›› 2013, Vol. 56 ›› Issue (1): 1-8.doi:

• 研究论文 •    下一篇

小菜蛾溴氰菊酯敏感和抗性品系蛋白质表达谱的比较分析

张娜娜1,#, 张宏1,#, 程晨1, 李凤良2, 高少奇1, 程罗根1,*   

  1. (1. 南京师范大学生命科学学院, 南京 210097; 2. 贵州省农业科学院植物保护研究所, 贵阳 550009)
  • 出版日期:2013-01-20 发布日期:2013-01-20

A comparative profiling of protein expression in the deltamethrin-sensitive and resistant strains of the diamondback moth (Plutella xylostella)

ZHANG Na-Na1,#, ZHANG Hong1,#, CHENG Chen1, LI Feng-Liang2, GAO Shao-Qi1, CHENG Luo-Gen1,*   

  1. (1. College of Life Sciences, Nanjing Normal University, Nanjing 210097, China; 2. Institute of Plant Protection, Guizhou Academy of Agricultural Sciences, Guiyang 550009, China)
  • Online:2013-01-20 Published:2013-01-20

摘要: 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 quantitative PCR, 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。

关键词: 小菜蛾, 溴氰菊酯, 蛋白质组, 选择压力, 杀虫剂抗性

Abstract: A major challenge in pest management is insecticide resistance. To effectively prevent and manage resistance, it is essential to understand the biochemical and physiological changes induced by insecticides. Some molecular mechanisms that are responsible for insecticide resistance were well known whereas others are to be discovered. So the comparative proteomic approach was employed to investigate the proteomic differences between the deltamethrin-sensitive strain (DS) and the deltamethrin-resistant strain (DR) of the diamondback moth (Plutella xylostella L.). In this study, about 300 differentially expressed protein spots were detected on each SDS-PAGE gel, among which 23 proteins had a 2.5-fold expression difference between the two strains. Eight proteins were successfully identified by MALDI-TOF-MS, including chemosensory protein CSP2, Cu/Zn superoxide dismutase (SOD1), peroxiredoxinlike protein, etc. Expression of the five proteins at the mRNA level was analyzed by real-time quantitative PCR, and the results showed that the mRNA expression level did not fully reflect the protein expression level. Western blotting validated the proteomic result of SOD1. This study provides strong evidence that deltamethrin can induce the proteomic change in adult diamondback moth, and is helpful to understanding the resistance development and selecting the action targets of deltamethrin.

Key words: Plutella xylostella, deltamethrin, proteomics, selection pressure, insecticide resistance