Regulation of Phosphorus Supply on Nodulation and Nitrogen Fixation in Soybean Plants with Dual-Root Systems
Round 1
Reviewer 1 Report
The manuscript “Regulation of phosphorus supply on nodulation and nitrogen fixation in soybean plants with dual-root systems” analyzes the effect of phosphorus on P application side and non-application side of dual roots created by grafting as well as on the shoot. The authors have carried out an impressive job of doing a difficult experiment like grafting to obtain related data. However, there needs to be thorough proofreading, some sentence restructuring to add ease of reading and coherency to the manuscript. Please find my specific comments below:
Edits to improve consistency and clarity
As mentioned before, thorough proofreading is required for grammatical errors, use of acronyms – there are many to be pointed out individually. Sentences that need to be considered for clarity: lines 34-37, line 47 name the compounds not ‘some substances’, lines 78-82, line 372 missing citation - please cite the studies which you found your result to be consistent with, lines 402-404. Line 338 - incomplete figure legend. Lines 433 – 436 ; two sentences can be joined using which to avoid using ‘this’. Again, the last sentence has ‘this’ in a way I cannot grasp what you are referring to. There are multiple instances of using ‘this’ in the manuscript – please try restructuring as much as possible because using too many ‘this’ adds confusion.
Major request
- Since the systemic effect of phosphorus is a major conclusion of the manuscript it would be better to be more quantitative on how much phosphorous, ureide content or gene expression change was there in the non-application side when one side was applied with phosphorous vs without phosphorous application. Percentage or fold change can be calculated instead of vaguely saying ‘much more or more marked’.
- Effect of phosphorous on ultrastructure – The result is not backed up by quantitative data for the number of infected /uninfected cells, area of nodule infection, and bacteroid count. The data can be added in the supplementary to back results on section 3.3 and figure 2.
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
In the present study, the effect of effects of P supply on soybean nodulation, including nitrogenase activity, ureide content, the number of bacteroids, the number of infected cells, and the key genes related to nodulation and nitrogen fixation, were determined.
The research manuscript presents an interesting research topic and good ideas, but there are some points that need to be addressed.
Comments
Lines 16 and 17: “number of bacteroids, number of infected cells,” please provide some data in the result section. Line 89: Please add the source of seed, add botanical name [Glycine max (L.) Merr.].
Lines 109-110: If P supplied nutrients added every day, then P concentration raised in pots? How were P rates maintained?
How were the nodule numbers counted, only pink and big nodules or total nodules?
Line 267: do you know the rhizobial density in a solution used for soybean inoculation?
Line 271:” the number of ICs” and Line 284: “The results show that the numbers of ICs and Bts “, how the number of infected cells was measured? No results were presented for the infected cells and Bts.
Line 276:; why Bradyrhizobium japonicum were not used for plant inoculation?
Line 346: “phosphorus-free side. We found..”
Since nitrogen fixation is the main process in the symbiotic association of bacteria with plants, why N concentrations in plant tissue were not measured?
Line 441: “number of Bts”, please clarify. In figure 2, infected cells and uninfected cells are shown; how the number of Bt was measured?
Author Response
Please see the attachment.
Author Response File: Author Response.docx
Reviewer 3 Report
Is two biological reps enough???
Line 21 omit above
Line 30. Omit inferior to nitrogen
Line 33 change affecting to affects
Line 33: should it be “nodule nitrogen fixation”
I think there is has been a spell check problem . Be sure than nodule and NOT module is used through out the paper
Line 67 should be “existing studies”
Line 173: In the R1 stage ….
Author Response
Please see the attachment.
Author Response File: Author Response.docx
