Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Original research covering the biocontrol potential of microbial consortia to suppress phytopathogens, while improving plant health parameters. The terms “forest” and “forestry” are used interchangeably throughout the manuscript, albeit incorrectly. Authors are strongly encouraged to revise the manuscript entirely and use the appropriate term. Species abbreviations can also be used after they are first referred in the text; use this consistently throughout the manuscript. I provide major and specific comments to improve the current version.

 

Major comments:

Abstract & Introduction: The abstract is concise and globally well-structured. The introduction’s structure can and must be improved (see specific comments).

Material & Methods: methods are adequate, but the structure can and should be improved. Subsections are too long and can be shortened to focus attention.

Results & Discussion: results and figures are nicely presented. The discussion can and should be more concise to focus attention.

 

Specific comments:

Abstract

Line 39: Candida albicans is a pathogenic yeast in humans, not plants. Also, order species name by alphabetical order.

Lines 40-41: authors are advised to order these 4 species alphabetically to facilitate reading.

Line 46: what does LRV stand for?

Line 49: remove the dash.

Line 50: compared to the control.

Line 53: which fungi cause powdery mildew and Schütte disease? If you want to refer these diseases, it should be clearly stated (at the beginning of the abstract).

Lines 54-55: “The Biogeosystem Technique methodology has been developed to improve a long-term forest growing.” – this sentence is irrelevant; I would remove it entirely.

 

Keywords: They need to be carefully considered and shorter in length. Change “microorganismal consortium” to microbial consortium.

 

Introduction

Lines 63-65: Authors are advised to rephrase the sentence to “Biological and ecological functions of forests in ensuring land restoration and conservation of biological resources are of vital importance to human well-being.”

Line 66-67: anthropogenically disturbed lands

Lines 71-74: “A forestry on unproductive lands unsuitable for agriculture, as well as the forest restoration and re-introduction of forest species, are of a paramount importance in the economic and recreational sphere of social development.” – this sentence is difficult to understand, and the beginning makes no sense; authors are advised to rephrase.

Line 75: re-afforestation?

Line 77: “launched in 2013” – is there a reference for this?

Lines 86-88: “Many authors have shown” – references are missing in this sentence.

Line 107: “equipment” is not an appropriate word for living organisms; authors are encouraged to change “biological protective equipment” to biopesticides or bioprotection products.

Lines 108-110: “The Moscow region is provided to the western part of the Volga basin. Due to its natural location, the Republic of Tatarstan is located in the zone of the Middle Volga upland.” – these two sentences are out of place.

Lines 117-123: “An aim of the study was as follows. An extraction of isolates of microorganism strains from natural sources in various ecological niches in the Moscow region and in the Republic of Tatarstan. A synthesis of a biological product, stimulating a Scots pine, pedunculate oak and small-leaved linden seedlings protection from diseases, caused by fungi Lophodermium pinastri Chev. and Microsphaera alphitoides. A Biogeosystem Technique methodology development to improve a long-term forest growing.” – the entire paragraph was copy-pasted here and makes no sense; authors are strongly advised to rewrite it.

 

Material and Methods

Lines 125-173: The subsection “Region of Investigations” is far too extensive and descriptive; it can and should be shortened to focus the reader’s attention.

Line 177: “Quercus robur” (without accents)

Lines 177-178: “Tilia cordata” (without accents)

Line 179: “bacterial isolates” which isolates were used?

Line 216: “[...] dry weight of root and [...]”

Lines 254, 259 and 263: why are there three different temperatures? Why were these in vitro experiments not subject to the same conditions?

Line 261: “specified medium” – which was...?

Lines 329-330: There are more robust statistical softwares than Microsoft Excel 10 available for free. I suggest Jamovi: https://www.jamovi.org/

 

Results

Line 343: Genera instead of “genus”

Line 344: “4 strains” – authors are listing four genera and not strains; please, rectify this!

Table 1: “Combination” – does this refer to a combination of all four isolates? Two of them? This needs to be clarified and clearly stated in the Table legend.

Comments on the Quality of English Language

Extensive editing of English language required.

Author Response

Dear Reviewer 1,

 

Thank you for your comments.

We accepted all comments with gratitude. The answers are presented below (highlighted in yellow) and in the text of the manuscript.

 

 

Comments and Suggestions for Authors

Original research covering the biocontrol potential of microbial consortia to suppress phytopathogens, while improving plant health parameters. The terms “forest” and “forestry” are used interchangeably throughout the manuscript, albeit incorrectly. Authors are strongly encouraged to revise the manuscript entirely and use the appropriate term. Species abbreviations can also be used after they are first referred in the text; use this consistently throughout the manuscript. I provide major and specific comments to improve the current version.

The term “forestry” was excluded from the text.

 

Major comments:

Abstract & Introduction: The abstract is concise and globally well-structured. The introduction’s structure can and must be improved (see specific comments).

Material & Methods: methods are adequate, but the structure can and should be improved. Subsections are too long and can be shortened to focus attention.

Results & Discussion: results and figures are nicely presented. The discussion can and should be more concise to focus attention.

Accepted

The introduction’s structure changed

Methods’ structure changed. Subsections are shortened.

The discussion was shortened and supplied with the links to the references to focus attention.

 

Specific comments:

Abstract

Line 39: Candida albicans is a pathogenic yeast in humans, not plants. Also, order species name by alphabetical order.

You are right. Candida albicans is a pathogenic yeast in humans. However, reducing the number of pathogens in the environment we make it more safe not only for a plant but also for a human.

The corresponding changes were done.

The species were listed by alphabetical order.

 

Lines 40-41: authors are advised to order these 4 species alphabetically to facilitate reading.

The species were ordered alphabetically

 

Line 46: what does LRV stand for?

LRV stands for the author’s full name – Lenar R. Valiullin.

 

Line 49: remove the dash.

The dash has been removed

 

Line 50: compared to the control

The text has been changed

 

Line 53: which fungi cause powdery mildew and Schütte disease? If you want to refer these diseases, it should be clearly stated (at the beginning of the abstract)

Powdery mildew and Schütte disease are caused by the group of molds (information is given in the Abstract and in the text).

 

Lines 54-55: “The Biogeosystem Technique methodology has been developed to improve a long-term forest growing.” – this sentence is irrelevant; I would remove it entirely.

The Biogeosystem Technique methodology allusion was excluded in the lines 54-55.

 

Keywords: They need to be carefully considered and shorter in length. Change “microorganismal consortium” to microbial consortium.

The keywords were shortened. The “microorganismal” were changed to “microbial”.

 

Introduction

Lines 63-65: Authors are advised to rephrase the sentence to “Biological and ecological functions of forests in ensuring land restoration and conservation of biological resources are of vital importance to human well-being.”

The sentence was rephrased.

 

Line 66-67: anthropogenically disturbed lands

The sentence was rephrased.

 

Lines 71-74: “A forestry on unproductive lands unsuitable for agriculture, as well as the forest restoration and re-introduction of forest species, are of a paramount importance in the economic and recreational sphere of social development.” – this sentence is difficult to understand, and the beginning makes no sense; authors are advised to rephrase.

The sentence was rephrased.

 

Line 75: re-afforestation?

Reforestation.

 

Line 77: “launched in 2013” – is there a reference for this?

The reference was added:

State program "Development of forestry in the Tatarstan Republic in 2014-2024", 2013 https://docs.cntd.ru/document/561428341/titles/2VJORFD (accessed May 29, 2023)

 

Lines 86-88: “Many authors have shown” – references are missing in this sentence.

Some text fragments were deleted. The links were added to the text.

 

Line 107: “equipment” is not an appropriate word for living organisms; authors are encouraged to change “biological protective equipment” to biopesticides or bioprotection products.

The term “equipment” was excluded.

Lines 108-110: “The Moscow region is provided to the western part of the Volga basin. Due to its natural location, the Republic of Tatarstan is located in the zone of the Middle Volga upland.” – these two sentences are out of place.

The two sentences in the lines 108-110 were deleted.

Lines 117-123: “An aim of the study was as follows. An extraction of isolates of microorganism strains from natural sources in various ecological niches in the Moscow region and in the Republic of Tatarstan. A synthesis of a biological product, stimulating a Scots pine, pedunculate oak and small-leaved linden seedlings protection from diseases, caused by fungi Lophodermium pinastri Chev. and Microsphaera alphitoides. A Biogeosystem Technique methodology development to improve a long-term forest growing.” – the entire paragraph was copy-pasted here and makes no sense; authors are strongly advised to rewrite it.

The lines 117-123 were re-written.

 

Material and Methods

Lines 125-173: The subsection “Region of Investigations” is far too extensive and descriptive; it can and should be shortened to focus the reader’s attention.

The subsection “Region of Investigations” was shortened.

 

Line 177: “Quercus robur” (without accents)

“Quercus robur” was spelled without accents.

 

Lines 177-178: “Tilia cordata” (without accents)

“Tilia cordata” was spelled without accents.

 

Line 179: “bacterial isolates” which isolates were used?

You are right, in the first paragraph of the Section 3 and in the Table 1 we mentioned the genera, the text is corrected. However, below into the text we talk about the four selected strains of the 46 isolated microorganisms.

Subsection 2.2.

The origin of Bacterial product LRV, its properties and test in the controlled environment conditions showed an effectiveness of Bacterial product LRV are presented in details below in the text.

 

 Line 216: “[...] dry weight of root and [...]”

Re-written

 

Lines 254, 259 and 263: why are there three different temperatures? Why were these in vitro experiments not subject to the same conditions?

Excuse our inattentiveness. The lines 254, 259 and 263 are about the same single experiment. A cause of your misunderstanding was that we were mistaken with the temperature. This is the same in every line. Corrected to 28–30 °C.

 

Line 261: “specified medium” – which was...?

In the line 261, we explained one more time that the specified medium was Czapek-Dox agar.

 

Lines 329-330: There are more robust statistical software than Microsoft Excel 10 available for free. I suggest Jamovi: https://www.jamovi.org/

The list of the statistical procedure used has been widened. 

 

Results

Line 343: Genera instead of “genus”

“Genera” was replaced by “genus”.

 

Line 344: “4 strains” – authors are listing four genera and not strains; please, rectify this!

You are right, in the first paragraph of the Section 3 and in the Table 1 we mentioned the genera, and the text was corrected correspondingly. However, below into the text we talk about the four selected strains of the 46 isolated microorganisms.

 

Table 1: “Combination” – does this refer to a combination of all four isolates? Two of them? This needs to be clarified and clearly stated in the Table legend.

“Combination” means a combination of four test cultures. We added this informantion to the Table 1.

 

 

 

Comments on the Quality of English Language

Extensive editing of English language required.

Extensive editing of English language provided. Excluding Authors, Affiliations, Tables, Author Contributions, Funding, Data Availability Statement, Conflicts of Interest and References, the manuscript AJE score is 8.9 https://secure.aje.com/en/researcher/grammar-check/details/7c111451-ea88-4abd-8bd4-350e8fdb641e. This score is higher than that the AJE platform could provide. Nevertheless, we will finish the English language editing at the final stage of review.

 

Regards,

From the name of the Authors,

Valery P. Kalinitchenko

 

29 June 2024

Reviewer 2 Report

Comments and Suggestions for Authors

I reviewed the article” Microorganism isolates stimulate forest crop protection growth and sustainability” but its needs major revision before publication. See some my suggestion below.

1.       The tittle is wrong, so its needs write a revise the tittle

2.       The article is full of English Grammatical and writing errors please revised carefully see below few examples.

Line 68-70 , . It is an important tool in a land 68 management that increases biodiversity, CO2 sequestration, carbon content 69 in the soil (as part of humus) and prevents an uncontrolled transport of 70 pollutants within the ecosystem.

 

Line 292-295  “To study the enzyme activity of the selected isolates, bacterial cultures 291 were grown on agar modified corn-lactose and MRS media, as well as on a 292 synthetic medium containing (g/l) sodium citrate – 1.29; (NH4)2HPO4 – 4.75; 293 K2HPO4 – 9.6; MgSO4•7H2O – 0.18 (рН 7.0±0.2) [36–39]. 294 Carboxymethylcellulose (CMC), water-soluble starch, and casein at a 295 concentration of 1.0% were used as sources of carbon and nitrogen.”

3.       All figures and tables need revision because its resolution and structure are not good.

4.       Conclusions section also have a lot of English writing mistakes

Comments on the Quality of English Language

Extensive editing of English language required

Author Response

Dear Reviewer 2,

 

Thank you for your comments.

We accepted all comments with gratitude. The answers are presented below (highlighted in yellow) and in the text of the manuscript.

 

 

 

Comments and Suggestions for Authors

I reviewed the article” Microorganism isolates stimulate forest crop protection growth and sustainability” but its needs major revision before publication. See some my suggestion below.

 

1. The tittle is wrong, so its needs write a revise the tittle

The tittle was revised and re-written

 

2. The article is full of English Grammatical and writing errors please revised carefully see below few examples.

The English grammatical and writing errors were revised and re-written.

Line 68-70 , . It is an important tool in a land 68 management that increases biodiversity, CO2 sequestration, carbon content 69 in the soil (as part of humus) and prevents an uncontrolled transport of 70 pollutants within the ecosystem.

Line 292-295  “To study the enzyme activity of the selected isolates, bacterial cultures 291 were grown on agar modified corn-lactose and MRS media, as well as on a 292 synthetic medium containing (g/l) sodium citrate – 1.29; (NH4)2HPO4 – 4.75; 293 K2HPO4 – 9.6; MgSO4•7H2O – 0.18 (рН 7.0±0.2) [36–39]. 294 Carboxymethylcellulose (CMC), water-soluble starch, and casein at a 295 concentration of 1.0% were used as sources of carbon and nitrogen.”

The formulae were corrected

 

3. All figures and tables need revision because its resolution and structure are not good.

All the tables were restructured. Concerning the figures, we used a 300 dpi photo image resolution according to the Sustainability author’s guide. Perhaps, the photos are perceived as too small in the text now, but let the editors choose the picture size when preparing the proof.

 

4. Conclusions section also have a lot of English writing mistakes

Conclusions section re-written, English writing mistakes were corrected.

 

 

Comments on the Quality of English Language

Extensive editing of English language required

Extensive editing of English language provided. Excluding Authors, Affiliations, Tables, Author Contributions, Funding, Data Availability Statement, Conflicts of Interest and References, the manuscript AJE score is 8.9 https://secure.aje.com/en/researcher/grammar-check/details/7c111451-ea88-4abd-8bd4-350e8fdb641e. This score is higher than that the AJE platform could provide. Nevertheless, we will finish the English language editing at the final stage of review.

 

Regards,

From the name of the Authors,

Valery P. Kalinitchenko

 

29 June 2024

Reviewer 3 Report

Comments and Suggestions for Authors

The topic of the manuscript is relevant to solving the problems of forest maintenance. Sustainable forestry requires a comprehensive approach to reforestation. Thus, these studies seek to investigate a very important problem - the control of fungal diseases.

The title reflects the content of the manuscript.

Abstract. The abstract corresponds to the research results. The obtained results are clearly and briefly presented in this chapter.

Keywords. The keywords reflect the content, but I'd suggest renouncing the long sentence "Scots pine, ...seedlings" and just using the terms.

Introduction. In my opinion, this section needs to be corrected by the authors. The quote (p.2, lines 75-85) is narrative, only one source is cited. The question also arises - are synthetic pesticides used in Russian forests (p. 2, lines 86-90)? If so, the negative impact on the forest ecosystem is unpredictable. Please clarify this issue. In this chapter, the problems of diseases spread on forest trees need to be revealed more.

 

Methods. The authors provide too broad a general characterization of the whole region.  Section 2.2, which characterizes the experimental site, is more important. So, I would suggest shortening section 2.1 significantly, because this information is redundant. On the other hand, I missed an assessment of how the influence of local environmental conditions was treated. Such experiments must first be conducted in a controlled environment. Only in a controlled environment conclusions can be drawn about the effectiveness of Bacterial product LRV.

In section 2.3. it is not clear how in what quantities the material was collected, for example, what were the farm birds, animals, etc.

Duncan’s or Tuckey’s tests should be used in the statistical analysis of such experiments.

 

Results. The results are presented in 6 Tables and 5 Figures. The first figure in the methods section is more appropriate for a popular article and is not necessary. As I mentioned, to compare the results, it is not enough to indicate the means, errors and Fisher's coefficients in the tables. Statistically significant differences should be demonstrated using Duncan's or Tuckey's tests.

Discussion. In this section, the authors evaluate their results by comparing them with the studies of other authors. The innovativeness of the research is presented. But again, there is a lot of narrative in this chapter as well (p. 14, lines 486-496). Here it is necessary to at least quote other authors, avoiding common reflections.

Conclusions focus on the issues raised in the manuscript, concentrate on the main findings, and present the state-of-the-art of this study.

Author Response

Dear Reviewer 3,

 

Thank you for your comments.

We accepted all comments with gratitude. The answers are presented below (highlighted in yellow) and in the text of the manuscript.

 

 

 

Comments and Suggestions for Authors

The topic of the manuscript is relevant to solving the problems of forest maintenance. Sustainable forestry requires a comprehensive approach to reforestation. Thus, these studies seek to investigate a very important problem - the control of fungal diseases.

The title reflects the content of the manuscript.

Abstract. The abstract corresponds to the research results. The obtained results are clearly and briefly presented in this chapter.

Keywords. The keywords reflect the content, but I'd suggest renouncing the long sentence "Scots pine, ...seedlings" and just using the terms.

The long sentence renounced.

 

Introduction. In my opinion, this section needs to be corrected by the authors. The quote (p.2, lines 75-85) is narrative, only one source is cited. The question also arises - are synthetic pesticides used in Russian forests (p. 2, lines 86-90)? If so, the negative impact on the forest ecosystem is unpredictable. Please clarify this issue. In this chapter, the problems of diseases spread on forest trees need to be revealed more.

The sources in the lines 75-85 were added.

The synthetic pesticides were not used in the study.

The problems of diseases spread on forest trees need were revealed in the first paragraph of Introduction: The tree diseases strongly limit a success of silviculture. Bacteria are an important biological control agents of forest plant diseases.

 

Methods. The authors provide too broad a general characterization of the whole region.  Section 2.2, which characterizes the experimental site, is more important. So, I would suggest shortening section 2.1 significantly, because this information is redundant. On the other hand, I missed an assessment of how the influence of local environmental conditions was treated. Such experiments must first be conducted in a controlled environment. Only in a controlled environment conclusions can be drawn about the effectiveness of Bacterial product LRV.

The subsection 2.1 has been shortened significantly.

Subsection 2.2.

The Bacterial product LRV was tested in the controlled environment conditions. The controlled environment tests showed an effectiveness of Bacterial product LRV presented in details in the text. Please see the Table 2 and discussion. After these tests, the experiment in the open environment was fulfilled. You are right, this is a too complicated task to full scale control the seedling growth in the open environment. However, we have no other environment for our experiment now. To overcome the discrepancy between the laboratory closed controlled environment and open uncontrolled environment we developed the Biogeosystem Technique (BGT*) methodology that is mentioned in the text briefly. The BGT* is an instrument to provide an open environment higher level control.

 

In section 2.3. it is not clear how in what quantities the material was collected, for example, what were the farm birds, animals, etc.

A material for experiment was collected in the following quantities: soil 1 kg, fermented dairy products 2 l, raw cow's milk 2 l, farm birds 0,5 g, animal’s gastrointestinal tract contents 0,15 kg, feces 0.300 kg, agricultural crops 3 kg.

In experiment, 60 g of every material were used for extraction.

 

Duncan’s or Tuckey’s tests should be used in the statistical analysis of such experiments.

Tukey test was used in the statistical analysis.

 

Results. The results are presented in 6 Tables and 5 Figures. The first figure in the methods section is more appropriate for a popular article and is not necessary. As I mentioned, to compare the results, it is not enough to indicate the means, errors and Fisher's coefficients in the tables. Statistically significant differences should be demonstrated using Duncan's or Tuckey's tests.

Figure 1 has been excluded.

Statistically significant differences were demonstrated using Tukey test.

 

Discussion. In this section, the authors evaluate their results by comparing them with the studies of other authors. The innovativeness of the research is presented. But again, there is a lot of narrative in this chapter as well (p. 14, lines 486-496). Here it is necessary to at least quote other authors, avoiding common reflections.

We weakened a narrative sound of Discussion. The links to the literature sources were added.

 

Conclusions focus on the issues raised in the manuscript, concentrate on the main findings, and present the state-of-the-art of this study.

Thank you for your high assessment of Conclusions.

 

 

Regards,

From the name of the Authors,

Valery P. Kalinitchenko

 

29 June 2024

 

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Review of “Ensuring tree protection, growth and sustainability by microbial isolates”

 

Original research covering the biocontrol potential of microbial consortia to suppress phytopathogens, while improving plant health parameters. I provide some more comments to further improve the manuscript.

 

Major comments:

Abstract & Introduction: The abstract is concise and globally well-structured. The introduction’s structure can and must be improved (see specific comments).

Material & Methods: methods are adequate, but the structure can and should be improved.

Results & Discussion: results and figures are nicely presented.

 

Specific comments:

Abstract

Line 39: You are right when you claim that “reducing the number of pathogens in the environment we make it safer not only for a plant but also for a human.” However, this is not what you imply when you wrote “[…] microscopic fungi that cause various plant diseases: Aspergillus flavus, Candida albicans, […]”; you are talking about plant pathogens, not the ones susceptible of causing harm in humans. Simply write something like: “[…] microscopic fungi that can cause animal and plant diseases: Aspergillus flavus, Candida albicans, […]”.

 

Introduction

Lines 62-63: “Currently, violations in forest cultivation have a negative impact on the climate and cause land anthropogenic disturbance.”

Lines 64-65: “[…] provide human well-being.”

Line 65: “Forest and agricultural lands […]”

Lines 69-70: “Bacteria are important biological control agents […]”

 

Material and Methods

Line 158: Since you are starting the sentence with a number, write it out: “Fifty seedlings were selected […]”

Line 264: I still think Microsoft Excel 10 should’ve been avoided altogether, when there are more robust statistical softwares available for free, like Jamovi: https://www.jamovi.org/

 

Results

 

Tables 3, 4, 5 and 6 do not mention what values followed by +/- numbers mean. Is it standard error? This should be clearly stated in all table legends.

Comments on the Quality of English Language

Extensive editing of English language required

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The authors revised the manuscript based on the comments.

Author Response

Dear Reviewer 3,

Thank you for your support of the manuscript.

Regards,

From the name of the Authors,

Valery P. Kalinitchenko

9 August 2024