Isolation of aphid-Potato leafroll virus (PLRV) proteins complexes from viruliferous insects using affinity purification-mass spectrometry

The control of insect borne disease is recognized as one of the major agricultural, animal and human health challenges of today. Viruses in the family Luteoviridae are phloem-limited, plant viruses that are vectored by aphids in a circulative manner. They are responsible for a wide-range of economically important disease in almost all staple food crops. In order to be transmitted, these viruses require species-specific passage of the pathogen across several membrane barriers within the insect. After the pathogen is ingested from the sap of an infected plant, the virus moves across and through the aphid gut into the hemoceol (insect blood) where it circulates until it reaches and enters the main or accessory salivary glands. From here, the pathogen is injected into a new plant host when the aphid feeds. The identification of insect proteins that interact with circulative plant viruses is technically challenging and a major goal for the plant vector biology field. Such information is critical to develop novel control strategies that block virus transmission by insects. In this study, we used affinity purification-high-resolution mass spectrometry (AP-MS) to rapidly capture and identify aphid proteins in complex with Potato leafroll virus (PLRV), a luteovirid transmitted by the green peach aphid (Myzus persicae), directly from viruliferous aphid tissue.