Investigation of the Impact of Soil Physicochemical Properties and Microbial Communities on the Successful Cultivation of Morchella in Greenhouses

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The abstract should be more informative and focus on specific findings, rather than containing general information that could be more appropriately included in the introduction. Line 18 requires punctuation correction. A significant part of the introduction relies on a single article [11]. Please expand the introduction to include additional relevant references. The physicochemical characteristics (such as concentrations and pH values) should be included in the main manuscript (section 3.1), rather than in the supplementary material. Phyla should not be italicized; only families, genera, and species should be. Section 3.2 is insufficient. It is necessary to include the relative abundances as well as the standard deviation. Please present the analysis of the results at both the phylum and genus levels. A shift in the diversity and relative abundance of the microbial community should be clearly highlighted and presented. The authors state, “We observed significant differences in microbial community structure and physicochemical properties between soil…” but this claim is not supported by the way the results are currently presented. Please include tables showing the dominant phyla and genera (relative abundance ± SD), as well as comparative figures displaying the main phyla and genera detected across the three experimental conditions.

Author Response

Dear reviewer:

Thank you for your decision and constructive comments on my manuscript.We agree with the reviewers' suggestions and will incorporate the recommended changes into the manuscript.

Reviewer1:The abstract should be more informative and focus on specific findings, rather than containing general information that could be more appropriately included in the introduction.

Response1:Thank you for your constructive feedback regarding the abstract.In response to your comment,we have expanded the abstract to include detailed findings related to specific results, e.g., microbial community dynamics, soil physicochemical properties, and their impact on morel growth], while removing general statements that are more appropriate for the introduction. All additions and modifications to the abstract have been highlighted in yellow to facilitate your review.

Reviewer2:Line 18 requires punctuation correction. 

Response2:We sincerely appreciate your careful attention to the punctuation issues. The necessary modifications have been made, and a thorough review of punctuation throughout the manuscript has been conducted.

Reviewer3:A significant part of the introduction relies on a single article [11]. Please expand the introduction to include additional relevant references. 

Response3:Thank you for your insightful comment regarding the introduction section.In response to your suggestion, we have expanded the introduction to include additional relevant references, and we have added a detailed discussion on the life cycle of Morchella,thereby providing a more comprehensive background for our study. 

Reviewer4:The physicochemical characteristics (such as concentrations and pH values) should be included in the main manuscript (section 3.1), rather than in the supplementary material. Phyla should not be italicized; only families, genera, and species should be. Section 3.2 is insufficient.

Response4:Thank you for your valuable comments and suggestions regarding the presentation of physicochemical characteristics and the formatting of taxonomic names in our manuscript. In response to your suggestion, we have moved the physicochemical characteristics (including concentrations and pH values) from the supplementary material to the main manuscript (Section 3.1).Additionally, we have carefully reviewed the entire manuscript to ensure that only family, genus, and species names are italicized, as per taxonomic conventions. All instances of "phyla" have been corrected to non-italicized form, and we have verified that the formatting of other taxonomic ranks adheres to standard guidelines.

Reviewer5:Section 3.2 is insufficient. It is necessary to include the relative abundances as well as the standard deviation. Please present the analysis of the results at both the phylum and genus levels. A shift in the diversity and relative abundance of the microbial community should be clearly highlighted and presented. The authors state, “We observed significant differences in microbial community structure and physicochemical properties between soil…” but this claim is not supported by the way the results are currently presented. Please include tables showing the dominant phyla and genera (relative abundance ± SD), as well as comparative figures displaying the main phyla and genera detected across the three experimental conditions.

Response5:Thank you for your constructive feedback and valuable suggestions regarding the presentation of our results. We greatly appreciate your attention to detail. As per your suggestion, we have added a new figure (Supplementary Figure 2) that illustrates the differences in the relative abundances of the top five phyla and genera across the three soil types, highlighting the shifts in microbial community diversity and relative abundance. Additionally, we have revised and expanded the Results section to better emphasize the correlation between the microbial community data and the growth and yield of Morchella, including a more detailed discussion of the observed differences in microbial community structure and physicochemical properties between soil types. Thank you again for your insightful comments, which have greatly improved the quality of our work.

We sincerely appreciate the time and effort invested by the reviewers in evaluating our manuscript. We look forward to any additional feedback or suggestions.

 

Yours sincerely

 

Xin-Hai Liu

 

8 March 2025

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Investigation of the Impact of Soil Physicochemical Properties  and Microbial Communities on the Successful Cultivation of  Morchella in Greenhouses

The name of the Corresponding author doesn’t appear as an author

The introduction is written very well, although a piece of general information regarding the life cycle of the Morchella is missing. The objective is clear to me as a reader.

Materials and Methods -: why only 1-5 cm soil layer were collected – request explanation. Not clear how many soil sample replicates? and how toward the end you have 9 samples. Soil moisture was not determined. why?

All the soil samples were collected at the same Morchella life cycle stage. At inoculation and the end of the growing?

One of the basic knowledge in general mushroom agriculture is soil moisture and air humidity – no data was collected about these two important abiotic variables.  It should be determined and added. As these two data are not available – the comparison of these sampling sites is not suitable.

Results

All the beautiful data collected with the new and nice figures are not relevant.

 

Discussion

The discussion is well written – however, missing the basic assumption. Soil physical and chemical analysis are of great importance IF they are related to the life cycle of Morchella.

The study is of great importance however the authors should ask the right questions. 

Author Response

Dear reviewer:

Thank you for your decision and constructive comments on my manuscript.Your feedback has been invaluable in improving the quality and clarity of our work. Below, we provide a point-by-point response to your comments and describe the revisions made to the manuscript.

Reviewer1:The name of the Corresponding author doesn’t appear as an author.

Response1:Thank you for pointing out this oversight. We sincerely apologize for the confusion caused by the missing corresponding author information in the manuscript.We have now corrected this issue by ensuring that the corresponding author’s name is included in the author list and clearly marked with an asterisk (*).

Reviewer2:The introduction is written very well, although a piece of general information regarding the life cycle of the Morchella is missing. The objective is clear to me as a reader.

Response2:Thank you for your positive feedback on the introduction and for pointing out the need for additional information regarding the life cycle of Morchella. In response to your comment, we have added a concise yet informative section in the introduction to describe the general life cycle of Morchella.As per your guidance, all the extended and revised sections in the manuscript, including the newly added content, have been highlighted in yellow for easy identification.Thank you again for your thoughtful review and constructive input, which have significantly strengthened the quality of our work.

Reviewer3:why only 1-5 cm soil layer were collected – request explanation. Not clear how many soil sample replicates? and how toward the end you have 9 samples. Soil moisture was not determined. why?All the soil samples were collected at the same Morchella life cycle stage. At inoculation and the end of the growing?One of the basic knowledge in general mushroom agriculture is soil moisture and air humidity – no data was collected about these two important abiotic variables.  It should be determined and added. As these two data are not available – the comparison of these sampling sites is not suitable.

Response3:Thank you for your thoughtful questions and for highlighting areas where additional clarification is needed. We appreciate the opportunity to address these points in detail.

Soil Layer Collection (1-5 cm):The 1-5 cm soil layer was specifically chosen because the primordia and fruiting bodies of Morchella are formed in the topsoil layer, which is primarily influenced by the microbial community in this region. This approach is consistent with previous studies, as referenced in our manuscript (References 30 and 42), where topsoil layers were also collected for similar analyses.

Soil Sample Replicates and Number of Samples:A total of 9 greenhouses were selected for soil sampling, categorized into three groups: 3 greenhouses with normal fruiting, 3 greenhouses with no fruiting, and 3 greenhouses with dead fruiting bodies. In each greenhouse, soil samples were collected using the five-point sampling method, and the five subsamples were mixed to form one composite sample per greenhouse. Consequently, a total of 9 composite samples (one per greenhouse) were subjected to high-throughput sequencing.

Timing of Soil Sample Collection:All soil samples were collected 90 days after the maturation of Morchella fruiting bodies, ensuring consistency in the sampling stage across all greenhouses.

Soil Moisture and Air Humidity:Soil Moisture and Air Humidity:During the primordium stage of Morchella, the greenhouses were subjected to heavy irrigation, maintaining soil moisture at approximately 30% until the maturation of the fruiting bodies. Additionally, Morchella cultivation in greenhouses involves regular air humidification after primordia formation, keeping air humidity consistently around 85%. Therefore, soil moisture and air humidity were relatively uniform across all greenhouses. This uniformity allowed us to focus on other potential factors influencing fruiting outcomes. However, temperature control in the greenhouses proved challenging. We hypothesize that high temperatures may have contributed to the occurrence of dead fruiting bodies and the absence of fruiting in some greenhouses. While soil moisture and air humidity are indeed critical factors in mushroom cultivation, the uniformity of these conditions across the greenhouses, as described above, suggests that they were not the primary variables influencing the observed differences in fruiting outcomes. Instead, temperature fluctuations likely played a more significant role.

Reviewer4:All the beautiful data collected with the new and nice figures are not relevant.

Response4:Thank you for your valuable feedback. We appreciate your suggestion to focus the results more directly on the core scientific questions of the study. In the revised manuscript, we have reorganized and expanded the results section to better highlight the relevance of the data to morel growth and yield. Specifically, we have emphasized how microbial community structure and soil physicochemical properties influence morel development, and how these factors may lead to successful cultivation or failure. We have also included additional interpretation of key findings, such as the role of nitrogen and organic matter in supporting mycelial growth, the impact of potassium and phosphorus depletion on fruiting body formation, and the inhibitory effects of pathogenic bacteria and unfavorable conditions.We believe these changes significantly improve the clarity and relevance of the results, ensuring that the data directly address the core scientific questions of the study. Thank you again for your insightful comments.

Reviewer5:The discussion is well written – however, missing the basic assumption. Soil physical and chemical analysis are of great importance IF they are related to the life cycle of Morchella.The study is of great importance however the authors should ask the right questions.

Response5:Thank you for your valuable feedback. We appreciate your suggestion to strengthen the discussion by linking soil physicochemical properties to the life cycle of Morchella. We agree that this connection is essential for a comprehensive understanding of the results.In the revised manuscript, we have added detailed explanations regarding how changes in soil nitrogen, organic matter, potassium, phosphorus, and pH are related to the growth and development of Morchella. Specifically, we have discussed how high nitrogen and organic matter levels support mycelial growth, potassium and phosphorus depletion may limit fruiting body formation, and pH changes influence microbial symbiosis and morel development. We have also proposed hypotheses to explain how these soil properties may impact the life cycle of Morchella.Thank you again for your insightful comments.

We sincerely appreciate the time and effort invested by the reviewers in evaluating our manuscript. We look forward to any additional feedback or suggestions.

 

Yours sincerely

 

Xin-Hai Liu

 

8 March 2025

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript could be accepted for publication