• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

西花蓟马不同RNA干扰技术比较研究

Artificial RNA Interference on Frankliniella occidentalis

    摘要
  • 摘要:
      目的  西花蓟马Frankliniella occidentalis (Pergande)是一种重要的外来入侵害虫，其繁殖能力强、寄主范围广、抗药性强，给我国蔬菜、花卉等经济作物造成了严重危害。本研究旨在评估膜饲喂和显微注射dsActin对西花蓟马Actin基因的沉默效率，以期为蓟马类等小型昆虫基因功能的研究提供方法和依据。
      方法  通过膜饲喂和显微注射的方法将体外合成的dsActin导入西花蓟马二龄若虫体内，用RT-qPCR方法检测Actin基因的mRNA表达；通过单头饲养方法观察统计西花蓟马成虫体长、成虫翅膀或胸腹部畸形率及死亡率。
      结果  dsActin膜饲喂后24 、48、72 h，Actin基因的相对表达量分别为对照组的97%、91%和98%；通过显微注射将dsActin注入西花蓟马体腔，在注射后24、48、72 h，Actin基因的表达量分别为对照组的68%、56%和53%。注射dsActin的西花蓟马在第24～120 h 死亡率为44%～98%，显著高于dsGFP对照组；此外，dsActin组个体体长仅为dsGFP对照组的90%，且翅膀或胸腹部出现畸形率为41%。
      结论  显微注射dsActin能显著沉默西花蓟马Actin基因mRNA水平的表达，并引起西花蓟马个体不正常发育和死亡，从而建立蓟马类小型昆虫RNAi体系。

     

    Abstract:
      Objective  Application of dsActin to control the proliferating, multiple-hosting, insecticide-resistant invasive western flower thrips on vegetables and ornamental crops in China was investigated.
      Method  Synthetic dsActin or dsGFP (as control) was introduced into the 2nd-instar nymphs of Frankliniella occidentalis (Pergande) by either membrane-feeding or microinjection. The treated thrips were collected for mRNA expression detection by RT-qPCR at 24, 48 and 72 h after the treatment. Effects of 0.5 μg·μL−1 dsRNA on the body length, wing or thorax malformation, and mortality of the insects were monitored in an individual culture experiment conducted in the laboratory.
      Result  The relative expression of Actin mRNA in the thrips was similar to that of control 24, 48 and 72 h after membrane-feeding. But the expressions in the injection group decreased to 68% in 24 h, 56% in 48 h, and 53% in 72 h. Significantly higher mortality rate (44%-98% in 24-120 h), shorter body length (90% of control), and higher bodily deformation rate (41%) were observed in the dsRNA-treated thrips than control.
      Conclusion  The Actin mRNA expression in F. occidentalis was significantly suppressed by dsActin injection at the test level to induce malformations or death in the insects. It suggested that an in vivo RNA interference method was made available for studying the gene functions and control of thrips.

     

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