Extracellular DNA of Fusarium oxysporum f. sp. cubense as a Priming Agent for Inducing the Resistance of Banana Plantlets

Round 1

Reviewer 1 Report

I really appreciate editor-in chief to invite me to review the manuscript (agronomy-2145652) entitled by ‘Extracellular DNA of Fusarium oxysporum f.sp. cubense as a priming agent for inducing the resistance of banana plantlets’. This manuscript presents an original and feasible idea to evaluated whether the eDNA from Fusarium oxysporum f.sp. cubense(Foc) could 14 limit the growth of Foc itself (self-inhibition test) while increasing the resistance of banana plant 15 (priming test)., but I feel that it would be better to improve some minor error a bit on this manuscript, please see the following below.

General comments:

1.      Line 31, Is the latest data available instead of 2019 ?

2.      Line 225 please uppercase “p”;

3.      Line 44-45, This sentence is a little unclear, please rewrite it;

4.      The reference format is not uniform, please revise;

5.      Fig.1, Day 0, Lack of significance analysis;

6.      Fig.1 “p” should revised as “P”

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

The paper presents an interesting study concerning the use of eDNA as an alternative method to counteract the fusarium wilt (Fusarium oxysporum f.sp. cubense) in banana, by promoting defence mechanisms in plants.

All the section are clearly presented. Small changes are required related to formatting and spelling.

Formatting:

-in the abstract several typing errors should be corrected: lines 14, 16, 18, 19, 20, 21 – spaces are required between values and symbols or elsewhere,

-line 40 – please revise the phrase,

-eliminate all unnecessary spaces between paragraphs,

- line 106 – please check and revise,

-line 168 – space required between value and symbol,

-line 330 – check and revise,

-line 345 – eliminate the additional space.

Figure 3, part A: the photographs would be more easily observed and analyses by readers if larger files are included in document. It is difficult to observe the differences between day 0 and 9 DPI.

How do authors consider the concentration of 200 μg/ml is? It is small or increased? At what concentration fungicides are usually applied? Please comment!

Author Response

Please see attached.

Author Response File: Author Response.pdf

Reviewer 3 Report

Dear Authors

Please check my review comments below:

Line 102 Page3: MS medium? Please when you use an abbreviation mention what it means.

Line 109 Page3: LCB? Please when you use an abbreviation mention what it's means.

Line 111 Page3: How the Foc TR4 DNA was extracted?

Line 113 Page3: (n = 5)?

Line 115 Page3: Are there any negative or positive control?

Line 120-121 Page3: what are the criteria used for selecting the different concentrations of eDNA of Foc TR4 suspension concentrations of 0, 40, 80, and 200 µg mL-1?

Line 127 Page3: “The Foc TR4 – infected” It should change to “The Foc TR4 – inoculated”

Line 164 Page4: Add reference for the primers that are used in this study.

Line 195 Page5: How the colonies of Foc has been counted with this high density of colonies? Add the colonies count as a supplemental table.

Line 195 Page5: Figure1 A, Why does the gel fragment for the eDNA of Foc TR4 10 µg mL look like a smear? Also, Add “e” to the DNA to the table title and figure 1A. Also, use numbers when you presented the results in paragraph 3.1.

Line 206 Page5: This wording is confusing “infected by eDNA Foc TR4 “ The eDNA doesn’t consider as infectious against the Foc TR4 as the pathogen. Please be careful when you descript in writing. And also you need to use inoculation whenever you do inoculate a host with a pathogen.

Line 224 Page6: in figure 2 what are the criteria that use in the section category to describe treatments as Highly Susceptible, Susceptible, Moderately Susceptible, and Tolerant? In the Rhizome discoloration index Salicylic Acid 5 uM shows 3,40 ± 0,55 BC compared to eDNA 80 µg mL-1 showed  3,80 ± 1,30 BC, its higher, which means it should be both “Susceptible” and described in the table as “Moderately Susceptible”

Line 261 Page7: In this figure, 2 methods are used to detect O2-  and H2O2. Why there are big differences between both? Either use one or repeat the experiment with the wet lab method which is more accurate.

Line 380 Page10: Figure 5, is the Foc TR4 on the left side of the figure should be - Foc TR4 NOT + Foc TR4?

The big question I want to ask here If the isolate of Foc TR4 was isolated from an infected banana plant which means it is pathogenic as I see the results of the control in the experiment. How can the author use a pathogenic isolate as a boifungicid agent against the disease? Even if it has the eDNA. Please explain.

Author Response

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Author Response File: Author Response.pdf

Reviewer 4 Report

This article submitted for revision (agronomy-2145652) presents a valuable research to enhance the knowledge of resistance breeding in banana plants against Fusarium oxysporum f.sp.cubense (Foc). Experiments revealed the suitability of eDNA FocTR4 as a growth inhibitor of Fusarium oxysporum f.sp. cubense and a priming agent to the banana plantlets. Results showed that this agent could be considered as a biofungicide candidate to induce the resistance against Fusarium wilt in banana plants. Regarding  the importance of this research for banana resistance breeding I recommend to publish the article in Agronomy journal.

I have noticed only one slight technical error in the text: in line 106 the word ''on'' is repeated twice. 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf