Myosin XI-2 is implicated in TuMV intercellular movement.

Abstract

<p>(A) Quantitative RT-PCR was used to determine the relative expression ratio of target genes (myosin VIII-1, myosin VIII-2, myosin XI-2, and myosin XI-F) in <i>N. benthamiana</i> infected with the indicated TRV silencing constructs versus a TRV control not expressing a myosin fragment. (B) <i>N. benthamiana</i> leaves silenced for individual myosin genes (myosin VIII-1, myosinVIII-2, myosin XI-2, and myosin XI-F) were agroinfiltrated with pCambiaTuMV/6K₂:mCherry//GFP-HDEL and surface area of red-only fluorescent foci was calculated and expressed in fluorescence units at 4 dpinf. Wild-type TRV (TRV) or buffer (Mock) were used as controls. Bars represent means and standard errors for 10 replicates per treatment. One-way analysis of variance calculation followed by Tukey's Multiple Comparison Test allowed analysis of differences between means: NS, not significant; *, P<0.05. (C) Level of expression of non-target myosins in <i>N. benthamiana</i> leaves silenced for myosin XI-2. The internal loading control for each sample was actin-2. Expression analysis was performed on extracts from systemic leaves at 20 dpinf with TRV constructs. Bars represent means and standard errors for three replicates per treatment. One-way analysis of variance calculation followed by Tukey's Multiple Comparison Test allowed analysis of differences between means: = NS, not significant; *, P<0.05. The experiment was repeated twice for each TRV silencing construct. (D) images of pCambiaTuMV/6K₂:mCherry//GFP-HDEL in <i>N. benthamiana</i> leaves silenced for individual myosin genes (VIII1, VIII-2, XI-2, XI-F). Scale bar = 200 µm.</p