Spray method application of transdermal dsRNA delivery system for efficient gene silencing and pest control on soybean aphid Aphis glycines
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RNA interference (RNAi) has been recognized as a novel and safe strategy in pest management due to its high sequence-dependent specificity. However, the existing dsRNA delivery methods largely restrict the application of the RNAi-based pest management strategy; thus, we previously constructed a nanocarrier-based transdermal dsRNA delivery system on the soybean aphid Aphis glycines with the help of nanocarrier and detergent. In the current study, we improved our transdermal dsRNA delivery system with a smaller and cheaper nanocarrier to investigate the efficacy of spraying aphid-infested soybean seedlings to apply our RNA pesticide. A dsRNA/nanocarrier/detergent formulation was performed, and the dsRNA could penetrate the aphid body wall within 4 h with the help of nanocarrier through the topical application. Four potential RNAi target genes (TREH, ATPD, ATPE and CHS1) were selected and cloned, and their dsRNA fragments were synthesized and tested through the transdermal dsRNA delivery system. The delivered dsRNA efficiently silenced the target gene expression with the knockdown effects ranging from 86.86 to 58.87% and resulted in a high mortality up to 81.67% (dsATPD + dsATPE) through the topical application, when through the spray method, with the highest percent mortality of 78.50% (dsATPD + dsCHS1). Our novel transdermal dsRNA delivery system not only provides a powerful tool for gene functional analysis in laboratory, but also shows a great potential for the pest management in the field, which will promote the practice and development of RNAi-based pest management.
KeywordsAphis glycines Nanocarrier Pest management RNA pesticide dsRNA delivery system
We would like to thank Dr. Dunlun Song and Dr. Zhen Li for providing the transcriptome data.
This research was supported by the Beijing Natural Science Foundation (6182020), National Key Research and Development Program (2017YFD0201200) and Natural Science Foundation of China (31900363).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Baum JA, Roberts JK (2014) Chapter five—progress towards RNAi-mediated insect pest management. Adv Insect Physiol 47:249–295Google Scholar
- Bolognesi R, Ramaseshadri P, Anderson J, Bachman P, Clinton W, Flannagan R, Ilagan O, Lawrence C, Levine S, Moar W (2012) Characterizing the mechanism of action of double-stranded RNA activity against western corn rootworm (Diabrotica virgifera virgifera LeConte). PLoS ONE 7:e47534PubMedPubMedCentralGoogle Scholar
- Chen J, Tang B, Chen H, Yao Q, Huang X, Chen J, Zhang D, Zhang W (2010) Different functions of the insect soluble and membrane-bound trehalase genes in chitin biosynthesis revealed by RNA interference. PLoS ONE 54:e10133Google Scholar
- Christiaens O, Smagghe G (2014) The challenge of RNAi-mediated control of hemipterans. Curr Opin Insect Sci 6:15–21Google Scholar
- Hofmann K, Stoffel W (1993) TMbase—a database of membrane spanning proteins segments. Biol Chem Hoppe-Seyler 374:166Google Scholar
- Li H, Guan R, Guo H, Xiao M (2015) New insights into an RNAi approach for plant defence against piercing-sucking and stem-borer insect pests. Plant Cell Eviron 38:2277–2285Google Scholar
- Li J, Qian J, Xu Y, Yan S, Shen J, Yin M (2019) A facile-synthesized star polycation constructed as a highly efficient gene vector in pest management. ACS Sustain Chem Eng 7:6316–6322Google Scholar
- Liu X, Zheng Y, Zhang S, Liu K, Zhang S, Yin M, Zhang L, Shen J (2016) Perylenediimide-cored cationic nanocarriers deliver virus DNA to kill insect pests. Polym Chem 7:3740–3746Google Scholar
- Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT method. Methods 25:402–408Google Scholar
- Shen D, Zhou F, Xu Z, He B, Li M, Shen J, Yin M, An C (2014) Systemically interfering with immune response by a fluorescent cationic dendrimer delivered gene suppression. J Mater Chem B 2:4653–4659Google Scholar
- Terenius O, Papanicolaou A, Garbutt JS, Eleftherianos I, Huvenne H, Kanginakudru S, Albrechtsen M, An C, Aymeric JL, Barthel A (2011) RNA interference in Lepidoptera: an overview of successful and unsuccessful studies and implications for experimental design. J Insect Physiol 57:231–245PubMedGoogle Scholar
- Wani SA, Ahmad ST (2014) Phenomenon of parthenogenesis, viviparity and endosymbiosis in aphids-a review. J Bio Innov 3:206–215Google Scholar
- Xu Z, He B, Shen J, Yang W, Yin M (2013) Fluorescent water-soluble perylenediimide-cored cationic dendrimers: synthesis, optical properties, and cell uptake. Chem Commun 49:3646Google Scholar
- Xu Z, He B, Wei W, Liu K, Yin M, Yang W, Shen J (2014) Highly water-soluble perylenediimide-cored poly(amido amine) vector for efficient gene transfection. J Mater Chem B 2:3079–3086Google Scholar
- Yan S, Zhu JL, Zhu WL, Qin M, Liu H, Zhao SQ, Wang L, Zhang J, Zhang QW, Liu XX (2017) The influences of wind speed on pollen-mediated gene flow from transgenic cotton. Chin J Ecol 36:2217–2223Google Scholar
- Zhang J, Khan SA, Hasse C, Ruf S, Heckel DG, Bock R (2015a) Full crop protection from an insect pest by expression of long double-stranded RNAs in plastids. Science 347:991–994Google Scholar
- Zhang J, Khan SA, Hasse C, Ruf S, Heckel DG, Bock R (2015b) Pest control. Full crop protection from an insect pest by expression of long double-stranded RNAs in plastids. Science 347:991–994Google Scholar