近日,西南大學于士將老師,使用IPHASE品牌產品:小鼠肝微粒體在《Journal of Hazardous Materials》全威期刊上發表文章《A down-regulated cytochrome P450 in Neoseiulus barkeri Hughes (Acari: Phytoseiidae) can dechlorinate and hydroxylate chlorpyrifos without producing chlorpyrifos-oxon》,影響因子12.2!
在有害昆蟲生物防治中農業系統中存在的天敵資源發揮著重要作用,常見的有機磷類殺蟲劑如毒死蜱等在農田噴灑作業時,對天敵的生存造成了負面影響,選育抗藥性天敵品系有助于生物防治和化學防治工作的協同開展。此前西南大學實驗室選育了一種捕食性天敵-巴氏新小綏螨毒死蜱中抗品系,在本研究中發現其抗性形成可能與P450解毒酶對毒死蜱代謝能力的改變有關。通過桿狀病毒蛋白表達系統,獲得了巴氏新小綏螨三種P450微粒體蛋白,其活性與陽性對照鼠肝微粒體P450活性類似,CO差光譜顯示在450nm處具有吸收峰,利用NADPH再生系統,建立了三種P450微粒體蛋白對毒死蜱體外代謝的反應體系,HPLC結合MS/MS分析發現,一個在抗性品系中下調表達的P450可以將毒死蜱吡啶環5號位的氯離子脫去并羥基化,而該蛋白下調表達產生抗性的原因我們推測其代謝產物為毒死蜱的毒性激活物,減少該成分產生有助于減緩其對作用靶標的蛋白活性抑制。
摘要
Selection of chemical-resistant predatory mites is a good alternative to balance the contradiction between chemical control and biological control. Previously, a resistant strain of Neoseiulus barkeri for chlorpyrifos was obtained. In the current study, two up-regulated (NbCYP3A6, NbCYP3A16) and one down-regulated (NbCYP3A24) P450s were screened through differential expression analysis and other detoxification-related genes such as CCEs, GST, etc. were not found. 3D modelling and molecular docking indicated that the chlorine at position 5 on the pyridine ring of chlorpyrifos, as well as a methyl group, were closest to the heme iron of the enzymes (less than 5 Å). Three active recombinant P450 proteins were heterologously expressed and metabolized with chlorpyrifos in vitro. HPLC assay showed that only NbCYP3A24 could metabolize chlorpyrifos, with a metabolism rate of 21.60 %. Analysis of the m/z of metabolites by LC-MS/MS showed that chlorine at the 5C position of chlorpyrifos was stripped and hydroxylated, whereas chlorpyrifos-oxon, a common product of oxidation by P450, was not found. Knockdown of the NbCYP3A24 gene in the susceptiblestrain did reduce the.
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