Cytochemistry of mature angiosperm pollen

The problems involved in applying histochemical and cytochemical methods to mature angiosperm pollen for bright light and fluorescence microscopy are discussed. These methods can be used for general examination or to reveal particular structures or groups of substances. The main methods of testing pollen viability and germinability based on stains and semiquantitative methods are also reviewed. The main methods of staining and their applications are summarised.     

Outward Extension of Spinules in Exine of Centrolepis aristata (Centrolepidaceae)

Spinules extend outwards 0.2 to 0.3 μm into the plasmodial tapetum during periods of rapid growth of microspores of Centrolepis. Before and after these intervals spinules commonly protrude only ca 0.05 μm above the surface of the tectum. The extended spinules have a rod shaped core ca 40 nm in diameter. The surface of the core is surrounded by loops ca 30 nm in diameter arranged in a regular pattern.     

Polarity, aperture condition and germination in pollen grains ofEphedra (Gnetales)

Pollen grain polarity, aperture condition and pollen tube formation were examined inEphedra americana, E. foliata, E. rupestris, E. distachya, andE. fragilis using LM, SEM and TEM. In the characteristic oblate pollen, as seen in situ in the tetrad configuration, the polar axis is the minor one and the equatorial plane runs between the two narrow ends of the microspore. The intine is thick in fresh fixed mature pollen but we have seen no indication of regions having an exceptionally thick intine that could be considered associated with an aperture or apertures. About three minutes after transferring fresh pollen to the germinating medium the ridged exine splits and twists away from the intine and its enclosed protoplast. The shed exine spreads out and curls into a scroll-like configuration that is as distinctive as that of the pollen shape had been but now having the ridges and valleys perpendicular to the long axis. The pollen tube develops, in our experience with more than a hundred germinating pollen grains, near one of the narrow tips of the pollen grain's equatorial plane. The location of the pollen tube initiation probably is related to the position of the tube cell nucleus. The pollen tube starts to grow about one hour after the exine was shed. The pollen tube emerges close to the narrow end (equator) of the gametophyte. This end emerged first as the exine is shed and is opposite to the prothallial cells. The stout pollen tube is c. 10µm in diameter grown in vitro on agar. In our germination medium the stout tube continued to elongate for about 24 hours reaching a length of c. 100 µm. With respect to exine morphology the aperture condition could be considered as inaperturate. The pollen tube, however, is formed in a germination area near one end of the exineless gametophyte.