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发布人:汇智和源生物技术(苏州)有限公司
发布日期:2024/8/27 11:56:48
近日,西南大学于士将老师,使用IPHASE品牌产品:小鼠肝微粒体在《Journal of Hazardous Materials》权威期刊上发表文章《A down-regulated cytochrome P450 in Neoseiulus barkeri Hughes (Acari: Phytoseiidae) can dechlorinate and hydroxylate chlorpyrifos without producing chlorpyrifos-oxon》,影响因子12.2!
在有害昆虫生物防治中农业系统中存在的天敌资源发挥着重要作用,常见的有机磷类杀虫剂如毒死蜱等在农田喷洒作业时,对天敌的生存造成了负面影响,选育抗药性天敌品系有助于生物防治和化学防治工作的协同开展。此前西南大学实验室选育了一种捕食性天敌-巴氏新小绥螨毒死蜱中抗品系,在本研究中发现其抗性形成可能与P450解毒酶对毒死蜱代谢能力的改变有关。通过杆状病毒蛋白表达系统,获得了巴氏新小绥螨三种P450微粒体蛋白,其活性与阳性对照鼠肝微粒体P450活性类似,CO差光谱显示在450nm处具有吸收峰,利用NADPH再生系统,建立了三种P450微粒体蛋白对毒死蜱体外代谢的反应体系,HPLC结合MS/MS分析发现,一个在抗性品系中下调表达的P450可以将毒死蜱吡啶环5号位的氯离子脱去并羟基化,而该蛋白下调表达产生抗性的原因我们推测其代谢产物为毒死蜱的毒性激活物,减少该成分产生有助于减缓其对作用靶标的蛋白活性抑制。
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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