Development of a New Genetic Transformation System for White and Green Ash Using Embryogenic Cultures

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

Comments and Suggestions for Authors

The manuscript presented for review is focused on the attempt to production of transgenic plants of multiple genotypes of white ash and green ash via Agrobacterium-mediated transformation of embryogenic cultures of these species. The importance of the production of such plants is determined by the fact that all North American ash (Fraxinus spp.) species are threatened by the emerald ash borer (EAB; Agrilus planipennis), an exotic beetle which has already destroyed millions of ash trees in the U.S. and Canada. Although both chemical insecticides and biological control can be effective, and host resistance appears possible, the speed of the invasion has defied traditional management approaches. The development of a host tree-induced gene silencing strategy using RNA interference (RNAi) constructs targeting EAB-specific genes is a potential, innovative approach to managing this destructive insect.

 

The manuscript is constructed according to requirements of “Forests” and is well written and presented. The research methods applied are appropriate, comprehensive and sufficient to achieve the objectives of the study. The illustrative material (tables and figures) are representative and with good quality.

A continuation, some recommendations are given:

Materials and Methods

• Its written: “Embryogenic cultures that were employed as target material for transformation included two white ash and six green ash genotypes, although all eight genotypes were not used in each experiment” In my opinion, it should be clarified here which of the genotypes in which of the experiments described below were used and on what basis it is determined which genotype in which experiment should participate.

Discussion

• The first sentence from „Discussion“ sounds better as a conclusion.
• The last part of „Discussion“ : “This work is foundational …” is the real conclusion to which the above cited sentence must be added, and the conclusion to be separated in a section „Conclusion“ that begins with the last sentence of the “Abstract”, and namely: In result of the study carried out transgenic somatic seedlings of two white ash and three green ash genotypes were produced and acclimated to greenhouse conditions.

 

In conclusion, this manuscript is recommended for publication in “Forests”, after consideration of the remarks shown.

 

 

Author Response

Please see attached file.

Author Response File: Author Response.docx

Reviewer 2 Report

This study outlines the development of a protocol for delivering double-stranded RNA into transgenic ash micropropagated material which will specifically disrupt gene function/cause mortality in emerald ash borer, resulting in a highly targetable tree protection strategy. Embryogenic cultures were used because previous work on adventitious shoot material indicated poor outcomes and for the advantages of scalability into clonal propagation systems. The Agrobacterium-based gene transfer system for white and green ash was successful as the EAB-specific GUS gene was detected at various developmental stages of regenerated ash genotypes -even to greenhouse-acclimatized plants.

This methodological study is an interesting one and contains much useful material and will make a very positive contribution to transgenic approaches to the control of xylophagous insects, altogether a highly topical issue in forestry and horticulture, and in a wider sense within crop science. As such it is worthy of publication, however I believe a number of areas could be improved to add greater clarity to the article, particularly to the less technical reader.

The article is very well written with excellent language construction throughout, however I think a little more emphasis might be devoted to describing hairpin gene silencing functionality in the introduction. It is a little briskly handled, considering the amount of text devoted to the general description of EAB and Fraxinous spp. Speaking of which, L80 mentions the lingering ash which may be targeted for resistance breeding, and later it turns out that one of the 8 genotypes used in this study came from such material. This is given very little air-time during the article; first there needs to be some description about how the very few genotypes were chosen (perhaps with regard to how intraspecific variation may affect the success of the transformation system), but also whether there is potential for confounding results in using apparently resistant material (I assume that the mode of potential resistance is not understood?).

L119 makes a declaration of the research intent of the article. If the authors will allow, this is a very passive statement -reporting production of transformed embryogenic cultured material etc. The overall and long-term goal as reported subsequently is useful and on target, but I think the specific aims of this study should be more actively stated at this point to identify more specifically where this study’s contribution lies. Reference to experiments 1 to 4 would be helpful.

L405 mentions arrows on image A but my version of the manuscript contains no arrows -please check image file versions perhaps.

I like the discussion which is nicely organized, short and to the point and spend just enough time on the important aspects of the study. The transformation efficiency section nicely sets the bigger-picture context before getting stuck into GUS expression. Likewise the last two paragraphs perfectly synthesis the outcomes of the work -why not insert a “Conclusion” heading at L640 and conform to journal convention?!

Author Response

Please see the attachment.

Author Response File: Author Response.docx