Putative gut receptors for Pea enation mosaic virus in the pea aphid, Acyrthosiphon pisum

Aphids and the plant viruses transmitted by aphids cause tremendous economic loss worldwide. The goal of this project is to increase our understanding of the molecular interactions that facilitate recognition and uptake of the model luteovirid, Pea enation mosaic virus (PEMV) into the pea aphid vector, Acyrthosiphon pisum. We are using PEMV virions and PEMV coat protein (CP)-eGFP fusions to determine whether glycans are involved in PEMV uptake into the aphid hemocoel and to screen for potential receptor proteins on the aphid gut epithelium. We used ligand blot analyses to show that these constructs bind to proteins in whole aphid brush border membrane vesicles (BBMV) and in total gut protein extracts. PEMV virions purified from plants as well as the CP-P-eGFP fusion peptide showed similar binding to whole aphid BBMV. Proteins that specifically bound PEMV and CP-P-eGFP were isolated from 2-D SDS-polyacrylamide gels with reference to the probed membranes and identified using MALDI-TOF MS/MS mass spectroscopy at the Iowa State University Protein Facility. Data on the identified proteins will be presented.