the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
Soil carbon accrual and biopore formation across a plant diversity gradient
Abstract. Plant diversity promotes soil organic carbon (SOC) gains through intricate changes in root-soil interactions and their subsequent influence on soil physical and biological processes. We assessed SOC and pore characteristics of soils under a range of switchgrass-based plant systems, representing a gradient of plant diversity with species richness ranging from 1 to 30 species 12 years after their establishment. We focused on soil biopores as indicators of root activity legacy, measured using X-ray computed micro-tomography scanning, and explored biopore relationships with SOC accumulation.
Plant functional richness explained 29 % of bioporosity and 36 % of SOC variation, while bioporosity itself explained 36 % of the variation in SOC. The most diverse plant system (30 species) had the highest SOC, while long-term bare soil fallow and monoculture switchgrass had the lowest. Of particular note was a two-species mixture of switchgrass (Panicum virgatum L.) and ryegrass (Elymus canadensis), which exhibited the highest bioporosity and achieved SOC levels comparable to those of the systems with 6 and 10 plant species, and were inferior only to the system with 30 species. We conclude that plant diversity may enhance SOC through biopore-mediated mechanisms and suggest a potential for identifying specific plant combinations that may be particularly efficient for fostering biopore formation and subsequently SOC sequestration.
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Status: final response (author comments only)
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RC1: 'Comment on egusphere-2025-2584', Shang Wang, 05 Aug 2025
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AC1: 'Reply on RC1', Kyungmin Kim, 18 Aug 2025
Dear Reviewer
We sincerely appreciate your valuable comments and suggetions on this manusript.
Please see the attached "Response to Reviewer" and our revised manuscript for SOIL.
If you require any further information or additional revisions, we will provide them promptly.
Sincerely,
Kyungmin Kim, on the behalf of all authors
Seoul National University, South Korea
km_kim@snu.ac.kr
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RC2: 'Reply on AC1', Shang Wang, 18 Aug 2025
Dear Alyssa, dear all,
Thank you for revising the manuscript. Your revisions have effectively improved it, and I do not have any further questions. I am happy to recommend this manuscript for publication.
Kind regards,
Citation: https://doi.org/10.5194/egusphere-2025-2584-RC2
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RC2: 'Reply on AC1', Shang Wang, 18 Aug 2025
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AC1: 'Reply on RC1', Kyungmin Kim, 18 Aug 2025
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RC3: 'Comment on egusphere-2025-2584', Anonymous Referee #2, 15 Sep 2025
The manuscript egusphere-2025-2584 titled “Soil carbon accrual and biopore formation across a plant diversity gradient” utilized a plant diversity gradient experiment (1-30 species) and employed X-ray computed micro-tomography scanning to quantitatively analyze soil pore structures (particularly biopores) under different plant systems, and examined their relationship with soil organic carbon (SOC) accumulation. The study suggests that plant diversity can enhance SOC content through biopore-mediated mechanisms. However, the conclusions lack generalizability due to the limited SOC data from only one year of measurement. More importantly, the interrelationships among plant diversity, biopores, and SOC remain unclear. The correlations among the three do not represent a causal relationship, and the driving process of diversity is also not deep enough. Although the authors repeatedly emphasize the importance of microorganisms in biopores, direct microbial measurements are lacking. It would be valuable to incorporate such data to substantiate the claims. Additionally, introduction lacks sufficient background and theoretical foundation, and results are not adequately contextualized within existing literature or mechanistic frameworks. It is recommended to strengthen the theoretical discussion and improve the scientific interpretation of the findings. Please see the specific comments below.
Introduction
The Introduction fails to clearly articulate the research gap or motivate the study’s focus on plant diversity effects on pore structure and SOC. The logic between paragraphs is relatively weak, and the importance of biopores in mediating plant-soil- carbon interactions is not sufficiently established.
Lines 34-37 and 41: The repeated emphasis on microbial processes is misleading since no microbial data were collected. This section should be refocused on mechanisms more directly related to the study
Lines 44-45: The claim that pore structure affects root-soil interactions and plant-derived carbon requires supporting references or clearer conceptual linkage.
Lines 46-50: Lack of references.
Lines 61-64: How are biopores and SOC related?
Lines 74-77: The research objective is the relationship between biopores and C. However, the significance of this part of the research is not elaborated in the Introduction. Instead, it has been constantly describing the spatial distribution and morphological characteristics of the root system. Here, no data on the morphology and distribution of the root system were measured in this study.
Materials and Methods
Lines 80-90: Key agronomic details are missing. i.e., sowing dates and frequency for switchgrass and Canadian rye.
Line 95: The 7-12 cm sampling depth may not capture the full root profile, especially for deep-rooted species like switchgrass. Justification for this depth is needed.
Line 104: The term “soil total C” is used in Methods, but “SOC” is used elsewhere. Ensure consistency
Line 106: Is the water content of all treatments the same? Or should the original moisture content of each treatment be maintained? Has the evaporation of soil moisture been taken into account? Is it necessary to weigh and replenish water daily?
Lines 138-141: The functional richness calculation method claims support from Mangan et al. (2011) and Spiesman et al. (2018), but these studies do not explicitly use the same method. Please provide a clearer rationale or additional citations.
Results
Line 166: The lack of correlation between aboveground biomass and SOC may be due to a temporal mismatch. SOC was measured in 2019, while biomass data came from 2018-2019. Aligning the data temporally or discussing this discrepancy is important.
Lines 185-190: Figure 2 lacks the statistical difference test between treatments. I suggest dividing the pore size into different size intervals and then comparing the differences between treatments.
Lines 208-216: The author needs to summarize the important correlation results instead of studying the correlation for each indicator.
Discussion
The Discussion should avoid referencing each figure individually. The authors are expected to focus on interpreting key findings and substantiating their arguments with relevant literature.
Lines 233-240: The variation in C mineralization across systems is not discussed.
Line 237: The non-significant relationships between aboveground biomass and SOC may be related to C mineralization rates.
Lines 237-240: Lack of explanation.
Lines 244-248: The results of the CE8 system were not explained. There is no logical connection between the two sentences.
Lines 273-276: Using legumes as an example is not convincing, as switchgrass and Canadian rye do not belong to the leguminous family.
Lines 276-278: Lack of literature.
Is a Conclusion section missing from the manuscript?
Citation: https://doi.org/10.5194/egusphere-2025-2584-RC3
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Review of the manuscript egusphere-2025-2584
This is a valuable study investigating the effects of plant diversity on soil carbon accrual and biopore formation. The topic is novel and highly relevant to the journal’s scope. The title is well-formulated, the experimental design is robust, and the dataset is comprehensive. However, there are a few minor issues, such as formatting errors, that should be addressed. Additionally, some methodological details in the Materials and Methods section need to be clarified or expanded. Overall, I believe the manuscript is suitable for publication after minor revisions.
Please review and feedback the general and specific comments as follows:
Specific remarks
Line 30: Please delete the extra space between including and grasslands.
Line 57: Please remove the extra space between Root-originated and biopores.
Lines 84–88: Could you clarify the rationale behind the assignment of treatment (CE) numbers? For instance, why is the bare system labeled as CE1, while the monoculture is labeled CE7?
Line 125: Please provide a citation to a published study that supports the biopore identification method used.
Line 129: Could you clarify what is meant by “distance average distance to biopore”? The phrasing is unclear.
Line 161: It would be helpful to include a brief summary sentence at the end of each paragraph or subsection to reinforce key points.
Line 181: Please format the R² correctly by placing the 2 as a superscript.
Lines 255–256: This sentence is unclear—“accumulated for 12 years” appears abrupt and could be better integrated.
Lines 255–262: These sentences are more appropriate for the Introduction section and would fit better there.
L284 It would be reasonable to include a clear conclusion section at the end of the main text.