Expression profile analysis of silkworm P450 family genes after phoxim induction
Expression profile analysis of silkworm P450 family genes after phoxim induction
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DOI：
10.1016/j.pestbp.2014.12.013
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发表时间：
2015-07-01
影响因子：
4.7
通讯作者：
Li, Bing
中科院分区：
文献类型：
--
作者：
Li, Fanchi;Ni, Min;Li, Bing
家蚕(Bombyx Mori)是重要的经济昆虫，也是鳞翅目昆虫的模式种。中国地区每年都发生O，O-二乙基-O-(α-氰基苯亚氨基)硫代硫酸酯(辛硫磷)农药中毒，给蚕业造成巨大的经济损失。家蚕脂肪体是营养物质储存、能量供应、中间代谢和解毒的主要器官。家蚕细胞色素P450解毒酶基因芯片分析表明，所有家蚕细胞色素P450解毒酶家族基因均在脂肪体中表达。采用实时定量聚合酶链式反应(QRT-PCR)检测辛硫磷染毒后0、24、48、72h家蚕脂肪体中细胞色素P4家族基因的表达。家蚕蜕皮激素合成相关基因的表达水平在辛硫磷暴露后24 h开始发生变化，其中细胞色素P302A1、细胞色素P306A1和细胞色素P314A1的表达分别增加1.38倍、1.33倍和2.10倍。参与类固醇激素失活的CYP18A1基因和参与金合欢酸甲酯(MF)合成保幼激素UFO环氧化反应的CYP15C1基因分别增加了3.85倍和7.82倍。系统发育分析表明，这些内源激素代谢相关基因属于CYP mito家族和家族2，辛硫磷暴露可能影响家蚕的发育和变态。其中，细胞色素P49A1、细胞色素P4L6、细胞色素P6AB4、细胞色素P9G3、细胞色素9A19和细胞色素P9A22的转录水平显著上调，分别为对照的12.77倍、2.64倍、2.42倍、4.06倍、3.32倍和2.98倍。在脂肪体中，经辛硫磷处理24、48和72h后，细胞色素P49A1、细胞色素P6AB4、细胞色素P9A19和细胞色素P9A22持续高水平表达；根据系统发育分析，这些基因属于与解毒相关的家族3和家族4。这些基因可能参与了辛硫磷在家蚕脂肪体中的代谢。本研究结果为进一步深入研究昆虫P450家族基因在代谢解毒中的作用奠定了基础。(C)2014 Elsevier Inc.保留所有权利。
Silkworm (Bombyx mori) is an important economic insect and a model species for Lepidopteran. Each year, O,O-diethyl O-(alpha-cyanobenzylideneamino) phosphorothioate (phoxim) pesticide poisoning in China results in huge economic losses in sericulture. Silkworm fat body is the main organ for nutrient storage, energy supply, intermediary metabolism, and detoxification. Microarray analysis of silkworm Cytochrome P450 detoxification enzyme genes revealed that all tested P450 4 (CYP4) family genes are expressed in the fat body. Quantitative Real-time PCR (QRT-PCR) was used to detect the expression of CYP4 family genes in silkworm fat body 0, 24, 48, and 72 h after phoxim exposure. The expression levels of silkworm molting hormone synthesis-related genes started to change 24 h after phoxim exposure, with those of CYP302A1, CYP306A1, and CYP314A1 being elevated by 1.38-, 1.33-, and 2.10-fold, respectively. The CYP18A1 gene that participates in steroid hormone inactivation and the CYP15C1 gene that participates in the epoxidation during the synthesis of juvenile hormone UFO from methyl farnesoate (MF) were increased by 3.85- and 7.82-fold, respectively. Phylogenetic analysis indicated that these endogenous hormone metabolism-related genes belong to CYP mito clan and clan 2, and that phoxim exposure may affect silkworm development and metamorphosis. The CYP4, CYP6, and CYP9 families all showed some degrees of increases in gene expression; among them, CYP49A1, CYP4L6, CYP6AB4, CYP9G3, CYP9A19, and CYP9A22's transcription levels were significantly upregulated to 12.77-, 2.64-, 2.42-, 4.06-, 3.32-, and 2.98-fold, respectively, of the control levels. In the fat body, CYP49A1, CYP6AB4, CYP9A19, and CYP9A22 were constantly expressed at high levels after 24, 48, and 72 h of phoxim treatments; according to phylogenetic analysis, these genes belong to detoxification-related clan 3 and clan 4 CYP families. These genes may participate in the metabolism of phoxim in silkworm fat body. The results obtained in this study provide a basis for future in-depth investigations of insect P450 family genes in metabolic detoxification. (C) 2014 Elsevier Inc. All rights reserved.