Exploring the Identity and Properties of Two Bacilli Strains and their Potential to Alleviate Drought and Heavy Metal Stress
Round 1
Reviewer 1 Report
Abstract
Add the important methods briefly in the Abstract.
Introduction
Line 41-52 in this para, the authors straight to the point about PGPR. Should the initial statement about the role and contribution of microbes in general for agriculture, need to be elaborated. There are rhizobium bacteria, mycorrhizal fungi, etc. which interact with PGPR (add references related to this)
Materials and Methods
Line 120-126 Please explain the history of land briefly. How long (years or months) the farmers planted Vigno mungo and Phaseoulus vulgari. Also, put in the text, the data of soil chemical conditions (pH, N, P, K, Ca, Mg, CEC, etc.) and some micro-nutrients, population of bacteria and fungi in the land.
Results
Could you provide some microscopic observations of Bacillus cereus and Bacillus haynesii? (including how to proceed with materials and methods)
Table 2. please make an open table
Discussion
what are the future prospects of these two species of bacteria for the utilization of environmental improvement? what processes are needed so that these two species of bacteria can be utilized on a business scale? Please add some references.
Author Response
Thanks for your suggestions. The revisions have been mentioned in the attached file and incorporated in the revised manuscript.
Regards
Author Response File: 
Author Response.docx
Reviewer 2 Report
General comments:
The manuscript describes the isolation and identification of bacterial strains from soil samples, planted with legume species. Analysis of EPS and ACC deaminase production hints to a potential as PGPRs. These might improve stress tolerance of legumes, especially cowpea. The English writing must be revised. The authors use extremely long sentences with errors in phrase structure and grammar. The methodology described in the Material and Methods section is very difficult to follow. Authors should avoid repetitions throughout the manuscript.
Detailed comments:
Abstract:
L13-16: I would suggest separation of this phrase into two sentences.
L16: Please correct - in the present study
L21-24: These bacterial strains exhibited higher…Please add some more information about the type of assay that was performed
L25: Please correct - These bacterial isolates with proven ability of an enhanced production of ACC deaminase
L26: Please correct - …. resistant strain can promise to be a potential heavy metal stress alleviator.
L27-29: Please include some details about the assay for heavy metal sensitivity (plate assay?)
Introduction:
L333-36: I would suggest: Human population keeps on increasing year by year and total population is expected to reach nine billion by 2050. This increase is accompanied by a rapid urbanization and industrialization, which inevitably leads to environmental stress and damage. Consequences are higher evaporation and transpiration, which eventually leads to global warming.
Please define “damage”
L77: Please correct - Finding ways to enable plants to adjust to environmental drought stress while maintaining normal plant growth is necessary in order to satisfy global food requirements.
L87: Literature reveals that chickpea plant also produces ethylene under stress condition - please mention the type of stress
L89: please correct - The outcome of this study was that one strain AV-7 out of the seven strains could be selected to carry out for a more in-depth study.
L100: Why did authors decide to work with AV-12? How was it identifies?
Title Table1: What means here “along with their soil sample sources”?
Table 1: In AV-12 – Morphology characteristics are displaced (Gram reaction – Bacillo (rod)
Table 1 could go to Results
Material and Methods:
122: I suggest: microbiological analysis including Gram staining and preliminary qualitative biochemical characteristics, were performed as describe previously (19).
Biochemical characteristics and Plant Growth Promoting Rhizobacterial traits evaluation: These paragraphs should be re-written. They seem to include aspects that should go to introduction (i.e., 138-140) and results (L128- Indole test demonstrated decomposition of amino acid and tryptophan to indole by certain bacteria which got accumulated in the media – is this a result?)
Authors should include number of repetitions.
L202: Please correct - collected from root zone
L229-234: This should go to discussion. Does this statement mean that a high ACC deaminase activity is not related to root growth and activity as PGPR. Why than was this parameter used here as a measure for PGPR activity?
L265: EPS – abbreviation needs to be explained
Heavy metal tolerance: How many repetitions? How was tolerance assayed? (Plate assay?)
Results:
L302: this is repetition from Material and methods
L317: Quantity of extracted DNA and gel picture (Fig. 1) are not relevant.
L330: repetition from L323
L335: repetition from L330
L371-373: should go to introduction
Discussion:
L446: acds gene was not detected? Does this refer to the expression of the gene?
L442-447: the statements here are very confusing. Are authors talking about mutations in the ACDs gene?
L454: genera
L481,482: identical gene size does not mean identical protein
The English writing must be revised. The authors use extremely long sentences with errors in phrase structure and grammar. The methodology described in the Material and Methods section is very difficult to follow. Authors should avoid repetitions throughout the manuscript. Discussion on ACC diaminase gene analysis in bacterial strains is not clear at all. The introduction focuses on the possible usage of the identified strains in cowpea cultivation, but no plant experiments were conducted. Results on the resistance to heavy metals are presented only superficially, also this topic is part of the title.
Author Response
The authors are grateful to the reviewer for very important suggestions. The suggested changes have been incorporated in the revised manuscript. The detail is given in the attached pointwise response.
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
this manuscript is suitable to publish
Author Response
Corrections have been incorporated and marked.
Reviewer 2 Report
The authors adressed the comments to the original manuscript and thereby improved the manuscript considerably. The English writing should be revised once again carefully.(I.e. In a recent study by Glick et al (2021) reported molecular weight of ACC deaminase from Pseudomonas strains.... - should be: A recent study by Glick et all reported a molecualr weight....)
Author Response
The suggested corrections have been incorporated and marked with track changes.
