Effect of Herbicide Clopyralid and Imazamox on Dehydrogenase Enzyme in Soil of Regenerated Pedunculate Oak Forests

Round 1

Reviewer 1 Report

Overall, the paper is well organized and structured. Requires some minor corrections. Please see attached for more comments.

Comments for author File: Comments.pdf

Author Response

The question of the reviewer: Was the study site free of weeds when the sites were seeded by acorn. Or there were natural weeds already growing. Need to mention.

Done

The question of the reviewer:  How was herbicides applied.....?

I explained it in 2.3. Experimental design and herbicide treatments Line: 141-143

The question of the reviewer:  If possible, the major weed species can be mentioned?

Done

The reviewer suggested to include the weather information of each herbicide application day

Done

 The reviewer: add comma after "added"

Done

 The reviewer states that I should restructure the sentence!

Done

The reviewer indicates an error ‘Figure 1’ should be write Figure 2

Done

 The reviewer suggested to use a better word ‘’reduction’’ in place of ‘’drop-in’’

Done

 

Reviewer 2 Report

Dear colleagues!

A good article, everything is clear, clear presentation, tables are numbered correctly, English is clear, scientific terms were used correctly. Unfortunately, the authors did not show the mechanism of action of the enzyme at the biochemical level, but it would be interesting for a narrow specialized audience. I would like to see some information about the practical use of the data obtained.

Please look at lines 264, 271...repeat text

Comments for author File: Comments.pdf

Author Response

The article is well structured, all the individual parts are well connected, so it is easy to read. The relevance of the article is not in doubt. Solving the food problem in the world is currently one of the pressing problems of our time. Insects, pathogenic microorganisms, diseases destroy crops. In this regard, it is necessary to use various "protectors" of plants. Herbicides are widely used in agriculture. When they enter the soil, they have a different effect on their properties. Enzymes are good indicators of changes, they reflect changes at the lowest level of the organization of matter – molecular and biochemical. They can be used for rapid assessment of the quality of the natural environment.

A short and educational overview of the paper from reviewer where I had nothing to correct.

 It should be noted that more than 100 different enzymes have been found in the soil. They are responsible for various stages of the transformation of organic matter, participate in soil-forming processes. Dehydrogenase is an enzyme that is responsible for the processes of oxidation, hydrogen transfer.

Done

The high expression of the enzyme and sensitivity is the right solution for the biological diagnosis of soils when applying herbicides. The paper presented model experiments. Due to the fact that its description in the article is given in great detail and well, there is no need to repeat. It is necessary to note only the very correct selection of the sampling interval (7 days, 14, 21, 30 and 60) of soils for analysis.

Done

As we know, uncontrolled growth of microorganisms on nutrient media under favorable conditions occurs within 72 hours (3 days). There are inhibitors of this process in the soil: pH, the content of trace elements (Cu, Ni, PB, etc.), the presence of biofilms on the soil surface can also be noted. The increase in the growth of microorganisms is stretched over time, corresponds no longer to 3 days, but to 7 days. At the same time, the functional response of microorganisms occurs later! Therefore, it was at this time that samples were taken for analysis.

The reviewer agreed when soil samples were taken for analysis. I had nothing to correct.

The uniqueness of the work lies in the fact that the study of such a level on soils with predetermined properties has not been carried out. It was at that moment at this point in space that the conditions for the synthesis of enzymes were formed, the activity of which was determined in the laboratory.

The reviewer indicate that uniqueness of the work lies in the fact that the study of such a level on soils with predetermined properties has not been carried out. I had nothing to correct.

It is possible that due to global climate warming, changes in nature, the activity of enzymes will be higher. Time will show the correctness of our and the world's reasoning!

I agree with reviewer.

The work uses statistical techniques and methods that adequately reflect changes in the microbiological and biochemical properties of soils. At the same time, it should be noted that many researchers are currently using new statistical methods in pursuit of fashionable trends, but biological methods do not interpret them correctly. This was not observed in this work.

The reviewer indicate that have been used adequately statistical techniques and methods which interpreted correctly. I had nothing to correct.

Everything is very logical, understandable, and most importantly biologically explicable. In the presented article, the drawings are very informative, the tabular data is clear. In the process of reading the manuscript, there were no questions about the subheadings of the tabdits, captions to the drawings.

The reviewer indicates that in the paper everything is very logical, understandable, and biologically explicable. I had nothing to correct.

The only small comment, or maybe a suggestion, was about discussing the results. I wanted to see a more in-depth discussion of the results: the effect of herbicides and enzyme at the enzyme + substrate (S+E) level. What happens during the interaction, what conformational changes occur during the formation of the enzyme-substrate complex, is it possible for the compounds in the herbicide to penetrate into the structure of the enzyme, etc. However, this may already be another article. In the end, I would like to wish good luck to the team in their difficult scientific work!

….the effect of herbicides and enzyme at the enzyme + substrate level and  is it possible for the compounds in the herbicide to penetrate into the structure of the enzyme, etc... I will do my best to investigate and write about these interesting topics soon!

Thank you very much for good wishes!