|Rodrigues, Thais - University Of Kentucky|
|Dhandapani Ramesh, Kumar - University Of Kentucky|
|Palli, Subba - University Of Kentucky|
Submitted to: Scientific Reports
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/18/2017
Publication Date: 8/21/2017
Citation: Rodrigues, T.B., Dhandapani Ramesh, K., Duan, J.J., Palli, S.R. 2017. RNA interference in the Asian Longhorned Beetle:Identification of Key RNAi Genes and Reference Genes for RT-qPCR. Scientific Reports. 7: 8913.
Interpretive Summary: Asian longhorned beetle (ALB) is a highly invasive forest pest that has established small populations in the United States, Canada and several countries in Europe. If not successfully eradicated or controlled in the newly invaded regions, this beetle has the potential to devastate the native forest ecosystems because it can feed on over 100 species of hardwood trees. Development of a new control method that specifically targets the pest but has no adverse impacts on environment and other organisms is ideal for integration with the current ALB eradication or control program. Scientists from the University of Kentucky and ARS in Newark, DE, identified ALB genes that could be used for developing control approaches that rely on RNAi-based inhibition of critical genes. Injection of double-stranded RNA (dsRNA) targeting a developmental gene into ALB larvae and adults resulted in a significant knockdown of this gene and caused the death of 90% of the larvae and 100% of adults. This suggests that RNAi pathways exist in ALB and a potential RNAi-based method could be developed to control this insect.
Technical Abstract: Asian longhorned beetle (ALB), Anoplophora glabripennis, is a serious invasive forest pest in several countries including the United States, Canada, and Europe. RNA interference (RNAi)technology is being developed as a novel method for pest management. Here, we identified the ALB core RNAi genes including those coding for Dicer, Argonaute, and double-stranded RNA-binding proteins (dsRBP) as well as for proteins involved in dsRNA transport and the systemic RNAi. We also compared expression of six potential reference genes that could be used to normalize gene expression and selected gapdh and rpl32 as the most reliable genes among different tissues and stages of ALB. Injection of double-stranded RNA (dsRNA) targeting the inhibitor of the apoptosis (IAP) gene into larvae and adults resulted in a significant knockdown of this gene and caused the death of 90% of the larvae and 100% of adults. No mortality of either larvae or adults injected with dsRNA targeting gene coding for green fluorescence protein (GFP, as a negative control) was observed. These data suggest that functional RNAi machinery exists in ALB and a potential RNAi-based method could be developed for controlling this insect.