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为了研究Interruptin B的杀虫活性,初步探讨其杀虫作用机理,本研究采用叶碟法和叶片浸渍法测定了Interruptin B对小菜蛾Plutella xylostella和亚洲玉米螟Ostrinia furnacalis3龄幼虫的拒食活性和毒杀活性;采用注射法和血球计数法研究对斜纹夜蛾Spodoptera litura幼虫血细胞数的影响;采用四甲基偶氮唑盐[3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide,MTT]法测定对斜纹夜蛾卵巢细胞(SL细胞)的增殖抑制作用;采用倒置显微镜观察对SL细胞形态、密度和贴壁率的影响。结果显示:处理后48h,Interruptin B对亚洲玉米螟和小菜蛾3龄幼虫的拒食中浓度(AFC50)分别为108.56和143.88μg/mL,致死中浓度(LC50)分别为346.23和194.32g/mL;处理后24h和48h,对SL细胞的抑制中浓度(IC50)分别为13.51和4.11g/mL。Interruptin B处理SL细胞后12-48h,细胞密度随时间的延长而下降,与对照差异显著(P<0.05);处理后3,6,12,24和48h,SL细胞变形率分别为28.18%,83.90%,85.18%,93.93%和100%,细胞贴壁率分别为96.59%,89.90%,83.18%,5.06%和1.09%,处理后6-48h,SL细胞变形率和贴壁率与对照差异显著(P<0.05)。Interruptin B对小菜蛾和亚洲玉米螟3龄幼虫具有良好的拒食和毒杀活性,能显著降低斜纹蛾幼虫血细胞数量,对SL细胞具有良好的增殖抑制作用,可显著降低SL细胞生长密度,改变细胞形态,降低贴壁率,表明该化合物可通过影响昆虫细胞活力毒杀活体昆虫。
In order to study the insecticidal activity of Interruptin B and its mechanism of insecticidal action, the antifeedant activity and toxicity of Interruptin B to 3rd instar larvae of Plutella xylostella and Ostrinia furnacalis were determined by leaf disc and leaf dipping method The effect on the number of blood cells of Spodoptera litura larvae was studied by injection and haemocytometry. The effects of the concentration of 3, 4,5-dimethylthiazole-2-yl-2,5-diphenyltetrazolium The MTT assay was used to determine the inhibitory effect on the proliferation of Spodoptera litura ovary cells (SL cells). The morphology, density and attachment rate of SL cells were observed by inverted microscope. The results showed that the antifeedant concentrations (AFC50) of Interruptin B to the 3rd instar larvae of Asian corn borer and Plutella xylostella were 108.56 and 143.88μg / mL respectively at 48h after treatment, with LC50 of 346.23 and 194.32g / mL, respectively. The inhibitory concentrations (IC50) on SL cells at 24 h and 48 h after treatment were 13.51 and 4.11 g / mL, respectively. After treated with Interruptin B for 12-48h, the cell density decreased with time, which was significantly different from the control (P <0.05). After 3, 6, 12, 24 and 48h treatment, the cell degeneration rate was 28.18% 83.90%, 85.18%, 93.93% and 100% respectively. The cell adherent rates were 96.59%, 89.90%, 83.18%, 5.06% and 1.09% Significant (P <0.05). Interruptin B had good antifeedant and toxin-killing activity against the 3rd instar larvae of Plutella xylostella and Asian corn borer, and could significantly reduce the number of hemocytes in the larvae of T. molitor and had a good proliferation inhibition on SL cells, significantly reducing the density of SL cells and changing the cell density Morphology, reducing the rate of attachment, indicating that the compound can kill live insects by affecting insect cell viability.