Ultrastructure of the Mesocarp of Mature Avocado Fruit and Changes Associated with Ripening

The mesocarp tissue of ripening avocado fruits was studied byfreeze fracture, thin section and scanning electron microscopy.Carbon dioxide and ethylene production by individual fruit weremonitored, and samples were analysed at several stages of theripening process. The tissue is composed primarily of large, isodiametric, lipid-containingparenchyma cells. At maturity these cells contain the normalcomplement of cell organelles, and all membranes appear intact.When ripening begins, several changes in the ultrastructureoccur. The most obvious changes are a loosening and eventualbreakdown of the cell wall, and swelling and vesiculation ofthe rough endoplasmic reticulum. In freeze fracture replicasa significant increase in the number of intramembranous particlesin the EF face of the plasmamembrane was observed at the climactericpeak. In post-climacteric, soft fruit the particle density of theEF face of the plasmamembrane decreased to the density observedin the membrane of pre-climacteric cells. All of the organellesand membranes appear whole and intact whether examined by thinsection, freeze fracture or scanning electron microscopy. However,the cell walls in post-climacteric fruit have almost completelydisappeared. These results indicated that the ripening process per se inavocados does not involve a complete loss of compartmentationnor a breakdown of organelle and membrane integrity. It may,however, lead to these or similar senescence changes as a resultof the loss of the cell walls. The variations in particle densityof the plasmamembrane during ripening may reflect one or moreof several structural, compositional, or functional membranephenomena, and this aspect of ripening warrants further study. Persea americana Mill., avocado pear, freeze fracture, fruit ripening, scanning electron microscopy, senescence, ultrastructure     

Stomatal Response to Humidity and Lanthanum

Lanthanum fed to the base of excised leaves of Sesamum indicum L. and Helianthus annuus L. was used as a tracer to investigate by electron microscopy the path of water in the apoplast of leaves. The generally random distribution of lanthanum in cell walls provided no support for the hypothesis that cuticular transpiration may be greater for guard cells than for adjacent epidermal cells. Occasionally, accumulations of lanthanum were observed in anticlinal walls of epidermal cells and at the outer surface of the plasma membrane but lanthanum was not observed in the symplast. The influx of ⁸⁶Rb to excised roots of sesame and sunflower was inhibited during incubation with 0.5 mM lanthanum or calcium for 15 or for 180 min. Stomata of sunflower partially closed when 2.5 mM lanthanum was supplied to the base of excised shoots in a potometer, whereas this treatment had little effect on stomatal conductance of sesame shoots maintained in a constant environment. Supplying 2.5 mM lanthanum to the base of sesame shoots strongly inhibited stomatal opening response to increase in ambient humidity but had little effect on stomatal opening response to light. It was concluded that stomatal opening response to increased humidity may be dependent upon some process, such as ion influx, that is inhibited by lanthanum, and that opening response to humidity may differ in mechanism from stomatal opening response to increased irradiance.     

Epidermal Peels of Avicennia germinans (L.) Stearn: A Useful System to Study the Function of Salt Glands

Adaxial peels were made from fully expanded leaves of Avicenniagerminans (L). Stearn. The peels consisted of the epidermis,epidermal salt glands, and underlying hypodermal cells. Electronmicroscopic examinations showed that the structural integrity(except for the innermost hypodermal cells) of all cell typeswas not altered in making the peels. When the peels were floatedon salt solutions, the glands were shown to be functionallycompetent in secretion. Secretion also occurred when the peelswere floated on distilled water, presumably from salts storedin the hypodermis. Secretion occurred in the dark and sinceall cell types in the peels lacked chloroplasts, glandular functioncould not be directly coupled energetically to photosynthesis.However, secretion was shown to be temperature dependent andinhibited by azide and dinitrophenol, which indicates that theenergy cost underlying secretion is mitochondrial and most likelycoming from the mitochondria-enriched gland cells. Inhibitorsknown to affect membrane H⁺ ATPase activity and membrane transportalso inhibited secretion, indicating that membrane transportis probably the primary mechanism underlying secretion. Lanthanum,a membrane calcium antagonist, also inhibited secretion. Avicennia germinans (Avicenniaccac), salt glands, metabolic inhibitors, ultrastructure, secretion     

The inhibitory effect of NaCl on barley germination

Abstract The possibility that the nature of the inhibitory effect of NaCl is different during imbibition compared to germination was investigated. Germination in both NaCl and betaine (a non-toxic solute) improved with pre-imbibition in water. Seeds imbibed in inhibitory concentrations of either solute could be induced to germinate by brief exposure to water. Electron micrographs of tissue from seeds imbibed in 0.5 kmol m^?3 NaCl for 25 h showed cells identical to those in seeds imbibed in water for only 1 h, but seeds imbibed for 6 h in water exhibited many changes in ultrastructure. These results are consistent with the hypothesis that seed hydration must reach a critical value before germination can proceed, and that the inhibitory effect of NaCl is primarily osmotic in barley seeds that have not reached this hydration threshold. Although isotonic solutions of betaine and NaCl were equally inhibitory to germination, isotonic solutions of betaine and NaCl were not equally inhibitory to continued development in seeds which had been pre-imbibed in water. Calcium ions improved both germination and plumule emergence of pre-imbibed seeds in NaCl solutions, but calcium had little effect on pre-imbibed seeds placed in betaine. Very high concentrations of NaCl or betaine inhibited germination, but did not kill dry seeds. Both solutes, on the other hand, were lethal at high concentrations to germinating seeds. NaCl killed germinating seeds more rapidly than betaine, but calcium reduced the rate of killing to nearly that of betaine. We conclude that hydrated seeds are sensitive to both osmotic and toxic effects of NaCl and that calcium mitigates the toxic effect of NaCl, but not the osmotic effect.     

Ultrastructure and membrane permeability in cowpea seeds

Abstract. The leakage of electrolytes and the localization of chloride within the cells of NaCl-imbibed seeds indicates the plasmalemma is quite permeable during the early stages of imbibition. However, lanthanum is not able to penetrate the plasmalemma, suggesting that the plasmalemma is not entirely porous. Freeze-fracture microscopy indicates that the plasmalemma is highly convoluted but reveals a fairly normal fracture plane. These observations suggest the membrane is a bilayer and leakiness may be more related to the degree of order within the bilayered membrane than to a primary restructuring and/or reorganization of the membrane components in the dry state.     

Changes in plasmalemma organization in cowpea radicle during imbibition in water and NaCl solutions

Abstract Freeze-fracture electron microscopy of the plasmalemma of dry cowpea radicle cells disclosed a normal-appearing membrane with a high area density of intramembraneous particles (IMPs). Seeds imbibed in either water or salt solution exhibited decreased area density of IMPs, but water-imbibed tissue showed the greatest decline. Mean particle size increased with hydration but not enough to suggest aggregation as the cause for this density decrease. Calculations of plasmalemma area expansion during imbibition show that such expansion can account for the decrease in IMPs per unit area in the cytoplasmic side (PF) of the membrane in water-imbibed, but not in salt-imbibed, tissue. During imbibition, there is a change in the ratio of IMPs per unit area of the PF versus EF (external) membrane faces, suggesting a relative increase in the number of EF particles. These changes in membrane structure are probably not related to any decrease in membrane permeability during the early phases of imbibition.     

Osmotic sensitivity in relation to salt sensitivity in germinating barley seeds

Abstract Cultivars of barley (Hordeum vulgare L.) were tested for germination sensitivity to progressively higher concentrations of salt, mannitol, and betaine. The three solutes were equally inhibitory at equal osmotic potential, but there was a consistent difference in osmotic sensitivity between two cultivars, CM-67 and Briggs (Briggs was the most sensitive). There was no difference between the two cultivars in salt or water uptake from salt solutions during imbibition. Brief presoaking in water did not improve salt resistance, indicating that a hydration-dependent decrease in membrane permeability is not involved in salt tolerance. The calcium content of Briggs was higher than CM-67. These results suggest that salt inhibits barley germination primarily by osmotic effects, and that salt influx during imbibition does not play a role in this inhibition. A hypothesis regarding salt effects on germination is discussed.