The Variations in Soil Microbial Communities and Their Mechanisms Along an Elevation Gradient in the Qilian Mountains, China
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis research article focuses on the microbial biodiversity of an extreme environment such as the Qilian mountains. The experiments performed are quite diverse and are thought to evaluate several aspects of five vegetation types in an elevation gradient. I think the Results and Discussion sections are well written and easy to understand. I just have some concerns about the methodology used, as stated in the point-by-point comments below.
In general, I recommend changing the size of the figures and graphs so that the words and numbers present (axis, legends, and so on) can be clearly seen without the necessity of zooming in.
Keywords: I suggest changing the keywords with others that are not already present in the title of the article (Qilian Mountains; Elevation gradient; Soil microbial community).
Lines 80-90: I suggest modifying this chapter to focus on the aim of the study, rather than the expected outcomes (as implied by the initial “We hypothesized”). Otherwise, change the verb so that it is clear that the following sentences are the main results obtained.
Line 95: The citation is in the wrong format.
Line 102: I suggest using the term “grassland” instead of “steppe”, to be more consistent.
Figure 1: What did you use to obtain these maps? Please add this information if necessary. Moreover, in the caption of the figure, add what is represented in the map on the left (I suppose the area of the national nature park). I suggest also adding where the mountain is located in a vaster region (for example, where it is located with respect to China).
Line 108: You state that “we established four plots area of 20 m×20 m at each vegetation type”. Please confirm that with this sentence you mean that you sampled four plots at the forest vegetation type, four plots at the meadow grassland, and so on for all the five vegetation types considered. Looking at the sampling points on the map in Figure 1, and the data presented in the Results, it seems that you only sampled one plot for each vegetation type. If that is the case, I do not think that the results obtained from a single plot could be representative of that type of vegetation in the entire mountain. Do you expect changes in the populations at different seasons? You sampled the soil in August, I guess, when the temperature is higher. Add, if possible, the temperature measured on the sampling days.
Lines 110-112: This method is not clear; please, rephrase the sentence to make it clearer. Perhaps a schematic representation, even in the supplementary materials, could be helpful.
Lines 113-117: The reference to the height is not present in any of the results, is this something that you actually used in the study? Clarify this aspect.
Line 120: Was the temperature of the samples maintained in some way? If that is the case, add the information.
Table 1: Looking at Figure 1 and at these geographic positions, it seems that G, S, M and R were sampled nearby with a change of elevation, while the forest was sampled at another site. Can you explain this choice?
Line 124: Rephrase the sentence to correct it: “The fresh soil samples collected were firstly used for soil microbe…”.
Chapter 2.3.1: How many replicates were performed for each experiment?
Line 129: Please add the reference to the gravimetric method.
Chapter 2.3.2: I recommend providing in the supplementary materials the data obtained from the analysis (OTUs with the corresponding sequences for each sample). Moreover, the information on the sequences must be deposited in a repository such as GeneBank.
Line 140: How many replicates were made for this analysis?
Chapter 2.3.3: What did you use to identify the species of plants? Implement the text with this information.
Line 204: Modify “meadow meadows” with “meadow grasslands”
Table 3: Are these indices the average obtained from the four plots studied for each vegetation type? Add that information to the table. Moreover, add the standard deviation. Have you verified that the results are statistically significant? Please add that information, or state that the diversity observed is nonetheless not significant.
Chapter 3.3: Have you performed the statistical analysis for these experiments? Add the results to the graphs or text.
Line 234: The concepts of “The richness, chao1, ace and shannon indices” were not introduced in the Materials and Methods section. Implement the text in that section with the information needed to understand these indices.
Figure 3: In the text only four out of the six graphs present for bacteria and fungi are described in the text, what about Simpson and Good coverage? As indicated in the previous comments, add the specifics of the two indices in the Materials and Methods section. To make the figure clearer, add a legend to indicate what F, G, S, M, and R stand for. The axes are missing the unit of measurement (this last comment is to be applied also for Figure 4).
Lines 242-243: I found this sentence very clear and helpful to the reader. I suggest using it as a model to explain in a few words the other parameters measured in previous paragraphs.
Lines 243-245: Rephrase this sentence because there are some errors.
Figure 4: If possible, change the color of Figures Ab and Bc so that they match the color of Figures Ab and Bd and those in the previous figure. For Ab and Bd, use the same names presented in the other figures (F, G, S, M, R instead of RvsF, etc.) or explain what RvsF stands for. Do not use letters A and B to differentiate between bacteria and fungi. It is only confusing. Use only a, b, c, and d to indicate the four graphs. The same applies to Figure 5.
Line 292: Correct the sentence, since plant diversity is not positively correlated with elevation.
Line 399: Add “between” after “understand the relationship”.
Conclusions: I suggest rephrasing the sentences of the whole paragraph since in the current form they are too much like a list. Making it more articulate will be helpful to highlight the key points and the main results obtained from the study.
Author Response
Dear Editors and Reviewers,
First of all, I sincerely thank you for your valuable comments and suggestions on this paper, and thank you very much for your valuable time and efforts. At the same time, I deeply respect your rigorous and serious academic accomplishment. According to your comments and suggestions, I have carefully modified and improved the paper, and the quality of the paper has been significantly improved. The revised part has been marked red in the revision draft, and if there are still shortcomings, I hope you can understand and criticize and correct. The following are the specific revisions to the manuscripts:
Comments 1：In general, I recommend changing the size of the figures and graphs so that the words and numbers present (axis, legends, and so on) can be clearly seen without the necessity of zooming in.
Responds1: According to the reviewer's comments, the numbers and legends in the figure have been enlarged.
Comments 2：Keywords: I suggest changing the keywords with others that are not already present in the title of the article (Qilian Mountains; Elevation gradient; Soil microbial community).
Responds2: According to the reviewer's comments, the original keywords“Qilian Mountains; Elevation gradient; Soil microbial community; Spatial patterns; Ecological drivers” has been revised to “Qilian Mountains; Soil bacteria and fungi; Diversity; Ecological niche differences; Ecological drivers”. Consulting experts on whether the revised were appropriate.
Comments 3：Lines 80-90: I suggest modifying this chapter to focus on the aim of the study, rather than the expected outcomes (as implied by the initial “We hypothesized”). Otherwise, change the verb so that it is clear that the following sentences are the main results obtained.
Responds 3: According to the reviewer's comments, this chapter has been modified. “In this study, we utilized 16S and ITS amplicon sequencing to determine the diversity and composition of bacterial and fungal communities along Qilian mountain elevation gradient. We hypothesized that (i) Soil bacterial and fungal communities would respond differently to elevation, and both exhibited spatial niche-differentiation; (ii) Environmental factors, especially plant communities and soil properties, would impact the soil microbial community, causing differences in composition and diversity along the elevation gradient, with fungi and bacteria responding differently according to their ecological traits and niche requirements. Therefore, five typical vegetation types (i.e., coniferous forest, meadow grassland, alpine shrub, alpine meadow, sparse vegetation of limestone flats) along the vertical belt of Qilian Mountains were selected to explore the elevational patters of bacterial and fungal communities and their driving ecological factors.”
Comments 4：Line 95: The citation is in the wrong format.
Responds4: According to the reviewer's comments, the citation “ (Zhang et al. 2021)” in line 95 is revised as “ [22]”.
Comments 5：Line 102: I suggest using the term “grassland” instead of “steppe”, to be more consistent.
Responds5: According to the reviewer's comments, revised “grassland” instead of “steppe”. Thanks to the expert reviewers for their careful review of this manuscript.
Comments 6：Figure 1: What did you use to obtain these maps? Please add this information if necessary. Moreover, in the caption of the figure, add what is represented in the map on the left (I suppose the area of the national nature park). I suggest also adding where the mountain is located in a vaster region (for example, where it is located with respect to China).
Responds 6: According to the reviewer's comments, Add the map source: The study was conducted at the Qilian Mountain National Nature Park of Xianmi Forest farm~Lenglongling (101°24'11"~101°57'29" E, 37°17'25"~37°41'30" N; http:www.gscloud.cn/; Figure1). To improve the perception of information, the quality of Figure 1 was enlarged and improved. Following picture:
Comments 7: Line 108: You state that “we established four plots area of 20 m×20 m at each vegetation type”. Please confirm that with this sentence you mean that you sampled four plots at the forest vegetation type, four plots at the meadow grassland, and so on for all the five vegetation types considered. Looking at the sampling points on the map in Figure 1, and the data presented in the Results, it seems that you only sampled one plot for each vegetation type. If that is the case, I do not think that the results obtained from a single plot could be representative of that type of vegetation in the entire mountain. Do you expect changes in the populations at different seasons? You sampled the soil in August, I guess, when the temperature is higher. Add, if possible, the temperature measured on the sampling days.
Responds 7: we randomly established four plot in each vegetation type. Each plot area of 20 m×20 m, of which plots of 4 tree (10 m×10 m), 4 shrub (5 m×5 m) and 8 herb (1 m×1 m) monitoring quadrats. Since the four plots are too close together, a study location is shown on the map. Plant community surveys and soil samples collected during plant growth season (August 2021). Unfortunately, the temperature was not measured at that time, and at a later stage, we will pay attention to the experimental design. According to the reviewer's comments, modified the text expression to make it clearer. I have drawn a schematic representation, to understand the monitoring quadrats. The detailed explanation is as follows picture:
Comments 8：Lines 110-112: This method is not clear; please, rephrase the sentence to make it clearer. Perhaps a schematic representation, even in the supplementary materials, could be helpful.
Responds 8: According to the reviewer's comments , modified the text expression to make it clearer. The specific modifications are as follows:
Plant community mainly investigate species composition and calculate plant diversity. Names, coverage and height of plant species were recorded within plots of 4 tree (10 m×10 m), 4 shrub (5 m×5 m) and 8 herb (1 m×1 m) monitoring quadrats. Plant species are identified by the plant professor through the method of plant morphology. Plant coverage was estimated visually, i.e., by percentage. For plant height, a steel tape measurer was used to measure the vertical height between the highest point and the ground in natural state of plants in the sample square, expressed in cm. Average height of species: 5 plants (clusters) of each species were randomly selected to measure the vertical height between the highest point and the ground in their natural state with a tape measure, which was represented by cm. Plant diversity was calculated based on the relative height and coverage of plant species[24]. The detailed calculation method is as follows:
(1) IV= (Rc + Rh) / 2×100;
(2) Species richness index (S’ plant) : the number, N, of total species in all plots;
(3) Shannon-Wiener index (H’ plant) = -∑(Pi × Ln Pi);
(4) Pielou evenness index(J’ plant) = H’ plant / ln S’ plant.
Where IV is the importance value of species; Rc is the relative coverage of species; Rh is the relative height of species; Pi is the importance value of the species i;
Comments 9：Lines 113-117: The reference to the height is not present in any of the results, is this something that you actually used in the study? Clarify this aspect.
Responds 9: Plant diversity was calculated based on the relative height and coverage of plant species. The detailed calculation method is as follows:
- IV= (Rc + Rh) / 2×100
(2) Species richness index (S’ plant) : the number, N, of total species in all plots
(3) Shannon-Wiener index (H’ plant) = -∑(Pi × Ln Pi)
(4) Pielou evenness index(J’ plant) = H’ plant / ln S’ plant
Where IV is the importance value of species; Rc is the relative coverage of species; Rh is the relative height of species; Pi is the importance value of the species i
Comments 10：Line 120: Was the temperature of the samples maintained in some way? If that is the case, add the information.
Responds 10: According to the reviewer's comments, add the information, “All soil samples were screened with a 2mm screen and then divided into two parts: one was stored in a refrigerator at −20°C for microbial studies, and the other was stored at 4°C for the determination of soil physiochemical properties.”
Comments 11：Table 1: Looking at Figure 1 and at these geographic positions, it seems that G, S, M and R were sampled nearby with a change of elevation, while the forest was sampled at another site. Can you explain this choice?
Responds 11: The Qilian Mountains continuely extend to northwest, where is a small hill and no forest. The frests are only distributed in the southast low elevations. I have drawn a schematic representation, to understand the research sites. The detailed explanation is as follows picture:
Comments 12：Line 124: Rephrase the sentence to correct it: “The fresh soil samples collected were firstly used for soil microbe…”.
Responds 12: It has been modified based on the reviewer's comments.“All soil samples were screened with a 2mm screen and then divided into two parts: one was stored in a refrigerator at −20°C for microbial studies, and the other was stored at 4°C for the determination of soil physiochemical properties.”
Comments 13：Chapter 2.3.1: How many replicates were performed for each experiment?
Responds 13: There were 4 replicates per sample, text table annotation “n=4”.
Comments 14：Line 129: Please add the reference to the gravimetric method.
Responds 14: According to the reviewer's comments, add the reference to the gravimetric method.
Comments 15：Chapter 2.3.2: I recommend providing in the supplementary materials the data obtained from the analysis (OTUs with the corresponding sequences for each sample). Moreover, the information on the sequences must be deposited in a repository such as GeneBank.
Responds 15: Add sequence information materials according to reviewer's comments. At present, we do not have OTUs corresponding to each sample in the gene bank, which will later supplement the gene bank information
Comments 16：Line 140: How many replicates were made for this analysis?
Responds 16: There were 4 replicates per sample, and add the information “There were 4 replicates per sample” to the text method.
Comments 17：Chapter 2.3.3: What did you use to identify the species of plants? Implement the text with this information.
Responds 17: Plant species are identified by the professor of plant classification through the method of plant morphology. Add this information to the text based on the reviewer's comments.
Comments 18：Line 204: Modify “meadow meadows” with “meadow grasslands”
Responds 18: According to the reviewer's comments, modified “meadow meadows” with “meadow grasslands”
Comments 19：Table 3: Are these indices the average obtained from the four plots studied for each vegetation type? Add that information to the table. Moreover, add the standard deviation. Have you verified that the results are statistically significant? Please add that information, or state that the diversity observed is nonetheless not significant.
Responds 19: This table has only numerical values, no standard errors or standard deviations. Because the species richness index refers to the number of all plant species present in the quadrat of the 4 plots, not the average number of plants present in the 4 plots. H’plant and J’ plant are one value. so, there is no standard deviation or standard error.
Comments 20：Chapter 3.3: Have you performed the statistical analysis for these experiments? Add the results to the graphs or text.
Responds 20: These experiments were performed using one-way ANOVA and multiple comparison (LSD) with SPSS 20.0，and add the information in the text (Fig 3 and Fig 4).
Fig 3. Alpha diversity index of soil bacteria along the elevation gradient.
Fig 4. Alpha diversity index of soil fungi along the elevation gradient.
Comments 21：Line 234: The concepts of “The richness, chao1, ace and shannon indices” were not introduced in the Materials and Methods section. Implement the text in that section with the information needed to understand these indices.
Responds 21: According to the reviewer's comments, this chapter has been modified.“ Using the mothur software (http://www.mothur.org/wiki/Calculators), to obtain richness, shannon, simpson, chao1, ace, good's coverage, and evaluate the alpha diversity of soil bacteria and soil fungi. A one-way analysis of variation (ANOVA) and a least significant difference (LSD) multiple comparison (P < 0.05) were used to assess the significant effects of soil properties, and soil microbial diversity (richness, shannon, simpson, chao1, ace, goods coverage).”
Comments 22：Figure 3: In the text only four out of the six graphs present for bacteria and fungi are described in the text, what about Simpson and Good coverage? As indicated in the previous comments, add the specifics of the two indices in the Materials and Methods section. To make the figure clearer, add a legend to indicate what F, G, S, M, and R stand for. The axes are missing the unit of measurement (this last comment is to be applied also for Figure 4).
Responds 22: Add the axes the unit of measurement in the text. The annotation of F, G, S, M, and R in Fig1. “Coniferous forests (F, 2700 m); Meadow grasslands (G, 3200 m); Alpine shrubs (S, 3500 m); Alpine meadows (M, 3700 m); Sparse vegetations of limestone flat (R, 4000 m). Same for follwoing text” .
Comments 23：Lines 242-243: I found this sentence very clear and helpful to the reader. I suggest using it as a model to explain in a few words the other parameters measured in previous paragraphs.
Responds 23: It has been modified according to the reviewer's comments, and standard sentences are used in the figure. “Fig 3. Alpha diversity index of soil bacteria along the elevation gradient.
Richness (a); Shannon (b); Simpson (c); Chao1 (d); Ace (e); Good's coverage (f). Fig 4. Alpha diversity index of soil fungi along the elevation gradient. Richness (a); Shannon (b); Simpson (c); Chao1 (d); Ace (e); Good's coverage (f). Similar sentence in other fig.
Comments 24：Lines 243-245: Rephrase this sentence because there are some errors.
Responds 24: The revision was based on the reviewer's comment that “Beta diversity refers to differences along environmental gradients of soil microbial communities”.
Comments 25：Figure 4: If possible, change the color of Figures Ab and Bc so that they match the color of Figures Ab and Bd and those in the previous figure. For Ab and Bd, use the same names presented in the other figures (F, G, S, M, R instead of RvsF, etc.) or explain what RvsF stands for. Do not use letters A and B to differentiate between bacteria and fungi. It is only confusing. Use only a, b, c, and d to indicate the four graphs. The same applies to Figure 5.
Responds 25: Based on the reviewer's comments, the original figures 4 and 5 has been modified.
Fig 4. Beta diversity of soil bacteria and fungi along the elevation gradient.
Soil bacterial NMDS (a); Soil fungal NMDS (b); Soil bacterial Bray-Curtis dissimilarity (c); Soil fungal Bray-Curtis dissimilarity (d).
Fig 5. Redundancy analysis of soil microorganisms and ecological factors.
Soil bacterial redundancy analysis (a) and ecological factors contrbution (b); Soil fungal redundancy analysis(c) and ecological factors contrbution (d).
Comments 26：Line 292: Correct the sentence, since plant diversity is not positively correlated with elevation.
Responds 26: According to the reviewer's comments, the sentence has been modified. “The result showed that elevation was negatively correlated with plant diversity and soil properties. Plant diversity was positively correlated with soil bacteria and fungi. Soil properties were positively correlated with fungi and negatively correlated with bacteria. In summary, elevation directly affects soil properties, and indirectly affects soil bacteria.”
Comments 27：Line 399: Add “between” after “understand the relationship”.
Responds 27: According to the reviewer's comments, “between” has been add after “understand the relationship”in line 399.
Comments 28：Conclusions: I suggest rephrasing the sentences of the whole paragraph since in the current form they are too much like a list. Making it more articulate will be helpful to highlight the key points and the main results obtained from the study.
Responds 28: According to the reviewer's comments, the conclusion has been modified. “Environmental factors, especially plant communities and soil properties, would impact the soil microbial community, causing differences in composition and diversity along the elevation gradient. Soil bacteria increased with increasing elevation, and soil fungi showed an obvious mid-elevation pattern. Fungi and bacteria responding differently according to their ecological traits and niche requirements, which likely due to their distinct ecological traits. All ecological factors showed that NO3--N, TN, SM and pH were the four most important contributors of soil bacteria phyla. For soil fungi, the most important factors were BD, H’plant and S’plant. Bacteria are mainly influenced by soil properties, while fungi prefer to be influenced by plant communities. Our study strengthens the current understanding the complex patterns and multiple drivers affected soil microbial communities along elevation gradients, and provides a theoretical basis for biodiversity research in the context of global change.”
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe article is devoted to the relevant topic of studying the influence of the altitude gradient on soil microbial communities. There is various information on changes in the diversity of microorganisms with increasing altitude, so it is necessary to study and clarify the environmental factors affecting the abundance and number of bacterial and fungal taxa in mountain soils. Apparently, vegetation and physicochemical properties of the soil have the greatest impact on the structure of the soil microbiome, so the authors of the article pay the greatest attention to them. Studying the response of the microbiome to changes in environmental factors will contribute to understanding the mechanisms of maintaining biodiversity and managing ecosystems. The authors formulated interesting scientific hypotheses and obtained several valuable results, so I consider the peer-reviewed article to be very relevant for this area of research. The article will be of interest to a wide range of researchers of soils, ecology and soil microorganisms. The authors' conclusions are based on the arguments of the results obtained and resolve the main issues posed in the study. Literature references are appropriate and adequate. Of course, the authors have done a tremendous job of analyzing samples, processing and interpreting data. I highly appreciate this article, but I have a few suggestions and comments to improve the article:
1 – The authors formulated 3 hypotheses, but there is no clear confirmation or refutation of them in the Conclusion. The authors should add this information.
2 – The authors should expand the description of the objects under study, add more characteristics, and also insert photos of the analyzed soils. The soil type according to various soil classifications is not indicated. There is no information on the type and history of land use of the areas from which the soil was collected. In addition, it is unclear which of the soil horizons was analyzed (The upper 10 cm of soil often contains: forest litter with several subhorizons and a humus layer. Which horizons were analyzed by the authors?). All these details are important for the study. The object of study may not be completely clear to readers, and therefore the obtained results are unclear.
3 – The number of physical replicates (the authors indicate that the soil was collected from three sampling points) of samples is too small even for agricultural ecosystems with controlled conditions. Forest and grassy soils are very heterogeneous, their chemical and physical properties vary greatly even over a small area. Therefore, the authors should have increased the number of replicates to at least 5 or 6.
4 – The abundance and diversity of vegetation was assessed by the authors without special geobotanical methods (such methods are not described in the article), so the results obtained for plants cause some mistrust. The authors should add a description of the geobotanical methods used or critically review the results obtained for vegetation.
5 – The results obtained by the authors on the dominance of Actinobacteriota and Proteobacteria among bacteria and Ascomycota and Basidiomycota among fungi are quite trivial. Usually, such results are characteristic of a very wide range of soils from different climatic zones and natural zones. Therefore, to identify the specifics of the objects under study, the authors should have focused on non-dominant taxa. I recommend that the authors revise the article and add information to the Discussion on rare taxa of bacteria and fungi.
6 – I recommend adding a figure with photographs of the objects of study - the soil and vegetation of the analyzed biomes. This will improve the perception of information.
7 – I recommend enlarging and improving the quality of Figure 1 to improve the perception of information.
8 – The authors should discuss in more detail the reasons why fungi are primarily affected by vegetation, and bacteria by the physical and chemical properties of the soil. This would be interesting to a wide range of readers.
9 – Perhaps the authors should reduce the number of literary references and at least refer to publications in the journal "Sustainability" several times.
Author Response
Dear Editors and Reviewers,
First of all, I sincerely thank you for your valuable comments and suggestions on this paper, and thank you very much for your valuable time and efforts. At the same time, I deeply respect your rigorous and serious academic accomplishment. According to your comments and suggestions, I have carefully modified and improved the paper, and the quality of the paper has been significantly improved. The revised part has been marked red in the revision draft, and if there are still shortcomings, I hope you can understand and correct. The following are the specific revisions to the manuscripts:
Comments 1：The authors formulated 3 hypotheses, but there is no clear confirmation or refutation of them in the Conclusion. The authors should add this information.
Responds 1: According to the reviewer's comments, I have revised and confirmed the assumptions in the conclusion. “In this study of the Qilian Mountain soil microbial community, we found the diversity of bacterial and fungal communities reflected a clear spatial niche-differentiation. Soil bacteria increased with increasing elevation, and soil fungi showed an obvious mid-elevation pattern, which likely due to their distinct ecological traits. Bacteria are mainly influenced by soil properties, while fungi prefer to be influenced by plant communities. These findings generally align with our hypothesis.”
Comments 2：The authors should expand the description of the objects under study, add more characteristics, and also insert photos of the analyzed soils. The soil type according to various soil classifications is not indicated. There is no information on the type and history of land use of the areas from which the soil was collected. In addition, it is unclear which of the soil horizons was analyzed (The upper 10 cm of soil often contains: forest litter with several subhorizons and a humus layer. Which horizons were analyzed by the authors?). All these details are important for the study. The object of study may not be completely clear to readers, and therefore the obtained results are unclear.
Responds 2: According to the reviewer's comments, the specific modifications are as follows:
(1) Insert photos into the text.
Coniferous forests (F, 2700 m); Meadow grasslands (G, 3200 m); Alpine shrubs (S, 3500 m); Alpine meadows (M, 3700 m); Sparse vegetations of limestone flat (R, 4000 m).
- The corresponding soil types of vegetation types were mountain gray brown soil, subalpine meadow soil, alpineshrub-meadow soil, alpine meadow soil and frigid desert soil.
- The 10cm soil analyzed in this paper includes only the humus layer. There is also no litter layer in the forest sample soil.
Comments 3：The number of physical replicates (the authors indicate that the soil was collected from three sampling points) of samples is too small even for agricultural ecosystems with controlled conditions. Forest and grassy soils are very heterogeneous, their chemical and physical properties vary greatly even over a small area. Therefore, the authors should have increased the number of replicates to at least 5 or 6.
Responds 3: We established four sample plots for each vegetation type, with an area of 20 m×20 m, and randomly set three sampling points for each plot. Therefore, the sampling points for our study were: 3×4=12.
Comments 4：The abundance and diversity of vegetation was assessed by the authors without special geobotanical methods (such methods are not described in the article), so the results obtained for plants cause some mistrust. The authors should add a description of the geobotanical methods used or critically review the results obtained for vegetation.
Responds 4: It has been modified based on the reviewer's comments. The specific modifications are as follows:
Plant community mainly investigate species composition and calculate plant species diversity. Species names, coverage and height of each plant species were recorded within plots of 4 tree (10 m×10 m), 4 shrub (5 m×5 m) and 8 herb (1 m×1 m) monitoring quadrats. Plant species are identified by the plant professor through the method of plant morphology. Plant coverage was estimated visually, i.e., by percentage. For plant height, a steel tape measurer was used to measure the vertical height between the highest point and the ground in natural state of plants in the sample square, expressed in cm. Average height of species: 5 plants (clusters) of each species were randomly selected to measure the vertical height between the highest point and the ground in their natural state with a tape measure, which was represented by cm. Plant diversity was calculated based on the relative height and coverage of plant species. The detailed calculation method is as follows:
(1) IV= (Rc + Rh) / 2×100;
(2) Species richness index (S’ plant) : the number, N, of total species in all plots;
(3) Shannon-Wiener index (H’ plant) = -∑(Pi × Ln Pi);
(4) Pielou evenness index(J’ plant) = H’ plant / ln S’ plant.
Where IV is the importance value of species; Rc is the relative coverage of species; Rh is the relative height of species; Pi is the importance value of the species i;
Comments 5：The results obtained by the authors on the dominance of Actinobacteriota and Proteobacteria among bacteria and Ascomycota and Basidiomycota among fungi are quite trivial. Usually, such results are characteristic of a very wide range of soils from different climatic zones and natural zones. Therefore, to identify the specifics of the objects under study, the authors should have focused on non-dominant taxa. I recommend that the authors revise the article and add information to the Discussion on rare taxa of bacteria and fungi.
Responds 5: The reviewer's comments are very valuable, and I have added the differences between non-dominant groups in different places in the discussion section. “(1)The high relative abundance of Basidiomycota in coniferous forests might be due to its strong ability to synthesize enzymes required for the degradation of complex polymers, thus increasing their relative abundance. In addition, tree communities (Picea crassifolia) have substantially greater rhizosphere resource input by root exudation, and therefore may promote Basidiomycota.(2)The relative abundance of Mortierellomycota was significantly higher in alpine scrub, which might be attributed to scrub rhizosphere resource input by root exudation. Olpidiomycota could adapt to acidic and resource-limited conditions and have higher relative abundances in the higher-elevation of sparse vegetations of limestone flats.”
Comments 6：I recommend adding a figure with photographs of the objects of study - the soil and vegetation of the analyzed biomes. This will improve the perception of information.
Responds 6: According to the reviewer's comments, enlarged and improved the quality of Figure 1 to improve the perception of information.
Comments 7：I recommend enlarging and improving the quality of Figure 1 to improve the perception of information.
Responds 7: According to the reviewer's comments, enlarged and improved the quality of Figure 1 to improve the perception of information.
Comments 8：The authors should discuss in more detail the reasons why fungi are primarily affected by vegetation, and bacteria by the physical and chemical properties of the soil. This would be interesting to a wide range of readers.
Responds 8: The results performed SEM, and the reasons covered the relationship between plant, soil and microorganisms in the discussion. According to the reviewer's comments this chapter has been modified.
Comments 9：Perhaps the authors should reduce the number of literary references and at least refer to publications in the journal "Sustainability" several times.
Responds 9: According to the reviewer's comments, references [20] and [34] were added.
- Zhao, W.; Yin, Y.; Li, S.; Liu, J.; Dong, Y.; Su, S. Soil Microbial Community Varied with Vegetation Types on a Small Regional Scale of the Qilian Mountains. Sustainability 2022, 14, 7910.
[34] Álvarez, V.E.; El Mujtar, V.A.; Falcão Salles, J.; Jia, X.; Castán, E.; Cardozo, A.G.; Tittonell, P.A. Micro-Environmental Variation in Soil Microbial Biodiversity in Forest Frontier Ecosystems—Implications for Sustainability Assessments. Sustainability 2024, 16, 1236.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript has improved; I have some additional comments.
Figures 3, 4, and 5: In the caption of the figure, please state what bars and lines indicate in the graphs.
Line 118: What do you mean by 'plant community'?
Line 121: I think the statement 'the plant professor' is too vague. Please be more specific.
Line 134: It is not clear whether the IV value is the same as the Pi; if not, where was the IV used and for what?
Author Response
Dear Editors and Reviewers,
First of all, I sincerely thank you for your valuable comments and suggestions, which have greatly improved the quality of this paper. According to your comments, I have carefully modified the manuscripts. The revised has been marked red in the revision draft, and if there are still shortcomings, I hope you can understand and criticize and correct. Thank you again for your hard work on this manuscript. The following are the specific revisions to the manuscripts:
Comments 1：Figures 3, 4, and 5: In the caption of the figure, please state what bars and lines indicate in the graphs. The specific modifications are as follows:
Fig 3. Alpha diversity index of soil bacteria along the elevation gradient.
Richness (a); Shannon (b); Simpson (c); Chao1 (d); Ace (e); Good's coverage (f); Bars represent the means ± SE (n = 4).
Fig 4. Alpha diversity index of soil fungi along the elevation gradient.
Richness (a); Shannon (b); Simpson (c); Chao1 (d); Ace (e); Good's coverage (f); Bars represent the means ± SE (n = 4).
Fig 5. Beta diversity of soil bacteria and fungi along the elevation gradient.
Soil bacterial NMDS (a); Soil fungal NMDS (b); Soil bacterial Bray-Curtis dissimilarity (c); Soil fungal Bray-Curtis dissimilarity (d); Bars represent the means ± SE (n = 4).
Comments 2：Line 118: What do you mean by “plant community” ?
Responds 2: According to the reviewer's comments, adjusted the sentence, “plant community” ? instead of “For the plant characteristics”.
Comments 3：Line 121: I think the statement “the plant professor”'' is too vague. Please be more specific.
Responds 3: According to the reviewer's comments, “the plant professor” ? term instead of “the plant taxonomist”.
Comments 4：Line 134: It is not clear whether the IV value is the same as the Pi; if not, where was the IV used and for what?
Responds 4: We didn't explain it clearly before and there was conceptual confusione. According to the reviewer's comments, the explanation of Pi has been added, “Pi =IV /100”.
Author Response File: Author Response.pdf
