Figure 5

Immunolocation of ZmTCRR-2 and ZmHPs on 10 DAP seeds. Areas squared in the low magnification image are shown on the left after detection with anti-ZmTCRR-2, ZmHP1/3 and ZmHP2 antisera and preimmune sera as indicated below the images. The signal appears as dark/light grey precipitates depending on its intensity. a upper endosperm and aleurone show detection of ZmTCRR-2 and a strong signal from the ZmHP1/3 serum, while ZmHP2 is only weakly detectable. No unspecific signal is visible in the presera treated sections. b transfer cells and transmitting tissue. ZmTCRR-2 is present in the transfer cells (black arrowheads) and in the transmitting tissue located above them, as is the case for ZmHP1/3. Anti-ZmHP2, on the other hand, produces a weaker signal in this area, still clearly distinguishable from the preserum. c placento-chalaza and pedicel. A dark precipitate appears in the crushed placento-chalaza in all sections shown (yellow arrowheads), including presera, which indicates its unspecific nature. Apart from this, no signal from ZmTCRR-2 is detected in the area. Anti-ZmHP1/3, on the other hand, broadly decorates the pedicel specially at the vascular bundles (blue circle), while anti-ZmHP2 labels (apart from the unspecific signal mentioned above) two distinctive patches at the edges of the placento-chalaza. This area at the adgerminal end is shown in the panels for immune serum and preserum (yellow circles). En, inner endosperm; Em, embryo; TCL, transfer cell layer; Pd, pedicel. The scale bars represent 1 mm in the low magnification image and 100 μm in the other images.