Identifying mosquito plant hosts from ingested nectar secondary metabolites

Establishing how plants contribute food and refuge to insects can be challenging for small species that are difficult to observe in their natural habitat, such as disease vectoring mosquitoes. Currently indirect methods of plant-host identification rely on DNA sequencing of ingested plant material but are often unsuccessful for small insects that feed primarily on plant sugars or have little contact with plant cells. Here we developed an innovative approach to determine species-specific phytophagy by detecting taxon-specific plant secondary metabolites (PSMs) in nectar. Two mosquito species were exposed to three PSMs, each present in the nectar of a known plant host, firstly from dosed sucrose solutions and secondly from flowers. Both experiments yielded high rates of PSM detection in mosquitoes using liquid chromatography-mass spectrometry (LC-MS). PSMs were consistently detected in mosquitoes up to 8 h post-ingestion. In experiments consisting of two or three plant species, multiple PSMs from different host plants could be detected. These positive results demonstrate that PSMs could be useful indicators of insect plant-hosts selection in the wild. With expanded knowledge of nectar-based PSMs across a landscape, improved knowledge of plant-host relationships could be achieved where direct observations in their natural habitat are lacking. Increasing understanding of vector insect ecology will have an important role in tackling vector-borne disease.