Liu Feng, Wang Xiao-Dong, Zhao Yi-Ying, Li Yan-Jun, Liu Yong-Chang, Sun Jie
Key Laboratory of Oasis Eco-agriculture, College of Agriculture, Shihezi University, Shihezi 832003, China.
Int J Biol Sci. 2015 Jan 1;11(1):67-74. doi: 10.7150/ijbs.10468. eCollection 2015.
Insect pests have caused noticeable economic losses in agriculture, and the heavy use of insecticide to control pests not only brings the threats of insecticide resistance but also causes the great pollution to foods and the environment. Transgenic plants producing double-stranded RNA (dsRNA) directed against insect genes have been is currently developed for protection against insect pests. In this study, we used this technology to silence the arginine kinase (AK) gene of Helicoverpa armigera (HaAK), encoding a phosphotransferase that plays a critical role in cellular energy metabolism in invertebrate. Transgenic Arabidopsis plants producing HaAK dsRNA were generated by Agrobacterium-mediated transformation. The maximal mortality rate of 55% was reached when H. armigera first-instar larvae were fed with transgenic plant leaves for 3 days, which was dramatically higher than the 18% mortality recorded in the control group. Moreover, the ingestion of transgenic plants significantly retarded larval growth, and the transcript levels of HaAK were also knocked down by up to 52%. The feeding bioassays further indicated that the inhibition efficiency was correlated with the integrity and concentration of the produced HaAK dsRNA in transgenic plants. These results strongly show that the resistance to H. armigera was improved in transgenic Arabidopsis plants, suggesting that the RNAi targeting of AK has the potential for the control of insect pests.
害虫给农业造成了显著的经济损失,大量使用杀虫剂防治害虫不仅带来了抗药性威胁,还对食品和环境造成了严重污染。目前已开发出产生针对昆虫基因的双链RNA(dsRNA)的转基因植物用于防治害虫。在本研究中,我们利用该技术沉默了棉铃虫精氨酸激酶(AK)基因(HaAK),该基因编码一种磷酸转移酶,在无脊椎动物的细胞能量代谢中起关键作用。通过农杆菌介导的转化产生了产生HaAK dsRNA的转基因拟南芥植物。用转基因植物叶片喂养棉铃虫一龄幼虫3天,最高死亡率达到55%,显著高于对照组记录的18%的死亡率。此外,取食转基因植物显著延缓了幼虫生长,HaAK的转录水平也被敲低了52%。取食生物测定进一步表明,抑制效率与转基因植物中产生的HaAK dsRNA的完整性和浓度相关。这些结果有力地表明转基因拟南芥植物对棉铃虫的抗性得到了提高,表明靶向AK的RNA干扰具有控制害虫的潜力。