A major stress-inducible Mr-42000 wall glycoprotein of French bean (Phaseolus vulgaris L.)

A major wall protein of suspension-cultured cells of French bean has been isolated and characterised. It can be prepared from walls or the culture filtrate and in composition it is particularly rich in proline, valine and glutamic acid/glutamine and contains appreciable amounts of hydroxyproline. The N-terminus shows some glycosylation, while following chemical deglycosylation the first 38 residues were found to be identical to those of proline-rich proteins from soybean. However, the composition of the highly purified M_r-42000 bean protein differs considerably from the soybean proteins and must contain its own specific domains. An antibody was raised and used to demonstrate the inducibility of the M_r-42000 bean protein in response to elicitor action. The protein was found to be mainly localised in the intercellular spaces of the cortical cells of bean hypocotyls and at the wall-plasmalemma interface of xylem vessels, another potentially accessible compartment for pathogens. Following wounding, the protein was found to be generally distributed in the wall of epidermal and cortical cells of the hypocotyls. The M_r-42000 protein is cross reactive with antibodies raised to glycoproteins of the Rhizobium infection thread and the chitin-binding hydroxyproline-rich glycoprotein, potato lectin. These common epitopes together with the previously demonstrated chitin-binding properties of the bean protein indicate a role in host-microbial interactions. Furthermore, the M_r-42000 protein itself bound to the growing hyphal tips of the bean pathogen, Colletotrichum lindemuthianum.Abbreviations FITC fluorescein isothiocyanate - IgG immunoglobulin G - PAL phenylalanine ammonia-lyase We thank Dr Nick Brewin for advice on interpretation of immunolocalisations and for the gift of MCA 265. We thank Dudley Fernandino for carrying out the confocal microscopy. GPB thanks the Science and Engineering Research Council for funding.     

Production and standing stocks of the kelp Macrocystis laevis Hay at the Prince Edward Islands,Subantarctic

Summary The recently described species Macrocystis laevis Hay is endemic to the Prince Edward Islands. Aerial photographs of Marion Island were used to outline the distribution of the kelp and to assess its cover. M. laevis occurs along the lee shore of the island, between the 5 and 20 m isobaths. Plant densities and gross plant morphology were measured by divers during April/May 1988. Net production was estimated from growth measurements taken in April/May 1988 and 1989 and again during August 1989. The mean biomass of kelp was 0.67 kgC·m^–2 within the kelp beds. Net production was estimated at 7.7 gC·m^–2·d^–1 and 11.5 gC·m^–2d^–1 during the months of April and August respectively. M. laevis had a uniform frond-length frequency distribution, which suggests that only the oldest fronds are lost by wave action or senescence. Based on calculations for M. laevis and Durvillaea antarctica (the two species making up most of the macrophyte biomass) macrophytes are more productive per unit area than the phytoplankton but contribute less to the seas around the Prince Edward Islands by virtue of their small spatial coverage. Neither of the kelps lose much material as particulate or dissolved organic carbon through fragmentation. The extent of grazing on live M. laevis fronds is unknown, and only D. antarctica contributes to a macrofaunal detrital community. The contribution of M. laevis production to the nearshore ecology of the islands seems limited, as we suspect that almost all of its production is exported to the open ocean pelagic system.     

POPULATION STRUCTURE AND KINSHIP IN POLISTES (HYMENOPTERA,VESPIDAE): AN ANALYSIS USING RIBOSOMAL DNA AND PROTEIN ELECTROPHORESIS

Six variable protein loci and one variable ribosomal DNA restriction site were used for an analysis of population structure in five species of Polistes from Texas. A sample-reuse algorithm was developed that estimated F_ST, F_IS, and ø (the coefficient of kinship) from probabilities of identity. Of the four species analyzed in detail only one, Polistes exclamans, had statistically significant values of F_ST. These values may reflect natural constraints on successful nesting for migrants of this species. Three of the four species had significant values of F_IS and three of the four species had significant values of ø. In many cases ø also differed from the expected value under haplodiploidy and random mating. Values of ø did not differ from expectations under haplodiploidy and local inbreeding. These results emphasize that theories of social behavior and evolution based on coefficients of kinship should include some explicit consideration of population structure.     

Growth of Octopine-Catabolizing Pseudomonas spp. under Octopine Limitation in Chemostats and Their Potential To Compete with Agrobacterium tumefaciens

The growth characteristics of five octopine-catabolizing pseudomonads have been determined in batch and continuous cultures. All five strains belonged to rRNA homology group I and showed a more psychrotrophic growth pattern than did Agrobacterium tumefaciens B6 and ATCC 15955. In chemostats limited by octopine, either as the source of carbon and nitrogen or the sole source of nitrogen, maximum specific growth rates and substrate affinities were lower than those in chemostats limited by glutamate. These growth dynamics were similar to those observed for Agrobacterium strains B6 and ATCC 15955 even though the catabolic genes and pathways are believed to be different in the two genera. An analysis of the yields in octopine-limited chemostats indicated that the use of octopine as the sole source of carbon and nitrogen was grossly inefficient. Octopine and presumably lysopine and octopinic acid provided a better source of nitrogen than of carbon. One of the Pseudomonas fluorescens strains, E175D, was able to produce its highest yield on octopine as a nitrogen source. Competition models formulated on pure culture parameters indicated that two of the Pseudomonas spp. would dominate A. tumefaciens B6 and ATCC 15955 when in simple competition for octopine as a limiting substrate.     

Ontogeny of an Extracellular Matrix Component of Sea Urchins and its Role in Morphogenesis

A monoclonal antibody, Sp14, recognizes fibers that form a complex meshwork within the blastocoel of embryos of the sea urchin, Strongylocentrotus purpuratus . The fibers first appear as the blastocoel begins to form and increase in density throughout development. Ultrastructural localizations using the immunoperoxidase method show bundles of 20 nm fibers that are continuous with the basal lamina and have an indistinct axial periodicity. Embryos treated with tunicamycin, β-D-xylopyranoside, β-aminoproprionitrile, proline analogues, or deprived of sulfate all form immunoreactive fibers although in some treatments the pattern formed is abnormal. Immunoreactivity of extracted fibers is not affected by digestion with chondroitinase ABC, hyaluronidase, collagenase or heparinase. However, proteinase K readily destroys immunoreactivity. Fibers will form in cultures of micromeres or mesenchyme 24 to 48 hr after plating with or without horse serum. In embryos in which the blastocoelar matrix has been altered by injection with Sp14, there is inhibition of the release of secondary mesenchyme from the tip of the archenteron and in some embryos supernumerary skeletal elements are formed. It is proposed that Sp14 recognizes a component of the blastocoelar extracellular matrix that is required for the migration of mesenchyme.     

Purification of 20α-hydroxysteroid oxidoreductase from bovine fetal erythrocytes

NADPH-dependent 20α-hydroxysteroid oxidoreductase (20α-HSD; EC 1.1.1.149) from bovine fetal erythrocytes was obtained for the first time free of hemoglobin by a new 2,500-fold purification scheme. This was achieved by a sequence of calcium phosphate gel adsorption, ammonium sulfate fractionation, and affinity chromatography. The present results lead us to believe that the NADPH-dependent 3β-hydroxysteroid oxidoreductase activity, which was co-purified with 20α-activity, may originate at the active site of 20α-HSD (2).     

A variation of the smooth septate junction in sea spiders (Pycnogonida)

A new type of septate junction considered to be a variation of the arthropod smooth septate junction is described in pycnogonid (sea spider) endothermal tissue based on the use of conventional thin-section, lanthanum tracer and freeze-fracture techniques. This new type of septate junction is apparently unique to the Pycnogonida but closely resembles septate junctions previously described in the Merostomata and Collembola. This work in conjunction with previous work suggests that the septa of smooth septate junctions may not be as ‘smooth’ as generally thought and probably have a complex substructure.