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杀虫剂抗性是害虫防治中的一个主要挑战。为有效治理抗性,我们必须了解杀虫剂诱导的害虫生理生化变化。目前害虫抗药性的一些机理已经清楚,但更多的相关机制还有待探究。本研究通过蛋白质组学方法检测了小菜蛾Plutella xylostella溴氰菊酯敏感和抗性品系间蛋白质组的表达差异。结果显示:SDS-PAGE胶上有大约300个蛋白差异点,其中23个蛋白点具2.5倍以上的表达差异,通过MALDI-TOF-MS,我们成功鉴定出8个蛋白,其中包括化感蛋白CSP2、铜锌超氧化物歧化酶和peroxiredoxin样蛋白。通过实时定量PCR(real-time quantitativePCR,qPCR)分析了其中5个蛋白的mRNA表达水平,结果表明mRNA表达水平不能真实反映蛋白的表达水平。免疫印迹验证了双向电泳中SOD1的表达差异。本研究有力地证明溴氰菊酯诱导小菜蛾成虫蛋白质组表达变化,这为进一步筛选抗性靶标提供很大的帮助。
Insecticide resistance is a major challenge in pest control. In order to effectively control the resistance, we must understand the insecticide-induced physiological and biochemical changes of pests. At present, some mechanisms of pest resistance are clear, but more relevant mechanisms have yet to be explored. In this study, proteomics was used to detect the differences of proteome between Plutella xylostella sensitive and resistant strains of Plutella xylostella. The results showed that there were about 300 protein spots on SDS-PAGE gel, among which 23 spots had more than 2.5-fold difference in expression. By MALDI-TOF-MS, we successfully identified 8 proteins, including CSP2 , Copper-zinc superoxide dismutase and peroxiredoxin-like protein. The mRNA expression levels of five proteins were analyzed by real-time quantitative PCR (qPCR). The results showed that the mRNA expression levels did not truly reflect the protein expression level. Immunoblotting validated the difference of SOD1 expression in two-dimensional electrophoresis. This study strongly demonstrated that deltamethrin induced changes in proteome expression in the diamondback moth, Plutella xylostella, which would be of great help to further screen the resistance targets.