Characterization of 33 HbbZIP Gene Family Members and Analysis of Their Expression Profiles in Rubber Tree in Response to ABA, Glyphosate and Powdery Mildew Treatment
and Yu Zhang 1,*Round 1
Reviewer 1 Report
Dear Authors
The present manuscript is very good. I appreciate its innovative character and the possibility of using the obtained results in other studies.
However, I would like to ask the authors to make two important additions to the text that currently make it impossible for me to call the manuscript ready for publication.
First, I lack information on the selection programmes for the species in question. The description of the molecular studies in the introduction misses the most important point of the paper: why the rubber tree was selected, how its selection is currently carried out, which traits are of high scientific interest, how the studied genes will improve the selection possibilities, etc.
Second, the methodology (lines 144-147) describes a ANOVA analysis and a post-hoc test; as a reviewer, I cannot indicate where in the results chapter I can find the results of this analysis. Please address the two issues described in your response to the review. If you think it is possible, please improve the text of the manuscript.
I am currently leaving the manuscript for minor revisions before I receive a response to the review.
Yours sincerely, and I look forward to correcting the text soon.
Author Response
Comment 1:-The present manuscript is very good. I appreciate its innovative character and the possibility of using the obtained results in other studies.
However, I would like to ask the authors to make two important additions to the text that currently make it impossible for me to call the manuscript ready for publication.
First, I lack information on the selection programmes for the species in question. The description of the molecular studies in the introduction misses the most important point of the paper: why the rubber tree was selected, how its selection is currently carried out, which traits are of high scientific interest, how the studied genes will improve the selection possibilities, etc.
Response:
Thank you for your suggestions. We add following sentences in the introduction section.
Natural rubber is an indispensable raw material for transportation, medicine and national defense industries, and has great economic value. Rubber trees (Hevea brasiliensis) are the main source of natural rubber and make great contributions to China's economic construction and social development. The Reyan73397, one of the varieties of rubber trees，has excellent rubber production performance and is one of the rubber tree varieties with a large planting area in the rubber planting area of China, and there are more studies on its variety characteristics. Therefore, Reyan 73397 variety was used as the research object in this study. Glyphosate is one of the most commonly used herbicides in the process of rubber tree planting, and its use often causes drug damage causing deformation or even loss of rubber tree leaves, which affects the normal growth of rubber seedlings and leads to lower yield of natural rubber and large economic losses. The results suggest that the rubber tree bZIP gene may play a key role in drought stress response, rubber tree powdery mildew resistance response and glyphosate damage relief, and the study of this gene can help improve rubber yield and reduce economic losses due to biotic and abiotic stresses such as drought and powdery mildew fungus, and facilitate the promotion of rubber tree breeding and cultivation.
Comment 2:- Second, the methodology (lines 144-147) describes an ANOVA analysis and a post-hoc test; as a reviewer, I cannot indicate where in the results chapter I can find the results of this analysis. Please address the two issues described in your response to the review. If you think it is possible, please improve the text of the manuscript.
Response: Thank you for your suggestions. The results section on analysis of HbbZIP expression in different tissues in response to ABA, powdery mildew, and glyphosate treatment（for example lines 251-252） involves ANOVA analysis and Tukey's test. In this paper, ANOVA analysis and Tukey's test were used in the statistical processing of the data.
Reviewer 2 Report
the proposed article further clarifies the functions of the 33 HbbZIP genes in rubber tree. Scientific method is classic and clearly presented but for me extrapolation about the importance of them for rubber breading should be be modulate.
Author Response
Comment 1:-The proposed article further clarifies the functions of the 33 HbbZIP genes in rubber tree. Scientific method is classic and clearly presented but for me extrapolation about the importance of them for rubber breading should be modulate.
Response: Thank you for your suggestions and affirmation of our work, and we revised to “in pesticide- and disease control”.
Reviewer 3 Report
The submitted manuscript is very current and interesting. The manuscript is written relatively neatly, with individual parts logically following each other. The introductory part is adequate. As part of the methodology, I recommend supplementing the conditions of plant cultivation in in vitro conditions. How was glyphosate spiked? I would also recommend adding the exact location of the sampling site. Have the plants been transferred from in vitro to ex vitro conditions? The results obtained are well described. Figure 3 is hard to read. I recommend doing correlation analyzes within the substances used and the occurrence of the pathogen. The discussion is sometimes rather descriptive. The authors also cite older literature. Are these older literary sources necessary?
Author Response
Comment 1:-The submitted manuscript is very current and interesting. The manuscript is written relatively neatly, with individual parts logically following each other. The introductory part is adequate. As part of the methodology, I recommend supplementing the conditions of plant cultivation in in vitro conditions. How was glyphosate spiked? I would also recommend adding the exact location of the sampling site.
Response: Thank you for your reminding! The conditions of plant cultivation in in vitro condition (lines 120-121) have been added and the exact location of the sampling site is described in lines 105-106. In this study, both glyphosate and ABA were treated on rubber seedlings by spraying, and are added in line 94 of the text.
Comment 2:-Have the plants been transferred from in vitro to ex vitro conditions?
Response: Yes. All the tissue culture-generated seedlings were first cultured in tube and transplant in plastic bag with soil about 1year in green house.
Comment 3:-The results obtained are well described. Figure 3 is hard to read. I recommend doing correlation analyzes within the substances used and the occurrence of the pathogen.
Response: Thank you for your suggestion. Figure 3 was modified according to the suggestions.
Comment 4:-The discussion is sometimes rather descriptive. The authors also cite older literature. Are these older literary sources necessary?
Response: Thank you for your suggestions. These older literary is closely related to the content of this study, and in the process of experimentation, we have gained a lot of inspiration from these literary, we thought that these older literary matters are much necessary. At the same time, we also consulted the newly published relevant literature according to the reviewer's comments, and updated and supplemented some literatures in the paper.
