussion on the economic aspects, costs, and potential benefits associated with different spore concentrations. This would add depth to the study's practical implications.

-While the study mentions the influence of NJAU 4742 spores on the rhizosphere fungal community, a more in-depth discussion on the specific changes, implications, and ecological aspects of the observed fungal community shifts would be valuable.

Further work for discussion:

https://www.sciencedirect.com/science/article/pii/S0924224423001929

https://www.mdpi.com/2311-5637/9/4/364

https://www.mdpi.com/2311-5637/9/4/322

Include a section or discussion on potential future directions for research in this area. Identifying any remaining questions or avenues for further exploration would enhance the study's contribution to the field.

 

Comments on the Quality of English Language

Moderate editing of English language required

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

I find this research to be interesting and relevant to the Journal of Agronomy; however, several modifications are needed for acceptance.

Below are the changes to consider:

Introduction

The introduction is satisfactory; however, it needs a more detailed description of high-throughput sequencing, including some examples in the text.

Materials and Methods

It's necessary to specify where the Trichoderma strains were collected.

Include their phylogenetic identification with Genbank accession numbers or similar databases.

Section 2.2:

Mention if the substrate was sterilized. 

Furthermore, the temperature and humidity conditions during the the experiments are not mentioned. Please consider these points.

Results and Discussion

The presented results are clear; however, the following changes are required:

In Figure 1, A and B are identified, but not mentioned in the figure caption. Please verify this.

In Figures 2 and 3, A, B, C, and D are identified, but not mentioned in the figure caption. Please verify.

Additionally, it's important to add: The analysis of the high-throughput sequencing product.

Conclusion

The conclusion is adequate; however, the results of fungal diversity and the correlation between the fresh weight of tea cuttings and Trichoderma are not mentioned.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The MS “Turning Waste into Wealth: Utilizing Trichoderma's Solid-State 2 Fermentation to Recycle Tea Residue for Tea 3 Cutting Production” report an important issue and can be published after following corrections.

·         Decimal should be only one point in abstract follow it through the whole MS.

·         In introduction write earlier study done on this issue and why your study is novel in this regard.

·         Line 54 L. should not italic

·         Line 90-94, if the Trichoderma species were characterized, can you please provide the accession numbers of all species.

·         Line 10 laboratory grade name of medium, provide for all media used under the study.

·         Line 103 latitude, longitude and elevation

·         Why did you used 5×10⁷ CFU/mL? not lower CFU level? It is quite high.

·         Line 112 mention the tea cultivar name

·         Line 145, why ITS1 and ITS2 it does not cover entire fragment of DNA, why not used ITS1 and ITS4 which cover a large fragment. Further for Trichoderma sequencing one gene is not acceptable you have to sequence at least one conserved gene like TEF or TUB.

·         Data analysis does not show whether experiments were repeated under same biological conditions, if repeated how did you deal with data from repeated experiments. Did you do homogeneity test? Further it is not clear whether you used raw data or transformed data, did you check assumption of ANIOVA to see whether your data was normal or not? These all things should be clearly described in the MS.

·         Each result section must be supported with ANOVA value like CV, SEm, F value.

·         Write separate discussion which recent papers.

·         Conclusion should provide recommendations and future directive.

Comments on the Quality of English Language

The MS “Turning Waste into Wealth: Utilizing Trichoderma's Solid-State 2 Fermentation to Recycle Tea Residue for Tea 3 Cutting Production” report an important issue and can be published after following corrections.

·         Decimal should be only one point in abstract follow it through the whole MS.

·         In introduction write earlier study done on this issue and why your study is novel in this regard.

·         Line 54 L. should not italic

·         Line 90-94, if the Trichoderma species were characterized, can you please provide the accession numbers of all species.

·         Line 10 laboratory grade name of medium, provide for all media used under the study.

·         Line 103 latitude, longitude and elevation

·         Why did you used 5×10⁷ CFU/mL? not lower CFU level? It is quite high.

·         Line 112 mention the tea cultivar name

·         Line 145, why ITS1 and ITS2 it does not cover entire fragment of DNA, why not used ITS1 and ITS4 which cover a large fragment. Further for Trichoderma sequencing one gene is not acceptable you have to sequence at least one conserved gene like TEF or TUB.

·         Data analysis does not show whether experiments were repeated under same biological conditions, if repeated how did you deal with data from repeated experiments. Did you do homogeneity test? Further it is not clear whether you used raw data or transformed data, did you check assumption of ANIOVA to see whether your data was normal or not? These all things should be clearly described in the MS.

·         Each result section must be supported with ANOVA value like CV, SEm, F value.

·         Write separate discussion which recent papers.

·         Conclusion should provide recommendations and future directive.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

I agree with the improved manuscript. 

Comments on the Quality of English Language

Minor editing of English language required

Reviewer 3 Report

Comments and Suggestions for Authors

The authors addressed my all comments, the MS can be accepted.

Comments on the Quality of English Language

The authors addressed my all comments, the MS can be accepted.