Tea Plantation Intercropping Legume Improves Soil Ecosystem Multifunctionality and Tea Quality by Regulating Rare Bacterial Taxa
Round 1
Reviewer 1 Report
Overall most of the literture is chinese. There intercropping studies from other crops in the world on incorporation of legume crops, that need to be included.
For other comments see the included file
Comments for author File: 
Comments.docx
Author Response
Response to reviewer
Dear reviewer, thank you very much for your suggestions and comments to improve our manuscript. All the comments/suggestions raised by you would significantly improve the quality of our manuscript. The following are the answers to the comments.
Some of the changes are red marked in the manuscript, please check the uploaded manuscript.
L43-45. The text will be more informative if you include names of the most common low-abundance species and the most common high-abundance species. There is a reference but the reader need clarification.
Response:
Classification of low-abundance species (rare taxa) and high-abundance species (abundant taxa) according to the relative abundance of each OTU in all samples. We combined always abundant taxa (AAT), conditionally abundant taxa (CAT), and conditionally rare and abundant taxa (CRAT) as abundant taxa and combined always rare taxa (ART) and conditionally rare taxa (CRT) as rare taxa. AAT: OTU with a relative abundance of ≥1% in all samples; CAT: OTU with a relative abundance of ≥1% in some samples and never ＜0.01%; ART: OTU with a relative abundance of ＜0.01% in all samples; conditionally rare taxa CRT: OTU with a relative abundance of ＜1% in all samples and ＜0.01% in some samples; moderate taxa MT: OTU with a relative abundance between 0.01% and 1% in all samples; and CRAT: OTU with a relative abundance ranging from rare (＜0.01%) to abundant (≥1%).
L49: Pleas explain what you mean by different structures and function. Include a few examples.
Response:
Structure refers to the relative abundance distribution of microbial communities at a taxonomic level. Function refers to soil ecological functions mediated by microorganisms such as climate regulation, nutrient cycling, plant growth promotion, and degradation of toxins and toxin and pollutant degradation. For example, biological nitrogen fixation is carried out by a wide range of diazotrophic bacteria and archaea, such as plant-symbionts like Rhizobium and Bradyrhizobium or free-living diazotrophs like Azospirillum and Azotobacter.
L71: Include a sentence or two that describe tea production. Row distances, management , age of tea plantation etc. Include the latin name Camellia sinensis sinensis
Response:
Relevant information such as row distances, management, age of tea plantation has been added in the materials and methods section (Line 123-124). Latin name of the tea plant has been added (Line 71).
L78: Please give more precise information on how the diversity declined.
Response:
Details on the decline of microbial diversity due to excessive fertilizer application have been added in the introduction section (Line 80-82).
In this section also include information on that the flavor and taste of tea is dependent on local soil conditions, weather, and cultivation methods.
Response:
Relevant information about the flavor and taste of tea is dependent on local soil conditions, weather, and cultivation methods have been added in the introduction section (Line 74-76).
L 113. Descibe the tea plantation, age, cultivar, row distance, soil type etc.
Response:
Relevant information about the tea plantation, age, cultivar, row distance, soil type have been added in the 2.1 section (Line 123-124 and Line 120-121).
L117: g kg -1
Response:
Unit g/kg has been uniformly changed to Unit g kg -1 (Line 122-123).
L118: Describe the plot size here. Did you establish the legume crops in 2018 and they lasted until 2021? When I read soybean+ milk vetch I interpret it as av mixture of the two specieces, but you first seeded soybean and then milk vetch. Please clarify the rotation and seeding techniques. Why di d you use the same density for all legume crops. You can express it as number per m2 or kilogrammes per hectar
Response:
Plot size has been described (Line 130).
There is a writing error here, the tea plantation intercropping legumious green manure trial started in 2017 and has been amended.
The tea plantation started intercropping legume crops in 2017 and lasted until 2021. To make it easier for the reader to understand, the description of confused “soybean + milk vetch” has been changed to “soybean - milk vetch” in the whole manuscript.
Crop rotation and seeding information has been added in Section 2.1 (Line 131-138).
The seeding density 120 kg ha -1 was the optimal seeding amount screened by the preliminary experiment. Unit kg hm -2 has been changed to Unit kg ha -1 (Line 137-138).
L121: Latin name for smoot vetch
Response:
The Latin names of red clover and smooth vetch, which did not appear in the previous manuscript, have been added (Line 128-129).
L118 and forward. You need to give the names of the cultivars seeded.
Response:
The names of the cultivars seeded have been added (Line 130-131).
How was the soil prepared prior to seeding?
Response:
Before sowing legume green manure, both sides of tea plants were pruned, and two trenches about 5 cm deep should be dug in the soil between the tea plant rows for sowing green manure seeds. The relevant description has been added in 2.1 section of manuscript (Line 135-137).
You must describe routine management according to the local convention.
Response:
Detailed routine management information on legume green manure has been added. Legume green manure should be irrigated in time after sowing, and growth period is not fertilized(Line 138-139).
Who was performing the experimental tasks? Staff from the university or local entrepreneurs?
Response:
University staff and research group student performing the experimental tasks.
How many leaves were harvested.
Response:
100 g of fresh tea leaves were harvested for the subsequent Indexes determination (Line 141).
From googling I understand that there are different harvest regimes during the year, and it is of interest for the reader to get a short introduction.
Response:
The experimental site in Jiangsu Province produces green tea, and fresh leaves are picked in Spring (Line 139-140).
Amount of biomass incorporated into the soil? Developmental stage of the intercrop when incorporated?
Response:
The green manure growing to full-bloom stage is all incorporated into the soil, and related descriptions have been added in the 2.1 section of manuscript (Line 132-135).
Tea leaves were sampled on xx May 2021.
Response:
Specific sampling dates have been added in the 2.1 section of manuscript (Line 140).
Were inoculants used for the legume species?
Response:
All legume plants were not inoculated with rhizobium.
L132: Do you have a reference for the five-point sampling method?
Response:
The five-point sampling method refers to the previous methods of soil sample collection, and the references following:
Zhang, C.; Xue, W.; Xue, J.; Zhang, J.; Qiu, L.; Chen, X.; Hu, F.; Kardol, P.; Liu, M. Leveraging functional traits of cover crops to coordinate crop productivity and soil health. Journal of Applied Ecology 2022, 59, 10. https://doi.org/10.1111/1365-2664.14264.
L133: You need to state the amount of soil that was drawn for DNA-extratcion and for measurements of enzyme activity
Response:
Soil sample quality for DNA extraction and enzyme activity determination has been added in 2.3 and 2.4 section of manuscript (Line 182 and Line 175).
L299: two legume species,
Response:
The description of confused “two kinds of legumes” has been changed to “two legume species” in 3.2 section of manuscript (Line 312).
L433: Do you have an explanation based on the crops to the decrease of pH? Was a sample taken across the experimental area. In that case there is an influence of different species an d cropping systems. I would have been better with plotwise sampling and statistics.
You should be careful drawing conclusions from one measurement. Was pH was measured in the TM plot after the experiment was conducted?
Response:
The decrease of soil pH value under intercropping legume green manure may be caused by organic acids secreted by leguminous plant roots. In addition, pH values of soil samples under different treatments were also determined at the same time in 2022, and the results were consistent with the results of this manuscript.
Overall most of the literture is chinese. There intercropping studies from other crops in the world on incorporation of legume crops, that need to be included.
Response:
Experimental cases of legume incorporating into soil from other countries have been added
[23], [24], and [26].
Thank you again for giving us an opportunity to address the concerns of yours. We hope, the manuscript is now much improved and suitable for acceptance in Agronomy.
Respectfully submitted by Wanping Fang.
Author Response File: Author Response.docx
Reviewer 2 Report
Comments and Suggestions for Authors:
Article ID: agronomy-2286338
Tea Plantation Intercropping Legume Improves Soil Ecosystem Multifunctionality and Tea Quality by Regulating Rare Bacterial Taxa
General comment:
I would like to stress that I support the potential publication of this paper due to its scientific interest.
The study is devoted to detecting the intercropping practice's effect (tea plant monoculture and tea plants respectively intercropped with soybean, soybean + milk vetch, soybean + red clover, and soybean + smooth vetch) on the soil bacterial and fungal communities' composition, pH of soil, ammonium and nitrate nitrogen content in the soil and quality improvement of tea leaves. Results showed that the tea intercropping with soybean + milk vetch and with soybean + red clover was more conducive to enhancing ecosystem multifunctionality and improving tea plant performance.
The "Abstract" is structured with a background, the main body of the abstract, and a short conclusion.
General comment to the "Introduction" section:
The content of the literature review chapter is related to the research topic. A systematic literature review identifies, selects, and critically appraises research.
General comment to the "Materials and Methods" section:
The methodology is adequate and give readers enough information.
In the chapter "Results",
The results are displayed correctly, supported by sufficient data, and arranged in a logical sequence.
Line 262. Please check fragment “Between 2 and 2”
In the chapter "Discussion",
The discussion builds on the results and is linked to relevant literature throughout the section. It is necessary to encourage further interest in the tea plant/legume intercropping system for its benefit for soil fertility, potential changes in beneficial bacteria, and the quality of tea leaves.
In the chapter "Conclusion",
The paper provides evidence for its conclusion.
Submission date:
21. March 2023
Comments for author File: Comments.doc
Author Response
Response to reviewer
Dear reviewer, thank you very much for your suggestions and comments to improve our manuscript. All the comments/suggestions raised by you would significantly improve the quality of our manuscript. The following are the answers to the comments.
Line 262. Please check fragment “Between 2 and 2”
Response：“Between 2 and 2” has been changed to “Between -2 and 2” (Line 275). Thank you for point out this error.
Thank you again for giving us an opportunity to address the concerns of yours. We hope, the manuscript is now much improved and suitable for acceptance in Agronomy.
Respectfully submitted by Wanping Fang.
Author Response File: Author Response.docx
