To determine appropriate leaves to use for the primary and secondary
systemically infected leaves, we inoculated the first true leaves
of tomato and pepper seedlings, the fifth true leaf of N. benthamiana,
and the cotyledons of squash seedlings with the CMV-Fny strain. The
inoculated leaves were detached at 24, 48, 72, and 96 h after inoculation,
using two plants per host at each of the four time points.
Plants were kept in the greenhouse to observe symptoms of systemically
infected leaves at 10 and 15 days post-inoculation (dpi).
Two seedlings of each host were kept without detaching the
inoculated leaves.