中国农业科学 ›› 2017, Vol. 50 ›› Issue (8): 1440-1451.doi: 10.3864/j.issn.0578-1752.2017.08.008

• 植物保护 • 上一篇    下一篇

氯虫苯甲酰胺亚致死剂量对甜菜夜蛾主要解毒酶活性与生长繁殖的影响

陈羿渠,向兴,贡常委,王学贵   

  1. 四川农业大学农学院无公害农药研究实验室,成都 611130
  • 收稿日期:2016-12-12 出版日期:2017-04-16 发布日期:2017-04-16
  • 通讯作者: 王学贵,Tel:028-86290977;E-mail:wangxuegui@sicau.edu.cn
  • 作者简介:陈羿渠,Tel:028-86290977;E-mail:chenyiqu123@163.com。
  • 基金资助:
    国家公益性行业(农业)科研专项(201203038)

Effects of Sublethal Doses of Chlorantraniliprole on the Detoxification Enzymes Activities and the Growth and Reproduction of Spodoptera exigua

CHEN YiQu, XIANG Xin, GONG ChangWei, WANG XueGui   

  1. Biorational Pesticide Research Laboratory, College of Agronomy, Sichuan Agricultural University, Chengdu 611130
  • Received:2016-12-12 Online:2017-04-16 Published:2017-04-16

摘要: 【目的】甜菜夜蛾(Spodoptera exigua)是一种杂食性害虫,在不同环境、药物等选择压力下表现出不同的生长发育特点。氯虫苯甲酰胺是一种新型广谱的鱼尼丁受体杀虫剂,对鳞翅目害虫杀虫活性强。本研究旨在探究氯虫苯甲酰胺亚致死剂量对甜菜夜蛾幼虫3种主要解毒酶——羧酸酯酶(CarE)、谷胱甘肽S-转移酶(GSTs)和多功能氧化酶(MFOs)活性以及对种群繁殖的影响。【方法】采用饲料混毒法测定氯虫苯甲酰胺对SE-Lab品系、SE-Sel品系的毒力,SE-Sel品系由SE-Lab品系经亚致死剂量LC25连续汰选6代得到;通过浸叶法测定磷酸三苯酯(TPP)、顺丁烯二酸二乙酯(DEM)、胡椒基丁醚(PBO)3种酶抑制剂与氯虫苯甲酰胺协同对SE-Lab和SE-Sel品系的毒力增效作用,提前12 h让试虫取食浸渍过酶抑制剂的叶片,对照组取食用0.1% TritonX-100浸渍后的叶片,再分别测定氯虫苯甲酰胺对使用酶抑制剂与未使用酶抑制剂试虫的毒力;在冰上解剖试虫的中肠和脂肪体,并通过离体酶活性测定,分析氯虫苯甲酰胺亚致死剂量与酶抑制剂对甜菜夜蛾体内的3种代谢解毒酶活力的影响;通过记录试虫各个年龄阶段的生长、死亡、产卵量等数据,参照两性生命表理论分析SE-Lab和SE-Sel品系的两性生命表参数差异。【结果】在3种酶抑制剂中PBO增效作用最强,其对甜菜夜蛾SE-Sel品系和SE-Lab品系对氯虫苯甲酰胺的毒力增效比分别为1.58和1.69。在氯虫苯甲酰胺亚致死剂量连续汰选下,甜菜夜蛾体内3种解毒酶活性均被诱导上升,其中MFOs酶活力上升最显著,SE-Sel品系中肠和脂肪体的MFOs活性相对于SE-Lab品系分别提高了2.07和2.10倍,而经氯虫苯甲酰胺亚致死剂量再次诱导的SE-Sel试虫的MFOs活性亦较SE-Lab品系上升4.02和3.44倍;在使用了酶抑制剂后3种解毒酶酶活力均有下降,其中MFOs活性下降最多,其酶比活力仅为未使用酶抑制剂处理的42.3%—44.8%。与SE-Lab品系相比,SE-Sel品系成虫的产卵前期和总产卵前期变长,而产卵量减少;SE-Sel品系的内禀增长率(r)、周限增长率(λ)和净增殖率(R0)均显著小于SE-Lab品系,SE-Lab品系与SE-Sel品系的r分别为0.18和0.16 d-1,λ为1.20和1.17 d-1, R0为358.42和203.12 d-1。尽管SE-Sel品系的平均世代周期(T)更长,但是与SE-Lab品系无显著差异。【结论】MFOs可能为甜菜夜蛾对氯虫苯甲酰胺解毒代谢过程中的主要解毒酶,在其后续抗性形成中起主要作用;甜菜夜蛾在氯虫苯甲酰胺亚致死剂量作用下,世代周期延长,繁殖力降低,种群增长减缓,氯虫苯甲酰胺亚致死剂量对甜菜夜蛾有持续控制作用。

关键词: 甜菜夜蛾, 氯虫苯甲酰胺, 亚致死剂量, 增效剂, 多功能氧化酶, 两性生命表

Abstract: 【Objective】 Spodoptera exigua is a polyphagous pest and shows different characteristics of growth and development under the selective pressure of different environments, drugs and so on. Chlorantraniliprole, a novel insecticide that acts on ryanodine receptors, has highly activity to those insects in Lepidopteran. The objective of this study is to explore the effects of S. exigua larvae treated by the sublethal doses of chlorantraniliprole on the toxicities, the activities of three main detoxifying enzymes, including carboxylesterase (CarE), glutathione S-transferase (GSTs), and mixed-function oxidase (MFOs), and population breeding.【Method】 The toxicities of chlorantraniliprole on SE-Lab and SE-Sel strains were detected by incorporation bioassay and the SE-Sel strain was achieved with continuous selecting with LC25 sublethal doses for six generations from SE-Lab strain, then the synergistic effects of enzyme inhibitors (TPP, DEM, PBO) with chlorantraniliprole on the SE-Lab and SE-Sel strains were assayed using the dip-leaf method. The toxicities of chlorantraniliprole on the insects which were fed the leaves soaked with the solution of enzyme inhibitor or 0.1% TritonX-100 as blank control before 12 h were assayed by the dip-leaf method. The midgut and fatbody of tested insects were dissected on the ice and the effects of the sublethal doses of chlorantraniliprole and enzyme inhibitors on three metabolic detoxification enzyme activities were analyzed by the determination of detoxification enzyme activities. According to the age-stage two-sex life tables theory, the growth, mortality, fecundity data of the tested insects were also recorded to analyze the differences of the age-stage two-sex life table parameters between the strains of SE-Lab and SE-Sel. 【Result】 The synergistic effect of the PBO was the strongest among the three enzyme inhibitors and the synergic ratios on the strains of SE-Sel and SE-Lab reached 1.58- and 1.69-fold, respectively. The activities of the three detoxification enzymes, which were induced by the continuous selection of the sublethal doses of chlorantraniliprole, were promoted and the MFOs activities were the most significant, which of the SE-Sel strain in the midgut and fatbody were enhanced by 2.07- and 2.10-fold, meanwhile, the MFOs activities of the insects of SE-Sel induced by the sublethal dose of chlorantraniliprole again were also promoted by 4.02- and 3.44-fold in the midgut and fatbody compared to those in SE-Lab strain, respectively. The enzyme activities of three detoxifying enzymes were decreased when the tested insects were treated with enzyme inhibitors and the descend range of MFOs activity among the three enzymes was maximum, which was only 42.3%-44.8% compared to the treatment not treated with the enzyme inhibitor. The adult preoviposition period and total preoviposition period of F1 generation of SE-Sel strains became shorter and spawning quantity of became higher compared to those in F1 generation of SE-Lab. SE-Sel strains had the longest mean generation time and least amount of eggs. The intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R0) of SE-Sel strain were significantly lower than those in SE-Lab strain. The r, λ and R0 values of SE-Lab and SE-Sel strains were 0.18 and 0.16 d-1, 1.20 and 1.17 d-1, 358.42 and 203.12 d-1, respectively. Even though the mean generation times of SE-Sel strain was longer than SE-Lab strain, there was no significant difference between the two strains.【Conclusion】MFOs may be the major detoxication enzyme on the metabolic detoxification of chlorantraniliprole in the S. exigua and involved in the formation of resistance. A longer generation period, lower fecundity and slower population growth of S. exigua were demonstrated when it was treated by a continuous selection of lethal dose of chlorantraniliprole. The sublethal doses of chlorantraniliprole have a continuous control effect on S. exigua.

Key words: Spodoptera exigua, chlorantraniliprole, sublethal doses, synergists, MFOs, age-stage two-sex life tables