Impact of Trichoderma asperellum on Chilling and Drought Stress in Tomato (Solanum lycopersicum)

Round 1

Reviewer 1 Report

This paper describes the use of Trichoderma asperellum to enhance tolerance to abiotic environmental stress.  In general the paper is well written.  However, there are a few sentences that require attention to correct spelling, grammar and clarity.   It is difficult to identify these without page and line numbers and so I provide only a few examples below:

Introduction - 2nd page 2nd paragraph - 'constrain' not constraint.

2nd page para 4 - please clarify what is meant by 'hardening'.  Expand this section of the introduction on priming for environmental stress as it is most relevant to the manuscript.

Methods - please provide more details on the method of inoculation with T. asperellum e.g inoculum per plant/pot?  The last sentence requires attention to improve clarity '..plants were grown into a greenhouse.."?

Please add a little more detail on experiment design to clarify the number of plants per treatment.

Statistics - SEM = standard error of the mean.

Results

Page 5 1st sentence - what is meant by 'due'

Figure 1 caption - please define the abbreviations

Section 3.2 header - change 'an enrichment of' to 'enhanced'

Page 6 - what is meant by 'lesser green leaves' ?  Is this fewer leaves or paler leaves?  If the latter, was this a visual assessment or quantitative (e.g SPAD meter).

Section 3.3 -  Please rewrite this section to clarify the main results.

Discussion - There is an assumption that T asperallum has colonised roots but no data to support this.  This should be acknowledged.  The treatment appears to be more effective against drought stress than chilling. There is not data to compare the relative populations of T asperellum in the droughted and chilled root zones.  Similarly, there is no data on root biomass which again is of importance. Is it possible that growth of the roots and the Trichoderma was affected by the lower temperature?  Please include some discussion of this.   

Author Response

Dear Sir

Thanks very much for your efforts and useful comments about our manuscript. We are very grateful and appreciate your good comments which help us to make the paper more quality and accurate.

Please see the attachment

Best regards

Author Response File: Author Response.pdf

Reviewer 2 Report

The current manuscript was written in very vague manner and many English grammatical errors. There reviewer suggest different suggestions in attached file.

Comments for author File: Comments.pdf

Author Response

Dear Sir

Thanks very much for your efforts and useful comments about our manuscript. We are very grateful and appreciate your good comments which help us to make the paper more quality and accurate.

Please see the attachment

Best regards

Author Response File: Author Response.pdf

Reviewer 3 Report

The manuscript is interesting and well drafted. However, I have some minor remarks.

Check the style and the grammar throughout the document.

Please add more information on the mechanisms of priming by T. asperellum.

Lines 40-42: Do you have a reference for this statement?

Line 100: Where did you get the substrates from?

Line 128: It should be 50 ml, I suppose.

Results section: Quite often results are presented as differences albeit they this is not proved by statistics (for example lines 188-189: it is not the lowest value; line 323).

Figure 2 and adjacent texts: Please change the formatting. The figure is splitted and the text is inserted within the figure.

Lines 325-326: Reference is needed.

Author Response

Dear Sir

Thanks very much for your efforts and useful comments about our manuscript. We are very grateful and appreciate your good comments which help us to make the paper more quality and accurate.

Please see the attachment

Best regards

Author Response File: Author Response.pdf

Reviewer 4 Report

 Dear Authors!

Please find below several comments and suggestions in order to improve the manuscript. 

General observations:

The whole manuscript should be carefully revised by the authors, because there are several grammar and other editing mistakes (eg. lines 45-47: sentence with no meaning/verb is missing, lines 68-70: space between words is missing, line 85: point is missing after spp., line 177: weird words are used, as example firstable, lines 227-228, 232-233 and the corresponding figures are not inserted correctly). 

Statistics: please indicate the number of repeats for each measurement. To use different number of measurements for the same parameter (n=3-6, line 200, n=12-37 in line 237) is wrong, it gives different degree of freedom is statistic tests that affects the result of the test. Also, in statistics we use SD and not SEM for standard deviation. 

Results: in this part only the results which are supported by the experimental data should be presented. In lines 204-205 the authors discussing about severity symptoms and wilting, line 227 lesser green leaves. If these were evaluated (manual inspection), please describe in the Material and methods part and also insert a table with the results obtained. 

Discussions: line 314: production of the new leaves. No data about new leaves production is presented in the manuscript. Please insert data or remove this information from discussion. 

Conclusions: no data about cold stress (as you use chilling) are presented. 

Detailed observations and suggestions:

Title: A question: why are you using chilling instead of cold or low temperature? 

Introduction:

Line 38: please replace a growing population with the growing population

Line 40: please replace then by 2100 with until 2100

Lines 45-47: there is a fragment of a sentence, a verb is needed

Lines 68-70, 80: spaces are missing: andprotein, factorsto, [7,26]and

The last sentence (lines 93-95) should be moved in front of the we investigated paragraph (line 87).

Materials and methods

Please indicate the number of plants per experiment. 

Line 112: How long were the plants exposed to 4C? 9 days constantly? 

Line 118: How many days lasted the experiment? When was the end of it?

Line 128: 50 l should be replaced with 50 ml plastic tube

Line 158: purified by filter paper should be replaced with filtrated through filter paper

Line 169-170: a space is missing: ofmean and SEM is not the proper abbreviation for standard deviation (SD)

Results:

Line 177: please replace firstable with the proper word

Lines 199-201: Figure 1. the legend should be completed with the explanation of abbreviations used (U, TI), SEM should be replaced with SD, number of repeats should be indicated for each experiment . 

Please consider the above suggestions also for the legend of Figure 2, 3.

Lines 242-245: in the sentence you compare free- and inoculated plants to their respective control. The controls are not the uninoculated plants? Please rethink and if necessarily reformulate. 

Discussion:

Several statements are not supported by the data presented in results. See comments above (wilting phenotype, much healthier plants, production of new leaves).

Line 332: please replace en with in

Line 343: please delete showed in Figure 2a. This is discussion, no need to mention the figures presented in Results section. 

Author Response

Dear Sir

Thanks very much for your efforts and useful comments about our manuscript. We are very grateful and appreciate your good comments which help us to make the paper more quality and accurate.

Please see the attachment

Best regards

Author Response File: Author Response.pdf

Round 2

Reviewer 4 Report

Thanks for corrections. Best wishes. 

Author Response

Dear Sir
Thanks very much for your efforts and useful comments about our manuscript. We are very grateful and appreciate your good comments which help us to make the paper more quality and accurate.
Please see the attachment
Best regards