The Role of Different Rhizobacteria in Mitigating Aluminum Stress in Rice (Oriza sativa L.)

Round 1

Reviewer 1 Report

Manuscript entitled: Rhizobacteria and their role in mitigating aluminum stress in Oryza sativa L., by Carranza-Patiño et al., demonstrates the potential of different PGPR strains in improving aluminum tolerance in rice, as well as their ability to promote growth and yield under Al stress conditions. This manuscript is of significant interest to the journal's readership. However, this manuscript needs revisions that could further improve its quality as stated in detail comments.

Manuscript entitled: Rhizobacteria and their role in mitigating aluminum stress in Oryza sativa L., by Carranza-Patiño et al., demonstrates the potential of different PGPR strains in improving aluminum tolerance in rice, as well as their ability to promote growth and yield under Al stress conditions. This manuscript is of significant interest to the journal's readership. However, this manuscript needs major revision that could further improve its quality as folows:

1.       Title: In order to present the subject of this research more clearly, the title should be modified to: The role of different rhizobacteria in mitigating aluminum stress in rice (Oriza sativa L.).

2.       Abstract: Correct the word „Pseduomonas“  to „Pseudomonas“  (Line 20).

3.       Keywords: Replace „Abiotic stres“ with „Aluminum stress“

4.       Introduction:

·         Explain in more detail the causes of Al toxicity in the soil (Lines 48-49);

·         The sentences which repeat mechanisms of PGPR should be combined into one paragraph (Lines 61-63, Lines 67-72);

·         PGPR mechanisms that are significant in mitigating Al toxicity should be included in the Introduction.

5.       Material and Methods:

·         The names and origins of the species, as well as the media and conditions used for cultivation, should be listed in section 2.1. (Lines 87-95);

·         Replace word „auxinic“ with „plant growth promoting (PGP)“ (Line 94, Table 1);

·         Title of table 1 shoud be modified to: „Characterization of rhizobacteria by their antagonistic and PGP properties“;

·         Table 1: Replace „Organism“ with „Rhizobacteria“; delete numbers „1-4“; delete „Antagonistic metabolites“; include „minus“ for negative reactions; check the names of strains (e.g., Pseduomonas);

·         Note of table 1: Delete „Metabolites“; Check the abbreviations (e.g., Pr or PR, DAPG or 2,4-DAPG); Delete „Sd“ and „IAA“ (these are not given in Table).

·         Include the origin of rice genotypes (Line 103);

·         Use and uniform abbreviations for the studied parameters throughout the text (e.g., Hypocotyl length (HL) (Line 112), Root Length (RL) (Line 114), Vigor Index (VI), Germination Percentage (GP) (Line 122), Relative Growth Index Germination Index (GI) (Line 131);

·         Replace AR and AH with RL and HL (Lines 118-121);

·         Include references for equations (Lines 118-121, Line 128, Lines 132-133);

·         If RLS is calculated using nine plants, how many plants were used for other parameters? (Line 129); this should be stated accurately in the materials and methods so that the results shown in Table 2 are clearer:

·         pH meter, spectrophotometer,  (Line 157, Line 166) – include model, manufacturer, city and country for all equipment used in this study

·         Check the codes for strains throughout the text (e.g., M-04, M -05 (Line 178), M-04, M0.5 (Line 197, 200))

6.       Results:

·         Title of table 2 shoud be modified to: „Aliminum tolerance of two rice varieties“;

·         Table 2: Delete treatment marks „T1-T5“, Delete „CA“, Use niform abbreviations for the studied parameters, replace minus with 0 and inlude letters for statistical significance for all parameters;

·         Table 2 Notes: Use niform abbreviations for the studied parameters.

·         Title of Figure 1 should be modified to avoid the repetiton of the methodology.

·         Figure 1: Mark aluminum concentrations on Figure 1, as on Figure 3; Check the red arrows that should indicate root necrosis at 8 and 16 mM (it seems like they currently point out 4 and 8 mM)

·         Al2 – Al₂ (Line 248);

·         Figure 2: UFC or CFU? veronni or veronii? Include the explanation for bars like in other figure notes; replace „milimoles“ with „Concentration of Al₂(SO₄)₃“.

·         The decription of the results presented on Figure 3 is missing in the text.

·         Figure 3: The red arrows indicate the degree of tolerance at a 16 mM concentration are not visible;

·         Figure 3: „Al concentration in X, while abundance of specific species in Y from the figure“ – It is not clear what this refers to;

·         Fugure 4: Optical density at 600 nm (Y); Concentration of Al₂(SO₄)₃ instead Concentrations (X);

·         Figure 4,5,6,7,8,9: P. veronni – correct the species name;

·         Figure 5: Alimunum concentrations - Concentration of Al₂(SO₄)₃;

·         Figure 6-9. Consorcio, P. Rhizobacterias – correct; the species names be italicized;

·         Figures 4,5,6,7,8,9: according to Tukey - according to Tukey's test;

·         Figures 7, 8: the title for other Y axis is missing

·         applied (Line 340, 352) – find better word;

·         Figure 8: leaf or aerial? Equalize terms

·         Line 358-359: Use symbol × instead X

·         Figure 9: UFC – CFU

·         Check citations in the text (e.g. ref. 52: Elisa Azura et al.)

7.       Discussion

·         A possible contribution of the specific rhizobacterial species/strain in mitigating aluminum stress should be discussed more thoroughly;

8.       Conclusion

·         P. veronni (line 475)

Author Response

Dear reviewer 

Thank you very much for your suggestions, each of your comments have been answered in the attached document.

Kind regards

Author Response File: Author Response.docx

Reviewer 2 Report

See the attachment for details.

The manuscript discusses four kinds of plant rhizotropic bacteria (PGPR) in alleviating the effects of aluminum toxicity on rice, which has certain academic value and practical significance. The experimental design is reasonable, and the results have important guiding effect, especially in improving crop productivity in acidic soil. But there are still some problems, as follows:

1. Preface: Please avoid sentences with the meaning of "filling a research gap". Alternative: Although PGPR is widely used to improve plant growth, its mechanism for mitigating aluminum toxicity remains limited, especially in rice, a global food security crop."

2. Why choose these four strains? Although the ability of the strain to antagonize and promote metabolites has been mentioned, the specific mechanism of its action in alleviating aluminum toxicity has been less discussed. It is suggested to supplement the mechanical research background related to the selected strains.

3. The results and analysis need to be more detailed: although ANOVA and Tukey tests were used, only the data were described during the analysis. It can be seen from the results in the figure that not every group presents a significant difference, but it does not mention the significance of the significant difference between specific groups. There is also no brief analysis, and this section is strongly recommended to be rewritten.

4. Chart improvement: The chart needs to be unified in style and clearly marked. The chart uses consistent colors or legends to identify different strains and treatment conditions to avoid confusion.

5. The discussion needs to be more in-depth: At present, the summary of the results in the discussion section is relatively comprehensive, but the elaboration of some key mechanisms (such as organic acid secretion, biofilm formation to alleviate aluminum toxicity) is too simple. It is recommended to explain in detail the specific biochemical effects of PGPR in the mitigation of aluminum toxicity in combination with the literature. Increased analysis of synergies between different PGPR strains to reveal the advantages of their combined application.

See the attachment for details.

Comments for author File: Comments.pdf

Author Response

Dear reviewer 

Thank you very much for your suggestions, each of your comments have been answered in the attached document.

Kind regards

Author Response File: Author Response.docx

Reviewer 3 Report

I have thoroughly reviewed the manuscript and found it necessary. Topic have be clearly explained and experiments and results are presented in a very well manner. overall manuscript quality is good, but some improvements are needed which are suggested below,

1- section wise comments are given in each section above

2- authors must add some limitations of the study and suggestion to overcome those especially in discussion in conclusion section. 

 

Comments have already been given above 

Author Response

Dear reviewer 

Thank you very much for your suggestions, each of your comments have been answered in the attached document.

Kind regards

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Thanks to the author for the revision.

Thanks to the author for the revision.

Reviewer 3 Report

authors have sufficiently improved the manuscript as per suggestions and now it cant be processed further 

authors have sufficiently improved the manuscript as per suggestions and now it cant be processed further