Fig 8. PVA^WD is defective in particle formation.

(A) PVA particle accumulation in PVA^WD and PVA^WT infected local leaves at 3 dpi, as measured by immunocapture qRT-PCR. N. benthamiana plants were agroinfiltrated with PVA^WD (OD₆₀₀ = 1) and PVA^WT (OD₆₀₀ = 0.1) to ensure comparable RLUC expression level between PVA^WD and PVA^WT (upper panel). The corresponding particle numbers and CP accumulation visualized by western blotting with anti-CP antibodies are presented in the middle and lower panels, respectively. (B) Electron microscopy images from the same samples as in A) showing normal virus particles formed by PVA^WT. No particles were found in PVA^WD, only few deformed particle-like structures were observed (marked by an arrow in the lower electron micrograph). The number of plants per experiment was 3. Statistical significance was assessed using student’s t-test (*P < 0.05). (C) Both PVA^WT and PVA^WD infections were allowed to spread in the local leaves until 9 dpi, and EM images of the infected tissues were taken.Pinwheel inclusions (marked as ‘CI’) were observed. However, the pinwheels from PVA^WT were associated with stacks of particles (marked as ‘PS’) in vicinity while those from PVA^WD were devoid of them.