Control
Systemic leaves were harvested at 21 days post-inoculation (dpi) from plants infected with PVY, PSTVd, or co-infected with both, as well as from mock-inoculated controls. Each biological replicate consisted of pooled leaves from five plants, with three biological replicates per treatment group. The total RNA was extracted from the leaves of N. benthamiana using TRIzol reagent. RNA purity and integrity were evaluated using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Only high-quality RNA samples (RIN > 7.0) were used for library construction.