Tannin accumulations in tissue culture cells of slash pine occurred in the smooth endoplasmic reticulum, in vesicles, and within cell vacuoles. Dense osmiophilic granules were observed also throughout the cytoplasm of many cells. Vacuolar deposits appeared as homogeneous spherules or as a froth-like material. Generalized degeneration of the cytoplasm and the organelles was observed in many of the cells synthesizing tannins. Accumulation of the tannin within the cell vacuoles did not necessarily result in cell degradation. Synthesis and deposition of tannin in slash pine cells closely resembles synthesis and storage of aromatic products in mammalian tissues.
The plasma membrane in immature cells is often irregular in contour. Some irregularities become conspicuous folds that continue to enlarge into the cytoplasm. These invaginations may continue to increase in size and typically expand into the central vacuole. Sections show two closely parallel membranes in areas where the invagination projects into the vacuole. A narrow layer of cytoplasm may traverse the intermembrane zone between the membranes. The interior of an invagination may lack obvious content, or may be occupied by a fibrous material, or vesicular and tubular structures. The small vesicles bound by a single membrane appear to be derived from a projection formed most frequently near the orifice of the invagination. The origin of large vesicles and tubules possessing one membrane is not certain although they may arise by the fusion of several small vesicles. Alternatively, tubules, once formed, often possess constrictions along their length which suggest that these structures may become divided into a series of smaller vesicles. There is some evidence that cytoplasmic vesicles may fold into the membrane of the invagination and subsequently be pinched off into the interior of the sac. These vesicles are bounded by two membranes. Vesicles frequently contain an electron‐dense content more or less homogeneous in composition and unlike the typical ribosomal character of the cytoplasm. The function of the invaginations or their content remains to be elucidated.
The plasma membrane of cultured cells of several plant species was observed to possess invaginations, or secondary vacuoles, of variable size in the adjacent cytoplasm. These structures, which occurred in cells at different phases in vacuolation, were very numerous in thin sections of some cells but fewer in others. In vacuolated cells enlarged secondary vacuoles protrude into the primary vacuole but are delimited from the tonoplast by an intermembrane zone of variable width. The plasma membrane at the orifice of an invagination may fuse and detach the secondary vacuole from the membrane to form in the cytoplasm a structure bounded by a single membrane. Complex accumulations of membranes consisting of spherical, tubular, and laminar structures, possibly containing cytoplasm, may develop within secondary vacuoles. Contents of many of these vacuoles arise from folds along its limiting membrane which pinch off into the interior of the secondary vacuole. A fibrous substance, possibly derived from the wall, is present in some secondary vacuoles. Observed folding of the plasma membrane and measurements of membrane width of various organelles and cytomembranes support an interpretation that endocytosis occurs in cultured cells.
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