扑草净胁迫对四角蛤蜊解毒相关基因表达的影响
Prometryn Stress in Expression of Detoxification-related Genes in Mactra veneriformis
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摘要: 扑草净是我国常用除草剂的主要成分,其药物残留在近岸海域广泛分布。在水温(15±1) ℃条件下,采用半静态水质接触染毒法开展胁迫实验。以β-肌动蛋白(β-Actin)基因为参比,分别研究了不同浓度扑草净胁迫对四角蛤蜊消化腺和鳃组织中超氧化物歧化酶(superoxide dismutase, SOD)、过氧化氢酶(catalase, CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)、谷胱甘肽S-转移酶(glutathione S-transferase, GST)、细胞色素P450第四亚族(cytochrome P450, CYP4)和金属硫蛋白(metallothionein, MT)6种解毒相关基因表达的影响规律。结果显示,6种基因表达调控对扑草净胁迫反应灵敏,在扑草净质量浓度分别为0.2、1.0和10 μg·L-1的养殖海水中,随着胁迫时间延长,四角蛤蜊消化腺呈现SOD、CAT、GSH-Px、GST和MT等5种基因表达均先上调、CYP4基因表达先下调,而后反复波动变化的趋势;鳃呈现SOD、GSH-Px和GST等3种基因表达均先上调,CYP4基因表达先下调,1.0 μg·L-1组的CAT基因表达先下调其他组先上调、MT基因表达先上调其他组先下调,而后反复波动变化的趋势。GST可作为扑草净对四角蛤蜊污染潜在的生物标志物。该结果为评价扑草净残留对海洋生态环境安全风险提供了参考依据。Abstract: Prometryn is a herbicide that is widely used in China and frequently detected in coastal environment. In this study, Mactra veneriformis were exposed to seawaters with prometryn at 0, 0.2, 1.0 μg·L-1, and up to 10.0 μg·L-1 by the semi-static method at (15±1) ℃. Digestive glands and gills in each group of Mactra veneriformis were collected at exposure time 6, 24, 48, 96, 144, 240, 360 and 504 h respectively. The expression of detoxification-related genes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), cytochrome P450 (CYP4) and metallothionein (MT), were analyzed in relative quantification by reverse transcription, quantitative real-time polymerase chain reaction (qPCR) technology and 2-△△ct method with β-Actin gene as the reference gene. The results showed that the regulation of expression of these genes was sensitive to the prometryn and fluctuated over time. The expression of SOD, CAT, GSH-Px, GST, MT mRNA was increased and CYP4 mRNA decreased in the digestive glands of Mactra veneriformis at exposure time 6 h. Subsequently, the expression of these genes was fluctuated in the digestive glands of Mactra veneriformis in all 3 treatment groups. In the gill of Mactra veneriformis, the expression levels of SOD, GSH-Px, GST mRNA increased and CYP4 mRNA decreased in all 3 treated groups at 6 h, when the expression levels of CAT mRNA decreased in the 1.0 μg·L-1 exposure group and increased in other groups. Meanwhile, the expression levels of MT mRNA was increased in the 1.0 μg·L-1 exposure group and decreased in other groups at 6 h, and then fluctuated. GST can be used as a potential mRNA biomarker for the impact of prometryn on Mactra veneriformis. The results provide a promising way for the evaluation of the risk of prometryn residue on marine ecological environment safety.
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Key words:
- prometryn /
- Mactra veneriformis /
- gene expression /
- detoxification
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