A path to defeating crop-killing graymold without toxicchemicals UCRiverside NatureComms
spores were cultured for 48 hours at room temperature with shaking on the rocker in 100 ml of YEPD medium.EVs were isolated and purified using sucrose gradients as described above, then EVs from fraction four were resuspended in 100 μl PBS. The spores were collected and diluted into 2 x inoculation medium for a final concentration of 2 × 10spores per mL. Equal volumes of EVs and spores from mutant strains were mixed.
AGO1 IP was performed with 5 g fresh leaves collected 24 hr after spray inoculation with. Tissues were collected and ground in liquid nitrogen before resuspension in 5 ml of extraction buffer . The extract was then precleared by incubation with 50 ul of Protein A-agarose beads at 4 °C for 30 min. The precleared extract was incubated with 10 ul of AGO1-specific antibody for 6 hours. 50 ul of Protein A-agarose beads was then added to the extract before overnight incubation.
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