Abstract
The two-spotted spider mite, Tetranychus urticae, is a chelicerate herbivore with an extremely wide host range and an extraordinary ability to develop pesticide resistance. Due to its responsiveness to natural and synthetic xenobiotics, the spider mite is becoming a prime pest herbivore model for studies of the evolution of host range, plant-herbivore interactions and mechanisms of xenobiotic resistance. The spider mite genome has been sequenced and its transcriptional responses to developmental and various biotic and abiotic cues have been documented. However, to identify biological and evolutionary roles of T. urticae genes and proteins, it is necessary to develop methods for the efficient manipulation of mite gene function or protein activity. Here, we describe protocols developed for the delivery of small molecules into spider mites. Starting with mite maintenance and the preparation of the experimental mite populations of developmentally synchronized larvae and adults, we describe 3 methods for delivery of small molecules including artificial diet, leaf coating, and soaking. The presented results define critical steps in these methods and demonstrate that they can successfully deliver tracer dyes into mites. Described protocols provide guidelines for high-throughput setups for delivery of experimental compounds that could be used in reverse genetics platforms to modulate gene expression or protein activity, or for screens focused on discovery of new molecules for mite control. In addition, described protocols could be adapted for other Tetranychidae and related species of economic importance such as Varroa, dust and poultry mites.
Highlights
The two-spotted spider mite (TSSM) Tetranychus urticae Koch (Acari: Tetranychidae) is an important agricultural pest worldwide
To fully exploit the potential of spider mite as a model experimental system, it is necessary to develop methods for the efficient manipulation of mite gene function. This includes the delivery of different types of molecules that modulate gene expression, such as double stranded RNAs, morpholinos, or transgenes, or molecules that alter protein activity, including natural or synthetic chemicals that act as agonists or antagonists
We report 3 different methods for the delivery of small molecules to T. urticae including artificial diet, leaf coating, and soaking protocols
Summary
The two-spotted spider mite (TSSM) Tetranychus urticae Koch (Acari: Tetranychidae) is an important agricultural pest worldwide. To fully exploit the potential of spider mite as a model experimental system, it is necessary to develop methods for the efficient manipulation of mite gene function This includes the delivery of different types of molecules that modulate gene expression, such as double stranded RNAs (dsRNA), morpholinos, or transgenes, or molecules that alter protein activity, including natural or synthetic chemicals that act as agonists or antagonists. Such methods must be robust, reproducible, cost-effective, and designed to record the effects of small molecules on otherwise physiologically normal mites. These bioassays should be applicable to a large population of mites, so that the effects of small molecules on mite physiology can be quantified phenotypically or using biochemical or molecular analyses
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