Influence of Biofungicides Containing Microorganisms Such as Pythium oligandrum and Bacillus subtilis on Yield, Morphological Parameters, and Pathogen Suppression in Six Winter Pea Cultivars

Round 1

Reviewer 1 Report

What is the determining factor in the selection of the cultivars used in the study? Brief information should be added in the Material / Method section.

When both the years, the varieties and the biopesticides factor were evaluated together, mixed results were obtained. For example, in 2018, Serenade was more effective in three cultivars and Polyversum was more effective in the remaining three cultivars for the disease agent Ascochyta blight. However, in 2019, the varieties in which biopesticides are effective have been replaced. This also applies to fusaium. Accordingly, it becomes very difficult to recommend biopesticides according to the variety.

This can be given by a simple chart (with all disease agents, pesticides, years, and varieties).

In the study, it is understood that the plants were grown without any external protective application as a -control.

In future studies, it will be useful to observe the effects of +control application, in which standard chemicals are used in addition to -control. This situation can be added to the conclusion part.

 

The writing of the author named (Line 332) Neugschwandtner in the list of references does not match the one in the text.

Author Response

We would like to thank for comments abd suggestion. We provided changes on manuscript. Changes are visible in text.

1. What is the determining factor in the selection of the cultivars used in the study? Brief information should be added in the Material / Method section.

A:  We did correction in text.  The information was provided to line 117:

 

2. When both the years, the varieties and the biopesticides factor were evaluated together, mixed results were obtained. For example, in 2018, Serenade was more effective in three cultivars and Polyversum was more effective in the remaining three cultivars for the disease agent Ascochyta blight. However, in 2019, the varieties in which biopesticides are effective have been replaced. This also applies to fusaium. Accordingly, it becomes very difficult to recommend biopesticides according to the variety.

A: This is a valid consideration. Pathogens and biopesticides are living organisms that respond to biotic and abiotic factors of the environment. In our study, the occurrence of diseases and the effectiveness of biopesticides were significantly affected by the course of the weather, as shown in Fig. 1 and Table 1.

3. This can be given by a simple chart (with all disease agents, pesticides, years, and varieties).

A: We considered such a way of presenting the data. However, we decided that presenting the data in the form of a graph is more readable.

4. In the study, it is understood that the plants were grown without any external protective application as a -control.

A: Yes, in our study we assumed that in the control facility: plants were grown without any external protective application.

5. In future studies, it will be useful to observe the effects of +control application, in which standard chemicals are used in addition to -control. This situation can be added to the conclusion part.

A: The information was added to line 462.

6. The writing of the author named (Line 332) Neugschwandtner in the list of references does not match the one in the text.

A: In the list of references there are 2 publications by the author Neugschwandtner et al. under numbers 28 (line 519) and 30 (line 524). Both are cited in the text.

 

 

 

 

 

Author Response File: Author Response.docx

Reviewer 2 Report

In this article, the morphological characteristics, yield and canopy health of selected cultivars of winter pea was evaluated. The study evaluated six cultivars of winter pea and two means of protecting the canopy, with biological products containing Pythium oligandrum (Polyversum WP) or Bacillus subtilis (Serenade ASO). Overall, authors did very well but I have few suggestions to improve the manuscript.

Aims and objectives, conclusion should be revised and must be in detail.

Biochemical parameters are missing, if authors have biochemical analysis, then must be added in the manuscript for cultivars comparison. This will add more value to the work and also strengthen the hypothesis.

If authors correlate the application with nitrogen metabolism

-In line 37, the full form of FAO must be provided at first use.

-Figure 1, provide vertical and horizontal axis titles with units.

-Figure 2, default error bar values should be replaced with statistically calculated standard error.

-Figure 3 and Figure 6, define the figure part (d) in figure caption.

-Table 2, 3 and 4. Provide standard error values with means.

Moderate editing is required

Author Response

We would like to thank for comments abd suggestion. We provided changes on manuscript. Changes are visible in text.

In this article, the morphological characteristics, yield and canopy health of selected cultivars of winter pea was evaluated. The study evaluated six cultivars of winter pea and two means of protecting the canopy, with biological products containing Pythium oligandrum (Polyversum WP) or Bacillus subtilis (Serenade ASO). Overall, authors did very well but I have few suggestions to improve the manuscript.

1. Aims and objectives, conclusion should be revised and must be in detail.

A: The aim was corrected. We included all issues which were taken to consideration

2. Biochemical parameters are missing, if authors have biochemical analysis, then must be added in the manuscript for cultivars comparison. This will add more value to the work and also strengthen the hypothesis. If authors correlate the application with nitrogen metabolism

A: The Reviewer's remark is interesting. However, in the study we evaluated the biometric parameters of the plants. In future studies, we will also do biochemical analyses of plants for comparison of varieties.

3.-In line 37, the full form of FAO must be provided at first use.

A: We provided required changes.

3. -Figure 1, provide vertical and horizontal axis titles with units.

A:  We did correction. Units were added.

3. -Figure 2, default error bar values should be replaced with statistically calculated standard error.

A:  We did not correction this issue, since meteoriological data, which were receive from meteorological station are not in replications.

4.-Figure 3 and Figure 6, define the figure part (d) in figure caption.

A:  We did required correction.

5.-Table 2, 3 and 4. Provide standard error values with means.

A: We did required correction.

Author Response File: Author Response.docx

Reviewer 3 Report

In this manuscript authors studied and compared some morphological and yield characteristics of six wintering pea cultivars for two seasons; in addition, they also evaluated the effect of two biofungicides on the level of pea protection against two economically important pathogens. The study and its results may be important for this climatic zone (Poland and other similar territories) since authors determined the most productive cultivars for these climatic conditions and evaluated the interaction of the tested preparations with the studied cultivars to determine the most effective combinations. Thus, the paper has a value and can be published; at the same time, I have some comments concerning the presentation of results and their discussion (see below).

 

Intro

 

Lines 38-39: it would be better to use ton/tons instead of t.

 

Results

 

Subsection 3.1: I suppose all data related to the definition, calculation, and interpretation of the Selyaninov’s coefficient should be moved to the Materials and Methods section.

 

Fig. 1: Please, add Y-axis titles for both pictures. Though authors indicated that (a) is the rainfall, and (b) is temperature sums, it is necessary to indicate at least measure units.

 

Table 2: I did not find any indications of significance of difference between the control and treated variants; this fact does not allow one to evaluate the real effect of treatment.

 

Table 3:

1) as authors mentioned, different cultivars are characterized by different stem length and other parameters. As I understand, the first two rows (2018 and 2019) contain data averaged for all cultivars. What is the sense of such averaging? These years significantly differed in weather conditions that could differently influence on these cultivars, so the averaged result could bring different relative contributions of each cultivar. Probably, it would be better to make two different tables, each for each year.

2) The next part of the table is “Cultivar”, and it describes the averaged indices for cultivars. The first question is whether this averaging includes only control variants for each cultivar, or the treated variants were also used for its calculation? The second question is about p-value. What should it indicate? If in the “Year” section I can suppose it indicates the significance of differences between the indices in 2018 and 2019, then in this part of the table we have 6 different numbers. Was the difference between the plant height values for cvs. Baltrap and Dexter (69.3 vs 69.6) significant at p = 0.001?? The same is for other columns; for example, the height until the first node is 38.3 and 38.2 for the same cvs., and the number of pods per plant is 13.1 for cvs. Baltrap and Myster, etc. What is the sense to use “p” values in this case? Please, explain. The same is for the third part of the table: say, was the difference between 54.2 and 54.3 (last column) significant? The whole table requires any clear indication, where the difference between the control and treated variant is significant, and where it is insignificant, since now it is difficult to see and understand this.

3) I suppose “ns” in Tables 3 and 4 means “insignificant difference”. It is necessary to add a note to the tables indicating the meaning of this abbreviation.

The same questions are for Table 4.

 

Lines 252-257: Authors wrote: “The biological agents strongly reduced the efficiency of some of the pea cultivars (Aviron, Baltrap, Myster and Dexter) in their uptake of nitrogen (Fig. 2 f).” At the same time, they also write in the same paragraph:  “A significant positive effect of the use of Polyversum was noted only for the Aviron cultivar, while Serenade ASO positively influenced only the Arkta cultivar.” Thus, biological agents strongly reduced the nitrogen uptake in the cv. Aviron, BUT a significant positive effect was observed for the Polyversum/Aviron pair. It seems to be controversial; moreover, Fig. 2f does not show any significant effect of any biological agent on cv. Averon. Please, check it.

 

Discussion

 

Lines 369-385: I suggest this part of text (description of diseases, symptoms, yield losses) should be shortened, probably via transfer of some of this information to the Intro section. The same comment concerning the description of microorganisms composing the tested biopreparations (lines 422-454) as this part of discussion does not relate to the obtained results.

Line 392, 405: please, check the names of the cited authors. Soova’’li? Or Soovӓli?

English language is quite fine and understandable, but in some places a slight grammar correction is required.

Author Response

We would like to thank for comments abd suggestion. We provided changes on manuscript. Changes are visible in text.

1. Lines 38-39: it would be better to use ton/tons instead of t.

A: We did required correction.

Results

2.  Subsection 3.1: I suppose all data related to the definition, calculation, and interpretation of the Selyaninov’s coefficient should be moved to the Materials and Methods section.

A:  We did required correction.

3. Fig. 1: Please, add Y-axis titles for both pictures. Though authors indicated that (a) is the rainfall, and (b) is temperature sums, it is necessary to indicate at least measure units.

A:  Below both pictures the information is provided.  Picture ‚a‘ is dedicated to reinfall, wheras picture ‚b‘ is dedicated to tempearture. The units were provided on the pictures.

4. Table 2: I did not find any indications of significance of difference between the control and treated variants; this fact does not allow one to evaluate the real effect of treatment. 

A: This table pesents only mean of plants denisty. The plant density was not treated as a core parameter.

5. Table 3: 

1) as authors mentioned, different cultivars are characterized by different stem length and other parameters. As I understand, the first two rows (2018 and 2019) contain data averaged for all cultivars. What is the sense of such averaging? These years significantly differed in weather conditions that could differently influence on these cultivars, so the averaged result could bring different relative contributions of each cultivar. Probably, it would be better to make two different tables, each for each year.

A:  Tables presents main effects of statistical analysis. The single tables would not presnt any extra results. Interactions between years and other factors were presented on figures.

 

2) The next part of the table is “Cultivar”, and it describes the averaged indices for cultivars. The first question is whether this averaging includes only control variants for each cultivar, or the treated variants were also used for its calculation? The second question is about p-value. What should it indicate? If in the “Year” section I can suppose it indicates the significance of differences between the indices in 2018 and 2019, then in this part of the table we have 6 different numbers. Was the difference between the plant height values for cvs. Baltrap and Dexter (69.3 vs 69.6) significant at p = 0.001?? The same is for other columns; for example, the height until the first node is 38.3 and 38.2 for the same cvs., and the number of pods per plant is 13.1 for cvs. Baltrap and Myster, etc. What is the sense to use “p” values in this case? Please, explain. The same is for the third part of the table: say, was the difference between 54.2 and 54.3 (last column) significant? The whole table requires any clear indication, where the difference between the control and treated variant is significant, and where it is insignificant, since now it is difficult to see and understand this.

A: P value presents significant difference among treatments if the value is below 0.05. This is output from Anova analysis.

3) I suppose “ns” in Tables 3 and 4 means “insignificant difference”. It is necessary to add a note to the tables indicating the meaning of this abbreviation.

A: The n.s. abreviation was explained below tables

6. The same questions are for Table 4.

A: as above. The n.s. abreviation was explained below tables

7. Lines 252-257: Authors wrote: “The biological agents strongly reduced the efficiency of some of the pea cultivars (Aviron, Baltrap, Myster and Dexter) in their uptake of nitrogen (Fig. 2 f).” At the same time, they also write in the same paragraph:  “A significant positive effect of the use of Polyversum was noted only for the Aviron cultivar, while Serenade ASO positively influenced only the Arkta cultivar.” Thus, biological agents strongly reduced the nitrogen uptake in the cv. Aviron, BUT a significant positive effect was observed for the Polyversum/Aviron pair. It seems to be controversial; moreover, Fig. 2f does not show any significant effect of any biological agent on cv. Averon. Please, check it.

A: I agree with comments that some cultivars such as Aviron and Baltrap were not strongly influenced by biological agents, and we did correction intext. Howeve I do not agree that results  Fig. 2 f do not presents significance. P – value is below 0.05.

Discussion

8. Lines 369-385: I suggest this part of text (description of diseases, symptoms, yield losses) should be shortened, probably via transfer of some of this information to the Intro section. The same comment concerning the description of microorganisms composing the tested biopreparations (lines 422-454) as this part of discussion does not relate to the obtained results.

A: We did corecction in text.

9. 9. Line 392, 405: please, check the names of the cited authors. Soova’’li? Or Soovӓli?

A: We did correction.

 

Comments on the Quality of English Language

English language is quite fine and understandable, but in some places a slight grammar correction is required.

A:  English correction was done

Author Response File: Author Response.docx

Reviewer 4 Report

 You may consider taken care of comments and suggestions in the attached file to make this report more appropriate.

Comments for author File: Comments.docx

 It is well presented in good English language except for some shade of meaning as pointed out in the attached file.

Author Response

We would like to thank for comments abd suggestion. We provided changes on manuscript. Changes are visible in text.

Proposed new title

Growth and yield of six winter Pea cultivars treated with two plant disease biocontrol fungicides.

Note the title is not in harmony with content.

A: Thank you for this suggestion. However proposed title is too descriptive. Our title includes biocontrol fungicides and their impact on yield, yield parameters and pathogen suppression as well.

 

2. Name of: bioagents” in title not same as inside the text

A:  Thank you for comment. We did correction in text.

 

3. Give complete definition of pesticides used, source(s) and producers and complete words abbreviated acronyms.

A: We did correction. The information was added to line 125.

 

4. Introduction is rather very wordy. Needs to be more concise and direct. However it miss information about the biocontrol agents

A:  We did correction of introduction. Some information was deleted.

 

5. There is no review of and about those biocontrol agents.

A:  The topic is new and there is only few references dedictaed to topic. However we did some information to introduction

 

6. It is “plant top of foliar..” not “plant canopy” ??

A:

 

7. The disease problem not completely covered as it was signified as Ascochyta leaf spot and Fusarium wilt? Lines 80-101.

A: We corrected as shown.

 

8. It is “Agricultural characteristic, values or parameters” not “Morphological” to describe growth and yield.

A: We did small correction, since we add “parameters” to better described morphological traits which were analyzed.

 

 

9. Fig 1, no value assigned for the “Y” axis.

A: We corrected as shown.

10. Table 2. Need to characterize the fungicides treatments by adding another title line inside the table over those two fungicides. The way it is now does not refer to the names as fungicides.

A: We added information in the heading of table

 

11. Table 2. Statistical significances not shown.

A:  We added standard deviation.

 

12. Table 3. The name of fungicide should be capitalized, also in Table 4.

A: We corrected as shown.

 

13. Repetitions in reporting data in more than one form, Figures 2 through 8.

A: Figs. 3 and 6 contain average statistical data showing the influence of the experimental factors in both years of the study. On contrast, Figs. 4, 5, 7 and 8 show the infestation index separately. for each year of the study.

 

14. Line 216. Nitrogen uptake ???

A: We corrected as shown.

 

15. Line 368 – 454. Very wordy and not significant to these results

A: We have corrected and shortened the discussion.

 

16. Not clearly dealt with the genetic resistance in those cultivars and being protected by the bio fungicide under different environmental conditions.

A: It was not a subject of our research.

 

Author Response File: Author Response.docx

Round 2

Reviewer 4 Report

It is good you made so many adjustments. However, in general it is needed to be further substantiated by repeating the same work over number of years because of the low differences recorded.  

I am sorry I still do not agree with your phrased tittle. I am surprised you said the suggested title is "too descriptive" white yours is !?

It is good but needs spelling and linguistic corrections such as Lines 261, 367, 383 and 535 as examples but not all what is needed to be observed.

Author Response

Dear Reviewer,

Thank you for aditional comments. We did some correction of title. We hope that it sounds good.

Conclusion includes information, that further studies are required in this topic.

Manucript was checked by english corrector service, and we belive that reach high quality.

Kind regards,

Authors

Author Response File: Author Response.docx