Downregulation of <i>TdPDS</i> and detection of TRV transcripts in VIGS photobleached leaves of <i>Thalictrum dioicum</i>.

<p><b>A: Expression of </b><b><i>TdACTIN</i></b><b> control, native </b><b><i>TdPDS</i></b><b> and viral transcripts in leaves by Reverse Transcriptase (RT)-PCR.</b> Untreated and mock-treated (empty TRV2) <i>T. dioicum</i> plants are compared to TRV2-<i>TdPDS</i> treated plants showing photobleached (white), variegated (green/white) and green leaf tissue. RT-PCR was performed with locus-specific primers to the housekeeping gene <i>ACTIN</i> (loading control), to endogenous <i>TdPDS</i> and to the viral transcripts TRV1/TRV2. Approximate band size indicated for TRV2: larger band results from the presence of the <i>TdPDS</i> insert, smaller band from an empty TRV2 (mock control). <b>B: Comparative expression of </b><b><i>TdPDS</i></b><b> normalized with </b><b><i>TdACTIN</i></b><b> among treatments and resulting phenotypes of </b><b><i>Thalictrum dioicum</i></b><b>.</b> Values based on quantification of RT-PCR gel bands in part A using ImageJ (see text for details). Different letters indicate statistical significance in a one-way ANOVA followed by Tuckey test (p<0.05), same letters indicate no statistical difference. Average and standard error bars are shown. Sample sizes are n = 4 for untreated and mock-treated, n = 6 for treated bleached and n = 3 for treated variegated or green.