An investigation was conducted into the isolation of plasma membrane vesicles from primary roots of corn (Zea mays L., WF9 x M14) by sucrose density gradient centrifugation. Identification of plasma membranes in cell fractions was by specific staining with the periodic-chromic-phosphotungstic acid procedure. Plasma membrane vescles were rich in K+-stimulated ATPase activity at pH 6.5, and equilibrated in linear gradients of sucrose at a peak density of about 1.165 g/cc. It was necessary to remove mitochondria (equilibrium density of 1.18 g/cc) from the homogenate before density gradient centrifugation to minimize mitochondrial contamination of the plasma membrane fraction. Endoplasmic reticulum (NADH-cytochrome c reductase) and Golgi apparatus (latent IDPase) had equilibrium densities in sucrose of about 1.10 g/cc and 1.12 to 1.15 g/cc, respectively. A correlation (r = 0.975) was observed between K+-stimulated ATPase activity at pH 6.5 and the content of plasma membranes in various cell fractions. ATPase activity at pH 9 and cytochrome c oxidase activity were also correlated.A major peak of ATPase activity at pH 6.5 was observed at low density in Ficoll after nonequilibrium centrifugation in a combination Ficollsucrose gradient. Twenty to forty percent of the vesicles in this ATPase fraction stained positively for plasma membranes, and with equilibrium centrifugation the major portion of the ATPase activity shifted to densities in sucrose which were characteristic of plasma membranes. All major vesicular ATPase activities observed in Ficoll or sucrose contained substantial amounts of plasma membranes. For unknown reasons, mitochondria and plasma membranes equilibrated over a broader density range and at lower peak densities in sucrose as a result of equilibrium centrifugation through Ficoll.

An ultrastructural study of the pollen tubes of Lilium and Clivia has demonstrated three different classes of longitudinal structures which could influence patterns of protoplasmic streaming and/or serve as "guide elements" in the vectorial migration of secretory vesicles: (a), cortical and noncortical microtubules; (b), microfilaments; and (c), subcortical tubules and cisternae of the endoplasmic reticulum ("subsurface cisternae"). Morphological details of these structures are described. Colchicine concentrations which lead to the complete disappearance of the microtubules affect neither germination of the pollen nor cytoplasmic streaming and tip growth of the elongating pollen tubes. Tip growth is initially uninhibited by cycloheximide, and cytoplasmic streaming is insensitive to this inhibitor. However, both of these processes are sensitive to cytochalasin B and vinblastine. Our results suggest that neither microtubules nor subsurface cisternae are essential for cytoplasmic streaming and directional secretion of cell surface materials in the pollen tube but would be consistent with an involvement of microfilamentous structures in these processes. Additionally, the possible importance of the lateral cross-link elements interconnecting all three types of structures is discussed.

Abstract— Sensitization of the phytochrome‐mediated germination at 20°C of lettuce seeds (Lactuca sativa L. cv. Grand Rapids) by pretreatment at 4°C, 28°C, or on 1% ethanol, was studied. The 660 nm fiuence‐response characteristics were similarly biphasic for all sensitizing treatments and displayed responses at very low fluences (VLFR) as well as responses characteristic of non‐sensitized seeds at 10000‐fold higher, low fluences (LFR). Maximum VLFR increased with the duration of sensitizing treatments. However, the fluence ranges required for the two types of responses remained relatively constant. These and additonal responses of sensitized seeds to 730 nm fluences were compared to simulations of a mechanism involving a receptor, X, and based on the dimeric structure of phytochrome in which each monomer is independently phototransformed from the inactive (Pr) to the active (Pfr) form. The fluence requirements for phytochrome photoconversion in seeds were determined to be similar to those of purified Avena phytochrome in vitro, on which photochemical parameters for the simulations were based. The analyses suggest that Pr:Pfr‐Xand Pfr:Pfr‐X are responsible, respectively, for the VLFR and the LFR, and that sensitization involves membrane influences on the activity of Pr:Pr‐X. They also suggest the concentration of X to be about 0.001 that of total phytochrome dimer in this system.