Bride J M, Cuany A, Amichot M, Brun A, Babault M, Mouël T L, De Sousa G, Rahmani R, Berge J B
Institut National de la Recherche Agronomique, Laboratoire de Biologie des Invertébrés, Antibes, France.
J Econ Entomol. 1997 Dec;90(6):1514-20. doi: 10.1093/jee/90.6.1514.
Studies were conducted between 1993 and 1996 using 3 natural grape vine populations, 1 susceptible laboratory strain, and 1 resistant selected strain of Drosophila melanogaster L. In vitro monooxygenase activity (ethoxycoumarine-O-deethylation) (ECOD) was recorded from microsomal fractions of all strains. Results varied over a 6-fold range between susceptible laboratory Canton and resistant selected RDDT strains and over a 2-fold range between the Canton strain and natural populations of flies. Few significant variations of ECOD activity were detected among the natural populations despite many insecticide treatments, but activities were significantly correlated with toxicological tolerance to 5 of the 15 insecticides (deltamethrin, fipronil, chlorpyriphos ethyl, DDT, and diazinon). Moreover, immunoblotting responses of microsomal protein encoded by Cyp6A2 showed that the levels of expression were quantitatively correlated with toxicological tolerance to almost the same group of insecticides (deltamethrin, fipronil, chlorpyriphos ethyl, DDT, fenvalerate, and fenthion). However, the level of CYP6A2 expression in some natural strains (still weakly resistant) was almost comparable with one of the resistant strains. In vivo monooxygenase activity recorded in individual abdomens of flies showed that frequency distributions of ECOD activity in natural populations overlapped those of the resistant and laboratory strains, which were much narrower. Substantial and fast frequency changes (of the narrowness) that obtained in laboratory were related to either the time of rearing of 1 of the natural populations or selecting this population with an insecticide that has a toxicology correlated with both of the monooxygenase signs measured. Perspectives on using the CYP6A2 expression and ECOD activity for detecting a resistance mechanism by cytochrome P450 in field populations are discussed.
在1993年至1996年间,利用3个天然葡萄藤果蝇种群、1个敏感的实验室品系以及1个经选育的抗性黑腹果蝇品系开展了研究。记录了所有品系微粒体部分的体外单加氧酶活性(乙氧基香豆素-O-脱乙基酶)(ECOD)。结果显示,敏感的实验室品系广州品系与经选育的抗性RDDT品系之间的活性变化范围达6倍,广州品系与果蝇天然种群之间的活性变化范围达2倍。尽管进行了多次杀虫剂处理,但在天然种群中未检测到ECOD活性的显著差异,不过其活性与15种杀虫剂中的5种(溴氰菊酯、氟虫腈、毒死蜱、滴滴涕和二嗪农)的毒理学耐受性显著相关。此外,由Cyp6A2编码的微粒体蛋白的免疫印迹反应表明,其表达水平与几乎同一组杀虫剂(溴氰菊酯、氟虫腈、毒死蜱、滴滴涕、氰戊菊酯和倍硫磷)的毒理学耐受性呈定量相关。然而,一些天然品系(仍具有弱抗性)中CYP6A2的表达水平几乎与抗性品系之一相当。在果蝇个体腹部记录的体内单加氧酶活性表明,天然种群中ECOD活性的频率分布与抗性品系和实验室品系的频率分布重叠,而后两者的频率分布范围要窄得多。在实验室中获得的(频率分布范围狭窄程度的)显著且快速的频率变化与其中一个天然种群的饲养时间有关,或者与用一种毒理学与所测两种单加氧酶指标均相关的杀虫剂对该种群进行选育有关。讨论了利用CYP6A2表达和ECOD活性检测田间种群中细胞色素P450抗性机制的前景。