Photosynthetic Nitrogen Metabolism in High and Low CO2-adapted Scenedesmus: I. INORGANIC CARBON-DEPENDENT O2 EVOLUTION, NITRATE UTILIZATION AND NITROGEN RECYCLING

Larsson, M., Larsson, C.-M. and Guerrero, M. G. 1985. Photosyntheticnitrogen metabolism in high and low CO₂-adapted Scenedesmus.I. Inorganic carbon-dependent O2 evolution, nitrate utilizationand nitrogen recycling.—J. exp Bot. 36: 1373–1386 Scenedesmus obtusiusculus Chod. was grown on an inorganic mediumflushed with either air or air supplemented with 3% CO₂. Inair-grown cells, O₂ evolution dependent on low, but not high,HCO^–₃ concentrations was strongly inhibited by the carbonicanhydrase inhibitor acetazolamide. Cells grown with 3% CO² exhibitedlow rates of O2 evolution at low external inorganic C; however,after 30 min in air O2 evolution rates at low inorganic C approachedthose of air-grown cells. These results are compatible withthe view that Scenedesmus develops a ‘CO² concentratingmechanism’ in air, with carbonic anhydrase as an importantconstituent When 3% CO²-grown cells were subjected to air-level of CO²,just a transient decline in NO^–₃ utilization was observed,but in the presence of acetazolamide the rate of the processdecreased drastically in response to the decrease in the CO²level. In CO²-free air NO₃^– was taken up at high ratesbut in a deregulated manner, leading to release of NH₄⁺. A portionof the NO₃^– taken up in the absence of CO² was apparentlyassimilated Cellular nitrate reductase (NR) activity initially decreasedbut subsequently recovered after a transition from 3% CO² toair. In the presence of acetazolamide, a persistent decreasein NR activity was observed. Cellular glutamine synthetase (GS)activity increased after transition from 3% CO² to air, theactivity increase being unaffected by acetazolamide. NH₄⁺ releaseto the medium in the presence of L-methionine-D, L-sulphoximine(MSO) transiently increased in 3% CO²-grown cells in responseto a transfer to air. MSO-induced NH₄⁺ release was in fact higherin air-grown cells than in 3% CO²-grown cells. Glycollate wasinitially released after transition from 3% CO² to air, butthere was no difference in glycollate release between MSO-treatedand untreated cells. In air-adapted Scenedesmus, N recyclingseems to be of minor importance in comparison to primary N assimilation Key words: CO²-fixation, N recycling, nitrate uptake, Scenedesmus     

Photosynthetic Nitrogen Metabolism in High and Low CO2-adapted Scenedesmus: II. EFFECT OF AMMONIUM AND METHIONINE SULPHOXIMINE ON NITRATE UTILIZATION

Larsson, C.-M., Larsson, M. and Guerrero, M. G. 1985. Photosyntheticnitrogen metabolism in high and low CO²-adapted Scenedesmus.II. Effect of ammonium and methionine sulphoximine on nitrateutilization.—J. exp. Bot. 36: 1387–1395 In 3% CO²-grown Scenedesmus obtusiusculus Chod. utilizing NO₃^–J as the N source, NH₄⁺ addition caused a prompt inhibitionof NO₃^– utilization. Nitrate reductase (NR) activity declinedrapidly in response to the presence of NO₄⁺, but the cessationof NO₃^– utilization was too rapid to be accounted forby the loss in NR activity. The first site of NO₄⁺ inhibitionin these cells seems to be the entrance of NO₃^– into thecells. Upon exhaustion of NO₄⁺ from the medium, NO₃^– utilizationwas rapidly restored and NR activity increased. Air-grown cellswere much less sensitive to the effect of NO₄⁺, more than 30min being required for added NO₄⁺ to cause complete inhibitionof NO₃^– utilization. In these cells, NO₃^– uptakeand NR activity decreased in parallel in response to NO₄⁺ addition.In 3% CO²-grown cells simultaneously subjected to NO₄⁺ and air-levelof CO², NO₄⁺ initially inhibited NO₃^– utilization completely,but a slight recovery took place after approximately 20 min The glutamine synthetase (GS) inhibitor L-methionine D, L-sulphoximine(MSO) behaved as a potent inhibitor of NO₃^– uptake in3% CO²-grown cells, but had considerably less effect in air-growncells, although the time-course of the MSO-induced inhibitionof GS was the same in both cases Key words: Ammonium, nitrate utilization, Scenedesmus     

Antigens of the Ii System on Lymphocytes

USUALLY Ii specificity is assigned to cold agglutinins on the basis of their reactions with the red cells of human adults (adult red cells) and newborn infants (cord red cells). Those which react more strongly with adult red cells are said to detect I antigen and are designated anti-I; those which react more strongly with cord red cells are said to detect i antigen and are designated anti-i^1,2. We have now found that I and i antigens can be easily detected on lymphocytes.     

Ecdysteroids During Embryogenesis in Locusta migratoria

SYNOPSIS. Ovaries of Locusta migratoria synthesize large amountsof ecdysteroids at the end of oöcyte maturation. The predominantecdysteroids in mature ovaries are conjugated 2-deoxyecdysone(100 µM) and conjugated ecdysone (50 µM) which outnumberthe corresponding free compounds by 50–100 fold. Thesevarious ecdysteroids persist during ovulation and are recoveredfrom newly-laid eggs. The conjugated maternal ecdysteroids aregradually metabolized as embryonic development proceeds; theyhave disappeared as such on day 6 after oviposition, that isafter blastokinesis and shortly after dorsal closure. Concomitantlyto this metabolism of the maternal conjugated ecdysteroids,other ecdysteroid conjugates appear in the eggs which have differentchromatographic behaviors and some of which are conjugates ofecdysone metabolites formed by the embryo. The data availableso far are compatible with the hypothesis that the maternalconjugates are hydrolysed to free 2-deoxyecdysone and ecdysoneby the embryo during early stages of development and subsequentlyconjugated to inactivation compounds. During the later stagesof embryonic development however, a de novo synthesis of ecdysoneis probable, the maternal conjugates having been metabolizedduring the earlier stages.     

Isoperoxidases in Epidermal Layers of Tobacco and Changes during Organ Formation in vitro

The content and pattern of soluble isoperoxidases were determined in epidermal explants taken from different internodes of tobacco plants in the vegetative and floral states. There were qualitative and quantitative differences in the isoperoxidases, with a decrease in content and fewer bands being observed acropetally, i.e., in going from the base of the stem towards the apex. Epidermal explants from floral branches were grown in in vitro culture, with various media moditications, to form de novo floral or vegetative buds, roots or callus. Changes in soluble isoperoxidases were followed electrophoretically in relation to these varying morphogenetic pathways. In each of them, the number of bands increased on both the anodic and cathodic sides with time in culture. Compared to each other these four morphogenetic programmes were different in their peroxidase zymograms, mainly through varying kinetics in the development of activity of the isoenzymes. The changes observed during root and vegetative bud formation agree with previously published data, and the changes during floral bud formation agree with those observed in vivo.     

An Inhibition and Kinetic Study of Acid Phosphatase in Paramecium caudatum and Paramecium tetraurelia1

ABSTRACT. Inhibition, inactivation, pH, and kinetic studies using both homogenates and purified lysosomal fractions of Paramecium caudalum and of P. tetraurelia were carried out to examine the lysosomal acid phosphatase (AcPase) and its relationship to p-nitrophenylphosphatase (pNPPase), glucose-6-phosphatase (G6Pase), and 5′-nucleotidase (AMPase). The results generally support the idea that Paramecium cells contain a distinct lysosomal AcPase with a broad substrate specificity. The hydrolysis of glucose-6-phosphate (G6P) and adenosine 5′-monophosphate (AMP) was shown to be due to this enzyme, suggesting that true G6Pase and AMPase may be lacking in these two species; however, some hydrolysis of AMP at pH 7.5 catalyzed by an unknown soluble enzyme distinct from alkaline phosphatase and Na⁺-K⁺-ATPase was observed. Since the hydrolysis of p-nitrophenylphosphate (pNPP) at acid pH was also shown to be due to AcPase alone, pNPPase could be used as a rapid assay for Paramecium AcPase. At an alkaline pH, however, this activity was catalyzed by an alkaline phosphatase located in the cytosol fraction. P. caudatum AcPase was shown to have kinetic properties similar to those of purified rat liver and human prostatic AcPase and to have relative substrate affinities in the order of G6P < β-glycerophosphate < pNPP < AMP. These different substrate affinities might account for the observed differences in the inhibition of the four lysosomal activities by NaF, L(+)-tartrate, and molybdate, all of which inhibited the hydrolysis of G6P, β-glycerophosphate, and pNPP competitively, but which exhibited a noncompetitive inhibition of a mixed type with the hydrolysis of AMP.     

The Correlation of Digestive Vacuole pH and Size with the Digestive Cycle in Paramecium caudatum1

ABSTRACT. The temporal changes in the size and pH of digestive vacuoles (DV) in Paramecium caudatum were reevaluated. Cells were pulsed briefly with polystyrene latex spheres or heat-killed yeast stained with three sulfonphthalein indicator dyes. Within 5 min of formation the intravacuolar pH declined from ～7 to 3. With the exception of a transient and early increase in vacuolar size, vacuole condensation occurred rapidly and paralleled the acidification so that vacuoles reached their lowest pH and minimal size simultaneously. Neutralization and expansion of vacuole size began when vacuoles were GT8 min old. No labeled vacuoles were defecated prior to 21 min after formation but almost all DV were defecated within 1 h so that the digestive cycle of individual vacuoles ranged from 21 to 60 min. Based on these size and pH changes, the presence of acid phosphatase activity, and membrane morphology, digestive vacuoles can be grouped into four stages of digestion. The DV-I are GT6 min old and undergo rapid condensation and acidification. The DV-II are between 4 to 10 min old and are the most condensed and acidic vacuoles. The DV-III range in age from 8 to ～20 min and include the expanding or expanded vacuoles that result from lysosomes fusing with DV-II. The DV-IV are GD21 min old, and since digestion is presumably completed, they can be defecated. The rise in intravacuolar pH that accompanies vacuole expansion suggests that lysosomes play a role in vacuole neutralization in addition to their degradative functions. The acidification and condensation processes in DV-I appear to be unrelated to lysosomal function, as no acid phosphaiase activity has been detected at this stage, but may be related to phagosomal functions important in killing food organisms, denaturing proteins prior to digestion, and preparing vacuole membrane for fusion with lysosomes.     

How river structure and biological traits influence gene flow: a population genetic study of two stream invertebrates with differing dispersal abilities

1. Determined by landscape structure as well as dispersal‐related traits of species, connectivity influences various key aspects of population biology, ranging from population persistence to genetic structure and diversity. Here, we investigated differences in small‐scale connectivity in terms of gene flow between populations of two ecologically important invertebrates with contrasting dispersal‐related traits: an amphipod (Gammarus fossarum) with a purely aquatic life cycle and a mayfly (Baetis rhodani) with a terrestrial adult stage. 2. We used highly polymorphic markers to estimate genetic differentiation between populations of both species within a Swiss pre‐alpine catchment and compared these results to the broader‐scale genetic structure within the Rhine drainage. Landscape genetic approaches were used to test for correlations of genetic and geographical structures and in‐stream barrier effects. 3. We found overall very weak genetic structure in populations of B. rhodani. In contrast, G. fossarum showed strong genetic differentiation, even at spatial scales of a few kilometres, and a clear pattern of isolation by distance. Genetic diversity decreased from downstream towards upstream populations of G. fossarum, suggesting asymmetric gene flow. Correlation of genetic structure with landscape topography was more pronounced in the amphipod. Our study also indicates that G. fossarum might be capable of dispersing overland in headwater regions and of crossing small in‐stream barriers. 4. We speculate that differences in dispersal capacity but also habitat specialisation and potentially the extent of local adaptation could be responsible for the differences in genetic differentiation found between the two species. These results highlight the importance of taking into account dispersal‐related traits when planning management and conservation strategies.     

Detection and validation of EST‐derived SNPs for poplar leaf rust Melampsora medusae f. sp. deltoidae

We report the discovery, characterization and validation of 118 single nucleotide polymorphisms (SNPs) for poplar leaf rust Melampsora medusae f. sp. deltoidae identified using a gene‐targeted approach in an expressed sequence tag (EST) library. We developed a genotyping assay using the iPLEX? primer extension method for two multiplex assays of 28 and 22 SNPs.