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利用化学合成的 siRNA 沉默 Rieske 铁硫蛋白,作为防治小菜蛾的潜在生物农药。

Silencing of Rieske iron-sulfur protein using chemically synthesised siRNA as a potential biopesticide against Plutella xylostella.

机构信息

Key Laboratory of Pesticide and Chemical Biology, College of Natural Resources and Environment, South China Agricultural University, Guangzhou, China.

出版信息

Pest Manag Sci. 2011 May;67(5):514-20. doi: 10.1002/ps.2086. Epub 2011 Jan 14.

Abstract

BACKGROUND

Extensive applications and frequent long-term use of pesticides can affect behavioural mechanisms and physiological and biochemical aspects of insects, leading to resistance. However, insect control strategies involving a different mode of action would be valuable for managing the emergence of insect resistance. In this context, the development of RNA interference technology has brought a turning point in the creation of new biopesticides.

RESULTS

Full-length cDNA of Rieske iron-sulfur protein (RISP) was cloned and characterised from Plutella xylostella L. Three siRNAs specific to RISP sequences were designed and chemically synthesised, and fed to P. xylostella larvae by coating cabbage leaves. This resulted in specific gene silencing of RISP, and consequently brought significant mortality of P. xylostella larvae compared with the control treatment. Silencing of RISP leads to significantly lower transcript levels of RISP compared with the control. In addition, the amount of ATP in the surviving larvae was lower than in the control. However, surviving larvae gradually recovered to normal transcript and protein levels.

CONCLUSION

This is the first demonstration of the potential use of chemically synthesised siRNA in the development of new biopesticides as a mitochondrial electron transport inhibitor.

摘要

背景

大量应用和频繁的长期使用农药会影响昆虫的行为机制以及生理和生化方面,导致其产生抗性。然而,涉及不同作用模式的昆虫控制策略对于管理昆虫抗性的出现将是有价值的。在这种情况下,RNA 干扰技术的发展为新型生物农药的创制带来了转折点。

结果

从小菜蛾中克隆并鉴定了 Rieske 铁硫蛋白(RISP)的全长 cDNA。针对 RISP 序列设计并化学合成了 3 条特异性 siRNA,并通过包被白菜叶片将其喂食给小菜蛾幼虫。这导致了 RISP 的特异性基因沉默,从而与对照处理相比,导致小菜蛾幼虫的死亡率显著增加。与对照相比,RISP 的沉默导致 RISP 的转录本水平显著降低。此外,存活幼虫中的 ATP 量低于对照。然而,存活的幼虫逐渐恢复到正常的转录本和蛋白质水平。

结论

这是首次证明化学合成的 siRNA 作为线粒体电子传递抑制剂在新型生物农药开发中的潜在用途。

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