Stabilized Double-Stranded RNA Strategy Improves Cotton Resistance to CBW (Anthonomus grandis)

Round 1

Reviewer 1 Report

 The manuscript entitled 'Stabilised double-stranded RNA improves resistance of cotton to CBW (Anthonomus grandis)' addresses a very important issue related to the attempt to obtain transgenic plants resistant to the adverse effects of the environment in this case of the pest Anthonomus grandis causing high losses in cotton crops. The authors present here a very interesting strategy based on expressing double-stranded RNA (dsRNA) molecules to trigger RNA interference-mediated gene silencing in CBW. This approach seems extremely interesting as it is an excellent alternative to traditional methods using chemical pesticides that show adverse effects on the environment while leading to contamination of food chains. The authors present the objectives of the work in a very correct and transparent way and interpret the results obtained in a correct manner. 

My only suggestion is that the authors present the transgenic plant lines themselves in more detail, indicating potential phenotypic differences from wild plants.

Author Response

IJMS 1820823 Manuscript

Response to Reviewer 1 Comments

Question 1: My only suggestion is that the authors present the transgenic plant lines themselves in more detail, indicating potential phenotypic differences from wild plants.

Thank you very much for all the comments. Supplemental Figure S1 has been added, which shows that there are no significant phenotypic differences between GM plants and the untransformed plant. Furthermore, flower bud, seed, and fiber weight production is similar between GM and WT plants, as shown in the table in Figure S1 (B). (See Lines 166-167 (pdf version),  Lines 197-199 - word version), and Supplementary Figure S1).

Author Response File: Author Response.pdf

Reviewer 2 Report

The literature references do not match the journal's format. Change capitals in paper titles to minuscles, delete "pp." in page, delete issue numbers. Give all publication years in bold characters if not done, e.g. line 637.

Be a bit more reserved mentioning the own merits in the introduction, e.g. lines 75-72.

As obviously growth is affected by disturbing PM, please consider the effects of food deprivation on JH level/activity, particularly regarding effects on oocyte growth and reproduction (see: Rankin, S.M. et al. (1997) Entomologia Experimentalis et Applicata 83(1), pp. 31-4)

Some minor comments can be seen in the attached ms. with comments

Comments for author File: Comments.pdf

Author Response

IJMS 1820823 Manuscript

Response to Reviewer 2 Comments

Question 1: The literature references do not match the journal's format. Change capitals in paper titles to minuscles, delete "pp." in page, delete issue numbers. Give all publication years in bold characters if not done, e.g. line 637.

Answer 1: Thanks for all the comments. All references have been revised and are in the format required by the IJMS. Reference 91 (Rankin et al., 1997) was added in the texte.

“Rankin, S.M.; Dossat, H.B.; Garcia, K.M. Effects of diet and mating status upon corpus allatum activity, oocyte growth, and salivary gland size in the ring-legged earwig. Entomol. Exp. Appl. 1997, 83, 31-40, https://doi.org/10.1046/j.1570-7458.1997.00154.x.”

Question 2: Be a bit more reserved mentioning the own merits in the introduction, e.g. lines 75-72.

Answer 2: The text has been modified (see lines 67-68 - pdf version, lines 87-91 - word version).

Question 3: As obviously, growth is affected by disturbing PM, please consider the effects of food deprivation on JH level/activity, particularly regarding effects on oocyte growth and reproduction (see: Rankin, S.M. et al. (1997) Entomologia Experimentalis et Applicata 83(1), pp. 31-4)

Answer 3: the point raised was considered and added in the text (see lines 394-395 - pdf version, see lines 460-463 – word version)

Question 4: Some minor comments can be seen in the attached ms. with comments

Answer 4: All suggested points have been corrected in the manuscript (see lines 204 and 399 – pdf version, see lines 242 and 467- word version).

Author Response File: Author Response.pdf