Supplementary Excel File including raw data.

Aphids are major agricultural pests, notably because they transmit nearly 30% of known plant viruses, including non-circulative ones. They can be collected and dispersed rapidly among crops while aphids feed on infected plants. Most of these viruses are retained on receptors located on the cuticle of the stylet tip. The acrostyle, a cuticular micro-territory at the apex of aphid stylets, has been identified for its ability to retain the cauliflower mosaic virus (CaMV). The acrostyle displays cuticular proteins, known as stylins, with exposed domains accessible at the virus-vector interface. RNAi-mediated silencing of Stylin-01 designated this protein as the prime candidate receptor of CaMV. However, the results were incomplete due to the transient effect and highlighted the need for stable mutants to advance our knowledge and validate these putative virus receptors. Here, we characterized the phenotype of two pea aphid Stylin-01 mutant lines, the first generated with CRISPR/Cas9 in this hemipteran. We showed that Stylin-01 mutations significantly disrupt CaMV transmission and impair the acrostyle’s ability to bind the CaMV helper protein P2. Stylin-01 mutations also reshape the distribution of other stylins on the surface of mutant aphid stylets. In addition, Stylin-02, the putative ortholog of Stylin-01, is overexpressed in the mutant lines, pointing out a potential partial complementation of Stylin-01 in its structural role but not for virus transmission. In conclusion, this study, using the first stable aphid mutant lines, allows the characterization of the central role of Stylin-01 virus receptor in CaMV transmission.