Activity of Hydrolases and Their Inhibitors in Potato Plants Treated with Bacillus subtilis, Salicylic, and Jasmonic Acids and Affected by the Combined Effect of the Late Blight and the Lack of Moisture
Round 1
Reviewer 1 Report
The paper is scientifically interesting but some consideration need to be addressed:
- In Material and Methods is better to specify relative humidity (RH) of the environment where the has been the cultivation.
- In Material and Methods the acronym ISM is not explicit, is better to do so.
- For the damage degree, is better to explicit the evaluation method and if possible refer a citation.
- For the enzymatic activity, for all of the applied methodologies is required to refer to a citation.
- For the histograms (Figure from 2 to 7) the legend should specify the type of enzymatic activity (eg. amylase, protease, ...).
Author Response
The authors thank the editor and reviewers for comments that helped improve the manuscript. Corresponding changes have been made to the article. Below are answers to questions and comments.
- In Material and Methods is better to specify relative humidity (RH) of the environment where the has been the cultivation.
Relative humidity was 50% ± 5%. Corresponding remark added in "Plant processing" section.
- In Material and Methods the acronym ISM is not explicit, is better to do so.
The "ISM" acronym was deleted, an "isolate" used instead.
- For the damage degree, is better to explicit the evaluation method and if possible refer a citation.
The description of the procedure for determining the degree of damage has been expanded and is now more detailed.
- For the enzymatic activity, for all of the applied methodologies is required to refer to a citation.
Methods for determining protein concentration and enzymatic activity were accompanied by references.
- For the histograms (Figure from 2 to 7) the legend should specify the type of enzymatic activity (eg. amylase, protease, ...).
Type of enzymatic activity was added to the legends of histograms.
With gratitude, authors .
Author Response File: 
Author Response.docx
Reviewer 2 Report
1- English must revise again, especially the title, abstract, and introduction.
2- Could the authors provide us with photo/s for the experiment?
3. My other comments are in the attached file.
Comments for author File: 
Comments.pdf
Author Response
The authors thank the editor and reviewers for comments that helped improve the manuscript. Corresponding changes have been made to the article. Below are answers to questions and comments.
1- English must revise again, especially the title, abstract, and introduction.
The text of the article has been revised, the errors found have been corrected.
2- Could the authors provide us with photo/s for the experiment?
Attached are photographs of reference leaves corresponding to different experimental options.
Why the authors selected potato?
According to the FAO, the potato (Solanum tuberosum L.) is the fourth largest food crop in the world. It serves as a valuable source of carbohydrates, antioxidants, vitamins and also as a raw material for the production of starch. Among the abiotic stresses that shape plant productivity, water scarcity is the most important in potato cultivation in many countries of the world. The growth and development of potato, the yield of tubers largely depend on soil moisture.
Why the authors selected any concentrations of SA and JA?
SA and JA were used at certain concentrations based on the literature data and the experience of both other researchers and our own. A link to the article with the corresponding research methodology is given in the "Materials and Methods" section.
Why after 3 days?
Plants after treatment with bacteria and signaling molecules were kept for three days, then they were infected with phytophthora. A pause of three days is necessary for the development of reactions to treatments. Again, we were guided by the literature data and the experience of other researchers, links to which are given in the corresponding section.
Why SA and JA are signaling molecules?
A feature of biopreparations based on microorganisms is the nonspecific activation of plant defense mechanisms. The protective effect of biological products based on Bacillus spp. may be due to the mediated participation of H2O2 in increased expression of PR protein genes [1]. Possibly, by promoting the generation of ROS, bacteria of the genus Bacillus induce the transmission of signals that trigger other defense mechanisms. It has been shown that the treatment of plants with Bacillus subtilis promotes the development of induced systemic resistance (ISR) mediated by the action of jasmonic acid [2]. However, the formation of plant resistance to pathogens under the influence of bacteria of the genus Bacillus can also develop through the salicylate signaling pathway according to the type of systemic acquired resistance (SAR) [3]. Despite the presence of a large amount of data on the suppression of the development of phytopathogens by bacteria of the genus Bacillus, the mechanisms of resistance formation under their influence remain not fully understood.
[1] Pfannschmidt T., Bräutigam K., Wagner R., Dietzel L., Schröter Y. Potential regulation of gene expression in photosynthetic cells by redox and energy state: Approaches towards better understanding // Ann. Bot. 2009. V. 103. P. 599.
[2] Veselova S.V., Nuzhnaya T.V., Maksimov I.V. Jasmonic Acid: Biosynthesis, Functions and Role in Plant Development, Series Plant Science Research and Practices. USA: Nova Sci. Publishers. 2015. Pp. 33.
[3] Cawoy H., Mariutto M., Henry G, Fisher C., Vasilyeva N., Thonart P., Dommes J., Ongena M. Plant defense stimulation by natural isolates of bacillus depends on efficient surfactin production // Molecular Plant- Microbe Interactions. 2014. V. 27. No 2. P. 87.
With gratitude, authors .
Author Response File: Author Response.docx
Round 2
Reviewer 2 Report
Accept in present form
