Re-examination ofPyrenomonas andRhodomonas (class cryptophyceae) through ultrastructural survey of red pigmented cryptomonads

Several taxa of cryptomonads, including species of marineChroomonas, Cryptomonas and freshwaterRhodomonas were examined using transmission electron microscopy. They have cellular structures fundamentally in common: a single bilobed chlorplast, a single pyrenoid between the chloroplast lobes, and a nuclemorph embedded within a cleft of the pyrenoidal matrix. These features are in accordance with the taxonomic characteristics of the recently established genusPyrenomonas. The algae also have similar pigmentation to that ofRhodomonas andPyrenomonas which is red or reddish-brown. On the basis of these observations, the genusRhodomonas Karsten (1898) is redescribed in this paper and the genusPyrenomonas Santore is considered to be synonymous withRhodomonas.     

Gymnodinium aeruginosum (Dinophyta): A blue-green dinoflagellate with a vestigial,anucleate, cryptophycean endosymbiont

Gymnodinium aeruginosum has the usual fine structure of a dinoflagellate but does not seem to contain a well elaborated peduncle or a microtubular basket. Naked cells are surrounded by a single large amphiesmal vesicle. It houses an endosymbiont with typical blue-green cryptophycean chloroplasts (generally only one), cryptophycean starch grains in the periplastidal cytoplasm without a nucleomorph, and two membranes separating the periplastidal cytoplasm from the cryptophycean cytoplasm which contains mitochondria, ER, vesicles and ribosomes, but no eukaryotic nucleus. The endosymbiont is surrounded by a single membrane. Possible ways of the acquisition of the endosymbiont and the problem of the existence of ribosomes within a compartment without nucleus are discussed.Devoted to Prof. Dr.L. Geitler, the Nestor of phycology and endosymbiosis research, on the occasion of the 90th anniversary of his birthday.     

Low temperature compartment formation in feline immunodeficiency virus-infected and uninfected feline kidney cells

Summary This study was to determine if feline immunodeficiency virus (FIV)-infected and uninfected Crandall feline kidney (CRFK) cells exhibited a low temperature (16°C) block in membrane trafficking between transitional endoplasmic reticulum and Golgi apparatus represented by intermediate compartment formation. Cells were cultured at different temperatures and membrane changes involving the Golgi apparatus and Golgi apparatus-associated membrane structures were monitored by electron microscopy and quantitated. With 30 min of incubation, membranes of the Golgi apparatus stack increased in amount at temperatures of 16°C and below compared to temperatures above 18°C. The increase was greatest along the major polarity axis as evidenced by an increased stack height. Neither the number of cisternae per stack nor the average stack diameter (width) was affected by temperature. The response was maximal between 15 and 30 min of low temperature treatment of the cells. Results with cells infected and uninfected with feline immunodeficiency virus were similar. The increase in stack height was due primarily to an increase of membranes at the cis face (cis Golgi apparatus network). At 18°C, membranes of the trans Golgi apparatus network accumulated suggesting that import from the cis Golgi network could proceed at this temperature, whereas exit from the trans Golgi network was still at least partially blocked. Also increased at 16°C and below were numbers of transition vesicles in the space between the Golgi apparatus and the transitional endoplasmic reticulum associated with the cis Golgi apparatus face. The results suggested interruption of the orderly flux of membranes into the Golgi apparatus at 16°C and below. Moreover, the block appeared to be reversible. Upon transfer from 16°C to 37°C, there was a time-dependent decrease in the accumulations of cis compartment membrane accompanied by a corresponding equivalent increase in the membranes of the trans Golgi apparatus compartment.     

Dielectric analysis of mitochondria isolated from rat liver II. Intact mitochondria as simulated by a double-shell model

The dielectric dispersion of isolated intact mitochondria in suspension has been measured between 10 kHz and 500 MHz. In isotonic KCI media at 4°C, the mitochondria maintained their characteristic ‘double membrane’ structure as examined by electron microscopy, and the observed dispersion curves were successfully simulated in terms of a superposition of two sub-dispersions having different characteristic frequencies and different permittivity magnitudes. Taking these observations into account we analyzed the dispersion data on the basis of a ‘double-shell’ model in which two concentric shells are meant to represent the mitochondrial outer and inner membranes. The analyses by a computerized curve-fitting method revealed that: (i) electric capacities for the outer and the inner membrane are 1.7 and 0.5 μF/cm², respectively, (ii) relative permittivity for the inner compartment (or the equivalent homogeneous matrical space) = 50–60, (iii) outer compartment-to-external conductivity ratio = 0.4–0.6, and (iv) inner compartment-to-external conductivity ratio = 0.14. The implications of these parameter values are discussed with due attention paid to the limitations inherent in our ‘double-shell’ model approach.     

Lysis of bacterioids in the vicinity of the host cell nucleus in an ineffective (fix-) root nodule of soybean (Glycine max)

In nodules of Glycine max cv. Mandarin infected with a nod ⁺fix^- mutant of Rhizobium japonicum (RH 31-Marburg), lysis of bacteroids was observed 20 d after infection, but occurred in the region around the host cell nucleus, where lytic compartments were formed. Bacteroids, and peribacteroid membranes in other parts of the host cell remained stable until senescence (40d after infection). With two other nod⁺ fix^- mutants of R. japonicum either stable bacteroids and peribacteroid membranes were observed throughout the cell (strain 61-A-165) or a rapid degeneration of bacteroids without an apparent lysis (strain USDA 24) occurred. The size distribution of RH 31-Marburg-infected nodules exhibited only two maxima compared with four in wild-type nodules and nodule leghaemoglobin content was found to be reduced to about one half that of the wild type. The RH 31-Marburg-nodule type is discussed in relation to the stability of the bacteroids and the peribacteroid membrane system in soybean.     

Scale‐down simulators for mammalian cell culture as tools to access the impact of inhomogeneities occurring in large‐scale bioreactors

During the scale‐up of a bioprocess, not all characteristics of the process can be kept constant throughout the different scales. This typically results in increased mixing times with increasing reactor volumes. The poor mixing leads in turn to the formation of concentration gradients throughout the reactor and exposes cells to varying external conditions based on their location in the bioreactor. This can affect process performance and complicate process scale‐up. Scale‐down simulators, which aim at replicating the large‐scale environment, expose the cells to changing environmental conditions. This has the potential to reveal adaptation mechanisms, which cells are using to adjust to rapidly fluctuating environmental conditions and can identify possible root causes for difficulties maintaining similar process performance at different scales. This understanding is of utmost importance in process validation. Additionally, these simulators also have the potential to be used for selecting cells, which are most robust when encountering changing extracellular conditions. The aim of this review is to summarize recent work in this interesting and promising area with the focus on mammalian bioprocesses, since microbial processes have been extensively reviewed.     

From the cytoplasm into the cilium: Bon voyage

The primary cilium compartmentalizes a tiny fraction of the cell surface and volume, yet many proteins are highly enriched in this area and so efficient mechanisms are necessary to concentrate them in the ciliary compartment. Here we review mechanisms that are thought to deliver protein cargo to the base of cilia and are likely to interact with ciliary gating mechanisms. Given the immense variety of ciliary cytosolic and transmembrane proteins, it is almost certain that multiple, albeit frequently interconnected, pathways mediate this process. It is also clear that none of these pathways is fully understood at the present time. Mechanisms that are discussed below facilitate ciliary localization of structural and signaling molecules, which include receptors, G-proteins, ion channels, and enzymes. These mechanisms form a basis for every aspect of cilia function in early embryonic patterning, organ morphogenesis, sensory perception and elsewhere.     

Peribacteroid space acidification: a marker of mature bacteroid functioning in Medicago truncatula nodules

Legumes form a symbiotic interaction with Rhizobiaceae bacteria, which differentiate into nitrogen‐fixing bacteroids within nodules. Here, we investigated in vivo the pH of the peribacteroid space (PBS) surrounding the bacteroid and pH variation throughout symbiosis. In vivo confocal microscopy investigations, using acidotropic probes, demonstrated the acidic state of the PBS. In planta analysis of nodule senescence induced by distinct biological processes drastically increased PBS pH in the N₂‐fixing zone (zone III). Therefore, the PBS acidification observed in mature bacteroids can be considered as a marker of bacteroid N₂ fixation. Using a pH‐sensitive ratiometric probe, PBS pH was measured in vivo during the whole symbiotic process. We showed a progressive acidification of the PBS from the bacteroid release up to the onset of N₂ fixation. Genetic and pharmacological approaches were conducted and led to disruption of the PBS acidification. Altogether, our findings shed light on the role of PBS pH of mature bacteroids in nodule functioning, providing new tools to monitor in vivo bacteroid physiology.     

Plant and soil effects on bacterial communities associated with Miscanthus × giganteus rhizosphere and rhizomes

Bacterial assemblages, especially diazotroph assemblages residing in the rhizomes and the rhizosphere soil of Miscanthus × giganteus, contribute to plant growth and nitrogen use efficiency. However, the composition of these microbial communities has not been adequately explored nor have the potential ecological drivers for these communities been sufficiently studied. This knowledge is needed for understanding and potentially improving M. × giganteus – microbe interactions, and further enhancing sustainability of M. × giganteus production. In this study, cultivated M. × giganteus from four sites in Illinois, Kentucky, Nebraska, and New Jersey were collected to examine the relative influences of soil conditions and plant compartments on assembly of the M. × giganteus‐associated microbiome. Automated ribosomal intergenic spacer (ARISA) and terminal restriction fragment length polymorphism (T‐RFLP) targeting the nifH gene were applied to examine the total bacterial communities and diazotroph assemblages that reside in the rhizomes and the rhizosphere. Distinct microbial assemblages were detected in the endophytic and rhizosphere compartments. Site soil conditions had strong correlation with both total bacterial and diazotroph assemblages, but in different ways. Nitrogen treatments showed no significant effect on the composition of diazotroph assemblages in most sites. Endophytic compartments of different M. × giganteus plants tended to harbor similar microbial communities across all sites, whereas the rhizosphere soil of different plant tended to harbor diverse microbial assemblages that were distinct among sites. These observations offer insight into better understanding of the associative interactions between M. × giganteus and diazotrophs, and how this relationship is influenced by agronomic and edaphic factors.