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The generative cell is initiated as a small, lenticular, unpolarized cell with a cell wall traceable to two origins: the external segment originates as intine, while an inner callose positive cell wall forms de novo. As the lenticular generative cell begins its migration into the pollen cytoplasm, the generative cell becomes polarized both externally and internally, displaying a characteristic shape and patterns of organelle distribution oriented with respect to the vegetative nucleus and independent of pollen aperture location. Separation of the generative cell from the pollen wall begins at the end opposite the vegetative nucleus and results in an elongating protuberance at the opposite end of the generative cell; this becomes associated with a preformed groove located on the surface of the vegetative nucleus. The generative cell subsequently separates from the intine near the vegetative nucleus and moves progressively toward the opposite end of the cell; during this separation, the edge of the wall facing the intine becomes callose-positive and remains so until separating from the intine. The generative cell becomes a free cell within the pollen, which is in physical association with the vegetative nucleus. Generative cell organization and organelle content become increasingly polarized during maturation, with microtubules evident both in the elongating protuberance of the generative cell and in association with organelles. The generative nucleus migrates away from the vegetative nucleus and toward the plastid-rich end of the generative cell, whereas mitochondria are more generally distributed within the cell. Generative cell polarization is made permanent during mitotic division and cytokinesis, i.e., two sperm cells differing in morphology are formed: the larger cell associated with the vegetative nucleus ([S.sub.vn]) contains a majority of the mitochondria, and the smaller, unassociated sperm cell ([S.sub.ua]) receives the plastids. Key words: generative cell; microgametogenesis (angiosperm); Plumbago; Plumbaginaceae; sperm cell.

The organization of isolated embryo sacs and eggs of Plumbago zeylanica was described before and after fertilization using microscopic cytochemistry and scanning electron microscopy. Major developmental events of fertilization, including preferential fertilization and early embryogenesis, are described in isolated embryo sacs. The two sperms, one unassociated with vegetative nucleus ([S.sub.ua]) and the other physically associated with the vegetative nucleus ([S.sub.vn]), fuse with nuclei of egg and central cell, respectively. The zygote divides asymmetrically to form a two-celled embryo, consisting of a massive suspensor occupying most of the micropylar portion of the embryo during early embryogenesis. Plastids are distributed in the perinuclear and micropylar regions of the egg cell and in cytoplasmic strands of the central cell before fertilization. Calcofluor white-positive fibrillar material in the filiform apparatus (presumed [beta]-1,4 linked glucans) was investigated using scanning electron microscopy. The egg of P. zeylanica can easily be divided into three cytologically distinct regions: 1) perinuclear cytoplasm, 2) lateral cytoplasm, and 3) micropylar cytoplasm. Cytological differences are evident in the organization of the cell walls, general degree of vacuolization, and the distribution of heritable organelles, storage bodies, and microtubules. The present study supports the concept that the egg of P. zeylanica plays combined synergid and gamete functions.