Menozzi Philippe, Shi Ming An, Lougarre Andrée, Tang Zhen Hua, Fournier Didier
Groupe de Biotechnologie des Protéines, IPBS-UMR 5089, F-31077 Toulouse, France.
BMC Evol Biol. 2004 Feb 5;4:4. doi: 10.1186/1471-2148-4-4.
Organophosphate and carbamate insecticides irreversibly inhibit acetylcholinesterase causing death of insects. Resistance-modified acetylcholinesterases(AChEs) have been described in many insect species and sequencing of their genes allowed several point mutations to be described. However, their relative frequency and their cartography had not yet been addressed.
To analyze the most frequent mutations providing insecticide resistance in Drosophila melanogaster acetylcholinesterase, the Ace gene was cloned and sequenced in several strains harvested from different parts of the world. Sequence comparison revealed four widespread mutations, I161V, G265A, F330Y and G368A. We confirm here that mutations are found either isolated or in combination in the same protein and we show that most natural populations are heterogeneous, composed of a mixture of different alleles. In vitro expression of mutated proteins showed that combining mutations in the same protein has two consequences: it increases resistance level and provides a wide spectrum of resistance.
The presence of several alleles in natural populations, offering various resistance to carbamate and organophosphate compounds will complicate the establishment of resistance management programs.
有机磷酸酯类和氨基甲酸酯类杀虫剂可不可逆地抑制乙酰胆碱酯酶,从而导致昆虫死亡。在许多昆虫物种中都发现了抗性修饰的乙酰胆碱酯酶(AChEs),对其基因进行测序后发现了多个点突变。然而,它们的相对频率及其图谱尚未得到研究。
为了分析在黑腹果蝇乙酰胆碱酯酶中提供杀虫剂抗性的最常见突变,我们从世界不同地区采集了多个品系,对Ace基因进行了克隆和测序。序列比较揭示了四个广泛存在的突变,即I161V、G265A、F330Y和G368A。我们在此证实,这些突变在同一蛋白质中可以单独出现或组合出现,并且我们发现大多数自然种群是异质的,由不同等位基因的混合物组成。突变蛋白的体外表达表明,同一蛋白质中组合突变有两个后果:它会提高抗性水平并提供广泛的抗性谱。
自然种群中存在多个等位基因,对氨基甲酸酯类和有机磷酸酯类化合物具有不同的抗性,这将使抗性管理计划的制定变得复杂。