Structure of the generative cell wall complex after freeze substitution in pollen tubes of Nicotiana and Impatiens

Abstract

The mature generative cell in pollen grains and pollen tubes is surrounded by a wall complex that includes two plasma membranes, one facing the generative cell cytoplasm and one facing the vegetative cell cytoplasm, and usually some intervening wall material. After conventional chemical fixation, the two plasma membranes are very uneven and often appear to be joined, giving the impression that numerous plasmodesmata connect the vegetative and generative cells. These areas alternate with swollen, distorted areas, which give the wall complex the appearance of being composed of a chain of vesicles. Utilizing rapid freeze fixation and freeze substitution, we have re-examined the ultrastructure of the generative cell wall complex from pollen tubes grown in vitro, and the differences are striking. The two plasma membranes are very smooth and closely appressed to a layer of wall material. Occasionally the wall complex contains swollen areas, or varicosities, and these may contain pockets of lightly stained material, but again the surrounding plasma membranes are tightly appressed to these areas. Plasmodesmata are not seen, but this does not eliminate the possibility that they may exist at an earlier stage of development.