| 摘要: |
| 【目的】研究骆驼蓬总碱对草地贪夜蛾幼虫的杀虫作用机理，为此类生物碱作为新型植物杀虫剂的开发应用提供理论依据。【方法】从骆驼蓬成熟种子中提取骆驼蓬总碱，采用MTT法测定0,1,5,10和20 μg/mL骆驼蓬总碱对草地贪夜蛾(Spodoptera frugiperda)卵巢细胞系(Sf9)的毒性；测定5 μg/mL 骆驼蓬总碱在处理10,30,50,70和90 min 时对Sf9细胞Na+和K+吸收的影响，并测定5 μg/mL骆驼蓬总碱在处理1,2,3,4,5 d 后对葡萄糖吸收的影响；最后采用叶碟法（叶碟在20 μg/mL 骆驼蓬总碱中浸3 s）和注射法（2 μg/头）分别测定处理1,8,16,24和48 h 时骆驼蓬总碱对草地贪夜蛾4龄幼虫血细胞数量的影响。【结果】10和20 μg/mL骆驼蓬总碱处理草地贪夜蛾Sf9细胞24 h后，对细胞的抑制率分别为62.04%和83.25%；骆驼蓬总碱处理后8，16，24和48 h，其对草地贪夜蛾Sf9细胞的LC50分别为35.32，23.21，14.87 和8.98 μg/mL，骆驼蓬总碱对Sf9细胞的毒性有时间滞后效应。5 μg/mL骆驼蓬总碱处理Sf9细胞后10 min，Sf9细胞对Na+、K+的吸收速率明显增加，处理后30 min Sf9细胞对Na+、K+的吸收逐渐减弱；处理后3 d Sf9细胞对葡萄糖的吸收迅速增加，处理后4 d Sf9细胞对葡萄糖的吸收趋于停止。以叶碟法和注射法处理草地贪夜蛾4龄幼虫，8 h后幼虫血细胞数量显著降低，但随时间延长幼虫血细胞数量又逐渐增加。【结论】骆驼蓬总碱对草地贪夜蛾卵巢细胞系Sf9有明显的毒杀活性，且对Sf9细胞中Na+、K+和葡萄糖的吸收具有抑制作用；对草地贪夜蛾4龄幼虫血细胞数量也有明显的抑制作用。 |
| 关键词: 骆驼蓬 生物总碱 草地贪夜蛾 卵巢细胞系 细胞毒性 |
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| 基金项目:安徽省自然科学基金面上项目(1308085MC39)；安徽省教育厅高校自然科学基金重点项目(KJ2013A207)；安徽省省级学科建设重大项目(2014-28-A12) |
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| Toxicity effects of total alkaloids of Peganum harmala against in vitro cultured Sf9 cells and larvae blood corpuscles of Spodoptera frugiperda |
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LI Jinmei,MU Kui,JIANG Shuanglin,et al
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| Abstract: |
| 【Objective】This study investigated the insecticidal mechanism of total alkaloids of Peganum harmala (TAPH) against larvae of Spodoptera frugiperda to provide basis for the development and application of alkaloids as a new plant insecticide.【Method】TAPH was isolated from mature seeds of P.harmala and its cytotoxicity (0,1,5,10,and 20 μg/mL) against S.frugiperda Sf9 cells was detected using MTT method.The effect of 50 μg/mL TAPH treatment of 10,30,50,70 and 90 min on absorption of Na+ and K+ by Sf9 cells was measured,and the effect of 50 μg/mL TAPH treatment of 1,2,3,4, 5 d on the absorption of glucose was determined.Finally,leaf disc method (leaves disc in 20 μg/mL TAPH dipping for 3 s) and injecting test (2 μg/head TAPH) were used to detect the effect on blood corpuscle of 4th instar larvae of S.frugiperda.【Result】After 24 h treatment with 20 μg/mL and 10 μg/mL TAPH,the inhibition rates were 83.25% and 62.04%,respectively.The LC50 to Sf9 cells after treatment of 8,16,24 and 48 h were 35.32,23.21,14.87 and 8.98 μg/mL,respectively.After 10 min treatment with TAPH,the absorption of Na+ and K+ increased rapidly,and the absorption decreased after 30 min.After 3 d treatment,the absorption of glucose increased rapidly, while the absorption stopped after 4 days.Treated by leaf disc and injection methods,the cell amounts of the 4th instar larvae of S.frugiperda decreased significantly,and recovered gradually as time extended. 【Conclusion】The total alkaloids had obvious toxicity against Sf9 cells.The absorption of Na+,K+,and glucose and the number of blood corpuscle of 4th instar larvae of S.frugiperda were significantly inhibited. |
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