Location: Temperate Tree Fruit and Vegetable Research
Project Number: 2092-30400-002-003-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jun 1, 2026
End Date: Dec 30, 2027
Objective:
1. Identify M. persicae stylins that bind potato virus Y (PVY) HC-Pro protein, the PVY protein that interacts with the aphid stylet.
2. Identify aphid salivary effectors that bind to M. persicae stylins allowing aphids to feed.
Approach:
Objective 1: Myzus persicae stylins binding (potato virus Y) PVY HC-Pro. Expression of stylins in M. persicae feeding on potato will be identified by q-RT-PCR (11 stylins were identified). PVY HC-Pro and the M. persicae described stylins will be cloned in yeast two-hybrid, bifunctional fluorescent complementation and co-immunoprecipitation vector to identify which stylins bind to PVY HC-Pro. These three complementary techniques allow to identify protein-protein interactions.
Objective 2: Identify M. persicae effectors interacting with stylins. Myzus persicae salivary glands will be dissected and RNA will be purified following the protocols developed at ARS Wapato, WA. We will use age-synchronized alates from a colony continuously maintained on Physalis pubescens in Wapato, WA. by isolating late-instar nymphs with wing buds and using fully sclerotized adults having emerged within 24 hours. Dissections will be conducted in 1X PBS buffer solution, and three replicates of 100 salivary gland pairs stored separately in RNALater at -80°C until RNA extraction. The obtained mRNA will be used to generate a yeast two-hybrid salivary gland library (Takara Bio Inc.). The library will be screened with M. persicae stylins to identify salivary proteins that are putative effectors.