GmDNAJC7 from Soybean Is Involved in Plant Tolerance to Alkaline-Salt, Salt, and Drought Stresses

Round 1

Reviewer 1 Report

The manuscript "GmDNAJC7 from Soybean is Involved in Plant Tolerance to Alkaline-salt, Salt, and Drought Stresses" focused on the tolerance to alkaline-salt, salt, and drought stresses in soybean, and identified a chaperone DNAJ protein gene GmDNAJC7. Both overexpression of GmDNAJC7 in soybean composite plants and ectopic expression of this gene in Arabidopsis could improve the alkaline-salt, salt, and drought tolerance. The study is of great interest to soybean breeders and provides new insights into the role of DNAJ in plant tolerance to abiotic stress. The manuscript is well organized and written, although some mistakes exist.

1. In the material and method part, I have some questions about the experiment design. First, since you select multiple concentrations for NaHCO3, NaCl or mannitol in Arabidopsis experiments, why do you select only NaHCO3 and only one concentration (90 mM) for the composite soybean plants? Furthermore, why do you select this concentration, please explain in the manuscript. Second, the hairy roots of the composite plants are usually chimeric. For the positive transgenic plants used for subsequent experiment, have you removed the negative transgenic hairy roots to make the results more accurate?
2. For Figure 6- 8, the germination rates at five days after NaHCO3 and NaCl treatments were shown. However, for mannitol treatment, the germination rates from 5 to 10 days after treatment were presented. Could you please explain why these results were shown in different ways?
3. The expression "overexpression of GmDNAJC7 in Arabidopsis" seems not suitable, because GmDNAJC7 is an exogenous gene for Arabidopsis. How about using "ectopic expression" instead.
4. The expressions of "DNAJ protein" are not consistent throughout the manuscript. "DNAJ protein" was present in some place, while "DnaJ protein" was present in other place. Please check it carefully.
5. Please provide the full name of SPAD value at its first appearance in the manuscript.
6. The resolution of figure in the manuscript is too low. Therefore, it is difficult to observe the differences for various genotypes in Figure 6-8.
7. Line 51, is the word "Cye" a spelling mistake for "Cys"?
8. Line 184, please provide the full name of “SMART”. Is it a software or an online website?
9. Line 194-196, the Table S3 should contain more detailed information for these 10 genes, such as the fold changes for each of them, and their corresponding abiotic stresses (alkali, salt, or drought). Moreover, the GmDNAJC7 indeed has the most gene ontology (GO) function annotations, however, how could you get the conclusion that “GmDNAJC7 is associated with abiotic stress” based on the information in Table S3?
10. Line 301, the “larger” should be replaced by “greater”?
11. Line 419, could you replace the “protein-protein interaction network analyses” with “protein-protein interaction network prediction analyses”, because the results were obtained by prediction not experiment.
12. Line 544, “arabidopsis” should be “Arabidopsis”.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Thank you for the opportunity to review the article "GmDNAJC7 from Soybean is Involved in Plant Tolerance to 2 Alkaline-salt, Salt, and Drought Stresses" . After reading it, however, numerous editorial, stylistic and substantive errors were noticed, which should be corrected in order to improve the quality of the work.

Main changes:

- abstract - written chaotically, it was not explained why Arabidopsis was analyzed since the title refers to soybeans. Maybe the title should be changed too

- the methodology needs to be improved, especially the paragraph on plant growth and growing conditions. The lack of how the soybeans were prepared and what kind they are, germination conditions, humidity, etc.

What was the research sample of how many plants

- confocal microscopy- what were the measurement parameters?

how the analysis was performed using Image J, which appears in the description under the figures, but there is nothing in the methodology

- explain the abbreviations of the media used for the first time, e.g. YEB, WT

- not explained why the stresses NaHCO3, PEG, Nacl were selected !  why this doses were used,  the references

Discussion -

- little up-to-date literature cited

- chaotically described 

There is no standardized way of spelling Arabidopsis names, the genes (italics) and proteins throughout the text,

editing errors (e.g. double spaces, line 469-470), lines 389,399,406

Entire sentences often incomprehensible, e.g. lines 406, 146-147 etc.

Abbreviations:

- repeated abbreviations twice, e.g. WT, ML, ROS etc.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Most of the comments were taken into account