Characterisation of a 34 kD protein from potato cyst nematodes, using monoclonal antibodies with potential for species diagnosis

Two monoclonal antibodies, which differentially recognise the two species of potato cyst nematodes (PCN), Globodera pallida and G. rostochiensis, are described. They have been shown to have potential for quantification of these two species, recognising proteins of the same molecular weight (34 kD) in both species. Further investigation showed these proteins to have isoelectric points at pH values of 5.7 in G. pallida and 5.9 in G. rostochiensis, in common with the proteins used by Fleming & Marks (1983) to differentiate the species of PCN. They are likely to be structurally very similar, with the same physiological function (and therefore similar concentrations) in the two species. In cross-reactivity tests with a wide range of soil nematode species, the antibodies reacted strongly only with species of the genus Globodera, and thereby confirmed their potential as the basis of a quantitative immunoassay likely to be useful in management of PCN populations.     

Gap junction structures: Analysis of the x-ray diffraction data

Models for the spatial distribution of protein, lipid and water in gap junction structures have been constructed from the results of the analysis of X-ray diffraction data described here and the electron microscope and chemical data presented in the preceding paper (Caspar, D. L. D., D. A. Goodenough, L. Makowski, and W.C. Phillips. 1977. 74:605-628). The continuous intensity distribution on the meridian of the X-ray diffraction pattern was measured, and corrected for the effects of the partially ordered stacking and partial orientation of the junctions in the X-ray specimens. The electron density distribution in the direction perpendicular to the plane of the junction was calculated from the meridional intensity data. Determination of the interference function for the stacking of the junctions improved the accuracy of the electron density profile. The pair-correlation function, which provides information about the packing of junctions in the specimen, was calculated from the interference function. The intensities of the hexagonal lattice reflections on the equator of the X-ray pattern were used in coordination with the electron microscope data to calculate to the two-dimensional electron density projection onto the plane of the membrane. Differences in the structure of the connexons as seen in the meridional profile and equatorial projections were shown to be correlated to changes in lattice constant. The parts of the junction structure which are variable have been distinguished from the invariant parts by comparison of the X-ray data from different specimens. The combination of these results with electron microscope and chemical data provides low resolution three- dimensional representations of the structures of gap junctions.     

Bead rings at the endoplasmic reticulum-golgi complex boundary: morphological changes accompanying inihibition of intracellular transport of secretory proteins in arthropod fat body tissue

Golgi complex beads are 10-nm particles arranged in rings on the smooth surface of rough endoplasmic reticulum (ER) makind the forming face of the Golgi complex (GC). In arthropod cells they stain specifically with bismuth. Their morphology has been studied after treatment with reagents known to interfere with GC function. Inhibitors of oxidative phosphorylation (antimycin A, cyanide, and anoxia), but not an inhibitor of glycolysis (iodoacetate), both cause the bead rings to collapse and the GC saccules to round up, and inhibit transition vesicle (TV) formation. Cycloheximide blocks protein synthesis on ribosomes but does not stop TV formation or disrupt bead rings, even after prolonged treatment (6 h) to allow emptying of the rough ER cisternae. Thus the collapse of bead rings is not attributable to inhibition of protein synthesis, and the ring structure of beads does not require continued protein synthesis and secretion for its maintenance. Valinomycin has effects on the GC similar to those of antimycin A, but {"type":"entrez-nucleotide","attrs":{"text":"A23187","term_id":"833253","term_text":"A23187"}}A23187, monensin, and lasalocid do not affect bead ring structure or TV formation. These results are consistent with valinomycin’s secondarily uncoupling mitochondria, which collapses bead rings and prevents TV formation. Thus inhibitors of oxidative phosphorylation do not influence the beads through cation movement. Because mononsin and lasalocid block secretion at the level of the condensing vacuoles, bead rings are not influenced by blocks in secretion distal to them or by the backup of secretory material. These experiments are consistent with inhibitors of oxidative phosphorylation collapsing bead rings by decreasing intracellular ATP. The concomitant block to TV formation and the collapse of bead rings suggests that integrity of the bead rings is essential for the transport of secretory material from the rough ER to the GC.     

Analysis of the Diffusion Symmetry Developed by the Alkaline and Acid Bands which form at the Surface of Chara corallina Cells

The diffusion symmetry developed by the alkaline and aeid bandsof Chara corallina was studied. The alkaline system developeda diffusion pattern which could not be fitted to the equationfor a continuous point-source efflux. However, good correlationwas obtained between experimental data and the diffusion equationfor a hollow sphere. The calculated OH- efflux values, obtainedusing the equation of a continuous spherical-surface source,were checked against the influx values of H₁₄ obtained under the same experimental conditions. IndividualOH– band efflux values ranged from 0.07 to 5.95 pmol s^–1and total cell fluxes of 25 pmol cm^–2 s^–1 for OH^-and H were obtained (in the presence of 0.5 mM NaHCO₃). The acid system developed a cylindrical diffusion pattern, butthis could not be fitted to a mathematical equation. Numericalanalysis will have to be employed to obtain values of H⁺ efflux.     

Cold-Inhibited Phloem Translocation in Sugar Beet: II. CHARACTERIZATION AND LOCALIZATION OF THE SLOW-COOLING RESPONSE

Experiments were undertaken on a simplified sugar beet systemto characterize the phloem translocation response to slow coolingtreatments that were applied to the source leaf petiole. Inthese experiments the temperature was decreased by 4°C every16 min, such that the tissue temperature was lowered from 25°Cto 1°C over a period of 80 min. Our results indicated thatan initial slow cooling treatment, on a given test plant, causedno change in the rate of translocation. However, all subsequentslow cooling regimes that were applied to the same petiole positionelicited a characteristic step-type inhibition. This inhibitionaveraged about 10% of the original translocation rate in allcases with no recovery being observed. The data suggest thatthe initial cooling treatment induced an alteration in the petioletissue which facilitated the inhibition phenomenon during subsequentslow coolings. This alteration was shown to be localized withinthe upstream region of the chilled petiole segment, followingan initial slow cooling, or throughout the chilled petiole segmentafter an initial quick cooling from 25°C to 1°C. Resultsalso show that the alteration is a long-lived phenomenon thathas no detectable influence on the quick-cooling induced transientinhibition of translocation. Key words: Phloem, Translocation, Cooling response, Petiole