Spermosphere Bacteria Promote Ormosia henryi Seed Germination by Activating Metabolic Pathways

Round 1

Reviewer 1 Report

The main goal of the research is to study the effect of spermospheric bacteria on the germination of seeds of Ormosia henryi, due to the activation of metabolic pathways.

The article presented by the authors is very relevant, as it is devoted to the issues of ensuring the germination of seeds of Ormosia Henry in natural conditions, which is a valuable tree species in China and has great economic and environmental potential.   But it is endangered.   Many researchers have studied the effect of spermosphere microorganisms on the germination of seeds of various tree species, but their effect on Ormosia Henry is studied by the authors for the first time.

To ensure seed germination under natural conditions, the authors investigated spermospheric microorganisms that promote seed germination of O. Henry, mainly by activating galactose metabolism and the lysine degradation pathway.   The main microbial groups of the spermosphere have been determined.   It has been established that differential microorganisms between the two groups can reduce the time of seed germination, increase the rate of germination and growth of seedlings.   The article widely used statistical and correlation methods of data processing.

The article is written in a clear,  a strict and competent scientific language, the authors use special terminology, the latest and authoritative literature.

In summary, the spermosphere bacteria of O. henryi seeds play a vital role in seed germination.   Under appropriate climatic conditions, at the early stage of germination, seeds may rely on the acid environment of the soil created by the bacteria themselves and on soil pH to promote germination.   At the later stages of germination, the spermosphere bacteria mainly maintain the development of seeds by activating more energy metabolism pathways.

Of course, a lot of research work has been done, but the illustrative material given in the article is cumbersome and poorly readable (for example, figures 1 and 5). It will be better to present them in the form of tables.

The article is recommended for publication, taking into account the comments.

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

Dear Authors

The paper submitted for review entitled "Spermosphere bacteria promote Ormosia henryi seed 2 germination by activating metabolic pathways" contains many interesting results, but requires a more complete discussion and interpretation. Here are the detailed notes:

1)     The abstract is not structured. Purpose is not explained, Summary should be the most important conclusion

2)     In the "Introduction" chapter, the purpose of the research should be better explained and an alternative research hypothesis should be put forward in relation to the null hypothesis, in order to then verify it

3)     in chapter 2.6, the description of the experiment is confused with the statistical analysis. What model was the experiment performed on? Was seed treatment (sterilized and unsterilized seed) the only variable? Means and standard deviations are not enough to determine statistically significant differences. To do this, you need to perform an ANOVA and apply confidence intervals (e.g., Student's t, Tukey). Power analysis is typically used to determine the smallest significant difference between data. Statistical power analysis estimates how large samples are needed to detect a specific difference between groups or variables at a given level of significance and test power.

a.      Here are some steps you can take to estimate the smallest significant difference between the data:

b.     Define research hypotheses: Define clearly your research hypotheses, i.e., what exactly do you want to compare and what is the expected difference between groups or variables.

c.      Select the significance level: Specify the significance level, which represents the maximum acceptable chance of making a Type I error (rejecting the null hypothesis when it is true). The standard significance levels used are 0.05 (5%) or 0.01 (1%).

d.     Determine test power: Test power is the ability of a statistical test to detect a difference if there is a difference (i.e., rejecting the null hypothesis when it is false). Typical test power values are 0.8 or 0.9, meaning that the test has an 80% or 90% chance of detecting a significant difference.

e.      Choose the right statistical test: Choose the right statistical test for your research project, for example, Student's t-test, analysis of variance (ANOVA) or other appropriate methods.

f.      Perform power analysis: Use available tools or statistical software that enables power analysis. Based on predetermined values (significance level, test power, sample size), the tool can estimate the smallest significant difference between the data.

g.     Statistical power analysis is important to estimate the appropriate sample size and potentially avoid statistical errors due to insufficient sample size. However, statistical power analysis can be more complicated and require advanced statistical knowledge. Consult a statistical expert or use available statistical tools to perform a power analysis in the context of your specific study.

h.     Was the number of results sufficient to calculate the correlation? (2 treatments x 3 repetitions = 6) and the minimum number of data needed to calculate the correlation is 12.

i.      To compute a simple Pearson correlation, you need two sets of data, usually of equal length. Let's denote one dataset as X = {x1, x2, ..., xn} and the other dataset as Y = {y1, y2, ..., yn}.

4)     The chapter "Research Results" is insufficiently covered:

a.      Table 1 does not show the significance of statistical differences - on what basis the differences between the objects were found. Standard deviations are not appropriate for discussing the significance of differences. Please perform ANOVA calculations and find confidence intervals (LSD) and these should be the basis for determining whether seeds germinate in sterilized and unsterilized soil

b.     Section 3.2, when considering the significance of dominant bacterial interactions, simple correlation coefficients should be used.

c.      Figure 1 is confusing and completely illegible, from which it is impossible to read any values or differences in this form of presenting the results. Please present these data in a legible table and provide the value of the smallest significant difference for each comparison of features to determine whether they really differed significantly. Similarly, figure 2 is not sufficiently described and interpreted. It seems that the authors do not fully understand the significance of the obtained results

d.     In Figure 3, the data values have been marginalized and reduced so that no significant differences can be seen at any significance level. The results at each stage of the research should be presented in a separate figure and discussed separately.

e.      Similarly, the results in Figure 4 are completely illegible. In this form, the presented results are not suitable for interpretation.

f.      The authors were not able to correctly, statistically interpret the results of the tests presented in Figure 5, although they refer to significant differences at p0.005 and p0.01, which are not visible at such a large reduced size of the figure.

g.     The analysis of the data presented in Figure 6, due to the large reduction in the scale of the figure, is impossible to interpret, and the authors are unable to statistically interpret the results of the study.

5)     The "Discussion" chapter should be divided into subchapters and the results should be thoroughly discussed.

6)     Conclusions are generalized and taken too far. There is insufficient evidence for some of them.

Comments for author File: Comments.pdf

Dear Editor,

The work in this form is not suitable for printing. Requires revision and re-review

Author Response

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Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Dear Authors,

The authors' answers, as well as the changes made to the manuscript and the materials included in the appendices, meet the requirements for statistical analysis of research results and its proper interpretation. The authors also included 2 alternative research hypotheses, which they then verified. They also corrected and supplemented the discussion of the results and verified the conclusions. I accept their explanations and answers. The manuscript with the materials included in the supplement is suitable for printing.

Comments for author File: Comments.pdf

Only a minor edition of the English language is required