Review Reports - Leguminous Green Manure Intercropping Promotes Soil Health in a Citrus (Citrus reticulata) Orchard

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

1. Abstract: line No. 25-26, Mention the numerical value or data of enzymatic activity of soil sucrase, and the β-1, 4-glucosidase activity in the soil compared to the monoculture treatment.

2. Rewrite the introduction with a proper hypothesis, as a lot of work has been carried out regarding intercropping with legume in agroforestry and horticultural system. So, please justify the novelty.

3. In conclusions, write 2-3 lines about the path ahead for the researchers in this field.

4. In conclusions: line No. 549-550, rewrite the sentence with appropriate words.

Comments on the Quality of English Language

Quality of english is adequate for a good quality research paper. However, review of more research works could help the authors to describe the importance of the paper more explicitly and should had been added in the introduction section to justify the hypothesis of the paper. Grammatical error was very less in the text

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

Below my comments.

Title: I suggest to remove the first word citrus in turn to have: Leguminous green manure intercropping promotes soil health in a citrus (Citrus reticulata B. ) orchard.

Lines 20-21: Specify the types of leguminous green manures and citrus species in Latin.

Lines 28-29: How much did the diversity and composition raise? Please specify in percentage.

Lines 37-40: Re-write this period. It is not understandable.

Lines 114-115: ratio plants:soil for the intercropping? Give more information

Line 116: how did you choose where to sample? According to which scheme? Describe more.

Line 125: soil chemical analysis is generally performed on air-dried samples. SpecifyLines 135-138: specify the acronyms AK and AP.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

This study demonstrates that intercropping leguminous green manure in citrus orchards significantly enhances soil health. In Sichuan, China, intercropping alfalfa and hairy vetch increased total nitrogen, nitrate nitrogen, and available phosphorus by up to 96.67%. Alfalfa boosted soil sucrase activity, while hairy vetch improved β-1,4-glucosidase activity. Both crops enriched the soil with beneficial microbes such as Mortierella and Streptomyces, resulting in improved soil nutrients, enzyme activity, and microbial diversity, thereby promoting healthier orchard soil. Below are the minor and major comments:

Abstract:

Lacks a clear novelty statement.

Keywords:

"Legume manure" should be added to the keywords.

Introduction:

Line 37: Remove the extra space before "....grown".
Line 58: Add a sentence and cite the following references: "Soil microbes also help in bioremediation by breaking down pollutants (https://pubs.acs.org/doi/full/10.1021/acs.est.3c01240) and promoting disease resistance in plants through beneficial microbial interactions (https://doi.org/10.3389/fsufs.2021.606454)."

Materials and Methods:

Section 2.2: The study mentions microbial diversity changes at a 0-15 cm soil depth. Which specific depths within this range were selected for analysis? Also, specify the amount of soil sampled for microbial study.
Section 2.4: The activity of the selected enzymes was determined in the study, please clarify why these particular enzymes were chosen. Additionally, detail the methods used to assess the activity of each enzyme.

Results:

Table 1: Define "control" clearly.
Figure 1: Define "a", "ab", and "b" in the figure legend.
Section 3.2: Why was the activity of the alkaline phosphatase enzyme not included in the study?
Figure 2: Indicate the color of the circles in the Venn diagram in the figure legend.
Table 2: The results show limited significant differences. Additionally, while species richness is discussed, species abundance is not. It is important to include the proportion of each species in the community.
Figure 3: The text in the figure is not very clear. Enlarge the plots and stack parts A and B for better visibility.

Discussion:

Discuss soil fungi, including Cunninghamella (https://link.springer.com/article/10.1007/s11356-022-23901-0), and bacteria (https://doi.org/10.3389/fbioe.2021.808075) in relation to plant pathogens and the biodegradation of soil organic matter.
Line 448: Correct the heading and remove the "".

Conclusion:

Revise the conclusion to better emphasize the practical benefits and broader implications of this study.

Comments on the Quality of English Language

Moderate English editing is needed to improve clarity, correct grammar, and enhance readability while maintaining the original meaning.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

1. Please mention aims of study in abstract.

2. The study highlights the positive effects of intercropping green manure but introduction does not clearly articulate the specific research gaps this study aims to address. Although there is a mention of inconclusive existing research, it would be helpful to specify what these inconsistencies are, particularly regarding soil chemical properties and microbial community responses in citrus orchards.

3. The introduction does not present a clear hypothesis or objectives of the study. Including a concise hypothesis would provide a clear focus for the research.

4. The introduction should clearly outline the specific research questions that the study intends to answer. This would help frame the significance of the research.

5. In whole manuscript, There are some grammatical issues and areas where sentence structure could be improved for clarity. For example, reducing passive voice usage can make statements more direct. Consider using more varied sentence lengths and structures to enhance readability.

6. While the introduction provides a good overview of the issues in citrus orchards, it could benefit from a more detailed explanation of the specific challenges posed by monoculture practices, such as the impact on soil health over time.

7. Ensure that all claims are properly cited, and consider adding recent studies to support statements about the benefits of green manure intercropping.

8. Was amplicon size selection performed following PCR amplification to ensure the removal of any undesired DNA fragments, and if so, what specific methods or criteria were used to achieve this?

9. Could you provide specific details on the parameters used for your analyses, such as the clustering parameters for defining operational taxonomic units (OTUs), the methods and thresholds for chimera removal, and any other relevant settings applied during data processing?

10. Which databases were used for taxonomic annotation (e.g., SILVA, GREENGENES, NCBI), specify and detail the methodology for taxonomic assignment, including the percentage coverage and identity thresholds applied?

11. 2.4. Activities of soil enzymes : Please elaborate the methodology of soil enzymes. Present presentation is not good.

12. Figures 1, 2, 3, 4, and 6 are not clearly visible in their current format. To enhance visibility and quality, I recommend separating the combined figures into a greater number of individual figures. This will allow for clearer presentation and better interpretation of the data.

13. While the Discussion section discusses various results, integrating findings more cohesively could enhance the narrative. Linking observations across different subsections can help emphasize the overarching themes and implications of the research.

14. Add some more citations claiming your study.

15. Discuss the biological mechanisms behind the observed increases in total nitrogen (TN) and available phosphorus (AP) in more detail.

16. Please discuss how your enzyme activity results align or differ from previous studies.

17. Elaborate on why alpha diversity did not significantly improve under intercropping conditions.

18. The role of root exudates in shaping microbial communities is interesting. Consider expanding on how these exudates may influence specific microbial interactions or nutrient cycling.

19. Discuss the implications of reduced abundance of phytopathogens for plant health and crop yield in more detail, considering how these changes might affect disease resistance in citrus.

20. When discussing environmental factors affecting Lysobacter abundance, consider including more specific data or examples from your study to reinforce the connection.