Influence of Heat Treatment and Solid-State Fermentation on the Lignocellulosic Fractions of Substrates Supporting Lentinula edodes (Berk.) Pegler Cultivation: Implications for Commercial Production

Round 1

Reviewer 1 Report

This manuscript deals with the cultivation of shiitake mushrooms on sawdust and wheat straw processed at different heat treatments. After a careful reading, I found this manuscript interesting and important for the Indian perspective of mushroom technology. The experiments are conducted with appropriate statistical design under suitable conditions. Also, the six tested strains suggest their suitability for commercial exploitation by farmers. However, I have observed some grey points in the manuscript which need to be carefully revised. I suggest minor revisions and my specific comments are:

1.      Add the scientific authority of L. edodes in the title and abstract sections.

2.      Remove the bracket from the last keyword.

3.      DMR-356 is a popular commercial strain of shiitake mushrooms from DMR, India. Why this strain was not tested?

4.      There are spacing, syntax, and grammatical problems in the paper which need to be corrected thoroughly.

5.      Material and method sections should be categorized into content-specific subheadings such as material collection, experimental treatments, design, and data analysis.

6.      Provide the malt extract agar composition and storage conditions of the culture.

7.      Any specific reason behind the selection of Toona sinensis? Also, provide the common name of it and the scientific name of wheat straw (Triticum aestivum) with the appropriate name of variety/botanical authorities.

8.      Provide the geocoordinates of sample/waste collection.

9.      Methods for QC and CRM are missing, also be specific in lignin (Klason?).

10.   A factorial randomized design with four replicates is fine but why only three replicates were used for analysis?

11.   Conduct an appropriate test of significance for the data given in Table 1.

12.   Create subheadings in the “Results and discussion” section too.

13.   Explanations for abbreviations used in the data tables are missing also. The unit of “production rate” is lacking. SE is strain specific or waste-specific?

14.   Axial legends in Fig. 1 are distorted.

Author Response

We are grateful to the reviewers for their insightful comments on our paper. Please see the attachment for point wise reply to the comments and suggestions.

Author Response File: Author Response.docx

Reviewer 2 Report

I congratulate authors for the relevant and very interesting work. In the current times when as a society we are increasingly conscious of the impact of own food choices (specially meat based) on climate, fungal/mushroom based protein rich food is the need of the hour and a study like this is a step ahead towards a solution. I however have a few comments, if authors can suitably address these, the manuscript should be ready to be accepted.

1. Both the substrates used for Lentinula production have been extensively researched and reported in literature, the same is with the mode of fermentation (solid state). It was hence difficult to fathom the novelty of this work merely based on the heat pre-treatment of the substrate which is anyhow mandatory for the sterilization. Authors should think over and state how this work makes a difference in field. The novel point of this work should be clearly mentioned in manuscript.

2. Line 111-114: The structure of hemicellulose (both C6 and C5 sugars) should be explained with more clarity and also a better explanation of the hemicellulose degradation by fungi (Hemicellulases (xylanase) is the major enzyme secreted by white rot fungi along with ligninases- is this the same case for this strain?). It might be well explained in the mentioned reference but diverging a bit on this part in text will be good and will help readers to understand more about lignin-hemicellulose complex and its dissolution by fungi.

3. Line: 151-153: Novelty of this work? Mention here. 

4. Line 165-167: I suppose this is the general perception of authors as the statement is not supported by any experimental work at molecular level in this work. Such an inference should be thought over before being mentioned in conclusion.

Author Response

We are grateful to the reviewers for their insightful comments on our paper. Please see the attachment for point wise reply to the comments and suggestions.

Author Response File: Author Response.docx

Reviewer 3 Report

I reviewed the manuscript entitled, Influence of heat treatment and solid-state fermentation on ligno-cellulosic fractions of substrate supporting Lentinula edodes cultivation- Implications on commercial production. The manuscript focused on the utilization of heat treatment and solid-state fermentation on ligno-cellulosic fractions in order to grow shiitake mushrooms. This study is innovative and contributes to the field. In my opinion, this manuscript can be accepted for publication after addressing the suggestion below. The methodology section must be improved. It is very poorly written.

Title should be revised as “Influence of heat treatment and solid-state fermentation on ligno-cellulosic fractions of substrate supporting Lentinula edodes cultivation: Implications on commercial production

Abstract:

Please provide the background of the study in abstract

Recommendations of the study should be added at the end of the abstract

Please keep the line numbers throughout the manuscript to facilitate easy review process

Provide the space between 2].Besides being very….

Introduction:

Objectives of the study must be clearly presented

Methodology is poorly written. Detailed methodology for each parameter must be provided with appropriate citation. How did the authors perform statistical analysis?

Heat treatment: There is no information available on heat treatment. How much heat is supplied?

Lignin (%), cellulose (%) and hemicellulose (%) were estimated … please provide detailed methodology

There is no information on pH and moisture content methodology

How did the authors calculate the yield?

Figure 1. a, b, c,… etc should be denoted in Figure. Statistical analysis must be performed to understand the significant differences. As such, it is difficult to understand the significant differences. Also, quote, what is S1, S2, S3 at the Figure footnote

Results and discussion are appropriate

Conclusions should be revised

References are not according to the journal format.

Author Response

We are grateful to the reviewers for their insightful comments on our paper. Please see the attachment for point wise reply to the comments and suggestions.

Author Response File: Author Response.docx

Reviewer 4 Report

Review for

 

 

 Influence of heat treatment and solid state fermentation on lignocellulosic fractions of substrate supporting Lentinula edodes cultivation- Implications on commercial production

 

 

In the present study authors investigate the possibility of growing shiitake mushroom on ligno-cellulosic biomass (saw dust and wheat straw) processed at different heat treatments.

 

I am not sure that using saw dust and wheat straw is truly innovative for shiitake

 

Assessing the nutritional quality of fungal treated wheat straw: Compounds formed after treatment with Ceriporiopsis subvermispora and Lentinula edodes

Mao, L., van Arkel, J., Hendriks, W.H., (...), de Vos, R.C.H., Sonnenberg, A.S.M.   2021      Animal Feed Science and Technology

276,114924

 

Wheat bran addition improves Ceriporiopsis subvermispora and Lentinula edodes growth on wheat straw, but not delignification   Mao, L., Cone, J.W., Hendriks, W.H., Sonnenberg, A.S.M.             2020      Animal Feed Science and Technology

259,114361

 

Authors attempted the cultivation of shiitake in both sawdust and wheat straw exposed to various degrees of substrate pre-heat treatment. The study also gives an insight into the chemical changes in the substrate due to the variation in heat treatments and its effect on mycelia growth and fruiting capacity.

 

Work not done with only one strain

A total no. of six strains (DMRO–35, 51, 297, 388s, 410, 412) ascribed to the genus Lentinula, were obtained from the culture bank of the ICAR-Directorate of Mushroom Research, Solan (HP), India.

 

Very short M&M, only one page, quite unusual for a scientific paper

 

The possibility of growing shiitake on WS either in pasteurized or sterilized substrate warrants more efforts to get economically viable returns.

 

not sure if this would be feasible on a large scale  (multi tons scale)

energy cost

 

 

 

Author Response

We are grateful to the reviewers for their insightful comments on our paper. Please see the attachment for point wise reply to the comments and suggestions.

Author Response File: Author Response.docx

Round 2

Reviewer 4 Report

Authors carefully revised their MS according to my comments/advices