Abstract
Phenol is one of the organic pollutants which can cause water environment pollution. It is not only enriched in aquatic organisms but is also a serious threat to human health. Chironomus kiiensis is very sensitive to the contaminants in water and its cytochrome P450s are usually chosen as biomarkers for water pollution. To examine whether CYP6EV11 plays a role in the oxidative metabolism of phenol, we measured the silencing efficiency of CYP6EV11 and evaluated larval susceptibility to sublethal phenol levels by RNA interference (RNAi) technology. The results showed that the transcription of CYP6EV11 was found significantly up-regulated when the 4th instar C. kiiensis larvae were exposed to three doses of phenol. However, the transcriptional levels of CYP6EV11 were significantly suppressed by 92.7% in the 4th instar C. kiiensis larvae soaked in dsCYP6EV11 compared with those soaked in dsGFP for 6 h. The CYP6EV11 expression and mortality of the 4th instar C. kiiensis larvae with CYP6EV11 silencing were mostly decreased under phenol stress. Therefore, the CYP6EV11 gene may be used as a molecular biomarker for earlier warning and monitoring for water pollution.
Highlights
IntroductionPhenol (carbolic acid, phenolic acid, phenylic acid or oxybenzene) consists of an aromatic ring linked a hydroxyl group and is widely used as the precursor to produce industrial compounds such as kerosene, phenolic resin and pesticides [1]
Phenol consists of an aromatic ring linked a hydroxyl group and is widely used as the precursor to produce industrial compounds such as kerosene, phenolic resin and pesticides [1]
To determine whether dsCYP6EV11 could inhibit the expression of CYP6EV11, the 4th instar larvae of C. kiiensis were soaked in dsCYP6EV11 and larvae soaked in dsGFP were chosen as the controls; qRT-PCR analyses showed that the CYPEV11 transcript level was reduced at three of four times, especially, the CYPEV11 transcript level was reduced by 92.7% at 6 h compared with those soaked in dsGFP (p < 0.05)
Summary
Phenol (carbolic acid, phenolic acid, phenylic acid or oxybenzene) consists of an aromatic ring linked a hydroxyl group and is widely used as the precursor to produce industrial compounds such as kerosene, phenolic resin and pesticides [1]. To provide molecular evidence of CYP gene detoxification that will be of benefit to further monitor water pollution, we: (1) examined the transcriptional responses of CYP6EV11 in C. kiiensis to the exposure of phenol at different concentration; (2) revealed the phenol-induced down-regulation of CYP6EV11 contributing to decreased toxicity of phenol to C. kiiensis using the RNA interference (RNAi) method. These results may potentially develop sensitive molecular markers of Chironomidae for monitoring pesticide exposures in non-target organisms in aquatic systems
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