Projects

Using RNA interference and systems biology approaches for validation of insecticide targets in colorado potato beetle guts

Project coordinator: PhD Marko Petek

Code: Z4-7068

Duration: 1. 1. 2016 - 31. 12. 2017

The authors acknowledge the project Z4-7068 was financially supported by the Slovenian Research Agency.

Abstract

The Colorado potato beetle (CPB) is an important agricultural pest. It is well known for its ability to rapidly evolve resistance to insecticides and is predicted to expand to the north of Europe due to the prospects of climate change. In the last decade double-stranded RNAs (dsRNAs) were shown to have the potential to become environmentally-acceptable insecticidal agents. These molecules activate the RNA interference (RNAi) mechanism that mediates a sequence-specific suppression of gene expression, also called gene silencing.
 
In CPB, gene silencing can be effectively triggered and spread systemically by ingestion of dsRNA. If the targeted gene silencing disrupts an essential life-sustaining process in CPB, the dsRNAs can act as an insecticide. The advantages of RNAi insecticides include high specificity for target species, low probability of resistance development and short environmental degradation period into nontoxic compounds. Crucial for the efficiency as well as the environmental safety of dsRNA insecticides is the selection of target genes.
 
This project's main objective is to identify and validate gene targets for CPB specific RNAi insecticides. To achieve that, the methodology for identification and validation of RNAi insecticide efficiency will be established at NIB. This will include a candidate gene selection procedure, establishment of dsRNA production methodology and the evaluation of dsRNAs' insecticidal potential in insect feeding trials. Additionally, a Next Generation Sequencing (NGS) RNA-seq analysis will be performed to explore the global changes induced by the RNAi insecticide on the CPB gut transcriptome. To explore how a RNAi insecticide changes the bacterial community structure in CPB guts, larval gut contents will be examined by NGS to obtain bacterial metagenomic profiles.
 
Available statistical tools will be used for evaluation of RNAi insecticide efficiency and already developed NGS pipelines for the analysis of RNA-seq data will be adopted, whereas for metagenomics data, a dedicated analysis pipeline will be established at NIB.
 
The project will contribute to science by:
- exploring the potential of selected candidate genes as new targets for RNAi insecticides,
- validating computationally-predicted gene functions in an agricultural pest,
- following the expression of selected target genes in different CPB tissues and life stages,
- studying the global transcriptional patterns by RNA-seq in CPB guts after RNAi insecticidal treatment,
- investigating the effect of an RNAi insecticide on CPB's gut metagenome.
 
In future, new discoveries based on this knowledge will lead to more sustainable agricultural practices. The developed gene silencing methodology could be adapted for other organisms and impact scientific disciplines beyond crop protection. Due to the prospects of RNAi technology in agriculture and biotechnology, this project can represent an opportunity for establishment of new companies that would commercialize the RNAi technologies.
 
This postdoctoral project will also be an opportunity to establish closer connections with other scientists working in the RNAi field and to join consortia for international project grant applications.
Information about the project - link to database SICRIS.

Project official side on - http://projects.nib.si/RNAinVAL.