Xu Lu, Wu Min, Han Zhaojun
Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing, China.
Pest Manag Sci. 2014 Jul;70(7):1118-29. doi: 10.1002/ps.3657. Epub 2013 Oct 25.
Laboratory selection is often employed in resistance mechanism studies because field-derived populations commonly do not have high enough resistance for such studies. In the present study, a field-collected Laodelphax striatellus population from eastern China was laboratory selected for chlorpyrifos resistance and susceptibility, and the developed strains, along with a field population, were studied for cross-resistance and resistance mechanisms at biochemical and molecular levels.
A 158.58-fold chlorpyrifos-resistant strain (JH-chl) and a chlorpyrifos-susceptible strain (JHS) were established after laboratory selection of 25 generations. Cross-resistance to deltamethrin, diazinon, methomyl, carbosulfan, acephate and imidacloprid were detected in JH-chl and a field-collected strain (JHF). Synergism and enzyme activity data suggested potential involvement of P450s and esterases in JH-chl as well as AChE alteration. Furthermore, CYP6AY3v2, CYP306A2v2, CYP353D1v2 and LSCE36 genes were significantly overexpressed in JH-chl (6.87-12.14-fold). Feeding of dsRNAs reduced the expression of the four target genes (35.6-56.8%) and caused significant adult mortality (75.21-88.45%), implying resistance reduction. However, mechanism(s) conferring chlorpyrifos resistance in JHF were unclear.
In contrast to previous reports, multiple overexpressed detoxification genes were potentially associated with chlorpyrifos resistance, as confirmed by RNAi feeding tests. Chlorpyrifos resistance exhibits cross-resistance with insecticides in the same and different classes.
在抗性机制研究中常采用实验室选育,因为从田间采集的种群通常抗性不足以进行此类研究。在本研究中,对从中国东部田间采集的灰飞虱种群进行了毒死蜱抗性和敏感性的实验室选育,并对选育出的品系以及一个田间种群在生化和分子水平上进行了交互抗性和抗性机制研究。
经过25代实验室选育,建立了一个对毒死蜱抗性达158.58倍的品系(JH-chl)和一个对毒死蜱敏感的品系(JHS)。在JH-chl和一个田间采集的品系(JHF)中检测到对溴氰菊酯、二嗪农、灭多威、丁硫克百威、乙酰甲胺磷和吡虫啉的交互抗性。增效作用和酶活性数据表明,P450s和酯酶可能参与了JH-chl的抗性,同时乙酰胆碱酯酶也发生了改变。此外,CYP6AY3v2、CYP306A2v2、CYP353D1v2和LSCE36基因在JH-chl中显著过表达(6.87 - 12.14倍)。饲喂dsRNAs降低了这四个靶基因的表达(35.6% - 56.8%),并导致显著的成虫死亡率(75.21% - 88.45%),这意味着抗性降低。然而,JHF中赋予毒死蜱抗性的机制尚不清楚。
与之前的报道不同,RNAi饲喂试验证实多个过表达的解毒基因可能与毒死蜱抗性相关。毒死蜱抗性与同一类和不同类别的杀虫剂存在交互抗性。