A Novel CYP4 Gene Identified in the Polychaete Sternaspis scutata and Its Transcriptional Levels along the Coasts of the Liaodong Peninsula
, Aimin Qi 2, Lili Wang 2,*, Encui Shan 1, Dongmei Li 2, Xiaolong Yang 2 and Anguo Zhang 2Round 1
Reviewer 1 Report
This study identified a new cytochrome CYP gene was from Sternaspis sculata, a polychaete distributed in coasts worldwide. The results indicated that the transcription and expression of SsCYP4V gene in the field environment was complex in response to PAHs. This study provides information into the use of polychaetes as biomarkers for monitoring PAHs pollution under field conditions. The idea of the article is clear and the experimental design is reasonable. After careful review of this study, I have listed several concerns in the comments to the authors.
Abstract:
1. In the abstract, the author can make a simple introduction to CYP gene.
2. The author can briefly introduce the importance of Sternaspis sculata for monitoring PAH pollution.
Introduction:
1. Line 42 “Pereineis nuntia” is misspelled and should be “Perinereis nuntia”.
2. Line 45-46: “Pereineis aibuhitensis” should be “Perinereis aibuhitensis”.
3. Line 55-59: The highlight of this manuscript is to prove the complexity of the response of the transcription and expression of SsCYP4V gene to PAHs in the field environment. It can be emphasized in this paragraph that the reason for conducting field research is that there are more laboratory research and less field related research.
Methods
1. Line 143-144: “one-way analysis of variance” change to “one-way analysis of variance (ANOVA)”.
2. Line 112: “Transcriptional expression of CYP4 Gene (SsCYP4V) in S. sculata”. The “SsCYP4V” is abrupt. It was named in the results. It is recommended to delete it here.
Results:
1. In Figure 4, please add more information in the note to illustrate the meaning of black part.
2. Figure 7, caption should be changed to “Relationship of the SsCYP4V mRNA relative expression level in S. scutata and concentration of sediment-borne PAHs”.
Discussion:
1. Line 235-236: “work related to sequencing and phylogenetic analysis of some CYP family genes has been carried out”. This sentence can refer to polychaetes in detail, because the following actually refers to the relevant research of polychaetes.
2. Line 247 and 278: “Perinereis aibuhitensis” has appeared in the previous article. It is suggested to change it to "P. aibuhitensis". Full text check to correct similar problems.
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The article „A novel CYP4 gene identified in the polychaete Sternaspis scutata and its transcriptional levels in coasts of Liaodong Peninsula” aroused my attention. It is written concisely and reads well. The following are my comments:
1. Abbreviations appear in the Introduction: 3-MC and B(a)P. Please include the whole names, and put the abbreviations in parentheses.
2. Materials and Methods: Isn't there an error in the description of "mean ± SE (standard error)" (146th line)? Shouldn't it be SD or SEM?
3 Results: This sentence „The relative expression of SsCYP4V gene was significantly different in the sampling sediments (F = 569.58, p < 0.001, Fig. 6) (201)”. suggests that the expression of the SsCYP4V gene was measured in the sediment. Please correct that it refers to polychaetes.
4. In Table 2 - please expand the abbreviations PAHs used in the table. In addition, please specify that the amount of PAHs was shown per amount of sediment dry weight.
5. In the supplementary materials, please provide the Ct of the SsCYP4V gene and actin gene in qPCR measurements. Please post a photo of example electrophoresis showing the degree of integrity of RNA isolated from polychaetes.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
