Li Xianchun, Berenbaum May R, Schuler Mary A
Department of Plant Protection, Nanjing Agricultural University, Nanjing, China.
Insect Mol Biol. 2002 Aug;11(4):343-51. doi: 10.1046/j.1365-2583.2002.00341.x.
Four cytochrome P450 genes, CYP6B8, CYP6B9, CYP6B27 and CYP6B28, exist in the Helicoverpa zea genome as two pairs of paralogs that evolved from gene duplication and 5'-polar gene conversion events. RT-PCR gel blot analyses have shown that all of these genes are expressed constitutively in midguts of all larval instars, suggesting that they have primary roles in the detoxification of plant allelochemicals. Among these, CYP6B9 is expressed only in midgut tissue whereas its paralog, CYP6B27, is expressed primarily in midgut and secondarily in fat body and ovary. CYP6B28 is expressed in midgut, fat body and, to lesser extents in ovary and integument whereas its paralog, CYP6B8, is expressed in midgut and to some extent in fat body. Comparison of the expression levels induced by eight plant allelochemicals, one drug (phenobarbital), and an insecticide (alpha-cypermethrin) indicates that, for the most part, the four P450s respond individually to these inducers, with all four induced strongly by chlorogenic acid, a shikimate pathway intermediate and a lignin biosynthesis intermediate present in a wide variety of plants, and indole-3-carbinol, a glucobrassicin breakdown product present in the Brassicaceae. The multiple levels at which these P450 genes are apparently diverging (e.g. transcriptional responses, protein sequences) support the suggestion that gene conversion events facilitate gene evolution by allowing duplicated copies greater time to acquire selectable differences in both coding and promoter sequences.
在棉铃虫基因组中存在四个细胞色素P450基因,即CYP6B8、CYP6B9、CYP6B27和CYP6B28,它们以两对旁系同源基因的形式存在,这些旁系同源基因是由基因复制和5'-极性基因转换事件进化而来的。逆转录聚合酶链反应(RT-PCR)凝胶印迹分析表明,所有这些基因在所有幼虫龄期的中肠中均组成性表达,这表明它们在植物化感物质的解毒过程中起主要作用。其中,CYP6B9仅在中肠组织中表达,而其旁系同源基因CYP6B27主要在中肠中表达,其次在脂肪体和卵巢中表达。CYP6B28在中肠、脂肪体中表达,在卵巢和体壁中的表达程度较低,而其旁系同源基因CYP6B8在中肠中表达,并在一定程度上在脂肪体中表达。对八种植物化感物质、一种药物(苯巴比妥)和一种杀虫剂(α-氯氰菊酯)诱导的表达水平进行比较表明,在大多数情况下,这四个细胞色素P450对这些诱导剂的反应是独立的,这四种细胞色素P450均被绿原酸强烈诱导,绿原酸是莽草酸途径中间体和木质素生物合成中间体,存在于多种植物中,以及吲哚-3-甲醇,它是十字花科植物中硫代葡萄糖苷的分解产物。这些细胞色素P450基因明显分化的多个层面(例如转录反应、蛋白质序列)支持了这样一种观点,即基因转换事件通过允许复制的基因拷贝有更多时间在编码和启动子序列中获得可选择的差异,从而促进了基因进化。