Figure 4.

CIP73 expression and subcellular localization. A, CIP73 and CCaMK expression was investigated by semiquantitative RT-PCR before and after M. loti inoculation using polyubiquitin as an internal control. Uninoculated roots (1D–12D) are shown as controls for the same periods post inoculation with M. loti. B, Nuclear localization of GFP::CIP73 in onion epidermal cells. The control GFP gene (a–c) and GFP::CIP73 fusion gene (d–f), delivered by particle bombardment, were transiently expressed in onion epidermal cells and observed with a laser-scanning confocal microscope 24 h after bombardment. Note that the control GFP is distributed both in the cytoplasmic and nuclear compartments, whereas GFP::CIP73 only localizes in the nucleus. Top images show GFP fluorescence of the cells (a and d); middle images show the cell architecture (b and e); bottom images show the overlays of GFP and bright-field images (c and f). C, The C terminus of CIP73 (486–691) shows strong nuclear localization in epidermal and cortex cells of transformed hairy roots. Bars = 50 μm for B and 20 μm for C. [See online article for color version of this figure.]