Establishment of an in vitro sciarid fly larvae assay to study plant resistance

Mechanisms underlying natural plant resistance to herbivorous invertebrates are still poorly understood in comparison with bacterial or fungal interactions. One reason is the difficulty in reliably and reproducibly assessing the effects under controlled conditions. This article describes a newly developed in vitro biological assay system that enables the interactions between sciarid larvae and plants, whose roots they feed on, to be studied under highly controlled conditions. The bioassay eliminates the problems created by the often variable environmental factors by providing an aseptic arena where experimental plants can be germinated and grown on agar within a Petri dish. Sciarid fly eggs are then collected, sterilised and added to the Petri dish. The system allows the eggs to hatch and the larvae to feed on the plant roots. A range of developmental parameters can then be recorded over time which can then be correlated with the experimental plant type. This assay system also allows a simultaneous comparison or 'choice chamber' between two (or more) different genotypes. The assay should greatly help to facilitate the identification of new components involved in insect resistance mediated pathway via the characterisation of mutant plants.