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. Author manuscript; available in PMC: 2016 Jul 6.
Published in final edited form as: Dev Cell. 2015 Jun 25;34(1):45–57. doi: 10.1016/j.devcel.2015.05.011

Figure 4. Induction of chloroplast stromule-to-nuclear connections during HR-PCD.

Figure 4

A. Transient expression of p50-Citrine (top panels) in N-containing NRIP1-Cerulean plants was used to monitor stromule-to-nuclear connections during HR-PCD. Expression of Citrine was used as a no HR-PCD control (bottom panels). Stromules were observed connecting to nuclei (N) with pad-like structures (P) at the points of contact (top panels) between stromules and nucleus. The larger chloroplasts (red) are located in mesophyll cells and contained very few stromules. Scale bars equal 10 μm.

B. During p50-mediated HR-PCD, confocal micrographs show stromules wrapped around nuclei (top left panel, maximum intensity projection of a z-stack). There were also clusters of stromule tip connections directly to the nucleus (top right panel, single plane of a z-stack) and nuclei with a mixture of tip or surrounding stromule connections (bottom panels, transparent projections of z-stacks). Scale bars equal 10 μm.

C. Stromule-to-nuclei connections were manually enumerated from 3D confocal microscopy z-stacks. There was an increase in stromule-to-nuclei connections per chloroplast during p50-Citrine-initiated HR-PCD compared to Citrine, the no HR-PCD control. Data represented as the mean ± SEM,***P<0.005 (Student’s t-test).