Effect of localized nitrogen availability to soybean half-root systems on photosynthate partitioning to roots and nodules

Soybean (Glycine max [L.] Merr. cv Davis) was grown in a split-root growth system designed to maintain control of the root atmosphere. Two experiments were conducted to examine how 80% Ar:20% O₂ (Ar:O₂) and air (Air) atmospheres affected N assimilation (NH₄NO₃ and N₂ fixation) and the partitioning of photosynthate to roots and nodules. Application of NH₄NO₃ to nonnodulated half-root systems enhanced root growth and root respiration at the site of application. A second experiment applied Ar:O₂ or air to the two sides of nodulated soybean half-root systems for 11 days in the following combinations: (a) Air to both sides (Air/Air); (b) Air to one side, Ar:O₂ to the other (Air/Ar:O₂), and (c) Ar:O₂ to both sides (Ar:O₂/Ar:O₂). Results indicated that dry matter and current photosynthate (¹⁴C) were selectively partitioned to nodules and roots where N₂ was available. Both root and nodule growth on the Air side of Air/Ar:O₂ plants was significantly greater than the Ar:O₂ side. The relative partitioning of carbon and current photosynthate between roots and nodules on a half-root system was also affected by N₂ availability. The Ar:O₂ sides partitioned relatively more current photosynthate to roots (57%) than nodules (43%), while N₂-fixing root systems partitioned 36 and 64% of the carbon to roots and nodules, respectively. The Ar:O₂ atmosphere decreased root and nodule respiration by 80% and nitrogenase activity by 85% compared to half-root systems in Air while specific nitrogenase activity of nodules in Ar:O₂ was 50% of nodules supplied Air. Results indicated that nitrogen assimilation, whether from N₂ fixation or inorganic sources, had a localized effect on root development. Nodule development accounted for the major decrease in total photosynthate partitioning to non-N₂-fixing nodules. Soybean compensates for ineffective nodulation by controlling the flux of carbon to ineffective nodules and their associated roots.     

The effect of glutaraldehyde fixation on the elucidation of the morphogenesis of annulate lamellae in oocytes ofRana pipiens

Summary Primary fixation of the frog oocyte with glutaraldehyde, compared to osmium tetroxide, alters the appearance of components involved in the morphogenesis of annulate lamellae. With glutaraldehyde, the outer layer of the nuclear envelope is connected with membranous laminae of variable length. Rounded blebs of the outer layer of the nuclear envelope are infrequently observed. Further, rather than clusters or rows of cytoplasmic vesicles as are observed in osmium tetroxide-fixed cells; numerous and long, smooth-surfaced lamellae are present in the ooplasm after glutaraldehyde fixation. The long membranous laminae then become concentrated in several ooplasmic packets. This is followed by the progressive alignment or orientation of the laminae within the packet. Eventually, those aligned and formerly smooth-surfaced lamellae are converted into annulate lamellae.This study was supported by research grants (HD-00699, GM-09229) and a Career Development Award from the National Institutes of Health, U.S. Public Health Service.     

Cytodifferentiation in the Rana pipiens oocyte

Summary In early diplotene frog oocytes incubated to illustrate thiamine pyrophosphatase (TPPase) activity, reaction product is uniformly distributed within the compartments of the endoplasmic reticulum and nuclear envelope as well as within the saccules and small vesicles comprising the dictyosomes. With continued oocyte development the reaction product becomes concentrated in localized regions of the dictyosome saccules. Eventually, the enzyme is no longer apparent within the endoplasmic reticulum, but is concentrated in the cisternae of the inner dictyosome saccules. The variations noted suggest that the enzyme is synthesized early in diplotene by the endoplasmic reticulum and is subsequently transported to the Golgi apparatus where it is consistently observed at later developmental stages. TPPase activity is also present in the Golgi apparatus of follicle and theca cells as well as in ovarian epithelial cells. The enzyme is also detected in micropinocytotic vesicles contained within the cells comprising the follicle envelope and in intercellular spaces of the follicle. Horseradish peroxidase injected into the coelomic cavity is transported via micropinocytotic vesicles into and through the cells comprising the follicle envelope and in intercellular spaces. The exogenous protein is not found even after a prolonged time period in early diplotene oocytes. The protein is, however, present in large spherical and tubular vesicles in the cortex of vitellogenic oocytes approximately 500 microns in diameter. The possible functional role of the enzyme TPPase during oogenesis is discussed.This investigation was supported by a research grant from the National Science Foundation (GB-8736).     

INTRANUCLEAR AND CYTOPLASMIC ANNULATE LAMELLAE IN TUNICATE OOCYTES

Electron microscope studies were made on various tunicate oocytes at different stages of growth and development. Both the inner and outer lamellae of the perforated nuclear envelope demonstrate considerable blebbing activity. The blebs of the inner lamella detach into the nucleoplasm where they undergo a special type of fusion process resulting in the formation of numerous, usually single, differentiated annulate lamellae of various lengths. The blebbing of the outer layer of the nuclear envelope contributes to the vesicular and granular endoplasmic reticulum characteristically present in the ooplasm and perhaps to the differentiation of cytoplasmic annulate lamellae as well. Cytoplasmic stacks of annulate lamellae frequently have ribosomes associated with them. In addition, granular accumulations are sometimes observed around or between the annuli. The morphological evidence suggests that, at least in many cases, the annuli in the annulate lamellae are patent.     

Relation of polysaccharide content to some biological properties of endotoxins from mutants of Salmonella typhimurium

Kessel, R. W. I. (Rutgers, The State University, New Brunswick, N.J.), Henry H. Freedman, and Werner Braun. Relation of polysaccharide content to some biological properties of endotoxins from mutants of Salmonella typhimurium. J. Bacteriol. 92:592-596. 1966.-Endotoxins were extracted by the phenol-water procedure from a variety of Salmonella typhimurium mutants with known differences in the composition of their cell wall polysaccharides. The lethality of these preparations for mice proved to be correlated with the complexity of the polysaccharide: endotoxin from the smooth parent strain and from rough strains with several sugars attached to the heptose-phosphate backbone were of high toxicity, whereas endotoxin from a mutant possessing only glucose attached to the heptose-phosphate backbone was less toxic, and endotoxin from a mutant possessing the backbone only was least toxic. All of these mutants yielded endotoxins that were equally capable of protecting mice against subsequent challenge with Pseudomonas aeruginosa. Material obtained from a heptoseless mutant by the phenol-water method proved to be neither toxic nor protective. The apparent dissociation of biological properties that can be achieved with the aid of endotoxin preparations from certain mutants is discussed in terms of possible mechanisms.     

Distribution of viruses in the Chesapeake Bay.

High virus counts were found in water samples collected from the Chesapeake Bay. Viruses were enumerated by ultracentrifugation of water samples onto grids which were visualized by transmission electron microscopy. Virus counts in September 1990, April 1991, June 1991, August 1991, and October 1991 ranged between 2.6 x 10(6) and 1.4 x 10(8) viruses ml-1 with a mean of 2.5 x 10(7) viruses ml-1. Virus counts were usually at least three times higher than direct bacterial counts in corresponding samples. Virus counts in August and October were significantly higher than at the other sampling times, whereas bacterial counts were significantly lower at that time, yielding mean virus-to-bacterium ratios of 12.6 and 25.6, respectively. From analysis of morphology of the virus particles, it is concluded that a large proportion of the viruses are bacteriophages. The high virus counts obtained in this study suggest that viruses may be an important factor affecting bacterial populations in the Chesapeake Bay, with implications for gene transfer in natural aquatic bacterial populations and release of genetically engineered microorganisms to estuarine and coastal environments.     

Homeotic transformations of murine vertebrae and concomitant alteration of Hox codes induced by retinoic acid.

Exposure of murine embryos to teratogenic doses of retinoic acid (RA) induced homeotic transformations of vertebrae. Posterior transformations occurred along the complete body axis after RA administration on day 7 of gestation and were accompanied by anterior shifts of Hox gene expression domains in embryos. Anterior transformations of vertebrae in the caudal half of the vertebral column were induced on day 8.5. We suggest that the identity of a vertebral segment is specified by a combination of functionally active Hox genes, a "Hox code." In this concept the sequential activation of Hox genes defines sequentially more posterior axial levels, while mesodermal cells leave the primitive streak. Exogenous RA interferes with the normal establishment of Hox codes and thus with axial specification.     

Inactivation of recombinant human tumor necrosis factor-alpha by proteolytic enzymes released from stimulated human neutrophils

Activated human neutrophils (PMN) degrade rTNF-alpha resulting in a loss of cytotoxic activity against murine L-929 cells (L cells). This inactivation is mediated through proteases released from activated PMN. Exposure of TNF to H2O2, glucose oxidase, xanthine oxidase, or myeloper-oxidase-H2O2-halide did not affect TNF cytotoxicity for L cells. Exposure to trypsin, chymotrypsin, pronase E, or elastase, however, did diminish TNF bioactivity. FMLP-stimulated PMN in the presence, but not in the absence, of cytochalasin B reduced TNF activity, whereas PMA-stimulated PMN did not affect TNF. Stimulation of PMN with opsonized bacteria also induced TNF inactivation as well as the supernatant of FMLP-stimulated cells. Addition of protease inhibitors to the FMLP-stimulated cytochalasin B-treated PMN abrogated the inactivation of TNF cytotoxicity for L cells, whereas scavengers were not protective. In addition, PMN from a chronic granulomatous disease patient also decreased TNF bioactivity. Inactivation of TNF by activated PMN correlated with granule release and not with superoxide production. Exposure of TNF to proteases and FMLP-activated PMN also resulted in a loss of reactivity with anti-TNF antibodies, as measured by ELISA, and in the formation of an approximately 10-kDa split product from the 17-kDa rTNF molecule. Partial degradation of TNF by proteases released from activated PMN may result in a diminished TNF bioactivity and thereby contribute to the regulation of local inflammatory reactions.     

The human transmembrane secretory component (poly-Ig receptor): molecular cloning,restriction fragment length polymorphism and chromosomal sublocalization

Summary The human transmembrane secretory component (SC) mediates glandular translocation of polymeric IgA and IgM into exocrine secretions. A 2898-bp cDNA clone, encoding the entire sequence of the human transmembrane SC, was isolated from a colonic adenocarcinoma cell line cDNA library. The deduced amino-acid sequence had a length of 764 residues and showed an overall similarity of 56% and 64% with the rabbit and rat counterpart, respectively. A restriction fragment length polymorphism (RFLP) was found with PvuII, revealing a two-alle RFLP with an autosomal codominant inheritance pattern and allele frequencies of 0.65 and 0.35. Southern blot analysis of human-rodent somatic hybrid panels, including hybrids with translocation chromosomes carrying different parts of chromosome 1, assigned the SC gene to 1q31-q42, thus confirming a previously reported provisional assignment.     

Inhibitory and stimulatory influences on mesodermal erythropoiesis in the early chick blastoderm

We use a standing-drop culturing method to investigate the effect on mesodermal erythropoiesis of ectoderm and endoderm from the area opaca vasculosa (AOV) and area pellucida (AP) of stage-4 chick blastoderms. We find that ectoderm from the AOV and ectoderm and endoderm from the AP exert an inhibitory influence on mesodermal erythropoiesis. This inhibitory influence is coupled with the tendency of the explants to spread out and become flattened in culture. In contrast, endoderm from the AOV is found to be stimulatory, in agreement with previous studies. We correlate these in vitro inhibitory and stimulatory influences with the morphogenetic patterns that occur during normal development.