The Secret Cemetery

The Secret Cemetery . Doris Francis, Leonie Kellaher, and Georgina Neophytou. New York: Berg, 2005. 298 pp.     

Localization of Glyoxylate Cycle Enzymes in Glyoxysomes in Euglena*

SYNOPSIS. We demonstrated previously microbodies in Euglena gracilis grown in the dark on 2-carbon substrates. We have now established in Euglena the particulate nature of enzymes known in other organisms to be localized in microbodies (glyoxysomes and leaf peroxisomes). On a linear sucrose gradient the glyoxylate cycle enzymes band together at a nigner equilibrium density (1.20 g/cm3) than mitochondrial marker enzymes (1.17 g/cm3), establishing the existence in Euglena of glyoxysomes similar to those of higher plants. Glyoxylate (hydroxypyruvate) reductase and, under certain conditions, also glycolate dehydrogenase co-band with the glyoxylate cycle enzymes, suggesting that Euglena glyoxysomes, like those of higher plants, may contain peroxisomal-type enzymes. Catalase, an enzyme characteristic of microbodies from a variety of sources, was not detected in Euglena.     

Interactions of Vesicular Stomatitis Virus Structural Proteins with HeLa Plasma Membranes

THE processes whereby nucleoprotein core particles of certain animal viruses become enveloped by and bud off from host cell membranes can be studied by preparing membrane^1,2 or “sedimentable”³ fractions from infected cells and examining them for the presence of virus proteins. We find that similar experiments designed to monitor assembly of vesicular stoma-titus virus (VSV) at sites along the plasma membranes of HeLa cells are best interpreted after first investigating the possibility that virus proteins adsorb to plasma membranes during cell fractionation and membrane isolation. In this report, we show that at 0° C the membrane protein of VSV, among other virus proteins, adsorbs to plasma membranes isolated from uninfected HeLa cells. With appropriate pulse-chase experiments, however, we are able to demonstrate the progressive association, in vivo, of VSV core protein with plasma membranes of infected HeLa cells.     

Stomata of Micropropagated DelphiniumPlants Respond to ABA, CO2, Light and Water Potential, but Fail to Close Fully

Morphological and physiological characteristics of micropropagatedplants of Delphinium cv. Princess Caroline were studied. Leavesproduced in vitro showed poor control of water loss which appearsto result from restricted responses by stomata and not frompoor cuticular development. Stomata of leaves produced in vitrowere larger and more frequent than those produced during acclimatization.Despite the fact that stomata from isolated epidermis of leavesproduced in vitro reduced their apertures when exposed to turgor-reducingtreatments, they did not close fully. This, together with highstomatal frequencies might explain the poor control of waterloss shown by intact leaves produced in culture when exposedto dry air. While leaves from acclimatized plants showed almostcomplete closure with ABA, low water potentials, darkness andCO₂, stomata from leaves produced in vitro reduced their apertureswhen exposed to those factors, but only to a limit. Therefore,stomata from leaves cultured in vitro seem to be partially functional,but some physiological or anatomical alteration prevents themfrom closing fully. Stomata from leaves produced in vitro wereparticularly insensitive to ABA which appears to be partly associatedwith the high cytokinin concentration in the culture medium.In the long-term, this stomatal insensitivity to ABA might contributeto plant losses when micropropagated plantlets are transferredto soil. Key words: Micropropagation, stomatal physiology, dehydration, PEG, ABA, BAP, darkness, CO₂, Delphinium