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为了探知基因组甲基化是否参与了昆虫抗药性,本研究在室内对褐飞虱Nilaparva talugens连续9个世代的3龄若虫施用吡虫啉,用AFLP检测褐飞虱抗性产生过程中DNA甲基化多态性的变化。利用25对AFLP引物共获得120个位点,其中15个位点呈现甲基化多态性,共获得78条多态性条带。根据多态性条带在不同世代样本中出现的多少计算多态性条带比例,其中最高比例出现在G5代(10.26%),最低比例出现在G6代(1.28%)。多态性条带在不同世代间比例的变化趋势表明,褐飞虱对吡虫啉的筛选产生快速应答。在筛选早期(G1,G2和G3)世代间DNA甲基化多态性比例差异相对较小,变化范围在3.85%～6.41%之间;在筛选中期(G4,G5和G6)世代间比例差异较大,变化范围在1.28%～10.26%之间;在筛选后期(G7,G8和G9)世代间比例差异相对较小,变化范围在5.13%～7.69%之间。结果说明,吡虫啉的连续施用能够诱导褐飞虱基因组产生甲基化变异,初步揭示甲基化在褐飞虱抗药性产生过程中参与了基因组的表达调控。
In order to find out whether methylation of the genome is involved in insect resistance, imidacloprid was applied indoors to the third instar nymphs of Nilaparva talugens for nine generations. AFLP was used to detect DNA methylation polymorphism in the development of N. lugens. . A total of 120 loci were obtained using 25 pairs of AFLP primers, of which 15 loci showed methylation polymorphism and 78 polymorphic bands were obtained. According to the number of polymorphic bands in different generation samples, the highest proportion appeared in G5 generation (10.26%) and the lowest occurred in G6 generation (1.28%). The trend of polymorphic bands in different generations showed that the brown planthopper had a rapid response to imidacloprid screening. The differences of DNA methylation polymorphisms in the early generations (G1, G2 and G3) were relatively small, ranging from 3.85% to 6.41%. In the middle of screening (G4, G5 and G6) The range of variation was between 1.28% and 10.26%. The difference in the percentage of the late generation (G7, G8 and G9) was relatively small, ranging from 5.13% to 7.69%. The results showed that the continuous application of imidacloprid could induce the methylation variation of the genome of BPH, and initially revealed that methylation was involved in the genome regulation during the development of BPH.