Development of reverse genetics tools in the two-spotted spider mite Tetranychus urticae
Abstract
Tetranychus urticae, commonly known as the two-spotted spider mite, poses a significant threat to agriculture due to its ability to feed on a diverse range of plant hosts and its strong detoxification abilities in overcoming xenobiotic response. With global warming projected to increase spider mite infestations, it is vital to study the detoxification genes that enable the mite to adapt and survive. The spider mite genome sequence reveals a unique set of detoxification genes that can be studied using RNAi as a promising reverse genetic tool. However, the current genetic toolkit requires improvement. This study examined the effectiveness of three precursor molecules (dsRNA, shRNA, and amiRNA) in inducing RNAi response in spider mites. While shRNA and amiRNA demonstrated variable responses, dsRNA significantly reduced transcript levels of all three target genes. The establishment of primary cell cultures from T. urticae embryos provides a valuable tool for investigating the uptake mechanism of different precursor RNAi molecules through 'RNAi of RNAi' experiments. Ultimately, this research could contribute to the development of RNAi-based pesticides that selectively target spider mites detoxification pathways, leading to more effective pest control in agriculture. Future research should aim to improve our understanding of the spider mite RNAi machinery, including the uptake mechanism and precursor molecule stability, to facilitate functional gene analysis and the development of new pest control strategies.