Effects of Organic Fertilizer and Biochar on Carbon Release and Microbial Communities in Saline–Alkaline Soil

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript is well written and sound in all scientific aspects. However few minor quaries are presented here to be varified for improvement.

The cumulative CO₂ in Fig 1 has been shown to decrease in SUB and SUH in comparison to SU. In the text, however this has not been added clearance.

 

“Different uppercase letters (organic matter 206 added) indicate significant differences (P < 0.05, Duncan’s multiple range test)”. Please correct that letters are not uppercase.

 

Line 232: CH4 uptake in saline soils (P < 0.01 Table 3)….is this uptake or emission.

The relative abundance % of Ascomycota has increased in SHU and SUB treatments and this group has outnumbered the rest of the fungal phyla. What can be possible cause for this?

In discussion section, it has been shown that CO2 and CH4 emissions decreased with addition of microbial agent and biochar but results are not showing such information.

Good luck

Comments on the Quality of English Language

The language of paper is good except few minor deficeinceis on typing sentence structure

Author Response

1. The cumulative CO₂ in Fig 1 has been shown to decrease in SUB and SUH in comparison to SU. In the text, however this has not been added clearance.

Response 1: Thank you for your comments. I acknowledge the significance of the issue you have raised, which concerns the insufficient clarification in the main text regarding the reduced CO₂ accumulation under SUB and SUH conditions as compared to the SU condition, as depicted in Fig 1. In response, I have emphasized the pertinent text in yellow within the abstract, discussion, and conclusion sections of the revised manuscript to provide clarity on this issue. (Line 22-25, Line 379-380, Line 412-414, Line 525-532,)

2. Different uppercase letters (organic matter 206 added) indicate significant differences (P < 0.05, Duncan’s multiple range test)”. Please correct that letters are not uppercase.

Response 2: Thank you very much for your meticulous review and valuable comments on my submitted paper. Following a rigorous examination, we have implemented the necessary correction: In the notes of Tables 2 and 3, we have altered the notation from uppercase letters to lowercase letters and emphasized them in yellow for enhanced clarity. (Line 229, Line 262)

3. Line 232: CH₄ uptake in saline soils (P < 0.01 Table 3)….is this uptake or emission.

Response 3: Thanks for your good comment. Recognizing the importance of this feedback, I have revised the manuscript to enhance its precision and clarity, thereby facilitating readers' understanding of the experimental outcomes and their implications. To avoid ambiguity, I have revised the results section as follows:

The ANOVA revealed that various organic amendments significantly impacted methane absorption in saline-alkali soil (P < 0.01; Table 3). Specifically, except for the U treatment group, the addition of all other organic materials increased methane absorption in saline-alkali soil. (Line 254-257)

4. The relative abundance % of Ascomycota has increased in SUH and SUB treatments and this group has outnumbered the rest of the fungal phyla. What can be possible cause for this.

Response 4: We greatly appreciate your insightful suggestions. We have conducted detailed discussions on the increased relative abundance of the Ascomycota phylum in the SUH and SUB treatment groups.

In the SUH and SUB treatment groups, the relative abundance of Ascomycota significantly increased in the soil. This phenomenon may be attributed to the enhanced stress effects of alkaline and saline components in saline-alkali soils on microorganisms, exacerbated by the release of alkaline substances from organic materials under semi-enclosed conditions. The study by Wu et al. revealed that Ascomycota fungi are the dominant group in degraded saline-alkali meadow soils in northeastern China and exhibit strong tolerance to saline-alkali environments[1]. Furthermore, Spearman correlation analysis confirmed a significant positive correlation between the relative abundance of Ascomycota and soil pH as well as electrical conductivity (EC) values. (Line 435-443)

5. In discussion section, it has been shown that CO₂ and CH₄ emissions decreased with addition of microbial agent and biochar but results are not showing such information.

Response 5: We greatly appreciate your thorough review and insightful comments. Your observation that the Discussion section highlights the reduction of CO₂ and CH₄ emissions with the addition of microbial agents and biochar, whereas this information is not directly stated in the Results section,We have thoroughly reviewed and reflected on our work and provided the following clarification:

Our study results clearly demonstrate that the addition of microbial agents and biochar significantly reduced CO₂ emissions compared to the straw plus urea (SU) treatment. Additionally, the addition of microbial agents alone under SU treatment conditions further reduced CH₄ emissions, as evidenced in Figure 1 and Table 3. (Line 241-243, Line 252-254)

In the Discussion section, we conducted a detailed analysis of these key findings and explicitly pointed out that these results were obtained under the same SU treatment, specifically referenced in Section 4.2 of the Discussion, where the highlighted yellow text clearly indicates this. (Line 379-380, Line 412-414)

Once again, I sincerely thank you for your thoughtful guidance and unwavering support of my work!

References

1. Wu, X.; Yang, J.; Ruan, H.; Wang, S.; Yang, Y.; Naeem, I.; Wang, L.; Liu, L.; Wang, D. The Diversity and Co-Occurrence Network of Soil Bacterial and Fungal Communities and Their Implications for a New Indicator of Grassland Degradation. Ecological Indicators 2021, 129, 107989, doi:10.1016/j.ecolind.2021.107989.

 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

I have been assigned to review the manuscript [Agronomy] Manuscript ID: agronomy-3114442 “Effects of organic fertilizer and biochar on carbon release and microbial communities in saline–alkaline soil”

I believe this topic is of great relevance and interest, and the current manuscript depicts that;

•            The current study has been presented by authors about the climate change and aridification that have increased the risk of salinization and organic carbon loss in dryland soils.

•            Authors have shown that enrichment using biochar and organic fertilizers has the potential to reduce salt toxicity and soil carbon loss. However, the effects of biochar and organic fertilizers on CO2 and CH4 emissions from saline soils in dryland areas, and their microbial mechanisms remain unelucidated.

•            To clarify these issues, authors performed a 5-month incubation experiment on typical soda-type saline soil from the western part of the Songnen Plain using five treatments: control treatment (CK), 5% urea (U), straw + 5% urea (SU), straw + 5% urea + microbial agent (SUH), and straw + 5% 21 urea + biochar (SUB).

•            Compared with the SU treatment, the SUH and SUB treatments reduced cumulative CO2 emissions by 14.85% and 35.19%, respectively.

•            The addition of a microbiological agent to the SU treatment reduced the cumulative CH4 emissions by 19.55%, whereas the addition of biochar to the SU treatment increased the cumulative CH4 emissions by 4.12%. CH4 emissions from the SU treatment.

•            These additions also increased the relative abundances of Proteobacteria, Planctomycetes, and Ascomycota.

•            Overall, the addition of biochar and organic fertilizer promoted CO2 emissions and CH4 uptake. This was mainly attributed to an improved soil gas diffusion rate due to the addition of organic materials and enhanced microbial stress due to soil salinity and alkalinity from the release of alkaline substances under closed-culture conditions. Our findings have positive implications for enhancing carbon storage in saline soils in arid regions

However, I have the following concerns given below, to be rectified by the authors

INTRODUCTION SECTION

1.         Introduction is not sufficiently addressed. It should cover a bit more of the latest scientific reports. For example, Microorganisms regulate carbon sequestration, CH4 metabolism, and carbon degradation and play a key role in the global carbon cycle and climate change[16]. This report is given in the introduction, however, reviewers and readers would like to know more background knowledge about this information. Which microorganisms (bacteria, fungi), names species, their traits, the mechanisms, etc.,

2.         Line 72. “Zhang et al. and 72 Li et al. reported that the combination of straw and a microbial agent increased the stability of soil carbon production and decreased carbon release during straw decomposition [23,24]”

Authors should write the content and information properly, what are the names of microbial agents?

3.         Line 75. “some studies have reported that microbial agent application increased soil microbial abundance and stimulated soil CO2 and CH4 emissions while promoting straw decomposition [25–27].”

4.         Authors should write the content and information properly, what are the names of microbial agents?

METHODS SECTION

 

1.         Line 126, Method sections the “Microbial agents based on the W-18 model of microbial decomposition” This information is not sufficiently complete. There is no reference to this information.

2.         In the methods section authors have presented that soil samples from each treatment group were sent to Shanghai “Payseno Biotechnology Co., Ltd. (Shanghai, China) for DNA extraction, polymerase chain reaction amplification, and sequencing using the methods of Zhang et al. [24].” However, I did not find any sequencing data sets in supplementary files or submitted to the database. This is a major concern.

RESULTS SECTION

1.         Table 1 does not have complete statistical analysis information.

2.         Table 2 has formatting issues; significant letters should be superscript.

3.         Column one should have units written accurately.

4.         Figure 1 data does not show any statistical analysis.

5.         Figure 3. “Predicted abundance of carbon degrading and CH4 oxidizing functional enzymes in soil”. Abundance of what? Scale is not showing any unit?

6.         Figure 4. What kind of correlation analysis is shown in this figure? Authors must include the name of the analysis.

7.         Figure 5. “SEM” authors should write the full name, not only the abbreviation in the figure legend.

 

General Comments

1.         Introduction is not sufficiently addressed. It should cover a bit more of the latest scientific reports.

2.         Experimental design does not accurately reflect the main objective and aims of the study.

3.         The method section has been explained poorly that is hard to achieve the reproducibility of experiments for researchers around the globe.

4.         Novelty and scope of current work are considered, however, there are several formatting issues, grammatical issues, and referencing wrongly placed.

5.         Grammatically there are several mistakes to be rectified by the authors.

3.         Statistical analysis is compromised. Biological and technical replicates are not described properly.

 

Comments on the Quality of English Language

Moderate editing of English language required

Author Response

1. Introduction is not sufficiently addressed. It should cover a bit more of the latest scientific reports. For example, Microorganisms regulate carbon sequestration, CH₄ metabolism, and carbon degradation and play a key role in the global carbon cycle and climate change[16]. This report is given in the introduction, however, reviewers and readers would like to know more background knowledge about this information. Which microorganisms (bacteria, fungi), names species, their traits, the mechanisms, etc?

Response 1: Thank you very much for reviewing our submission and providing valuable comments and suggestions. We fully concur with your observation of insufficient coverage of certain pertinent issues in the introduction section. Guided by your insightful advice, we have thoroughly revised the Introduction section, incorporating more detailed content on the crucial roles of microorganisms in carbon sequestration, methane (CH₄) metabolism, and carbon degradation. In the revised version of our manuscript, we have color-coded the newly added content in green for clear and effortless identification. The following lists the changes that have been made to the manuscript:

Soil microorganisms play a pivotal role in regulating the turnover of soil organic matter pools and contribute significantly to climate feedback mechanisms through various ecological processes, including the sequestration and release of carbon in diverse forms [1,2]. The application of organic fertilizers such as biochar affects microbial diversity and community composition by altering the supply of carbon and nitrogen substrates[3], decreasing salt stress[4], and regulating the soil microenvironment[5,6]. Microbial community composition, diversity, and biomass are important indicators for predicting soil carbon emissions [7]. Zhao et al. found a positive correlation between the relative abundance of bacterial phyla such as Proteobacteria and Firmicutes, as well as the fungal phylum Ascomycota, in long-term fertilized soils and straw-derived carbon dioxide emissions [8]. The microbial species composition regulates the abundance of functional genes, subsequently affecting enzymatic activities associated with carbon degradation and methane oxidation [9,10]. Thus, the enzyme activities of soil microorganisms are important limiting factors that mediate carbon degradation and CH₄ metabolism[11,12].The research conducted by He et al. reveals that incorporating both straw and biochar leads to an increase in the relative abundance of the PMOA gene, concurrently resulting in a significant reduction in CH₄ emissions [13]. Yu et al. demonstrated that incorporating straw and biochar into saline-alkali wetlands stimulates the opportunistic bacterial subgroups (e.g., Bacillaceae and Cellvibrionaceae), enhancing bacterial carbon metabolism and concurrently augmenting both carbon storage and mineralization processes [14].( Line 87-107)

2. Line 72. “Zhang et al. and 72 Li et al. reported that the combination of straw and a microbial agent increased the stability of soil carbon production and decreased carbon release during straw decomposition [23,24]. Authors should write the content and information properly, what are the names of microbial agents?

Response 2: Thanks for your good comment. We have made correction according to your comment. The following lists the changes that have been made to the manuscript:

Li et al. specifically reported a significant reduction in carbon release during organic matter decomposition when using a bacterial mixture (BM) consisting of Citrobacter freundii, Arthrobacter woluwensis, and Bacillus licheniformis, in combination with straw [15].Additionally, Zhang et al. noted that the application of commercial microbial agents comprising Bacillus subtilis, Bacillus megaterium, and gelatinous Bacillus mixtures can enhance soil carbon stability and effectively mitigate CO₂ emissions during straw incorporation into saline-alkali soil [5]. (Line 71-77)

3. Line 75. “some studies have reported that microbial agent application increased soil microbial abundance and stimulated soil CO₂ and CH₄ emissions while promoting straw decomposition [25–27].”  Authors should write the content and information properly, what are the names of microbial agents?.

Response 3: Thanks for your good comment. We have made correction according to your comment. The following lists the changes that have been made to the manuscript:

However, various research studies have demonstrated that the introduction of commercial microbial inoculants, comprising a blend of bacteria and fungi, notably Bacillus subtilis and lignin-degrading, thermophilic, and heat-tolerant bacteria, into soil leads to a substantial increase in soil microbial abundance. These microorganisms not only facilitate straw decomposition but also elicit emissions of CO₂ and CH₄ from the soil, as reported in [16–18]. (Line 77-82)

4. Line 126, Method sections the “Microbial agents based on the W-18 model of microbial decomposition” This information is not sufficiently complete. There is no reference to this information.

Response 4: Thanks for your good comment. We have made correction according to your comment. The following lists the changes that have been made to the manuscript:

The microbial agent utilized is W-18 straw decomposer, made of Bacillus subtilis, Bacillus megaterium, and Bacillus jelly-like, with an effective viable count ≥ 2.0 ´ 10⁸ mL^-1, created by Heilongjiang Huxufeng Ecological Technology Company[5]. (Line 150-152)

5. In the methods section authors have presented that soil samples from each treatment group were sent to Shanghai “Payseno Biotechnology Co., Ltd. (Shanghai, China) for DNA extraction, polymerase chain reaction amplification, and sequencing using the methods of Zhang et al. [24].” However, I did not find any sequencing data sets in supplementary files or submitted to the database. This is a major concern.

Response 5: Thank you very much for your detailed and insightful review of my submitted paper, as well as for pointing out the shortcomings and providing valuable suggestions for improvement. In particular, I take seriously the issue you raised regarding the mention in the Methods section that soil samples were sent to "Persono Bio-Tech Co., Ltd." in Shanghai for DNA extraction, PCR amplification, and sequencing, but the corresponding sequencing datasets are not found in the supplementary materials or databases. I would like to provide a detailed explanation and response to this matter.

Firstly, I fully recognize the seriousness of this issue, which is the failure to comply with the standard requirements of academic journals to present key experimental data comprehensively and clearly. I sincerely apologize for this and pledge to pay greater attention to the integrity and transparency of data in my future work to ensure the verifiability and reproducibility of research results.

Regarding the absence of sequencing datasets in the supplementary materials or databases, the primary reason is that these raw sequencing data involve unpublished research findings, and some data are subject to confidentiality requirements. Therefore, I did not include the complete raw sequencing data in the current paper submission. However, I fully understand the importance of data sharing in promoting scientific research communication and advancing disciplinary development. Consequently, appropriate measures have been taken. Readers can contact the corresponding author of this paper to request access to relevant data, and I will make every effort to cooperate to ensure the reasonable sharing and use of the data.

6. Table 1 does not have complete statistical analysis information.

Response 6: Thank you very much for your meticulous review of my submitted paper and for providing valuable feedback. I have thoroughly reviewed your comments, with particular attention to the observation that Table 1 lacks comprehensive statistical analysis details. In response to this specific suggestion, we have revised Table 1 by removing redundant data pertaining to biochar properties while ensuring consistency in data presentation and prioritizing key information. This modification intends to improve the clarity of Table 1 and highlight the direct relevance of the presented data. (Line 158)

7. Table 2 has formatting issues; significant letters should be superscript.

Response 7: Thank you very much for your thorough review and valuable comments on my submitted paper. I have carefully read your feedback, particularly the suggestion that there is a formatting issue with Table 2, specifically that "significant letters should be superscript." I wholeheartedly agree with your observation, and I deeply recognize the importance of this revision in enhancing the academic rigor of my paper. Hereby, I confirm that I have made the necessary changes by superscripting the relevant significant letters in Table 2, as per your request. (Line 226-228)

8. Column one should have units written accurately

Response 8: Thanks for your good comment. We have made correction according to your comment. (Line 226-228)

9. Figure 1 data does not show any statistical analysis

Response 9: Thank you very much for your valuable suggestion regarding the absence of any statistical analysis results in Figure 1. We have carefully considered your comment and would like to provide the following response:
The primary aim of Figure 1 is to visually depict the overall trend and the variations among distinct treatment groups in carbon emissions throughout the cultivation phase. The detailed statistical analysis results pertaining to these carbon emission data are comprehensively presented in Table 3. This arrangement ensures that Figure 1 remains centered on showcasing data trends, whereas Table 3 offers a more comprehensive and detailed quantitative analysis. We gratefully acknowledge your insightful feedback, which has significantly contributed to enhancing the clarity and presentation of our paper.

10. Figure 3. “Predicted abundance of carbon degrading and CH₄ oxidizing functional enzymes in soil”. Abundance of what? Scale is not showing any unit?

Response 10: Thank you very much for your thorough review and valuable comments on my submitted paper. I have carefully read your feedback and provided the following responses: The heatmap analysis of predicted abundance of carbon degradation and methane oxidation functional enzymes was achieved by normalizing the abundance of metabolic pathways in non-hierarchically clustered samples based on EC-normalized data. Thank you once again for your suggestions. I have supplemented the title of Figure 3 in the newly submitted manuscript and highlighted the revised text in green. The specific changes are as follows:

Normalized Heatmap Analysis of Predicted Abundances of Carbon Degradation and CH₄ Oxidation Functional Enzymes Derived from Soil Bacterial (a) and Fungal (b) Sequencing Data Following Cultivation Experiments (Line 293-295)

11. Figure 4. What kind of correlation analysis is shown in this figure? Authors must include the name of the analysis.

Response 11: Thank you very much for your meticulous review and valuable suggestions on my submitted paper. In response to your inquiry about the type of correlation analysis shown in Figure 4 and the necessity to specify the analysis method, we have thoroughly reviewed the matter and offer the following clarification:In Figure 4, we utilized Spearman's correlation analysis. We recognize the paramount importance of clearly labeling analytical methods in academic writing, as it facilitates accurate interpretation of research findings by readers. Consequently, we have incorporated a clarifying note beneath Figure 4 in the revised manuscript, precisely identifying the employed analysis method. We once again express our sincere appreciation for your invaluable advice, which has significantly enhanced the quality of our paper. The specific changes are as follows:

Heatmap of Spearman's Correlation Analysis between CO₂ emissions (a) and CH₄ emissions (b) with Microbial Diversity Indices and Soil Physicochemical Properties at the Phylum Level (Line 325-326)

12. Figure 5. “SEM” authors should write the full name, not only the abbreviation in the figure legend.

Response 12: Thank you very much for your meticulous review and valuable suggestions on my submitted paper. I have carefully read your feedback, and in particular, I deeply appreciate the importance of your suggestion to replace the abbreviation "SEM" in Figure 5 with its full name. This change, I believe, will significantly enhance the clarity and readability of the paper. Hereby, I solemnly commit to making the modification strictly according to your requirements:

In the caption of Figure 5, we substituted "SEM" with its full designation, "Structural Equation Modeling," to facilitate comprehension without reliance on external references. Furthermore, to explicitly highlight this alteration, we emphasized the amended portion using green text. (Lines 345)

Once again, I sincerely thank you for your thoughtful guidance and unwavering support of my work!

References:

1. Cavicchioli, R.; Ripple, W.J.; Timmis, K.N.; Azam, F.; Bakken, L.R.; Baylis, M.; Behrenfeld, M.J.; Boetius, A.; Boyd, P.W.; Classen, A.T.; et al. Scientists’ Warning to Humanity: Microorganisms and Climate Change. Nat Rev Microbiol 2019, 17, 569–586, doi:10.1038/s41579-019-0222-5.
2. Crowther, T.W.; van den Hoogen, J.; Wan, J.; Mayes, M.A.; Keiser, A.D.; Mo, L.; Averill, C.; Maynard, D.S. The Global Soil Community and Its Influence on Biogeochemistry. Science 2019, 365, eaav0550, doi:10.1126/science.aav0550.
3. Yao, R.; Li, H.; Yang, J.; Zhu, W.; Yin, C.; Wang, X.; Xie, W.; Zhang, X. Combined Application of Biochar and N Fertilizer Shifted Nitrification Rate and amoA Gene Abundance of Ammonia-Oxidizing Microorganisms in Salt-Affected Anthropogenic-Alluvial Soil. Applied Soil Ecology 2022, 171, doi:10/gpc8dv.
4. Cui, Q.; Xia, J.; Yang, H.; Liu, J.; Shao, P. Biochar and Effective Microorganisms Promote Sesbania Cannabina Growth and Soil Quality in the Coastal Saline-Alkali Soil of the Yellow River Delta, China. Science of the Total Environment 2021, 756, 143801, doi:10/gjjx2n.
5. Zhang, P.; Jiang, Z.; Wu, X.; Qian Lu; Yue Lin; Yanyu Zhang; Xin Zhang; Yi Liu; Siyu Wang; Shuying Zang Effects of Biochar and Organic Additives on CO2 Emissions and the Microbial Community at Two Water Saturations in Saline–Alkaline Soil. Agronomy 2023, 13, 1745, doi:10.3390/agronomy13071745.
6. Jiang, Z.; Zhang, P.; Wu, Y.; Wu, X.; Ni, H.; Lu, Q.; Zang, S. Long-Term Surface Composts Application Enhances Saline-Alkali Soil Carbon Sequestration and Increases Bacterial Community Stability and Complexity. Environ. Res. 2024, 240, 117425, doi:10.1016/j.envres.2023.117425.
7. Liu, Y.-R.; Delgado-Baquerizo, M.; Wang, J.-T.; Hu, H.-W.; Yang, Z.; He, J.-Z. New Insights into the Role of Microbial Community Composition in Driving Soil Respiration Rates. Soil Biology and Biochemistry 2018, 118, 35–41, doi:10.1016/j.soilbio.2017.12.003.
8. Zhao, S.; Qiu, S.; Xu, X.; Ciampitti, I.A.; Zhang, S.; He, P. Change in Straw Decomposition Rate and Soil Microbial Community Composition after Straw Addition in Different Long-Term Fertilization Soils. Appl Soil Ecol 2019, 138, 123–133, doi:10.1016/j.apsoil.2019.02.018.
9. Trivedi, P.; Delgado-Baquerizo, M.; Trivedi, C.; Hu, H.; Anderson, I.C.; Jeffries, T.C.; Zhou, J.; Singh, B.K. Microbial Regulation of the Soil Carbon Cycle: Evidence from Gene–Enzyme Relationships. The ISME journal 2016, 10, 2593–2604, doi:10.1038/ismej.2016.65.
10. Kalyuzhnaya, M.G.; Gomez, O.A.; Murrell, J.C. The Methane-Oxidizing Bacteria (Methanotrophs). Taxonomy, genomics and ecophysiology of hydrocarbon-degrading microbes 2019, 245–278, doi:10.1007/978-3-030-14796-9_10.
11. Burns, R.G.; DeForest, J.L.; Marxsen, J.; Sinsabaugh, R.L.; Stromberger, M.E.; Wallenstein, M.D.; Weintraub, M.N.; Zoppini, A. Soil Enzymes in a Changing Environment: Current Knowledge and Future Directions. Soil Biology and Biochemistry 2013, 58, 216–234, doi:10.1016/j.soilbio.2012.11.009.
12. Liu, Y.; Wang, S.; Li, S.; Deng, Y. Advances in Molecular Ecology on Microbial Functional Genes of Carbo n c Ycle. Microbiology China 2017, 44, 1676–1689, doi:10.13344/j.microbiol.china.160941.
13. He, T.; Yun, F.; Liu, T.; Jin, J.; Yang, Y.; Fu, Y.; Wang, J. Differentiated Mechanisms of Biochar- and Straw-Induced Greenhouse Gas Emissions in Tobacco Fields. Applied Soil Ecology 2021, 166, doi:10.1016/j.apsoil.2021.103996.
14. Yu, L.; Bai, J.; Huang, L.; Zhang, G.; Wang, W.; Wang, X.; Yu, Z. Carbon-Rich Substrates Altered Microbial Communities with Indication of Carbon Metabolism Functional Shifting in a Degraded Salt Marsh of the Yellow River Delta, China. Journal of Cleaner Production 2022, 331, doi:10/gpx277.
15. Li, M.; Tang, C.; Chen, X.; Huang, S.; Zhao, W.; Cai, D.; Wu, Z.; Wu, L. High Performance Bacteria Anchored by Nanoclay to Boost Straw Degradation. Materials (Basel) 2019, 12, doi:10.3390/ma12071148.
16. Kalkhajeh, Y.K.; He, Z.; Yang, X.; Lu, Y.; Zhou, J.; Gao, H.; Ma, C. Co-Application of Nitrogen and Straw-Decomposing Microbial Inoculant Enhanced Wheat Straw Decomposition and Rice Yield in a Paddy Soil. Journal of Agriculture and Food Research 2021, 4, 100134, doi:10.1016/j.jafr.2021.100134.
17. Liu, G.; Yu, H.; Ma, J.; Xu, H.; Wu, Q.; Yang, J.; Zhuang, Y. Effects of Straw Incorporation along with Microbial Inoculant on Methane and Nitrous Oxide Emissions from Rice Fields. Science of The Total Environment 2015, 518–519, 209–216, doi:10.1016/j.scitotenv.2015.02.028.
18. Zhang, S.; Hussain, H.A.; Wang, L.; Hussain, S.; Li, B.; Zhou, H.; Luo, H.; Zhang, X.; Ma, Z.; Long, L.; et al. Responses of Soil Respiration and Organic Carbon to Straw Mulching and Ridge Tillage in Maize Field of a Triple Cropping System in the Hilly Region of Southwest China. Sustainability 2019, 11, 3068, doi:10.3390/su11113068.

 

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The paper presents a valuable study on the effects of organic fertilizer and biochar on carbon release and microbial communities in saline-alkaline soil. The findings provide new insights and contribute to existing knowledge in soil carbon management, especially in arid regions.

Besides very minor typos observed (see below), I recommend this manuscript for publication.

Line 25- 25 - Incomplete sentence. Please check

Line 124 - 'Rice straw' is not a product of pyrolysis at 500°C. Correct to "rice-straw biochar" as used elsewhere in the manuscript.

Line 195 - Did the authors mean 'Table 2' instead of Table 1?

 

Author Response

1. Line 25- 25 - Incomplete sentence. Please check.

Response 1: Thank you very much for your thorough review and valuable comments. Regarding your observation that "Line 25 is an incomplete sentence, Please check." I have revised the text by removing the incomplete sentence on Line 25. Your valuable suggestions are much appreciated once again!

2. Line 124 - 'Rice straw' is not a product of pyrolysis at 500°C. Correct to "rice-straw biochar" as used elsewhere in the manuscript.

Response 2: Thank you for your comment. We have rewritten it in line 144 of the newly uploaded manuscript based on your suggestion and highlighted the text in red for clarity. The following lists the changes that have been made to the manuscript:

The rice straw samples were initially air-dried in preparation for subsequent experimentation. Biochar was generated through the pyrolysis of the rice straw at a temperature of 500 °C. Subsequently, both the air-dried rice straw and the rice biochar were individually crushed, sieved using a 2 mm sieve, and then stored separately for future experimentation.  (line 144-147)

3. Line 195 - Did the authors mean 'Table 2' instead of Table 1?

Response 3: Thank you very much for your suggestion! We have replaced Table 1 with Table 2 in line 218 of the revised manuscript, and highlighted the text in red for clarity

 

Once again, I sincerely thank you for your thoughtful guidance and unwavering support of my work!

Author Response File: Author Response.pdf