Mechanisms by Which Soil Microbial Communities Regulate Ecosystem Multifunctionality in Tea Gardens of Longnan City, China

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript of “Spatial Variation of Soil Microbial Contributions to Ecosystem Multifunctionality in Tea Gardens of Longnan City, China” did many works. The manuscript is interesting and systematic, but there were many questions existed in the present study.

1. The title emphasizes "spatial variation", but the study only covered three counties in Longnan City (Wen County, Wudu District, and Kang County), and did not clarify whether it represents a larger-scale spatial variation. It is suggested to limit it to "tea gardens in Longnan City" or explain the geographical limitations in the discussion.
2. In materials and methods, the results show that Figure 1 presents the microbial composition and Alpha diversity. However, the text merely describes the data (such as "the bacterial richness in Kang County is higher"), without analyzing the trend of the chart.
3. The results show that Figure 1 presents the microbial composition and Alpha diversity. However, the text merely describes the data (such as "the bacterial richness in Kang County is higher"), without analyzing the trend of the chart.
4. The discussion attributed it to "accelerating organic matter mineralization", but did not explain why the mineralization dominated by fungi would reduce carbon fixation/nutrient storage (further explanation is needed in combination with the substrate C/N ratio, etc.).
5. It was mentioned that "bacteria are deterministic processes while fungi are random processes", but no specific environmental factors (such as how soil pH drives bacterial heterogeneous selection) were associated.
6. The results support a positive correlation (high diversity in Kang County → strong multifunctionality), but do not discuss the reasons for the differences (such as ecosystem types, selection of functional indicators) with contradictory conclusions (such as the negative correlation in Wang et al. 2021).

Author Response

Dear Editor:

Thanks for your letter and for reviewer's comments concern our manuscript entitled “Spatial Variation of Soil Microbial Contributions to Ecosystem Multifunctionality in Tea Gardens of Longnan City, China” (Manuscript ID: 3775500). Those comments are valuable and helpful for revising and improving our paper. We have studied all comments carefully and have made conscientious correction. Revised portion are marked in blue in the paper. The main corrections in the paper and the responds to the reviewer comments are as flowing.

Reviewer 1

1.The title emphasizes "spatial variation", but the study only covered three counties in Longnan City (Wen County, Wudu District, and Kang County), and did not clarify whether it represents a larger-scale spatial variation. It is suggested to limit it to "tea gardens in Longnan City" or explain the geographical limitations in the discussion.

Response: Thank you for your valuable comment. We agree that the original title may imply a broader spatial scope than our actual study area. To address this issue, we have revised the title to specify the study region more accurately as “tea gardens in Longnan City.” The revised title now appears on Lines 2–4 of the manuscript.

2.In materials and methods, the results show that Figure 1 presents the microbial composition and Alpha diversity. However, the text merely describes the data (such as "the bacterial richness in Kang County is higher"), without analyzing the trend of the chart.

Response: Thank you for pointing this out. We agree that the previous description of Figure 1 was overly descriptive and lacked sufficient analysis. In the revised manuscript, we have added more detailed interpretation of the trends shown in Figure 1, including the spatial differences in microbial composition and potential ecological implications. These additions can be found in the Results section (Lines 246-269). We believe this enhancement provides a more meaningful understanding of the data beyond simple description.

3.The results show that Figure 1 presents the microbial composition and Alpha diversity. However, the text merely describes the data (such as "the bacterial richness in Kang County is higher"), without analyzing the trend of the chart.

Response: Thank you for your valuable comment. We agree that the original text was largely descriptive and lacked in-depth analysis of the observed patterns. In the revised manuscript, we have added a detailed interpretation of the trends presented in Figure 1. Specifically, we observed a potential trade-off between the total relative abundance of dominant microbial phyla and the overall microbial diversity. For example, Wudu County exhibited the highest relative abundance of dominant bacterial phyla (Actinobacteriota, Acidobacteriota, and Proteobacteria), yet it did not have the highest bacterial diversity—this was found in Kang County, where the relative abundance of these phyla was the lowest. A similar pattern was observed in fungal communities: Wen County showed the highest dominance of major fungal phyla but the lowest fungal richness and diversity. These findings suggest that the over-dominance of certain microbial groups may reduce overall community diversity, possibly due to competitive suppression. The revised content can be found in the Results section (Lines 246–269).

4.The discussion attributed it to "accelerating organic matter mineralization", but did not explain why the mineralization dominated by fungi would reduce carbon fixation/nutrient storage (further explanation is needed in combination with the substrate C/N ratio, etc.).

Response: Thank you for this insightful comment. We agree that the initial explanation of fungal-driven mineralization was insufficient. In the revised Discussion section (Lines 518–536), we have provided a more detailed mechanistic explanation. Specifically, we discuss how fungal-dominated decomposition tends to accelerate organic matter mineralization. Fungi often allocate more carbon substrates toward respiration to fuel nitrogen-mineralizing enzyme systems, which can result in 60–85% of organic carbon being released as CO₂. This leads to rapid carbon loss and reduced carbon sequestration. Furthermore, fungal decomposition is often associated with high C/N substrates and intense nutrient competition, both of which can limit the storage of available soil nutrients. These mechanisms are consistent with our observations in Wen County, where fungal dominance was highest (92.86%) and bacterial diversity was lowest. This indicates that fungal-dominated mineralization may enhance short-term nutrient availability, but at the cost of long-term carbon and nitrogen retention. This revision also incorporates discussion of the role of substrate C/N ratio, as suggested.

5.It was mentioned that "bacteria are deterministic processes while fungi are random processes", but no specific environmental factors (such as how soil pH drives bacterial heterogeneous selection) were associated.

Response: Thank you for your insightful comment on the mechanisms of microbial community assembly. We fully agree with your point regarding the need to associate the observed deterministic processes in bacterial communities and stochastic processes in fungal communities with specific environmental factors, such as the role of soil pH in driving bacterial heterogeneous selection. In response, we have generated a new correlation heatmap illustrating the relationships between microbial assembly processes and environmental variables. Furthermore, we have expanded the Discussion section to analyze how key factors (e.g., soil pH and nutrient availability) influence deterministic or stochastic assembly mechanisms. These revisions can be found in the revised manuscript at Figure3 and Lines 293–296, 467–472.

6.The results support a positive correlation (high diversity in Kang County → strong multifunctionality), but do not discuss the reasons for the differences (such as ecosystem types, selection of functional indicators) with contradictory conclusions (such as the negative correlation in Wang et al. 2021).

Response: Thank you for your valuable comment. We have added a discussion in the revised manuscript (Lines 474–490) to explain the observed positive relationship between microbial diversity and ecosystem multifunctionality (EMF) in Kang County.

This result may be due to enhanced microbial interactions and enzyme activities associated with high microbial diversity, which improve resource use efficiency and soil resilience. Additionally, our study was conducted in intensively managed tea gardens with relatively uniform vegetation and focused on nutrient-related functions, while Wang et al. (2021) investigated broader ecosystems with different environmental gradients and functional indicators. These differences may explain the contrasting results. We have now included this explanation in the Discussion section.

Reviewer 2 Report

Comments and Suggestions for Authors

The authors of the manuscript ID microbiolres-3775500 presented the results of a study in which they verified two research hypotheses:

(1) soil C sequestration, nutrient storage and nutrient delivery functions show synergistic relationships with ecosystem multifunctionality,

(2) soil nutrients shaped by tea gardens in different regions indirectly affect microbial diversity, which in turn affects ecosystem functions.

To verify the research hypotheses, they determined the predominant types of bacteria and fungi in soils taken from 3 localities in Wenxian, Wudu, and Kangxian,

Soil multifunctionality (SMF) was assessed by dividing soil functions into four subsets: carbon sequestration (organic carbon content), nutrient storage (nitrogen, phosphorus and potassium content), nutrient supply (assimilable phosphorus and potassium content) and nutrient cycling (activity of five soil enzymes). A total of one bulk sample from each locality was analyzed in detail.

The title of the manuscript seems very interesting, but the scope of the research performed does not entitle it to such a formulation.

First of all, there was a lack of information about the soil types in each locality, their parent formations granulometric composition and their use.

Original results of determinations of soil physicochemical properties and soil enzyme activity were not included in the supplement.

No information was provided on which publicly available database the sequences of microbiological determinations are deposited in.

In addition:
The abstract is too long. It should contain about 200 words. Currently, it consists of 325 words.

Subsection 3.2 has the title Characterization of soil heavy metal content, but the chapter did not discuss heavy metal content. The authors did not label the heavy metal content at all.

Figure 3 - the letters defining homogeneous groups prove the high error of the study for VL,VA, CS, EMF and C/P. Such an error disqualifies the interpretation of results and inference.

References are not collated as required by the journal.

In conclusion, I state that in order for the manuscript to be published, it needs to be thoroughly revised taking into account the full characterization of the soil, since soil functions depend on its structure, physical properties, chemical properties and biological properties, among others.

Author Response

Dear Editor:

Thanks for your letter and for reviewer's comments concern our manuscript entitled “Spatial Variation of Soil Microbial Contributions to Ecosystem Multifunctionality in Tea Gardens of Longnan City, China” (Manuscript ID: 3775500). Those comments are valuable and helpful for revising and improving our paper. We have studied all comments carefully and have made conscientious correction. Revised portion are marked in blue in the paper. The main corrections in the paper and the responds to the reviewer comments are as flowing.

Reviewer 2

The authors of the manuscript ID microbiolres-3775500 presented the results of a study in which they verified two research hypotheses: soil C sequestration, nutrient storage and nutrient delivery functions show synergistic relationships with ecosystem multifunctionality, soil nutrients shaped by tea gardens in different regions indirectly affect microbial diversity, which in turn affects ecosystem functions.To verify the research hypotheses, they determined the predominant types of bacteria and fungi in soils taken from 3 localities in Wenxian, Wudu, and Kangxian,Soil multifunctionality (SMF) was assessed by dividing soil functions into four subsets: carbon sequestration (organic carbon content), nutrient storage (nitrogen, phosphorus and potassium content), nutrient supply (assimilable phosphorus and potassium content) and nutrient cycling (activity of five soil enzymes). A total of one bulk sample from each locality was analyzed in detail.The title of the manuscript seems very interesting, but the scope of the research performed does not entitle it to such a formulation.

1.First of all, there was a lack of information about the soil types in each locality, their parent formations granulometric composition and their use.

Response: Thank you for your helpful suggestion. In the revised manuscript (Lines 124-128, 133-135, 142-144), we have added detailed descriptions of the soil types, parent materials, and textural characteristics for each study site. Specifically, we noted that: Bikou (Wenxian County) soils are mainly red clay and yellow-brown soils, classified as transitional yellow-brown soils, with strong aluminization and clay accumulation. The parent material is weathered rock, and clay particles are translocated downward, resulting in heavy clay texture. Yangba (Kangxian County) soils consist mainly of black loam, yellow-black soils, and sandstone soils, classified as acidic red-yellow soils with higher sand content. Yuhe (Wudu District) soils are acidic red-yellow soils derived from alluvial valley deposits (clay loam) and limestone residuals (sandy clay). These additions provide a clearer context for interpreting soil microbial and functional variations across regions.

2.Original results of determinations of soil physicochemical properties and soil enzyme activity were not included in the supplement.

Response: Thank you for your valuable comment. We have added the results of soil physicochemical properties and soil enzyme activities in the revised manuscript. Please refer to Lines 375–383 and Figure 6 for the updated content.

3.No information was provided on which publicly available database the sequences of microbiological determinations are deposited in.

Response: Thank you for your valuable comment. The raw sequencing data of the microbial communities have been deposited in the NCBI Sequence Read Archive (SRA) under the project accession number PRJNA1272242. This information has been added to the revised manuscript (Lines 603–604).

4.The abstract is too long. It should contain about 200 words. Currently, it consists of 325 words.

Response: Thank you for your suggestion. In accordance with the journal's word limit requirements, we have revised and shortened the abstract to approximately 200 words while retaining the key objectives, methods, results, and conclusions of the study. Please refer to the revised abstract in the manuscript.

5.Subsection 3.2 has the title Characterization of soil heavy metal content, but the chapter did not discuss heavy metal content. The authors did not label the heavy metal content at all.

Response: Thank you for pointing this out. We acknowledge that the original title of Subsection 3.2 was misleading, as the section did not include any analysis or discussion of soil heavy metal content. To avoid confusion, we have revised the subsection title accordingly. The updated title now reflects the actual content of the section (Line 322).

6.Figure 3 - the letters defining homogeneous groups prove the high error of the study for VL,VA, CS, EMF and C/P. Such an error disqualifies the interpretation of results and inference.

Response: Thank you for your valuable comment. We acknowledge the concern regarding the high variability observed in some indicators, particularly VL and VA. To address this issue and improve the reliability of the statistical analysis, we applied a logarithmic transformation to VL and VA during data processing. This transformation effectively reduced the standard errors and improved data distribution. The corresponding revisions have been made in the manuscript (Lines 329–332). We also rechecked the statistical methods used for CS, EMF, and C/P, and confirmed that appropriate tests (ANOVA with Tukey’s HSD) were applied, with all necessary assumptions examined.

References are not collated as required by the journal.

In conclusion, I state that in order for the manuscript to be published, it needs to be thoroughly revised taking into account the full characterization of the soil, since soil functions depend on its structure, physical properties, chemical properties and biological properties, among others.

Response: Thank you for your thorough review and constructive suggestions. We have carefully revised the manuscript based on your recommendations. All references have been reformatted and collated according to the journal’s guidelines. In addition, we have added detailed descriptions of the soil's physical, chemical, and biological properties in the revised manuscript to ensure a more comprehensive characterization. We sincerely appreciate your comments, which helped us significantly improve the clarity, completeness, and scientific rigor of the manuscript.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors have revised all the comments.

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript entitled 'Mechanisms by Which Soil Microbial Communities Regulate Ecosystem Multifunctionality in Tea Gardens of Longnan City, China' (microbiolres-3775500), which was previously submitted, has been revised in accordance with the comments provided. Thank you to the authors for all the improvements they have made to the article. In its current version, the article has been significantly enhanced and, in my opinion, now meets the criteria for publication in Microbiology Research. Its content is consistent with the journal's research profile.